Nematology,2002,V ol.4(2), 123-314 Symposiumabstracts

001 Bursaphelenchusxylophilus and B.mucronatus untilthe recent identiŽ cation in Portugal. It is felt that if inJapan: where arethey from? introducedthe nematode would establish populations or interbreedwith endemic non-virulent species. This ban 1; 2 Hideaki IWAHORI ¤, Natsumi KANZAKI and hashadmajorconsequences on theNorth American forest 2 Kazuyoshi FUTAI industry.Recently many new species of Bursaphelenchus 1NationalAgricultural Research Center for Kyushu Okinawa havebeen described from deador dyingpines throughout Region,Nishigoushi, Kumamoto 861-1192, Japan Europe.Because morphological characters are limited 2 KyotoUniversity, Kyoto 606-8502, Japan inusefulness for speciesdescriptions and cannot be ¤[email protected] usedto differentiate populations, molecular taxonomy hasbecome important. W ewilllook at the accuracy Geographicaldistribution and speciation of Bursaphelen- ofmethods used for speciesidentiŽ cation and at what chusxylophilus (pinewoodnematode) and B. mucrona- criteriamight be used to deŽ ne and differentiate species tus were inferredfrom molecularphylogenetic analysis of Bursaphelenchus whenconsidering import and export andchromosomal number .Severalisolates of B. xylop- bans. hilus and B.mucronatus inJapan and from someother countrieswere usedfor DNA sequencingof the ITS re- 003Mitigating the pinewoodnematode and its gionsin ribosomalDNA. Publishedresearch on thenum- vectorsin transported coniferous wood berof chromosomesof selectedisolates was usedto iden- tifya relationshipin speciation of these nematodes. W e David DWINELL identiŽed three (or four) groupsof B.xylophilus and two B.mucronatus groups of basedon molecularphylogenetic USDA ForestService, 320 Green St., Athens, GA 30602,USA information.These groups corresponded to differences in [email protected] chromosomalnumber .Wehypothesiseda routeof geo- graphicalmigration and speciation of these two nema- Thepinewood nematode ( Bursaphelenchusxylophilus ) todesby estimatingthe chronology of speciation.The re- (PWN) hasbeen intercepted in pine chips, unseasoned sultssupport a previoushypothesis that B.xylophilus and lumberand packing-case wood. Likewise, PWN insect B.mucronatus divergedwith the separation of theEuro- vectors, Monochamus spp.,have been found in pallets, peanand North American continents. In conclusion, our crates,and dunnage. As aresultof these interceptions resultsconŽ rmed that Japanese B.xylophilus was intro- andthe known consequences of the introduction of the ducedrecently to Japan ( ca 100years ago) from North PWN, manycountries regulate the import of coniferous America,and suggested that both Japanese B.xylophilus wood.Science-based procedures are needed to ensurethat B.mucronatus and mighthave two origins in NorthAmer- globallytransported wood is free ofthe PWN andits icaand Eurasia, respectively. vectors.Mitigation measures that have been investigated includeprevention, host selection, and treatment by 002Molecular taxonomy as amethodfor describing fumigation,irradiation, chemical dips, pressure treatment populationsand species within the pinewood withpreservatives, and elevated temperature. A mill nematodespecies complex certiŽcation program for lumber(no bark, no grubholes) isrational, but has not got much support. Air-drying Robert I. BOLLA woodto its equilibrium moisture content will eliminate Departmentof Biology,Saint Louis University, 3507 Laclede, thePWN. Switchingto hardwood lumber for wood St.Louis, MO 63103-2010,USA packingmaterial is a short-termsolution. Irradiation [email protected] anddipping wood in borates have shown little promise. TheefŽ cacy of pressure-treatinggreen pine lumber with Shipmentof untreatedpinewood from NorthAmerica and chromatedcopper arsenate has been shown. Pine chips severalAsian countries into Europe has beenbanned. This canbe fumigatedwith phosphine. Logs, lumber, and wood banhas arisen because of the potential for introduction packingmaterial can be fumigatedwith methyl bromide. ofspecies of Bursaphelenchus pathogenicto pine into Heatingconiferous wood to a coretemperature of 60 ±C Europewith the consequent effect comparable to thatseen willeradicate the PWN andits vectors. inJapan, China and Korea. This ban is basedon thelack ofidentiŽcation of Bursaphelenchusxylophilus in Europe

© KoninklijkeBrill NV ,Leiden,2002 123 Epidemiologyof PineWilt disease (1-7) and Plenary Session (8)

004Occurrence of the pinewoodnematode, iscaused by the pinewood nematode, Bursaphelenchus Bursaphelenchusxylophilus inPortugal and xylophilus,withthe pine sawyer, Monochamusalternatus , perspectivesof the diseasespread in Europe asvector .Wepresenta mathematicalmodel to describe thehost-parasite interaction between pines and pine Manuel MOTA sawyerscarrying nematodes on thebasis of detaileddata NemaLab,ICAM, Dept.de Biologia,Universidade de Évora, takenfrom theincidence of pine wilt disease at a study 7000É vora,P ortugal sitelocated on the northwest coast of Japan. With this [email protected] modelwe simulatethe temporal change in the incidence ofthe disease and predict how the epidemic could be In1999, the pinewood nematode (PWN), Bursaphelen- controlledby eradicationof thepine sawyer. Furthermore, chusxylophilus, was foundand identiŽ ed for theŽ rst themodelis extended to studythe spatialspread of disease timein Portugaland in Europe. Following detection, Por- ona largescale, by incorporating short-range dispersal tugueseauthorities initiated the implementation of erad- ofthe pine sawyer, together with long-range dispersal icationmeasures during 1999 and 2000, following an throughair convection or transportation of logs infested alertprovided to European Community ofŽ cials; as a re- withnematodes. W eestimatethat more than 10% of sult,the nematode was conŽrmed to be conŽ ned in the sawyers undergolong-distance dispersal in areas where Setúbal region, near Lisbon.A taskforce from thefollow- rapidspread of thedisease is observed. upgroup (GANP) createdby the Secretary of State for theRural Development established a nationaleradication programme(PROLUNP) to i)containPWN withinthe 006Fluctuation in genetic structure andvirulence initialgeographic limits; ii)implementeradication mea- ofpinewoodnematode in host pinetrees and insect sures; and iii)monitorPWN atanationallevel. Research vectorsat the end ofapinewilt epidemic ispresently being conducted both at universities as well Katsumi TOGASHI1; , Sachie ENOKI 1, Yuji ISAGI 1 and asresearch institutes, focusing on thecharacterisation of ¤ Takuya AIKAWA 2 Bursaphelenchus speciesassociated with maritime pine, aswell as ontheinsect vector, Monochamusgalloprovin- 1 Facultyof IntegratedArts and Sciences, Hiroshima University, cialis.Recentreports indicate that the nematode may be Kagamiyama1-7-1, Higashi-Hiroshima 739-8521, Japan 2 presentin Siberia (Russia), which would present a threat Forestryand F orestProducts Research Institute, Kukisaki, toEastern European forestry. Efforts arepresently being Ibaraki305-8687, Japan [email protected] developedby severalEuropean countries to establisha re- ¤ searchconsortium to detect and study the possible pres- Thepinewood nematode, the causative agent of pinewilt enceof PWN, for anewPRA. Arecentworkshop held disease,is harboured as a subpopulationin each dead inPortugal in 2001 was anopportunity for sharingexpe- pinetree and then in each vector beetle of Monochamus riencesand techniques on detectionand control. There is alternatus inJapan. The transmission is limited during clearlya greaterawareness of thisissue in Europe. earlyand mid-summer. T odeterminea temporalchange ingeneticstructure and virulence of pinewoodnematode, 005Mathematical models for spatial pattern of thenematode was sampledfrom dead Pinusdensi ora the spreadof pinewilt disease treesand from M.alternatus beetlesappearing in a P. densiora standbetween 1996 and 1998 after a heavy 1; 1 Nanako SHIGESADA ¤, Yukie MIMURA , infestation.Alleles of 30 nematodes from eachsample Fugo TAKASU 1, Kohkichi KAWASAKI 2 were determinedfor fourmicrosatellite loci. A nematode and Kazumi TOGASHI 3 isolatewas establishedfrom eachsample and inoculated on 30 P.densiora seedlingsto determine the virulence. 1 NaraW omen’s University,Kita-Uoya Nishimachi, Nara Consequently,the gene frequency was similarbetween a 630-8506,Japan nematodesubpopulation in adeadtree and those in beetles 2 DoshishaUniversity, Kyo-T anabe610-0321, Japan emergingfrom itin many cases. A veragegene diversity 3 HiroshimaUniversity, Hiroshima 793-8521, Japan [email protected] betweennematode subpopulations was smallerthan the ¤ averagegene diversity within subpopulations. Genetic Anepidemic of pine wilt disease has been spreading in distancebetween nematode subpopulations harboured in wideareas of Japan for nearlya century.The disease beetlesemerging from thesame trees was oftensmaller

124 Nematology Symposiumabstracts thanthat between those from differenttrees. The virulence thephenomenon of ‘ tiredsoils’ [ Boden Muedigkeit ] in ofnematode was low.Multiple infection may not induce a Germanyled to the isolation of fungal species from largevariation of virulenceincluding high virulent strains withincysts and females of Heteroderaschachtii from atthe end of infestation. sugarbeetŽ elds.It was thoughtthen that some of these fungiwere pathogensof the nematodes. Throughout the 007Nematode candidates for the biologicalcontrol 20thcentury additional examples of soils suppressive ofpinewilt disease toother pathogens were described.This led to the ideathat the phenomenon of suppressiveness was Hajime KOSAKA linkedto the presence of one or a limitednumber ofmicrobial species antagonistic to the pathogens. ForestMicrobiology Division, F orestryand F orestProducts Researchefforts duringthe century led to the successful ResearchInstitute, Tsukuba, Ibaraki 305-8687, Japan [email protected] developmentof several schemes and cultural practices thatgenerate or increasesuppressiveness of soilsagainst Environmentallyfriendly control methods are being individualpathogens. These efforts evidencedthat the demandedfor pestcontrol. There is undoubtedlythe same phenomenonof suppressiveness is closely linked with demandfor thecontrol of pinewilt disease. The disease biologicaldiversity of soils.Also, that suppressiveness or iscaused by thepine wood nematode, Burusaphelenchus ‘resistance’of soilsagainst pathogens is acontinuum.It xylophilus,whichis transmitted by the Japanese pine isdemonstrable that a soilseverely affected by disease sawyer, Monochamusalternatus .Inthis presentation, canbe convertedgradually into a suppressivesoil where recentstudies on three kinds of beneŽ cial nematodes diseaseincidence becomes unimportant. There are in arereported for thebiological control of the disease; soilsdegrees of ‘ resistance’against pathogens. Non- theentomopathogen icnematodes, Steinernema spp.; the phytopathogenicnematodes play an integral role in the tylenchidinsect parasitic nematode, Controtylenchus phenomenonof suppressiveness of soils against plant genitalicola ;andavirulent isolates of B.xylophilus . pathogens.These nematodes can be directlyactive against Steinernematidscan kill the vector insects in the dead pathogensand can also disseminate microbial species pinelogs by spraying nematode suspension on the throughoutthe soil and increase microbial activity, hence surfaceof logs. Controtylenchusgenitalicola has the ‘resistance’to pathogens. potentialto reduce the fecundity of the vector insects and propagatesby feedingon afungusas wellas in the host 009Molecular diagnostics: going towards new insects.This mycetophagy could allow mass production frontiers ofthe nematodes. A virulentisolates of B.xylophilus caninduce the resistance of pine trees against the C. André LEVESQUE diseaseby inoculation of pinetrees with the nematodes. However,there are also many barriers to develop the Agricultureand Agri-F oodCanada, Eastern Cereal and practicalcontrol of the disease using these nematodes. OilseedResearch Centre, K.W .NeatbyBuilding, 960 Carling AvenueOttawa, ON, K1A 0C6,Canada Thedirection of furtherstudies to cross these barriers is [email protected] discussed. Moleculardiagnostic tests developed and used in the 008Suppressiveness of soilsagainst pathogens: pastfew decadeshave been relying on antibodies for biodiversityand nematology techniquessuch as ELISA andon DNA basedassays, inparticular PCR. A newrevolution is in the making R. RODRIGUEZ-KABANA whichis generated by the synergy between genomics andmicrofabrication, two of themost important research Departmentof Entomology& PlantP athology,Auburn Želdsworldwide. Genomics tools can already be tapped University,Auburn, AL 36849,USA directlyfor environmentalresearch by using existing [email protected] DNA microarrays, e.g.,ageneexpression proŽ le of asoil ScientiŽc recognitionof theexistenceof soilssuppressive samplecan be determinedby usingbacterial microarrays. ofplant diseases is over a centuryold and was Severalmolecular and computer companies have made basedoriginally on the Fusarium-root-knotcomplex in ‘bio-chip’and are already producing a completelynew Alabamacotton Ž elds.Prior tothis, researches into generationof instrumentsfor thelife sciences. It is already

Vol.4(2), 2002 125 Moleculardiagnostics (9-13) possibleto combine all the steps of complex molecular 011Real time PCR forquantitative detection biologyprotocols within devices of the size of a credit card.The competition is Ž erceto see which company John MARSHALL, Sandi KEENAN and Simon BULMAN willcome up with the Ž rst hand-helddevice to perform New ZealandInstitute for Crop & FoodResearch Ltd, Private standardmolecular diagnostic tests for geneticdisorders Bag4704 Christchurch, New Zealand orinfectiousdiseases at thepointof care.The competitive [email protected] researchon thesemolecular diagnostic devices is goingat afastpace which has been accelerated further by recent PCRassayfor thesimultaneous detection and identiŽ - bioterrorismevents. In the near future, soldiers are likely cation of Globoderarostochiensis and G. pallida (PCN) tocarry specialised miniature molecular diagnostic labs as hasbeen developed but cannot be used to determine the partof theirequipment against biological warfare. When levelof infestation in the soil as the standard PCR sig- DNA orbiomolecules meet the microchip in silica, the nalis not quantitative. Direct single step quantitative de- possibilitiesare endless and science Ž ctionmovies might tectionis possibleusing T aqmantechnology as the inten- beoutdonesoon. sityof the  uorescentsignal generated during the PCR reactionis quantitative. Using this system, simultaneous 010Progress in developing nematode diagnostics detectionand quantiŽ cation of the target DNA ofboth G.rostochiensis (Ro) and G. pallida (Pa) was optimised. Vivian C. BLOK QuantitativePCR ofPCN DNA templateconcentrations demonstratedthat pure DNA from bothRo and Pa gave ScottishCrop Research Institute, Invergowrie, Dundee, DD2 regressionco-efŽ cient of the standard Ct curve against 5DA,Scotland, UK templateconcentration on average better than 0.95. This [email protected] resultwas repeatedusing second stage juvenile individu- alsand whole PCN cysts.Duplex primers for Roand Pa Distinguishingpathogenic nematodes at the species or were developedand we were ableto detect both species subspecieslevels is relying increasingly on molecular andalso determine the ratio of both species in the mix- tools.Morphological differences may be absent or difŽ - ture.Other applications of thistechnology, problems and cultto observe. PCR-based diagnostics offer possibilities limitationswill be discussed. for precision,sensitivity and quantiŽ cation. For example, ribosomalDNA hasbeen widely exploited to distinguish 012Quantifying nematodes and other root diseases manyspecies of plant-parasiticnematodes, to investigate inthe Australianwheat belt intraspeciŽc variationand to examine evolutionary rela- tionships.Further reŽ nements are in progressto use rDNA John CURRAN toquantify and determine the relative proportions in mix- turesof speciesfor usein riskassessments. Mitochondrial CSIRO EntomologyP .O.Box 1700, Canberra ACT 2601, DNA sizepolymorphisms have also been used to distin- Australia [email protected] guishmany of theeconomically important root-knot ne- matodes;however, for manygenera this molecular target Rootlesion and cereal cyst nematodes are the major plant- remainsunexplored or may be problematic. Where the parasiticnematodes to infest grain crops in Australia. ‘conventional’targets have lacked sufŽ cient variability Theyrarely occur in isolation and several major fungal todistinguish closely related species, randomly ampliŽ ed pathogenssuch as take-all, Rhizoctonia and crown polymorphismsfrom Žngerprintingpatterns have been ex- rotcontribute to signiŽ cant soil borne disease. A ploited.Challenges remain, however. Honing molecular comprehensivetesting system has been developed that diagnosticsto distinguish between pathogenic and non- quantiŽes DNA extractedfrom soilusing a platebased pathogenicvariants within species will require identifying hybridisationassay to identify the major pathogens moleculartargets which are either closely linked to or de- presentin a pooledsoil sample. This data is then fed Žnethe functional pathogenicity. Incorporating diagnos- intoa DecisionSupport System that takes into account ticsinto management programmes to more effectively de- thepathogen levels, environmental and other agronomic ployresistance, pesticide use or croprotation will require factorsand provides advise on riskcategories to growers. improvingthe simplicity, reproducibility and portability ofthese diagnostics to encouragetheir uptake.

126 Nematology Symposiumabstracts

013Nematodes, quarantine pests andpathogen developedand are increasingly utilised to manage crops in detectionusing rapid automated sampling of bulk zoneswhich are smaller than the total Ž eldarea. The Ž rst cargoshipments toolwas theyield monitor. Y ieldmaps provide growth patternsthat may either be consistent over time (often 1; 2 Shimon BAREL ¤, Sam S. BUECHLER and basedon soilphysical properties), or bevariablebecause 3 Colin C. FLEMING ofan unusual or sporadic stress. Grid soil sampling for 1Ministryof Agriculture,National Residue Control Labs fertilitybased on global positioning systems (GPS) was NRCL-KVI, P.O.Box 12 Bet Dagan, Israel anotherearly tool. However, this has been superseded by 2 RAY DetectionT echnologies,Holon, Israel zonesampling in some areas. V ariablerate application 3 DANI Departmentof Agriculture& RuralDevelopment equipmenthas been most heavily utilised in fertiliser NorthernIreland and The Queen’ s Universityof Belfast, UK application.However, other uses include seeding rates, ¤[email protected] pesticides,and plant growth regulators. Guidance systems Nematodesare usually not evenly dispersed. Therefore, arenow used to improveprecision of applicationsand row testingof bulk cargo for thepresence of nematodes is positioning.Precision agriculture is highly dependent on onlyas good as the methodology and sampling strat- informationand its success may depend on theability to egyapplied. Limiting factors for takinglots of samples gatherinexpensive, yet accurate information. arethe produce bulk size, laborious and costly process- ing.While there have been signiŽ cant advances in pro- 015Using electrical conductivity to predict ceduresfor invertebratepest identiŽ cation, the method- nematodedistribution in cottonŽ elds ologyand tools for samplingthese organisms are less welldeveloped. Effective sampling procedures are criti- John MUELLER 1, Ahmad KHALILIAN 1, calto successful implementation of quarantinemeasures. Terry KIRKPATRICK 2, Allen WRATHER 3 ™ The Discovery CERT technologywas originallydevel- andStephen L EWIS 4 opedby RA Yfor detectionof explosives and chemical- 1 biologicalwarfare agentsin bulk luggage, mail and cargo. ClemsonUniversity, 64 ResearchRd., Blackville, SC 29817, USA Inspectionprocess lasts 8 minfor anULD aviationpal- 2 SouthwestResearch and Education Center ,Hope,AR 71801, let2449 kg checked as a wholein one batch. The sys- USA temallows selective collection of ultratrace and particles 3 Universityof Missouri-Delta Center ,P.O.Box160, ofadistributionsize of 0.5-800 ¹m.Testscarried out by Portageville,MO 63873,USA DANI/ QUB andNRCL using spiked bacterial/ fungaland 4 108Long Hall, Clemson University, Clemson, SC 29634,USA quarantinepests, combinations of invertebrate/ substrate e.g.,non-viablenematode cysts ( Globoderarostochien- Meloidogyneincognita and Hoplolaimuscolumbus are sis)placedin boxes, lemons infested by live thrips ( He- controlledin cotton by the uniform application of liothripshaemorrhoidalis ),specimensof leafminer( Liri- nematicidesacross a Želd.In this system a signiŽcant omyzasativae )andwhite y ( Trialeurodesvaporariorum ), portionof the nematicide is applied where nematode inoculatedonto bulk cabbages, have shown that Discov- densitiesare below treatment thresholds. This is due to ery™ cansuccessfully recover the target organisms for thespatial aggregation patterns these nematode species identiŽcation. typicallyexhibit in agricultural Ž elds.V ariablerate applicationsof nematicidewould allow applications only toareas of the Ž eldwhere treatment thresholds are 014Precision agriculture: tools of the trade exceeded.However, generating the nematode distribution mapsrequired for variablerate applications by grid Terry WHEELER orzone sampling is cost prohibitive. Soil composition, TexasAgricultural Experiment Station, Rt. 3, Box219, especiallypercentage of sand, is one of the primary Lubbock,TX 79403,USA factorsin uencing nematode distribution. Soil electrical [email protected] conductivity(SEC) canpredict the sand content of asandy Precisionagriculture is the optimisation of management loamsoil with a veryhigh degree of accuracy.A 9%drop ofagricultural crops. Crop management is often affected insand content can result in a 57%drop in population byphysical, chemical, and biological factors which are densitiesof H. columbus.Use ofacommercially-available non-uniformin their impact. Therefore, tools have been SECmetercoupled with a GPS systemallows mapping

Vol.4(2), 2002 127 Precisionagriculture and GPS (14-17)and Global comparative nematode management (18-26) ofa tenha Ž eldin several hours. These maps can then thelargest potential cost savings from spatiallyvariable beused for cost-effectivevariable rate applications of application,and these savings would be accompaniedby nematicides. environmentalbeneŽ ts. The potential beneŽ t:cost ratios for sucha strategywill be considered,along with some of 016Remote sensing and precision nematicide therisks. Maps of realPCN infestationsin potatoŽ eldsin applicationsfor reniform nematode management in whichsusceptible potato crops were grownwill be usedto Mississippicotton demonstratesome of theprinciples and risks. The inverse relationshipbetween population density before planting 1; 1 2 G.W. LAWRENCE ¤, A.T. KELLEY , R.L. KING , andmultiplication rate of PCN makesit difŽ cult to J. VICKERY 2, H.K. LEE 1 and K.S. MCLEAN 3 devisereliable spatial nematicide application procedures, especiallywhen the pre-planting population density is 1 Departmentof Entomologyand Plant P athologyMississippi StateUniversity, Mississippi State, MS, USA justless than the detection threshold. Also, the spatial 2 Departmentof Electricaland Computer Engineering dependencefound suggests that the use of sampling MississippiState, MS 39762,USA gridsthat are too coarse is likely to produce misleading 3 Departmentof Entomology and Plant P athology,Auburn distributionmaps. Coarse grids can miss patches of PCN, University,Auburn University, AL 36849,USA butan economic analysis of therisks of missingpatches ¤[email protected] can be made. Site-speciŽc managementof cotton pests is practised in Mississippiusing remote sensing to direct spatially vari- 018Management strategies for nematode control in ablepesticide applications. Remotely sensed imagery is Europe correlatedwith crop growth parameters using Normalized 1; 2 DifferenceV egetationIndex (NDVI) valuesto createpre- Nicola GRECO ¤ and Daniel ESMENJAUD scriptionmaps for bi-modalor multi-modaldistributions. 1CNR, Istitutoper la Protezione delle Piante, Sezione di Bari, Thistechnology is beingexamined using the spatial dis- ViaG.Amendola165/ A,70126Bari, Italy tributionof the reniform nematode. Re ectance proper- 2 INRA, Unité‘Interactions Plants-Micro-organismes et Santé tiesexhibited by reniform-infected cotton plants are be- Végé tale’ , B.P.2078,06 606Antibes cedex, France ingacquired with hyperspectral spectroradiometers using ¤[email protected] bothairborne and ground platforms. SpeciŽ c reectance Theimprovement of food and environment quality is curves,collected with the spectroradiometers, can be cor- apriorityof the European Union. Therefore, funds for relatedwith nematode numbers and used to generate ne- agriculturalresearch and management aim at achieving matodecontour maps for globalpositioning system (GPS) thisgoal and alternative strategies to chemicals are directedvariable rate technology (VRT) nematicideappli- promoted.In Europe, the most severe nematode pests cations.Hyperspectral data may provide useful remote- arecyst, root-lesion, root-knot (RKN), stemand bulb, sensingtools reducing the timeconsuming and costly grid citrusand virus vector nematodes. Classical control of samplingprocess required in precision nematicide appli- Želdcrop nematodes relies on crop sequences, resistant cations. cultivarswhen available and clean planting materials. Nematicidesare used to control nematodes of high- 017Economies in nematodemanagement from valuecrops (vegetables,  owers,nurseries) for which precisionagriculture – limitationsand possibilities the ban of e.g.,methylbromide is a concern.Soil solarisation,new resistant cultivars or rootstocksand soil- Ken EVANS and Andy BARKER ¤ lesscrops are gaining interest as alternative methods. NematodeInteractions Unit, IACR-Rothamsted, Harpenden, Inperennials, management strategies may be preventive Herts,AL5 2JQ,UK byusing nematode-free plant material obtained through ¤[email protected] certiŽcation schemes or curativewith tolerant rootstocks. Thepotato cyst nematodes (PCN), Globoderapallida Pre-plantsoil fumigation is limited to severe nematode and G.rostochiensis ,representa severeconstraint to attacks.Research for geneticsolutions is in progress potatoproduction in the United Kingdom. Management for variouscrops, such as for RKNin Prunus spp. and ofPCN dependsheavily on nematicides,which are costly. Xiphinemaindex /GFLVingrape.For thislatter problem, Of allthe inputs in UK agriculture,nematicides offer anoriginalalternative to nematicides is theapplication of

128 Nematology Symposiumabstracts systemicherbicides to kill roots before pulling out the Instituteof Biology,Karelian Research Centre RAS, cropcombined with fallow to eliminate the virus from Pushkinskayast, 11, P etrozavodsk,185610, Karelia, Russia survivingnematodes. [email protected] InRussia and neighbouringcountries, of tenimportant ge- 019Nematode management practices in African neraof plant parasitic nematodes, six have a greateco- agriculture nomicimpact. They are Heterodera,Globodera, Meloi- dogyne,Ditylenchus, Pratylenchus and Helicotylenchus. E.E.A. OYEDUNMADE Thedivision of USSR intomany independent states and recenteconomic reforms inRussia led to deep changes Departmentof Crop Production, University of Ilorin,Ilorin, inagricultural practices. These changes increase dam- Nigeria ageof main crops caused by plant-parasitic nematodes, [email protected] andprovide the opportunity for somedangerous nema- todesto come back and new species to appearand spread. TheAfrican continent is mostly exposed to tropical Thesecountries are characterised by zones specialising climates,which favour the reproduction and feeding inmonocrop growing ( e.g.,cereals,potato, sugar beet, activityof plant-parasitic nematodes all the year round. grapevineand cotton-sowing areas). Hence a closerela- Invarious parts of Africa, manynematodes are parasitic tionshipexists between the degree of cropconcentration oneconomic and food crops, but the three most inhigh-specialistcollective farms andthe acuteness of ne- importantplant-parasitic nematodes are the root-knot matodeproblems. Damage caused by nematodesis oneof nematodes (Meloidogyne spp.),the root lesion nematodes thelimiting factors in cropproduction. Annually farmers (Pratylenchus spp.)and the cyst nematodes ( Heterodera losea signiŽcant part of cropyields due topoorcontrolof spp.)which attack and cause severe yield reductions theseed materials, low agricultural technologies and ab- incrops such as cowpea, soybean, maize, yam, cotton, senceof nematicides.Traditional nematode management coffee,sugarcane and several vegetables. Agriculture includesthe followingmeasures: prophylactic (prevention inAfrica ischaracterised by small sized farms and ofthenematode establishment in agricultural Ž eldsdue to low-leveltechnology. As aresultof these, the most quarantineregulations and cleaning of propagation ma- commonlyadopted nematode management practices in terials),physical (control by heating and vapour), chem- severalparts of Africa arecultural control measures which ical(nematicides), crop rotations, resistant cultivars and arenot capital intensive and which require little or no somecultural practices (soil amendments). At present,al- technicalknow-how on the part of the mostly illiterate ternativenematode control strategies are being developed. peasantfarmers. Toalargeextent, the use of synthetic Theyare based on theuse of biologicalproducts, wastes nematicidesis restrictedto largeand experimental farms, from loggingand the timber industry, new resistant culti- butthe recent awareness of environmentalsafety coupled varswith complex resistance to plantpathogens, and stim- withthe prohibitive costs of synthetic pesticides has ulationand modulation of hostresistance by usingof phy- encouragedresearch into safer andcheaper alternatives. tohormonaland adaptogenic phytoregulators and phyto- Themost commonly used cultural methods of nematode controltechnologies. managementin Africa are: i)shiftingcultivation and bush fallowing(until very recently due to land limitation, this 021Nematode management strategies in EastAsian methodhas played a verysigniŽ cant role in the control of countries manyeconomically important plant parasitic nematodes); ii)mixedcropping; iii)useof soil amendments; iv) Zen-ichi SANO soiltillage; v)useof resistant varieties and healthy propagationmaterials; vi)croprotation; vii)roguingand NationalAgricultural Research Center for Kyushu Okinawa Region,Nishigoshi, Kumamoto 861-1192, Japan farm sanitation;and viii)soilsolarisation. EastAsian countries have diverse agricultural systems. 020Nematode management in Russiaand However,root-knot nematodes, cyst nematodes and root- neighbouringcountries of the formerUSSR lesionnematodes are important pests throughout this area.Extremely intensive farming systems are prevalent E.M. MATVEEVA inJapan. Many farmers continuouslycultivate high

Vol.4(2), 2002 129 Precisionagriculture and GPS (14-17)and Global comparative nematode management (18-26) valuecrops such as tomato or melon for yearsin certaincropping systems to provideintegrated nematode thesame plastic greenhouses. Currently similar farming managementmodules to Ž tintoplant protection packages. practicesare increasing in Korea and other countries. Insuch intensive farming, control of soil-borne pests is 023Nematode management strategies in major inevitableto maintain proŽ table yields. Fumigation with cropsin the Australasianregion nematicidesis the most reliable and practical control measure.1,3-dichloroprop eneis often applied with Mike HODDA granularnematicides. Methyl bromide or chloropicrin is appliedin Ž eldswhere both nematodes and soil-borne CSIRO Entomology,GPO Box1700, Canberra ACT 2601, diseasesoccur .Theprohibition of methyl bromide is a Australia [email protected] seriousproblem for pestcontrol. Solarisation of sealed plasticgreenhouses for about30 daysinmid-summer is an TheAustralasian region encompasses a hugerange effectivemethod with few adverseenvironmental effects. ofagricultural systems, from subsistenceto highly Covercrops such as Crotalaria spp.are also used to mechanisedcommercial farms. Thereare many different reducenematode populations and improvesoil conditions. nematodepests and management strategies, but similar Tomatocultivars with resistance to root-knot nematodes approachesin broad agricultural systems. Heterodera has arecommercially available. However, resistant breaking beenthe major pests of broadacre low intensity farming racesare present in manyŽ eldsin Japan. ofgrains in the region, but its successful management bycrop rotation and resistance have seen Pratylenchus emergingas major pests of this cropping system. 022Management practices in South Asian Inpasture, nematode management is directed largely agriculture towardsresistance, with lesser efforts inagronomic Hari S. GAUR practicesand a rangeof emerging techniques including mulchingand soil amendments. In intensive horticulture, Divisionof Nematology, Indian Agricultural Research Institute, Meloidogyne , Radopholus, Heterodera and, in New New Delhi-110012, India Zealand, Globodera aremanaged by phytosanitary [email protected] controls,chemical methods, and emerging techniques. In SouthernAsia comprises very diverse and intensively cul- woodycrops, Meloidogyne , Tylenchulus , Xiphinema and tivatedareas, characterised by small farm holdingsand criconematidsare all important, and controlled mostly economicallypoor, and often illiterate or poorly edu- chemically.In small-scale horticulture Meloidogyne , cated,farmers. Thenematode management practised over Radopholus and Pratylenchus aremost important, and largeareas, often inadvertently, depends upon crop ro- controlledby enforced crop rotations. Quarantine and tation,tillage, solar heat and organic amendments. Use phytosanitationare also common themes in nematode ofnematicidal pesticides is very limited. Some organo- managementbecause a numberof pestspecies common in phosphatesand carbamates are applied for thecontrol of therest of theworld are absent from countriesor certain insectpests and rarely against nematodes. Small doses areas.Trends are for increasinguse of plantresistance, but ofpesticidesare advised for applicationto nursery-beds alliedwith emerging techniques and greater knowledge of orto seed of direct seeded crops. Research efforts have thepest species systematics and biology. beenmade for Žndingsources of nematoderesistance in cropsand a few fairlyresistant varieties or crops like 024Nematode management in North American rice,tomato, chillies, cowpea, mungbean, cotton, grapes, crops etc.,havebeen selected or speciŽ cally bred. Use ofge- neticengineering for nematoderesistant crops is being D.P. SCHMITT¤ and B.S. SIPES attempted.Biological control agents including imported andindigenous strains of Paecilomyceslilacinus , Tricho- Universityof Hawaii, Plant and Environmental Protection dermaviride , T.harzianum , Aspergillusniger and VAM, Sciences,3190 Maile W ay,Honolulu, HI 96822,USA Glomus spp., Pasteuriapenetrans , Pseudomonas uo- ¤[email protected] rescens, Bacillussubtilis etc .,havebeen found effective Thecrop and the associated nematodes in NorthAmerica buttheir production and use are still very limited. Combi- inuence the type of management used. Control of nationsof morethan one practice have been designed for nematodesin low cash value crops depends heavily on

130 Nematology Symposiumabstracts hostplant resistance and crop rotation. For example, atthecommercial farm level.The distinct socioeconomic thesoybean cyst nematode is largely controlled with a worldof MesoAmerican producers is a goalfor future combinationof crop rotation, often the non-host corn, generationsof nematologistsand throughINM techniques andnematode race speciŽ c cultivars.In contrast, Ž elds theyshould develop different INM systemsappropriate to designatedfor plantingwith high cash value crops are theneeds and socioeconomic realities of producers. frequentlyfumigated or treated with a nonfumigant nematicide.Treatment with dichloropropene is a common 026The Mediterranean agrosystem as practicefor manyvegetable crops and fruit crops. anagroecologicalmodel Methylbromide fumigation is used on crops such as strawberrybecause beneŽ ts beyond nematode control, 1; 1 J. Antonio LÓPEZ-PÉREZ ¤, Antonio BELLO , suchas control of numerousother pathogens and weeds, Javier TELLO 2, Avelino GARCÍA-ÁLVAREZ 1 areachieved with a singletreatment. For nematode and Maria ARIAS 1 managementin some perennial tree crops, resistant rootstocksare employed. A casein point is the grafting 1 DptoAgroecologí a, Centro de CienciasMedioambientales, ofpeach scions onto ‘ Nemaguard’rootstocks for areas CSIC,Serrano 115 dpdo, 28006 Madrid, Spain 2 whereroot-knot nematode is a problem.Sanitation is a DptoProducció n Vegetal,Escuela P olitécnica Superior , CITEB II,Univ.Almerí a, Cañada de S. Urbanos/ n,04120 crucialmanagement component for potatoproduction in Almería, Spain thePaciŽ c Northwestin theUSA. Cashvalue of thecrop [email protected] andthe longevity of the crop are major factors in the ¤ selectionof managementtactics. Thenecessity of designing a newagricultural model is proposed,which would take into account the demands ofconsumers,who are concerned not only about greater 025Nematode management practices in foodsafety, but also about the environmental and MesoAmerica socialimpact of agricultural production. New models Nahum MARBAN-MENDOZA ofagriculture are emerging, which use the ecological managementof agricultural systems as a reference, Dptode Parasitología Agrícola, Universidad Autó noma basedon the management of biological, environmental Chapingo,Chapingo, Mé xico andcultural diversity among territories. Environmental andsocial aspects of Mediterranean agriculture are Thecrops of greatest socioeconomic importance in analysedas a diversiŽed agroecological model, which MesoAmerica(southern Mexico and Central America) couldserve as a referencefor thedevelopment of arecoffee, banana, cucurbits and tomato. V arious newproductive systems, integrating proposals from the phytoparasiticnematodes are associated with them but EuropeanCommunity agricultural policy. It is also withoutdoubt the gall inducers ( Meloidogyne spp. and indicatedthat the environmental characteristics of the Nacobbusaberrans ),alongwith nematodes causing root Mediterraneanenvironment permit the design of an lesions (Pratylenchus spp. and Radopholussimilis ), are integratedproduction system based fundamentally on the widelyconsidered by producers to be factors limiting useof non-chemical alternatives for nematodecontrol, productivity.In this talk, we willdiscuss efforts that whichprevent pests and disease from becomingserious arebeing made to attack nematode problems by crop problems.The deŽ nition of ‘ Mediterranean’refers toa andby geographic zone. W ewilltry to distinguish that conceptthat describes a regionwith its own identity, whichis academic from thatwhich has been adopted emergingwith characteristics from aspeciŽc climateand byproducers. In this process, we willtry to put into environmentand high agroecological values produced perspectivethe difŽculties of makingintegrated nematode whenvarious cultures interacted and the outcome of management(INM) arealityin the Ž eld.Based on integrationby the people of the Mediterranean basin. reportsin the literature the following generalisations can Moreover,the contributions must be emphasised of the be made: i)thereare few studies; ii)mostdeal with greatbiotechnologic alrevolution and the impact of the chemicalcontrol; iii)non-chemicalalternatives for the discoveryof Americaon diversiŽcation in theproduction mostpart are based on excellent investigations but have ofhighquality agricultural systems. notbeen adopted by producers; and iv)thereare very few demonstrationstudies on non-chemical alternatives

Vol.4(2), 2002 131 Marineand fresh water nematodes (27-30)

027Re ections on the systematicsof onnematode diversity through macrobenthic predators, the Dorylaimoidea,a groupof terrestrialand predaciousnematodes and the diversity of trophicgroups freshwater nematodes willbe presented.W ediscussthe relevance of theseresults for themaintenance of biodiversity. P.A.A. LOOF Departmentof Nematology, W ageningenUniversity, P .O.Box 029Status quoof limno-nematology: how closeare 8123,6700 ES Wageningen,The Netherlands we tounderstanding the ecologyof nematodesin [email protected] .wau.nl inlandwater bodies? Thesystem of Dorylaimoidea is (some well-deŽ ned Abebe EYUALEM familiesexcepted) unsatisfactory. It is difŽcult to establish groupson thebasisof synapomorphies, the morphological Instituteof Cell, Animal and P opulationBiology, Ashworth charactersbeing distributed erratically, many species Laboratories,King’ s Buildings,W estMains Rd, The University were discoveredand described in the 19th century when ofEdinburgh,Edinburgh EH9 3JT,Scotland,UK knowledgeof the morphologywas almostnil (itremained Despitean early start, limno-nematology can safely be inthis state until 1936); moreover the importance of tail calleda sciencein its infancy compared to marine shape(very easy to observe) has been overrated strongly. nematologyor other aspects of limno-bentholog y.Past Furthermore,the literature is full of misidentiŽcations and limitedinterest in free-living limnetic nematodes has shouldbe consultedvery critically. The author gives some givenway to an upsurge of interest in the past two suggestionsthat in future might lead to developmentof a decades.Most nematological research in limnetic habitats bettersystem. onfree-living forms stillhas a lotto achieve compared tostudies in the marine meiobenthos. Lotic free-living 028Maintenance of biodiversitythrough predation nematodesare relatively less well studied than those from andcompetition in freshwater nematodes? lentichabitats. T axonomicreports continue to appear,but anoticeableproportion of the recently published work Iris VORNDRAN¤ and Walter TRAUNSPURGER was devotedto theirecology, i.e.,mainlyto distribution, spatialand temporal variation, abundance, diversity, and Deptof Animal Ecology, University Bielefeld, Morgenbreede 45,33615 Bielefeld, Germany therelation of these phenomena with some biotic and abioticfactors. Experimental data that revitalises our ¤[email protected] notionof their ecological role has started to appear . Whydoes a particularhabitat harbour a certainnumber Theirpotential use as indicators of water pollutionwas ofspecies and how is this species diversity maintained? alsostrengthened. Hitherto ignored tropical habitats were Theorypredicts that biotic interactions may give rise studied,albeit to a limitedextent. Lentic nematodes toan arms racethat favours variability. Nematodes are seemnot to conform to the general understanding of idealto study ®-diversity.They show high abundance biodiversitygradient with latitude. Gaps are indicated and andspecies richness and play a centralrole in soil recommendationsmade. foodwebs. Species within a genus( e.g., Eumonhystera ) showdifferences in the width of their buccal openings, 030Molecular studies ofnematodediversity: whichcould be interpreted as mutual exclusion through past,present andfuture competition.In this study we reviewthe species richness offreshwater nematodesand ask whether predators and Alex ROGERS 1 andP .JohnD. L AMBSHEAD 2 competitorscan explain species richness in the Ž eld.The 1 numberof freshwater nematodespecies in 28 different BritishAntarctic Survey, High Cross, Madingley Road, lakesranges between three and 120. Within 1 year, ®- CambridgeCB3 0ET,UK 2 TheNatural History Museum, London SW7 5BD, UK diversityremains at a constantlevel within a habitat inLake Brunnsee and Kö nigssee. T ostudythe effect Marinenematodes are an important component of the ofpredators we startedmesocosm experiments with ecosystemand their ecological and biodiversity properties differentmacrobenthic taxa and analysed soil samples of makethem a potentiallyideal taxocene for testing 12alpinelakes. Plathelminthes, chironomids and annelids ecologicaltheory and biomonitoring. W orkat the species donot in uence nematode abundance. The in uence levelwill yield signiŽ cantly greater information than

132 Nematology Symposiumabstracts workat higher taxonomic levels. However, identifying controlmethods. Our dataand analyses are available at samplesof nematodes is difŽ cult, time consuming and http://www.nematodes.org . expensive.In addition, the morphological species concept usedfor classiŽcation may be unreliable, and known 032On the evolutionof parasitismgenes species,especially those used for biomonitoring,may be complexesof siblingswith different functional responses, Geert SMANT especiallywhen animals are examined across wide geographicranges. Molecular methods offer amore Laboratoryfor Nematology, W ageningenUniversity, Binnenhaven5, Wageningen,The Netherlands objectiveestimation of speciesidentity. These techniques [email protected] rangefrom sequencinginformative segments of DNA and usingthem to deŽ ne ‘ molecularoperational taxonomic Inthe last 5 yearsconsiderable effort has been put units’(M-OTU) toassessment of species diversity in intothe identiŽ cation of parasitismgenes from sedentary samplesthrough techniques such as denaturing gradient plant-parasiticnematodes. Thousands of (partial) DNA gelelectrophoresis (DGGE). IdentiŽcation of species in sequencesare now available from severalnematode bulksamples offers arapidand comparative means to species,which allows comparison of gene expression in analysesamples from differentsites, even over large plant-parasiticand other nematode species. From this geographicranges. In the future, genomic technologies comparisonit is evident that plant-parasitic nematodes mayoffer theprospect of developing ‘ identiŽcation haveevolved a setof genes that have no homologues chips’or means of identifying speciŽ c geneexpression inother nematodes or even in other phyla. Similarly, patternsassociated with activities of nematodesrelated to plant-parasiticnematodes have genes in common with environmentalmonitoring. otherplant-pathogens, which are absent in other groups ofnematodes. And Ž nally,plant-parasites and animal parasitesshare a uniqueset of homologues,too. Pieces of 031Comparative nematode genomics thisevolutionary puzzle will be illustratedwith examples. Mark BLAXTER 1, John PARKINSON 1, Jen DAUB 1, Claire WHITTON 1, Marian THOMSON 1, 033Parasitism gene discovery in sedentary David GUILIANO 1, Neil HALL 2, Mike QUAIL 2 phytonematodes and Bart BARRELL 2 Thomas J. BAUM 1 Instituteof Cell, Animal and P opulationBiology, University of Edinburgh,Ashworth Laboratories, Kings Buildings, Departmentof PlantP athology,Iowa State University, Ames, EdinburghEH9 3JTUK IA50011,USA 2 TheSanger Institute, W ellcomeTrust Genome Campus, [email protected] Hinxton,Cambridge, UK Parasitismgenes expressed in phytonematodepharyngeal Thecompletion of thesequencing of the Caenorhabditis glandcells code for thestylet secretions that control the elegans genomein 1998 was amilestonein biology. processof plant parasitism. In sedentary endoparasitic Thegenome sequence has spurred discoveries in basic nematodes,the products of parasitism genes induce the biomedicalscience, but is of added importance to us elaboratemodiŽ cations of plant cells into feeding cells asnematologists because, of course, C. elegans is a requiredto support growth and development of the nematode.W e,and others, are engaged in large scale nematode.Efforts toidentify and characterise parasitism projectswhich aim to gather genomics data from a geneshave been accelerated by several molecular phylogeneticallyand biologically diverse selection of approaches,including direct analyses of secretedproteins nematodes.Due to funding constraints, most of thiseffort andcloning of genes differentially expressed in the isfocussed on parasites of humans and our domestic pharyngealgland cells. For differentialgene expression, animals.This presentation highlights Ž ndingsfrom these comparativeanalyses have been conducted among ongoingnematode genome projects, illustrating topics nematodelife stages and among nematode tissues. The suchas theconservation of syntenyand linkage between mostdirect approach for cloningparasitism genes, C. elegans andother nematodes, the nematode-speciŽ c however,involves the microaspiration of pharyngeal andparasite-speciŽ c geneswe haveidentiŽ ed, and the glandcell cytoplasm from multipleparasitic stages prospectsfor usingcomparative genomics to identify new for cDNA libraryconstruction followed by expressed

Vol.4(2), 2002 133 Parasitismin Nematoda (31-34) and Current trends and future directions in entomopathogenic nematode research (35-38) sequencetag sequencing and bioinformatic analyses. 2 Deptof Plant P athology,University of Arizona,Tucson, AZ Currentresults reveal a largenumber of novelparasitism 85721-0036,USA genesand paint a complexpicture of cellularevents under ¤[email protected] speciŽc controlby thenematode. Furthermore, only very Assimilationof molecular approaches into nematode few parasitismgenes have homologues in Caenorhabditis systematicshas escalated dramatically in the last few elegans,limitingthe usefulnessof thismodel nematode in years.With respect to entomopathogeni cnematodes studyingnematode adaptations for plantparasitism. (EPN), variousmolecular techniques have been used for diagnosticpurposes and for separatingcryptic species, 034Adaptations of parasiticnematodes to populationsand strains. Another valuable contribution to mammalianhost immunity EPNsystematicshas been the discovery and description ofnew species in a phylogeneticcontext. This new Murray E. SELKIRK¤ andKleoniki G OUNARIS approachhas been applied in taxonomic decisions, includingre-evaluation of previously described species Departmentof Biological Sciences, Biochemistry Building, andtheirrelationships. In this presentation we willdiscuss ImperialCollege of Science,T echnology& Medicine,London SW7 2AY, UK theimportance of several molecular markers and their currentuse for diagnosisand identiŽ cation of EPN at ¤[email protected] differenttaxonomic levels. Additionally, we willdiscuss Nematodeparasites of man and animals are typiŽ ed by theapplication of combined evidence datasets ( i.e., theirlongevity in the mammalian host, although little morphological molecular)to interpret phylogenetic C isknown about molecular mechanisms of establishment relationshipsamong EPN speciesand as tools to make andsurvival of theseorganisms in theface of aggressive predictionsof ecologicaltraits that will have signiŽ cance immuneresponses. In addition to a robustarsenal of for theapplication of EPNinbiocontrol programs. anti-oxidants,there is growing evidence that parasite- secretedproducts include a panelof anti-in ammatory agentswhich act in concert to suppress the recruitment 036Advances in molecularbiology of andactivation of effectorleukocytes. These products may entomopathogenicnematodes constituteenzymes which hydrolyse pro-in ammatory Itamar GLAZER signallingmolecules such as chemokines,lipid mediators, andnucleotides released from damagedtissues, or Departmentof Nematology, ARO, TheV olcaniCenter , proteinswhich antagonise receptors involved in this BetDagan, 50250 Israel process.In addition, a numberof cytokine homologues [email protected] areexpressed which have the potential to modulate Entomopathogenicsteinernematidand heterorhabditid immuneresponses. Recent EST projectshave highlighted nematodeshave been used for almosta decadeas abundantlyexpressed genes which encode secreted biologicalcontrol agents of insect pests. Despite the proteinsof unknownfunction, although the lack of genetic impressiveprogress in propagation and application little knockouttechniques has resulted in a bottleneckin hasbeen done, so far, toutilisethe advances in molecular theassessment of these proteins as potential mediators biologytechniques to advance the useof thesenematodes. ofpersistence and the mechanisms involved. RNA Moleculartools have been use mainly for identiŽcation interference(RNAi) offers anopportunityto addresssome andclassiŽ cation of entomopathogen icnematodes. In oftheseproblems, and we areapplying this technique to recentyears, molecular approaches have been employed addressthis speciŽ c question.Progress and current data in tostudy key processes in development and survival of thisarea will be reviewedin thispresentation. steinernematidsand heterorhabditids. These included: i)isolationand characterisation of genesinvolved in the 035Biosystematics of entomopathogenic initiationof developmentfrom theinfective juvenile stage nematodes:current status andfuture directions and ii)usingcDNA subtractivehybridisation to identify genesthat are differentially expressed during exposure to 1; 2 Alex P. REID ¤ andS. PatriciaS TOCK desiccationstress. The advances in the molecular research arereviewed. 1CABI Bioscience(UK Centre),Bakeham Lane, Egham, Surrey TW20 9TY, UK

134 Nematology Symposiumabstracts

037EPN product development:from strain Weareusing microscopic and molecular biological tech- discoveryto commercial Ž eldapplication niquesto investigate the cellular and molecular interac- tionsbetween nematode and bacteria with the follow- Ralf-Udo EHLERS ing goals: i)developinga detailedmodel of nematode- Institutefor Phytopathology, University of Kiel,Dept of bacteriainteractions throughout the course of theirassoci- Biotechnologyand Biological Control, Klausdorfer Str .28-36, ation and ii)deŽning bacterial factors that are important to 24223Raisdorf, Germany thismutualism. Achieving these goals will yield general [email protected] insightinto mechanisms of microbe-hostassociations, as Entomopathogenicnematodes(EPN) havean enormous wellas provide information important to thefurtherdevel- potentialto serve as biocontrol products in protected opmentof theseorganisms as biocontrolagents. Construc- andoutdoor crops. There are several examples of tionof aGFP-labeled X.nematophila strainhas allowed thesuccessful introduction of EPN productsinto the directobservation of bacteria-nematodeassociations, re- ornamentaland turf care market. However, promising sultingin the description of distinctstages of themutual- EPN strainsoften sit on the shelf of highly motivated isticinteraction. Additionally, bacterial mutants have been scientistsand cannot easily be transferred to product generatedthat are either completely or partiallydefective development.After thecontrol potential of a strain inthe ability to colonise nematodes. W earenow char- hasbeen demonstrated under Ž eldconditions, research acterisingthe genetic lesions in thesemutants to identify intobiotechnology has to solve the problem of mass genesand their products that are necessary for colonisa- production,down- and upstream processing, storage and tion. formulation.Quality control procedures are needed to predictŽ eldperformance of production batches. Some 039Nematode management in subsistence farming countriesrequire registration for EPNcausingsigniŽ cant inAfricawith emphasison bananas,plantains and costsand delays in product introduction. Prior tothis yamcropping systems development,information is neededon alternativecontrol 1; 2 strategies,the commercial signiŽ cance of the target pest Roger FOGAIN ¤, Charles KWOSEH and 3 andan approximate cost-beneŽ t analysisfor theuse of Mieke DANEEL EPN. Marketingneeds to motivate potential users to try 1CARBAP (CentreAfricain de Recherchesur bananiers et alternative,environmentally friendly control strategies. Plantains)BP 832,Douala, Cameroon Productintroduction is accompanied by intensive trials 2 Departmentof Crop Science & Technology,Kumasi, Ghana underdifferent production and environmental conditions 3 ITSC,Private Bag X11208, Nelspruit 1200, South Africa inorder to investigate the ecological potential and limits ¤[email protected] oftheEPN strain. Bananas (Musa AAA), plantains( Musa AAB) andfood yamsof the genus Dioscorea playa majorrole in the 038Investigating mutualism between dietof many people in Africa. Thesecrops are also entomopathogenicbacteria and nematodes animportant source of revenues to farmers inrural

† areas.W orldproduction is estimated at about 86 m t Eric MARTENS¤, Eugenio VIVAS, Kurt HEUNGENS , for bananasand plantains and about 34.5 m tfor food Charles COWLES and Heidi GOODRICH-BLAIR yams.Nematodes are among the major biotic constraints Deptof Bacteriology University of Wisconsin-Madison,1550 tothese crops in Africa, causingsigniŽ cant yield losses. LindenDrive, Madison, WI 53706,USA Themajor species attacking bananas and plantains in †Presentaddress: CLO-Gent-DGB, Burg.V anGansberghelaan Africa are Radopholussimilis and Pratylenchus spp. 96,B-9860 Merelbeke, Belgium (P. goodeyi and P. coffeae). Scutellonemabradys, P . ¤[email protected] coffeae andnematodes of thegenus Meloidogyne are the Theentomopathogeni cnematode Steinernemacarpocap- damagingspecies on yams.This report presents progress sae forms amutualisticassociation with the bacterium onthemanagement of thesepests on banana,plantain and Xenorhabdusnematophila .At leastone nematode devel- yamsin subsistence farming systems where these crops opmentalstage, the infectivejuvenile (IJ), iscolonisedat a areoften grown in association with tree crops such as discreteintestinal location by multiplebacteria and is re- coffeeand cocoa and other food crops, including cassava sponsiblefor transportingbacteria between insect hosts. cocoyamsand maize.

Vol.4(2), 2002 135 Nematodemanagement in subsistencefarming (39-42)

040Nematode management in subsistence farming bananaand plantains, fruits and vegetables. From a in Mexico strictnematological viewpoint, all these crops can suffer severelyfrom nematodedamage e.g., Ditylenchusdipsaci, Rosa H. MANZANILLA-LÓPEZ Meloidogyne spp., Pratylenchus spp.,Radopholussimilis, NematodeInteractions Unit, Harpenden, Herts. AL5 2JQ,UK Scutellonemabradys butoften the expertise of farmers, [email protected] theextensive way the lands are cultivated, and the mixed-cropproduction systems let these crops escape Factorssuch as geography, climate, vegetation, biogeog- nematodedamage. Numerous nematological studies have raphyand human settlement have contributed in deŽ n- beenconducted in the Caribbean with different levels of ingMexican rural and socio-economic scenarios. Mexi- knowledge,technical skill and success in orderto improve canagriculture has two main approaches: the campesino nematodemanagement on subsistence crops through andthe agro-industrial, both with contrasting character- fallowand crop rotation, organic soil amendment, use of isticsrelated to production and management strategies resistantcultivars, non-host and antagonistic plants. The againstpests and diseases. In campesino and subsistence experienceof Cuba, Dominican Republic, Guadeloupe, systems,low external-input strategies that have proved ef- Jamaicaand Martinique would be a goodexample of fectivein the management of themost common and im- whatcan be achieved with low inputs within the scope portantplant-pathogeni cspeciesof nematodesin Mexico ofsustainableagriculture. (i.e., Meloidogyne spp., Nacobbusaberrans , Pratylenchus spp. and Punctoderachalcoensis ),includeproduction of 042Nematode management in subsistence farming plantand tree seedlings free ofnematodes,physical con- inBangladesh,Nepal and Pakistan trolmethods, rotations, organic amendments, antagonistic plants,suppressive soils and maintenance of biodiversity. Anjum MUNIR 1,RameshRaj P OKHAREL 2 and Mostof thesepractices are considered to beenvironmen- 3; John BRIDGE ¤ tallyfriendly and, although subsistence agriculture can produceenough food to feed families or smallcommuni- 1 CropDiseases Research Institute, NARC, ParkRoad, ties,it may be adverselyaffected by socio-economicfac- Islamabad-45500,P akistan 2 torssuch as availability of goodland, adequate water, mi- Instituteof Agriculture and Animal Science, Rampur , gration,pressures on land use, shortened rotations, crop Chitwan,Nepal 3CABI Bioscience,UK Centre,Bakeham Lane, Egham, valueand changes in cultural patterns, thus compromis- TW20 9TY, UK ingsustainability. ¤[email protected] Theeconomy of Bangladesh, Nepal and Pakistan is 041Nematode management in subsistence farming agriculturallybased. The main crops of the region are inthe Caribbean wheat,rice, sugarcane, cotton, maize, fruit orchards 1; 2 andvegetables. In Bangladesh, Ditylenchusangustus, E. FERNÁNDEZ ¤, P. QUÉNÉHERVÉ and P. DE LA CRUZ 3 Meloidogynegraminicola , and Hirschmanniellaoryzae areall major pests of rice. Management of ufra is 1Institutode Investigaciones de Sanidad V egetal,Calle 110 bythe burning of stubble. Root-knot can be prevented #514,Miramar ,Habana,Cuba onrice by the transplanting of healthy seedlings and 2 IRD, BP8006,97259, F ort-de-FranceCedex, Martinique suitablerotations. No methods to successfully manage 3 InstitutoDominicano de InvestigacionesAgropecuarias y Hirschmanniella areknown. The important nematodes ForestalesIDIAF ,DominicanRepublic M. [email protected] ofvegetables are root-knot species. In Nepal, ¤ graminicola,H. oryzae and H.mucronata areassociated Thekey characteristics of Caribbeanislands are insularity withrice when followed by wheat, winter legumes, anddiversity: diversity in size, climate, vegetation, cornand summer legumes. In upland systems, where crops,economy and human history. Subsistence farming corn,wheat, different legumes and vegetables are grown, alsore ects the diversity from islandto island. It is differentnematode species are reported. Meloidogyne possibleto look at these differences in considering isreported from almostvegetables, legumes, fruit, someimportant subsistence crops and related nematode ornamentalsand weeds. M.aranaria,M. incognita problems e.g.,smallgrain cereals, roots and tubers, and M. javanica arelimited to Terai (plains) and M.

136 Nematology Symposiumabstracts hapla tothe hills. Tylenchulussemipenetrans is found encodesa mitoticinhibitor responsible for nuclear associatedwith citrus crops causing slow decline. Soil endoreduplication,and ENOD40,whichencodes an early, solarisationand long-term use of farmyard manure nodulationmitogen, suggest a modelin which inputs from showencouraging results, while compost prepared from nematodesand rhizobia elicit a developmentalpathway. manuresof different animal species, host resistance and Thisdevelopmental pathway converges at PHAN and chemicalsin bringal, tomato, gram and pigeon pea are KNOX tomodulate phytohormones, and subsequently usedto control Meloidogynespecies .InPakistan, root- divergesto form giantcells, nodules and meristems. Gene knotis the major problem. Root-knot nematodes are ablationexperiments in tomatofurther showed that PHAN damagingvegetables and M.graminicola isassociated isnecessary for giantcell formation, and is epistatic to withrice and wheat. Potato cyst nematode ( Globodera KNOX.Our recentŽ ndingof a stronghomologue of S. rostochiensis )isrestricted to northern Pakistan. Some melilotinodL inroot-knot nematode suggests similarities successin the management of thisnematode is achieved betweenthe endosymbiontsthemselves, and an intriguing bygrowing turnips, carrots and peas before potato possibilityis that nematode and rhizobia produce similar crop,thereby lowering the nematodes population almost signallingmolecules to suppress host defence responses 70%.Trap cropping is tested and tobacco is reported and/ortoinducenovel plant structures. toinduce signiŽ cant amount of hatch. Other important nematodesare R. similis inbanana and T.semipenetrans 044Syncytium development: the resultof incitrus. In vegetables root-knot is one of the major asophisticatedmanipulation of plantcells by cyst problemin almost all the vegetable growing areas of nematodes Pakistan, M.incognita,M. javanica,M. arenaria are foundin the plains whereas M. hapla is found in 1; 1 Aska GOVERSE ¤, Aneta KARCZMAREK , Magdalena thehills. Most farmers havesmall land holdings and PIERZGALSKA 2, Hein OVERMARS 1, Marcin cannotafford chemical controls. Paecilomyceslilacinus, FILIPECKI 2, Jaap BAKKER 1 andJohannes H ELDER 1 Pasteuriapenetrans, and Verticillium sp.are being tested, 1 aswell as neem products for themanagement of plant- Laboratoryof Nematology, Department of PlantScience, parasiticnematodes, particularly root-knot nematode. WageningenUniversity, Binnenhaven 5, 6709 PD Wageningen, TheNetherlands 2 Departmentof PlantGenetic Breeding and Biotechnology, 043Shared signal transduction pathways lead to WarsawAgricultural University, Nowoursynowska 166, nematodeparasitism, rhizobial symbiosis and PL-02-787W arsaw,Poland programmedplant development ¤[email protected] Potatocyst nematodes have evolved a sophisticatedway Hinanit KOLTAI1; , Jennifer SCHAFF 2 and David BIRD 2 ¤ toparasitisetheir host plants. They migrate intracellularly 1Departmentof Genomics,A.R.O. VolcaniCenter ,BetDagan throughthe root, and an outercortex cell is selectedas a 50250,Israel startingpoint for syncytiumformation. The recruitment 2 Departmentof Plant P athology,North Carolina State ofplant cell wall-degrading enzymes by the nematode University,Raleigh 27695, NC, USA resultsin an expansion of the syncytium towards the [email protected] ¤ vascularbundle via aso-calledcortical bridge. A Duringthe compatible parasitic interaction, root-knot localaccumulation of auxin is crucial for syncytium nematodes (Meloidogyneincognita )facemany challenges development.Analysis of cellulase expression patterns in tosuccessfully infect their host, including avoiding tomatorevealed that two auxin-inducible members ( LE- hostdefences, and redirecting host gene expression Cel7 and LE-Cel8)were speciŽcally up-regulated during toestablish the complex structures of giant cells, theonset of syncytiumdevelopment. Intriguingly, nodule centralto the parasiticrelationship. Conceivably, rhizobial formationin legumes starts in the cortex, proliferates bacteria (Sinorhizobiummeliloti )symbiontsface similar towardsthe stele and is also accompanied by a local challengesduring establishment of nitrogen Ž xing accumulationof auxin. This process is preceded by the nodules.Our Žndingsof similar pattern of gene expressionof ENOD40.Toseewhether this gene is expressionin giant cells and nodules of twotranscription alsoessential for syncytiumdevelopment, the role of regulatorsrequired for theestablishment of shoot and ENOD40 was investigatedin thenon-legumestomato and rootmeristems (PHAN andKNOX) of ccs52, which Arabidopsis uponcyst nematode infection. T ounravelthe

Vol.4(2), 2002 137 Compatibleparasitic interactions (43-46) and Resistance breeding against sedentary nematodes (47-52) molecularmechanisms that underlie syncytium induction Our resultsshow that endoreduplicatio nandmitotic morethoroughly, a highthroughput approach is needed. cyclesplay a roleduring the root cell de-differentiation cDNA-AFLP was usedto monitor the expression of processcaused by nematodeinfection. auxin-regulatedgenes in cyst nematode-infected tomato roots,and some preliminary results will be presented. 046Characterisation of plantgenes involved in giantcell formation induced byroot-knot nematodes 045Cell cycle genes as markers to study the in Arabidopsisthaliana and Medicagotruncatula ontogenyof nematodefeeding sites in plantroots 1; 1 Bruno FAVERY ¤, Fabien JAMMES , Philippe 1; 1;2 1 2 Janice DE ALMEIDA-ENGLER ¤, Gilbert ENGLER LECOMTE , Arnaud COMPLAINVILLE , Jose Maria andGodelieve G HEYSEN 1;3 VINARDELL 2, Danièle VAUBERT 2, Peter MERGAERT 2, Adam KONDOROSI 2, Eva KONDOROSI 2, Martin 1Departmentsof Molecular Genetics and Plant Genetics, CRESPI 2 and Pierre ABAD 1 FlandersInteruniversity Institute for Biotechnology (VIB), GhentUniversity, K.L. Ledeganckstraat35, B-9000 Gent, 1UnitéInteractions Plantes-Microorganismes et Santé Belgium Végé tale, INRA, 123,Bld F .Meilland,BP 2078,06606 Antibes 2 LaboratoireAssocié de l’ Institut National de laRecherche Cedex,France Agronomique(France), Ghent University, B-9000 Gent, 2 Institutdes Sciences Vé gé tales, CNRS, Avenuede la Terrasse, Belgium 91198Gif sur Yvette, France 3 VakgroepMoleculaire Biotechnologie, F aculteit ¤[email protected] Landbouwkundigeen ToegepasteBiologische W etenschappen, UniversiteitGent, Coupure links 653, B-9000 Gent, Belgium Toidentifyplant genes involved in giant cell formation ¤[email protected] inducedby the root-knot nematode Meloidogyneinco- gnita,apromotertrapping strategy was developedin Ara- Researchon the ontogeny of feeding sites induced by bidopsisthaliana .WeidentiŽed a gene, NEC, early up- sedentaryplant endoparasitic nematodes has been in regulatedin the giant cells. During root development, this focusalready for manydecades. Nematodes induce root geneis expressed in meristems including initiation sites cellsto de-differentiate into large multinucleated and oflateralroots. This gene encodes a calcium-bindingpro- cytoplasm-densefeeding cells. W ehaveused Arabidopsis teinwith an EF-hand motif. In order to investigate its thaliana asa modelhost to study cell cycle progression function,analysis of homozygous nec plantsand plants innematode feeding sites induced by the root-knot over-expressingsense or anti-sensegene were carriedout. nematode Meloidogyneincognita andthe cyst nematode The role of NEC insignal transduction will be discussed. Heteroderaschachtii .Cellcycle markers were usedto Secondly,we investigatedwhether nitrogen-Ž xing nod- monitorhow nematode feeding sites develop and what ulesand giant cell formation induce distinct or overlap- theinitial signals are that trigger certain root cells to pingregulatory pathways in Medicagotruncatula . We evolveinto giant cells embedded in a gall,or into a studiedexpression patterns of nodule-expressedgenes af- syncytium.Tritiated thymidine incorporation experiments terinfection with M.incognita. Analysesof promoter- were appliedto monitor DNA synthesis,which was used GUS fusionsindicate an accumulation of the cell cycle asa markerfor theS phaseof the cell cycle. Extending gene CCS52a ingiant cells while the early nodulin gene ouranalysis, the expression pattern of two CDKs and ENOD40 expressionwas detectedonly in cellssurround- twomitotic cyclins were examined.The cycB1;1 gene ingthegiant cells. Interestingly, on transgenicplants over- was usedas amarkerfor theG2 phase, the cdc2bAt and expressing enod40,asigniŽcantly higher number of galls cycA2;1 for theS andG2 phases whereas the cdc2a gene was formed.In addition, results obtained on macroarray was usedas amarkerfor allcell cycle phases. Nematode analysiswill be discussed. T akentogether, these results infectedseedlings were alsotreated with two cell cycle suggestthat certain events might be commonbetween gi- inhibitors(hydroxy-urea and oryzalin) to investigate antcell formation and nodule development. therelevance of DNA synthesisand mitosis on the developmentof thesefeeding sites. A strongcorrelation was observedbetween initiation of feedingcells by both 047Molecular markers for resistance to cyst root-knotand cyst nematodes and the induction of DNA nematodesin potato synthesisand theexpressionof particularcell cycle genes.

138 Nematology Symposiumabstracts

1; 1 Erin BAKKER ¤, Jeroen ROUPPEVANDER VOORT , cultivars,but in non-infested Ž eldsyield of COAN was Herman VAN ECK 2, Jaap BAKKER 1 and only80-90% of the susceptible cultivars. The cultivar Aska GOVERSE 1 NemaTAMwas releasedfrom theseventh backcross generationand has yields equal to that of susceptible 1Laboratoryof Nematology, W ageningenUniversity, cultivarsin non-infested Ž elds.In Ž eldsinfested with M. Binnenhaven5, 6709 PD Wageningen,The Netherlands 2 Lab.Plant Breeding, W ageningenUniversity, The Netherlands arenaria,yieldsof NemaT AMaregreater than those of COAN. Resistancein COAN andNemaT AMisinherited ¤[email protected] asasingledominant gene. The resistance in the synthetic Amappingstrategy based on catalogued, chromosome- triplespecies hybrid is inherited as one dominantgene and speciŽc AFLP markersfacilitated the localisation of onerecessivegene. Thus, at leastone additionalresistance genesfor resistanceagainst potato cyst nematodes. geneappears to existin agenotypecross-compatible with The genes Gpa2, Grp1, Gpa3, Gpa5 and Gpa6 could cultivatedgroundnut that can be used to increase the relativelyeasily be mapped with this AFLP catalogue. durabilityof resistance. Finemapping showed that most of these genes are locatedin genomic regions harbouring also resistance 049Global importance of cyst( Heterodera spp.) and genesfor otherpathogens. Especially, Grp1 and Gpa5 lesionnematodes ( Pratylenchus spp.) oncereals: onchromosome V and Gpa3 onchromosome 11 seem areviewof yieldloss studies, populationsdynamics tobe located in ‘ hotspots’for resistance.Remarkably, andprogress of the use ofhost resistancefor bothqualitative and quantitative resistances map to these nematodecontrol using traditional methods with resistancegene clusters indicating that they might be the applicationmolecular tools controlledby similar molecular mechanisms. Dissecting resistancein potatowill in the near future be accelerated 1; 2 3 Julie NICOL ¤, Roger RIVOAL , Sharyn TAYLOR and bythe availability of an ultra dense genetic map. Using Maria ZAHARIEVA 4 anoffspring of about100individuals,approximate 10 000 AFLP markershave beenplaced on the mapof potato.The 1InternationalWheat and Maize Improvement Center possibilitiesfor marker-assistedbreeding in potato will be (CIMMYT), WheatProgram, P .O.Box 39, Emek, 06511, discussed. Ankara,Turkey 2 UMR INRA/ENSAR, Biologiedes Organismes et des Populationsappliqué e àlaProtectiondes Plantes (BiO3P), 048The identiŽ cation and development of BP35327,35653 Le Rheu,France resistanceto Meloidogynearenaria ingroundnut 3 SouthAustralian Research and Development Institute (SARDI),Field Crops P athologyUnit, GPO Box397 Adelaide, 1; 2 2 J.L. STARR ¤, G. CHURCH and C.E. SIMPSON SA5001,Australia 4 1Departmentof Plant P athology& Microbiology,T exasA&M CIMMYT,WheatProgram, A.P .6-641,06600 Mexico D.F ., University,College Station, TX 77843,USA Mexico 2 TexasAgricultural Experiment Station, Stephenville, TX ¤[email protected] 76401, USA Heterodera [email protected] Thesedentary cereal cyst nematode (CCN, ¤ spp.)and the migratory endoparasitic root lesion nema- Highlyeffective resistance to M. arenaria ingroundnut tode (RLN, Pratylenchus spp.)have a globaldistribution was Žrst discoveredin diploid wild Arachis spp. andare economically important to cereal production sys- Developmentof asynthetictetraploid interspeciŽ c hybrid temsworldwide. Several species of CCNandRLN cause (A.batizocoi (A. diogoi A.cardenasii )) that yieldloss on cereals;however, morphologically these ge- was cross-compatible£ with cultivated £ groundnut was neraare difŽ cult to identify at the species level. The po- theŽ rst steptowards achieving a resistantcultivar. A pulationdynamics and plant damage thresholds in rela- backcrossbreeding programme was usedto introgress tionto yieldloss will be discussedand compared for both theresistance into agronomically acceptable groundnut typesof nematodes.Resistance is one of themajor con- genotypes.The Ž rst cultivar(COAN) releasedfrom this trolmeasures and offers thepossibility to reducepopula- breedingprogramme was selectedfrom theŽ fthbackcross tionsof nematodesbelow damaging thresholds. Through generation.Yields of COAN inŽ eldsinfested with M. extensivescreening work, several sources of plantresis- arenaria were 20-120%greater than those of susceptible tanceat various levels have now been identiŽ ed for both

Vol.4(2), 2002 139 Compatibleparasitic interactions (43-46) and Resistance breeding against sedentary nematodes (47-52)

CCNandRLN from landracematerials and conventional Instituteof Grasslandand Environmental Research, germplasm.CCN hasbeen more extensively studied and Aberystwyth,Ceredigion mostof theresistant sources currently identiŽ ed possess SY233EB, Wales,UK singledominant genes. The introgression of thismaterial [email protected] intowheat breeding programmes will be discussed, par- Evidencefrom threecyst nematode crop pests (of small ticularlywith reference to the application of molecular graincereals, Heterodera spp.;soybeans, H. glycines and tools.With further understanding of thedistribution and potatoes, Globodera spp.)will be used to emphasise the importanceof bothCCN andRLN, the usability of these signiŽcance of therelationship between the zone of plant resistantsources should reduce losses caused by thesene- nematodeco-evolution and the cropping areas in which matodes. resistanceis used. The impacts of these relationships ondurability of resistance provide important lessons for 050Resistance in soybean to soybean cyst thetwo approaches likely to be of value in enhancing nematode, Heteroderaglycines durabilityin agriculture. These are either seeking to imitatesustainable natural metapopulation dynamics or Greg NOEL anintensive on-going management of gene deployment. Bothare technically feasible with modern plant breeding UnitedStates Department of Agriculture,Agriculture Research technologies,but the latter requires substantial continuing Service,Department of CropSciences, University of Illinois, R&Dinvestments,only likely to bejustiŽable in intensive Urbana,IL 61801,USA agriculturalsystems where nematodes cause prominent [email protected] andeconomically signiŽ cant crop loss. This latter situationis likely to be more widespread in relation to Soybeancyst nematode (SCN), Heteroderaglycines , was root-knotnematodes, and an attempt will be made to foundin the USA in1954, and breeding programs considerhow the biology and co-evolutionary histories soonwere initiated.The source of resistance in the ofthese nematodes may indicate approaches to use and Žrst resistantcultivars was Peking,which was obtained managementof resistanceso as toenhanceits durability. inChina. Subsequent sources of resistance incorporated intocommercial cultivars include, PI88.788, PI90.763, 052Breeding for durable resistance to RKN in PI209.332,all of which also originated in China. The perennials:a Europeaninitiative for Prunus highlevel of genetic variability within and among rootstocks populationsof SCN hasproved problematic to soybean breedersand to farmers. In1991, germplasm was Daniel ESMENJAUD releasedthat incorporated resistance from PI437.654, INRA, UnitéInteractions Plantes-Micro-organismes et Santé whichwas collectedin the Amur regionof Russia and Végé tale, B.P .2078,06 606Antibes cedex, France highlyresistant to most populations of SCN. Classical [email protected] breedingtechniques were usedexclusively until the late 1990swhen marker-assisted selection using PI437.654 Breedingfor durableresistance in Prunus rootstocksis a resistancewas developedand is now being utilised in challengingtask since the long generation time increases severalbreeding programs. Genes, rhg1 – rhg3, and Rhg3 therisk for resistancebreaking. The breeding strategy is – Rhg5,havebeen reported to beinvolvedin expressing basedon the selection and pyramiding of major genes resistancein soybeanto SCN. Somegenes are linked and conferringwhen possible a high-level,wide-spectrum and multiplealleles may be involvedin expressing resistance. heat-stableresistance. Screening among Prunus species Monocultureof resistance has not proven durable, but andaccessions for appropriateresistance sources was genedeployment may offer long-termsustainable control performedthrough a Žrst EUprojectfrom 1993to 1997. of SCN. SomeMyrobalan (M) plumaccessions resisted all RKN species,even under durable inoculum pressure and high 051Sustainability and mechanisms for durable use temperatures,due to single major genes with a complete ofresistance to control sedentary nematodes spectrum,termed Ma,recentlyconsidered as allelic. By contrast,peach (P) andalmond (A) sourcesexpressed Roger COOK near-completeor incomplete spectra. In a secondEU

140 Nematology Symposiumabstracts project(1999-2003), breeding for anewgeneration of 054Effect ofOxycomon growthof tomatoand interspeciŽc rootstocks,of the type M P, M A and reproductionof Meloidogyneincognita £ £ M (P A), basedon thepyramiding of one Ma allele £ £ andR genesfrom peachor almond is being performed Safdar A. ANWAR¤ and M.V. MCKENRY Prunus for all species.In order to develop marker-assisted Universityof California,Riverside, CA 92521,USA selection(MAS), locationof the R genesusing SSR ¤[email protected] markersfrom aEuropeanreference Prunus map and a BSA positionalapproach are carried out. Ma (LG7) and Singleor multipleapplications of Oxycom,peroxyacetic atleast one peach gene (LG2) areindependent. Several acidplus various biocontrol agents, were comparedon 15- SCARor AFLP markerslocated less than 0.5 cM or day-oldsusceptible tomato inoculated with 1500 second cosegregatingwith the Ma1 alleleare available for MAS stagejuvenile (J2) M.incognita. Fortypots were dipped ofall interspeciŽ c crosses. into2500 ppm (v/ v)Oxycom,and 20 inwater .Twentyof the40 potsreceived four additional Oxycom applications at10-day intervals. Plants were harvestedat 60 days 053Molecular phylogenies of plantand afterinoculation to assess the effect of treatments on entomoparasiticnematodes: congruence and plantgrowth, nematode development and reproduction. incongruencewith morphologicaland biological data Asingletreatment of Oxycom just prior to nematode inoculationsigniŽ cantly increased tomato top weight Sergei SUBBOTIN1; , Dieter STURHAN 2 and ¤ butnot other growth parameters. Associated with plant Maurice MOENS 3 growthbeneŽ t was asigniŽcant increase in the number ofroot-knot females. Multiple treatments of Oxycom 1Instituteof Parasitologyof RAS, Leninskiiprospect 33, Moscow,117071,Russia signiŽcantly reduced leaf area, top weightand root weight 2 BiologischeBundesanstalt, Institut fü r Nematologieund whilesigniŽ cantly increasing the number of galls,females Wirbeltierkunde,T oppheideweg88, 48161, Mü nster ,Germany andJ2 per plant or per g root.This study demonstrates 3 CropProtection Department, Agricultural Research Centre, thatOxycom stimulates plant growth and earlier fruiting Burg.V anGansberghelaan96, 9820 Merelbeke, Belgium whilehastening nematode development. Proper timing ¤[email protected] andfrequency of applicationsis important. No treatment reducednematode population levels in this60-day trial. Oneof the exciting developments in nematology during thelast years has been the application of nucleic acid 055A new systemfor Nematoda: combining dataanalysis to problems of systematicsof certaingroups. morphologicalcharacters with moleculartrees, and Inmany cases, molecular data conŽ rm themonophyly translatingclades into ranks and taxa ofspecies groups recognised by morphological studies. 1; 2 Inother cases, the molecular data reveal taxa of which Paul DE LEY ¤ and Mark BLAXTER thephylogenetic relationship based on the morphology 1 seemsto be problematic. Molecular data put more light Departmentof Nematology, University of California – onproblems of species boundaries and co-evolution of Riverside,Riverside, CA 92521,USA 2 nematodeswith their hosts. The early presumption that Instituteof Cell,Animal and P opulationBiology, University of Edinburgh,Ashworth Laboratories, Kings Buildings, phylogenetictrees based on molecular data would more EdinburghEH9 3JT,Scotland,UK preciselyre ect the true phylogeny than morphological ¤[email protected] dataseems to be wrong. Analyses of in uence of alignmentor tree building methods on phylogenies Werecentlyproposed a newclassiŽ cation for thephylum showedthat molecular data could be subjected to many Nematoda,based primarily on phylogenetic hypotheses problems.Congruence and incongruence of results of resultingfrom smallsubunit rDNA sequenceanalysis. analysesof molecular and morphological data sets Thestrength and weaknesses of this new system are andthe possible reasons for conicts are discussed discussedwith particular regards to ranks and names of withexamples of plant-parasitic nematodes from the nematodeclades. As almost2 yearshave passed since we familyHeteroderidae, Anguinidae, Longidoridae, and Žnishedwriting the original manuscript, we alsoprovide entomoparasiticnematodes of the T ylenchidaand the atimelyupdate on the implications of phylogenetic familiesSteinernematidae and Heterorhabditidae. analysesconducted in the past 2 years,and improve

Vol.4(2), 2002 141 QuarantineNematodes (56-58) and Organic amendments for the management of plantparasitic nematodes (59-65) orcorrect our system where appropriate. W eemphasise sourcesfor curationhave led to concerns for thewhole theimportance of disentangling the earliest branches of basisof the quarantine identiŽ cation, despite the recent thenematode tree, and discuss various scenarios with developmentof molecular techniques which have been sometantalising implications for ourunderstandingof the hailedas the resolution to this problem. However, there morphologyand ecology of theearliest nematodes. remainsa lackof robust,reliable methods using DNA for theimmediate distinction of listed species from alloth- ers thatmight reasonably occur in world trade. Increasing 056Measures to avoidnew introductionof demandsfor qualityaccreditation procedures and the re- quarantinednematodes into Brazil and the spreadof Žnementof computer-basedimagery are stimulating new non-regulatedquarantine nematodes waysof providingevidence of statutoryŽ ndings,and may OdilsonLuiz R IBEIRO E SILVA proveto be the salvation of many national collections, turningmany into a world-wideresource. NationalPlant Protection Organization of Brazil,MAP A/SDA, AnnexoB, Sala307, CEP 70043-900Brasilia, DF ,Brazil [email protected] 058Quarantine nematodes or invasivealien species?

TheBrazilian National Plant Protection Organization is David MCNAMARA issuinga numberof new actions and regulations to avoidintroductions of quarantinepests in thecountry and Europeanand Mediterranean Plant Protection Organization, amongthem quarantinenematodes. A newquarantine pest Paris,France listbased on technicaljustiŽ cation and pest risk analysis [email protected] isavailable; an integrated electronic system to detect TheConvention on Biodiversity (CBD) hasrecently pestsis starting to be used in the main points of entry beendeveloped as a globalagreement with the overall inthe country; new laboratories, quarantine stations and aimof ensuring the maintenance of biological diversity centresfor conductingpest risk analysis are in theprocess throughoutthe world. One of themain thrusts of theCBD ofaccreditation to facilitate inspections at the points of istopreventthe transfer, by humanactivity, of speciesof entry.All the 27 ruleswith general requirements to import plantsor animalsfrom onepart of the world to another. commoditiesfrom allcountries are in theprocess of being Speciesthat are transported (or transportable)are referred replacedby measures that will be based on pest risk toas ‘ invasivealien species’ and can be any type of analysisrelated to the origin and to the product. Besides plantor animal species that could disrupt ecosystems, that,an expert group has been working for morethan 3 habitatsor species. As such,the concept of invasive yearsto determine the tolerance levels for non-quarantine speciesoverlaps with the long-established ‘ quarantine regulatedpests for plantsfor planting,and new regulations pest’. Plantquarantine services in many countries are onthatmatter will be inforcesoon. reactingto thesedevelopments by expandingthe range of organismsfor whichthey have responsibility, beyond the 057Dilemmas for the quarantinediagnostician and pestsof economicallyimportant crops. The consequences taxonomistin the 21 st Century for nematologywill be discussed and could include the needfor studyof nematode pests of wild plants, the Sue HOCKLAND ecologyof non-plant-parasitic nematodes and the effects InvertebrateIdentiŽ cation T eam,Plant Health Group, Central ofbiocontrolnematodes on naturalecosystems. ScienceLaboratory, Sand Hutton, Y ork,YO41 1LZ, UK [email protected] 059Organic amendments for the managementof Duringthe last century, many nematologists were in- phytonematodes:enhancing natural biological control volvedin the golden era of identiŽcation and many type 1; 2 specimenswere storedin key institutes to serve as ref- José A. CHAVARRÍA-CARVAJAL ¤ , Edwin ACEVEDO , Carlos FLORES 3, Nidia VYCENTE 1, erencepoints for futurediagnosticians and taxonomists. 1 1 Thebasis of identiŽ cation was theability to distinguish Luis SILVA-NEGRÓN and Evelyn ROSA anewspecies from allothers that had preceded it by 1Departmentof Crop Protection, Mayagü ez Campus, recognisingsigniŽ cant variation of criticalmorphological Universityof PuertoRico, P .O.Box9030, Mayagü ez, characters.The demise of suchskills and the lack of re- 00681-9030,Puerto Rico

142 Nematology Symposiumabstracts

2 Departmentof Agronomy and Soils, Isabela Substation, andthe method of application. Cost of transportation is Isabela,00662, Puerto Rico shownas its principal limiting factor. Differences are 3 Departmentof Horticulture,Isabela Substation, Isabela, establishedbetween it andother control alternatives such 00662,Puerto Rico assolarisation,  oodingand biosterilisation, although ¤[email protected] theycan be consideredcomplementary. The effectiveness Organicamendments are useful in plantdisease manage- ofbiofumigation increases with time, when included mentbecause they suppress pathogen populations through inan integrated production system, which should take productionof toxicsubstances during decomposition, and intoaccount the speciŽ c characteristicsof areas with a byalterations on soil microbial ecology. However, little Mediterraneanclimate. It is concludedthat biofumigation informationis available on the mode of action of these isacontrolalternative based on theuse of localresources materials,especially how they affect activities and popu- andthat it reducesenvironmental impact from agriculture lationsof soil microorganisms. Interactions between an- andincreases the quality of agriculturalproduction. tagonisticmicroorganisms and plant pathogens are wide- spreadin nature and organic amendments may be useful 061Soil solarisation, nematicide and chicken toenhance naturally occurring biological control and re- compostfor root-knot nematode control in carrot ducediseases caused by nematodes. Experiments were cropsin Brazil conductedduring the last 5 yearsin Puerto Rico that 1; 1 showedthe value of organic matter to suppress nema- ClaudioM.G. O LIVEIRA ¤,RobertoK. K UBO , todepopulations under tropical conditions. Changes in Mario M. INOMOTO 2,Flávia R.A. P ATRÍCIO 1, populationsof microorganismsassociated with use of or- Osvaldo CABRAL 3 andBenedito C. B ARROS 1 ganicmatter were studied.The use of organic amend- mentsimproved the activity of microorganisms associated 1InstitutoBioló gico, Caixa P ostal70, Campinas, SP , withthe rhizosphere of tropicalcrops. W earefocusing on 13001-970,Brazil 2 theexploitation of microorganisms promoted by organic Universidadede São Paulo,Escola Superior de Agricultura ‘Luizde Queiroz’ , CaixaP ostal09, Piracicaba, SP ,13418-900, amendmentsas sources of biological control agents for Brazil phytonematodes.This approach may be apracticaltool to 3 EMBRAPAMeioAmbiente, Caixa P ostal69, Jaguariú na, SP , reducethe dependenceon syntheticpesticides and tomiti- 13820-000,Brazil gatethe environmental contamination produced by thein- ¤[email protected] tensiveuse of chemicals. AŽeldexperiment was performedin sandy soil, naturally infestedwith Meloidogyneincognita and M.javanica, 060Biofumigation and nematode control in the inPiracicaba, SP ,Brazil,to assess the effectiveness of Mediterraneanregion soilsolarisation, chicken compost (20 t/ ha),carbofuran 100G(40 kg/ ha),and combinations of these treatments Antonio BELLO¤,J.AntonioL ÓPEZ-PÉREZ, Avelino GARCÍA-ÁLVAREZ and Rafael SANZ for thecontrol of root-knotnematodes in carrotcv. Aline. Soilsolarisation was performedover 69 days during DptoAgroecologia, Centro de CienciasMedioambientales, thewarmest monthsof summer of 2000/ 2001,using a CSIC,Serrano 115 dpdo, 28006 Madrid, Spain 100 ¹mpolyethyleneplastic Ž lmas soil covering. The ¤[email protected] soiltemperature in solarised plots was 8-12 ±C higher Biofumigationis deŽ ned as the effect from thegases thanin the non-solarised. Maximum soil temperatures producedduring the biodecompositio noforganicmatter were recordedin solarised soil combined with chicken for controllingplant pathogens. Biofumigant action is compostand reached 77, 56 and 46 ±C,atdepths of 5, indicatedas a generalproperty of organic matter, of 10and 20 cm. Population densities of M.incognita and animalmanure as wellas green manure or agroindustrial M. javanica were controlledby both soil solarisation remains.Its effectivenessis similar to conventional aloneand combined with chicken compost or carbofuran. fumigants,at the same time as it improvessoil chemical, Carrotcv. Aline yield was signiŽcantly increased by physicaland biological characteristics. The scientiŽ c combinationof solarisation and chicken compost plus basesfor biofumigationare established, differentiating nematicideapplication before planting. itfrom theuse of organic matter as an amendment, withregard to the characteristics of the biofumigant

Vol.4(2), 2002 143 QuarantineNematodes (56-58) and Organic amendments for the management of plantparasitic nematodes (59-65)

062Green manure amendments and management ¤[email protected] ofroot-knotnematode on potatoin the PaciŽc Northwest ofUSA Amendingsoil with paper mill residuals is a strategyto improvesoil fertility and minimise the loss of nutrients Ekaterini RIGA 1, Hassan MOJTAHEDI 1, andpesticides. W einitiatedan experiment in 1998 to Russell E. INGHAM 2 andAndrew M. M CGUIRE 3 studyshort- and long-term changes in the chemical, physicaland biological properties of soil after adding 1 WashingtonState University, IAREC, 24106N. BunnRd., papermill residual amendments to the plots every spring. Prosser,WA 99350,USA Thetreatments were tworates of fresh papermill 2 Departmentof Botany and Plant P athology,Oregon State residuals,paper mill residuals composted alone, paper University,Corvallis, OR 97331,USA 3 WashingtonState University Cooperative Extension, P .O.Box millresiduals composted with bark, and a non-amended 37,Ephrata, WA 98823,USA control.The experiment, a randomisedcomplete block ¤[email protected] withŽ vereplicationsof each amendment treatment, was plantedwith potato in 1998, snap bean in 1999,cucumber Meloidogynechitwoodi, aseriouspest of potatoesin the in2000, and potato in 2001. Population densities of PaciŽc Northwest(PNW) ofthe USA, blemishespotato Pratylenchuspenetrans were 88nematodesper 100 cm 3 tubersand renders them unmarketable. Currently, this soilat the start of theexperiment. Root lesion population nematodeis effectivelycontrolled by anenvironmentally densitiesdeclined in the control plots and stayed the hazardousand costly fumigant, Telone II. As an sameor increased in amended plots from 1998to 2001. alternativeto fumigation, green manure cover crops InNovember 2001 P.penetrans was mostabundant in includingrapeseed, white mustard, and sudangrass, and the‘ highrate-composted alone’ treatment. Incidence of theorganic amendments canola seed meal, and milkweed symptomsindicative of the potato early dying disease seedmeal, have been used to control M.chitwoodi in the were alsomost severe for thistreatment. Amended plots PNW.Greenmanures and seed meals alone have reduced hada highercapacity for waterretention and increased thenematode impact on potatoesby 50-80%.In addition, organicmatter . theabove amendments have been used successfully in combinationwith contact nematicides, such as ethoprop, andprovided control against M.chitwoodi at a level 064Nonchemical control of root-knotnematodes comparableto soil fumigation. Our studiesdemonstrated (Meloidogynecomplex )inglasshouseconditions and thatthe use of green manures together with 2 yearsof Xiphinemaindex inthe vineyard croprotation controlled M.chitwoodi toacceptablelevels 1; 1 Boryana CHOLEVA ¤, Emil MOLLE and withoutthe use of a nematicide.However, studies from 2 otherregions of the USA haveshown that theamendments Jordan TZVETKOV failedto control other species of root-knot nematodes. 1Facultyof Biology, University of SoŽa ‘St.Kliment Ochridski’ , Thesecon icting results indicate that amendments should 8DraganTzankov blvd., 1164 SoŽ a, Bulgaria beutilised according to local environmental conditions. 2 ExperimentalStation, Septemvri, Bulgaria Managementwith green manures may be moresuccessful ¤[email protected]Ža.bg inshort or coolgrowing seasons where nematode damage Agroupof scientistsfrom theuniversity of SoŽa andthe potentialis lower .Inaddition, we foundthat green ExperimentalStation in Septemvriinvestigated more than manuresimprove soil physical characteristics, especially 10yearsplan residues and manufacturing industries waste waterinŽ ltration and penetration resistance. productsas means to avoid agrochemical for controlof soilpathogens (nematodes). The last 3 yearswe continued 063Impact of papermill residual amendments on theexperiments with substrates and extracts from waste Pratylenchuspenetrans ina vegetablerotation productsof microbialindustry (Biovet factory – Peshtera). Theinvestigated products are obtained on the base of an A. MACGUIDWIN¤, A. REID-RICE, L. COOPERBAND originalrecycling technology. The results showed a very and D. ROTENBERG favorableeffect and advantages for Meloidogynecomplex Departmentof Plant P athology,University of Wisconsin, asa signiŽcant reduction of L 2,alaterand weaker Madison,WI, USA;Department of Soil Science, University of infectionof the tomato and the cucumber roots and the Wisconsin,Madison, WI, USA totalyield increased up to62%.These substrates reduced

144 Nematology Symposiumabstracts verystrongly the population density of Xiphinemaindex [email protected] andstimulated several time the plant crowing and the yield.The investigated products pointed to the new Wolbachia bacteriaappear to have evolved a mutualistic possibilitiesof the nonchemical control of the parasitic symbiosiswith their nematode hosts. This is in contrast nematodes. tothe closely related Wolbachia endosymbiontsof arthropods,which display a diverserange of associations. Withinthe Nematoda, Wolbachia appearto be restricted 065Organic amendments as therapeutictreatment toŽ larialparasites, and molecular phylogenetic analysis ofguava trees ( Psidiumguajava L.)infested with suggestsnematodes probably acquired their symbionts Meloidogyneincognita inZulia State, V enezuela dueto their intimate association with their arthropod vectorssome 100 million years ago. Since then, however, Ana Maria CASASSA-PADRÓN 1;2, Evelyn thereis no evidence for recenthorizontal transmission PÉREZ-PÉREZ, Merylin MARÍN 2, Casilda GONZÁLEZ, betweennematodes and arthropods. Wolbachiahave so far Dubia CHIRINOS and Luis SANDOVAL 1 beenfound in morethan 20 speciesof Žlarialnematode, 1 Universidaddel Zulia, F acultadde Agronomí a, Institutode withonly two species appearing to be uninfected. InvestigacionesAgronó micas, Maracaibo, Apto 15205, Of thosewith Wolbachia, theinfection appears to be Venezuela ubiquitousin all individuals, developmental stages and 2 Departamentode Botánica, Ciudad Universitaria, Av .Ziruma, populationsthroughout their global distribution. The Maracaibo,Apto 15205, V enezuela bacteriareside in vacuolesand are restricted to the lateral [email protected] ¤ cordcells and developmental stages within the female Thebiofumigant effect of organic amendments as reproductiveorgans and intrauterinedevelopmental stages atherapeutictreatment on 6-year-old guava trees asa consequenceof their vertical transmission via the infestedwith root-knot nematode was evaluatedat the egg.As Žlarialnematodes are major pathogens of humans CentroFrutí cola del Zulia-CORPOZULIA. Treatments throughoutthe tropics, our research has focused on the includedcontrol (T1), 30 kg (T2), 60 kg (T3) of contributionof Wolbachia todiseasepathogenesis and as goatmanure per tree, 30 kg (T4) and60 kg (T5) anoveltarget for antibiotictherapy. These studies show ofcomposted sugarcane bagasse, and combinations of that Wolbachia endotoxin-likeactivity is the principal 15kg (T6) and30 kg (T7) ofeach, applied every 3 causeof inammatory Ž larialdisease. Antibiotic therapy months.Results after 10 monthsof evaluationsindicated ofŽ larialnematodes and removal of Wolbachia results nostatistically signiŽ cant differences in Meloidogyne inacompleteand long-term block of embryogenesisand incognita populations.However, comparing the Ž rst and theeventual death of thenematode. Recent studies in our thirdsampling dates, a reductionin populations could laboratorysuggest that the symbionts also contribute to beseen, with T7 having the fewest (827 second stage theevasion of immunityin mammalian and vector hosts juveniles(J2)/ 100g soiland 10 g roots)and T1 having andso may be responsible for thelong-term survival of themost. Regarding phenological characteristics, the Žlarialnematodes. highestvalue for leafpresence was observedin T3 (60%),branching and  ower budsin T7 (25 and 14%, 067Coevolution between Fergusobia and respectively),and fruiting in T5 (24%). Preliminary Fergusonina mutualists resultsof this study suggest we shouldcontinue the evaluationof these organic amendments as an efŽ cient 1; 2 Robin M. GIBLIN-DAVIS ¤, Sonja J. SCHEFFER , alternativefor integratedpathogen control in fruit Kerrie A. DAVIES 3 andW .KelleyT HOMAS 4 horticulture. 1FortLauderdale Research and Education Center ,Universityof Florida/IFAS,3205College Ave, Davie, FL 33314,USA 066The symbiosis of Wolbachia bacteriaand Ž larial 2 USDA-ARS SystematicEntomology Lab, 10300 Baltimore nematodes Ave,Beltsville, MD, USA 3 Departmentof Applied and Molecular Ecology, Adelaide Mark J. TAYLOR University,PMB 1GlenOsmond, SA, Australia5064 FilariasisResearch Laboratory, Division of Molecularand 4 HubbardCenter for Genome Studies, University of New BiochemicalP arasitology,Liverpool School of Tropical Hampshire,35 Colovos Road, Durham, NH 03820,USA Medicine,P embrokePlace, Liverpool L3 5QA, UK ¤giblin@u.edu

Vol.4(2), 2002 145 Nematodesas mutualists(66-68) and Incompatible parasitic interactions (69-72)

Thecontemporary associations between Fergusobia ne- malefertility, however, depend on both the host and matodes(Neotylenchidae) and Fergusonina ies(Fergu- theparasite. In particular, survival of two Drosophila soninidae)represent the only putative example of nema- specieswas drasticallyreduced when infected with todeand arthropod-associated mutualism. The nematode anallopatric parasite. Thus, virulence is evolutionarily appearsto induce a budor leaf gall that both organisms labilein associationsbetween Drosophilatestacea group usewhile the  yprovidesgall maintenance, dispersion, speciesand their Howardula parasites.The high level of andsustenance for thenematode. Based upon molecular virulencemanifested in somehost-parasite combinations analysis,this is a potentiallylarge monophyletic radia- isdue to a lackof resistance in the hosts, perhaps tionof more than 50 mostly undescribed species of ne- asa resultof recent host colonisation by Howardula. matodesand  iesthat exhibit a highdegree of host speci- Theseresults provide evidence for acontinuumof host- Žcitywithin the Australasian Myrtaceae (mostly Lep- parasiteinteraction, with virulence initially high, followed tospermoideae; e.g., Eucalyptus, Corymbia, Angophora byevolution of the parasite to reduce the rate of host and Melaleuca).TheAgromyziidae (with no known ne- mortality,and evolution of the host to resist parasite- matodeassociates) is the putative sister group to theFer- inducedsterility. gusoninidae. Fergusobia couldhave evolved from para- siticnematodes similar to present day Howardula that 069R-gene homologuesin potatoconfer resistance parasitisedthe cyclorrhaphan stem ancestor of Ferguson- againstdistinct pathogens:a virusand a nematode ina iesand developed a plant-parasiticassociation that provideda mutualbeneŽ t to yhostand nematode. Al- 1; 1 1 Erin BAKKER ¤, Joke VAN VLIET , Hein OVERMARS , ternatively, Fergusobia couldbe related to present day Geert SMANT 1, Hans SANDBRINK 2, Edwin VAN DER anguinidsthat produced aboveground galls and devel- VOSSEN 2, Jaap BAKKER 1 and Aska GOVERSE 1 opedan association with an agromyzid ancestor .Inei- thercase, the evolution of the host-parasite interaction 1Laboratoryof Nematology, W ageningenUniversity, requiresthat host resistance and virulence be moderated. Binnenhaven5, 6709PD Wageningen, Contemporaryexamples of commensaland parasitic asso- TheNetherlands ciationsbetween nematodes and insects are used to gener- 2 BU Genomics,Plant Research International, The Netherlands atehypotheses about how such an associationcould have ¤[email protected] evolvedand is maintained. Genomeanalysis of a singleR-gene cluster in potato revealedtwo highly homologous resistance genes, Gpa2 068Evolutionary attenuation of virulencein Rx1 Drosophila-parasiticnematodes and .Thetwo genes confer resistance to fully unrelatedpathogens, namely the potato cyst nematode 1; 2 Globoderapallida andthe potato virus X. Togetmore John JAENIKE ¤ and Steve PERLMAN insightinto the evolutionary relationship between the 1 Departmentof Biology,University of Rochester ,Rochester, tworesistance genes, three new homologues mapping New York14627, USA 2 tothe same cluster on chromosome 12 of the diploid Departmentof Ecology and Evolutionary Biology, University potatoclone SH83 were identiŽed by using a single ofArizona, Tucson, Arizona 85721, USA LRR-basedprimer pair. cDNA analysisresulted in the [email protected] ¤ identiŽcation of the ORFs for twoout of the three new Virulenceis of central importance in host-parasite homologues.T oinvestigatethe diversiŽ cation process interactions,yet little is known about how it changes withinthis cluster, DNA sequencedata were analysed;the overextended evolutionary periods. Four species in the remarkablesequence conservation between the effector testacea speciesgroup of Drosophila were experimentally domains of Gpa2 and Rx1 pointsat the activation of infectedwith sympatric and allopatric nematodes in asimilarresistance mechanism. On the other hand, the Howardulaaoronymphium speciescomplex, and the infectionstudies and histological observations revealed effectof parasiteinfection on hostŽ tnesswas quantiŽed. arelativelyslow resistance response for Gpa2, while Thefertility of infected Drosophila femalesappears to Rx1 resultsin an extreme resistance. T ogainmore bedetermined solely by the host, with some species insightin themechanism underlying nematode and virus beingsterilised and others not, regardless of the source resistancestructure-function analysis of GP A2and RX1 ofthe nematodes. Reductions in host adult lifespan and arecurrently under investigation.

146 Nematology Symposiumabstracts

070Resistance to soybean cyst nematode: changesbetween Mi-1.2 and Mi-1.1,ahomologue an Xa21-likegene family that requirespossible thatdoes not confer resistance against nematodes, dimerisationfor signal transduction havebeen tested in two assays. Nematode resistance was testedin transgenic roots, and ability to induce Khalid MEKSEM, Aziz JAMAI, Eliza RUBEN, ahypersensitiveresponse was testedafter transient Kanokporn TRIWITAKORN, Hirofumi ISHIHARA, expressionin Nicotianabenthamiana leaves.Analysis Kimberly ZOBRIST andDavid A. L IGHTFOOT ofreciprocal swaps has identiŽ ed regions of the gene Departmentof Plant,Soil and General Agriculture, Room 176, thatare important for nematoderecognition and for SouthernIllinois University at Carbondale,Carbondale, IL signalingthe defence response. Our resultsindicate 62901-4415,USA thatintramolecular interactions of the Mi-proteinare Inthe soybean ( Glycine max (L.) Merr.) cv.Forrest, involvedin itsregulation. W ehavealso used the transient diseaseresistance inheritance to the soybean cyst expressionsystem to investigate the role of signaling nematode Heteroderaglycines Ichinohe(SCN) race3 moleculesand plant hormones in theresistance response. (Hg0)was demonstratedto be digenic, and conditioned bythe presence of both the Rhg1 and Rhg4 genes. A 072Understanding the geneticaland molecular high-densitygenetic map was constructedfor thetwo basisof (a)virulencein the root-knotnematode chromosomalregions carrying resistance loci to SCN. An Meloidogyneincognita integratedphysical genetic map was constructedaround the Rhg1 and Rhg4loci.Candidate gene sequences for Philippe CASTAGNONE-SERENO Rhg1 and Rhg4havebeen isolated from soybeancv. ForrestBAC clone 73P6 and 100B10. Shotgun gene INRA UnitéIPMSV ,BP2078,06606 Antibes cé dex, France [email protected] sequencingand cDNA hybridisationidentiŽ ed twogenes withhigh homology to the Xa21rice disease resistance Plantresistance is currently the most effective and envi- geneand to an Arabidopsis receptor-likekinase family, ronmentallysafe methodto control root-knot nematodes. whichcontain three functional domains each. Domains Resistancegenes generally act by inducingan hypersen- withinthe 854 residues of Rhg1showthe presence of sitivereaction at or near the infection site, that prevents 12extracellular leucine rich repeats, a trans-membrane theparasiteinstallation and/ orreproduction.However, the spanningdomain and a kinasedomain. Rhg4 shares emergenceof virulentbiotypes able to overcomethe plant thesame structure, with a longpeptide sequence of resistancegenes may constitute a severelimitation to this 894residues and has low amino acid homology to the controlstrategy. T oinvestigatethe moleculardeterminants Rhg1peptide. An active-dimerisation form oftheRhg1- of(a)virulencein root-knot nematodes, we havebeen de- Rhg4protein is suggested from theinteraction studies velopingdifferential strategies based on theselection and for theactivation of thesignal transduction pathways for subsequentcomparative analysis of pairsof Meloidogyne resistanceto SCN. incognita near-isogeniclines, just differing in their abil- ityto reproduceor noton tomatoesbearing the Mi resis- 071Functional analysis of the Mi-1 gene tancegene. The objective of theseapproaches is toiden- tifygenes differential or differentiallyexpressed between V.M. WILLIAMSON theavirulent and virulent lines. Recent results will be pre- Departmentof Nematology, University of California, Davis, sented,and discussed in relation with the mode of re- CA95616,USA productionof thenematode, i.e.,mitoticparthenogenesis. [email protected] Elucidatingthe genetical and molecular mechanisms in- Thetomato gene Mi-1 confersresistance against several volvedin theselection process of virulentnematodes from speciesof root-knot nematodes and against speciŽ c avirulentones shouldhave important consequences for the isolatesof potato aphid. Mi,likemany plant resistance managementand durability of naturalresistance genes in genes,encodes a proteinwith an apparent nucleotide the Ž eld. bindingsite and a C-terminalstring of looselyconserved leucine-richrepeats. Resistance is characterised by 073Nematode biodiversity research – current status alocalisednecrosis or hypersensitive response. The andfuture promise phenotypesof domain swaps and single nucleotide

Vol.4(2), 2002 147 Nematodesas mutualists(66-68) and Incompatible parasitic interactions (69-72)

Klemens EKSCHMITT 14are new to science. The interaction with international taxonomistsis essential to both train INBio staff and JustusLiebig University, IFZ –Departmentof Animal Ecology, generateknowledge related to tropicalnematode diversity. H.-Buff-Ring26-32, D-35392 Giessen, Germany [email protected] 075Nematode diversity in Dutch soils,from Rio to Thepresentation attempts an evaluation of the current Dutch SoilQuality Network (DSQN) statusof nematodebiodiversity research. Starting from an overviewof hownematology is integratedin international Ton SCHOUTEN biodiversityprogrammes, and from asynopsisof biodiversity-relatedinformation which has been made Laboratoryfor Ecotoxicology, National Institute for Public generallyaccessible to the scientiŽ c public,three major Healthand the Environment, P .O.Box 1, 3720BA Bilthoven, questionsare addressed: i)whatare the drivers of TheNetherlands [email protected] nematodediversity; ii)howtightly is nematodediversity linkedto environmental factors; and iii) what can Analysisof nematode fauna has been part of a Dutch we thereforeexpect to be the information content monitoringnetwork since 1993. It has the advantage ofnematode diversity? Knowledge gaps and research thatavailable comprehensive abiotic measures can be prioritiesare speciŽ ed anddiscussed. combinedwith biological characteristics. The monitoring networkconsists of 200 locations, representing ten 074Nematode survey in Costa Rican conservation majortypes of soiltype/ soiluse combinations. Sampling areas locationsare chosen in proportion to the form ofland- usebecause the network aims to give a generalpicture Alejandro ESQUIVEL ofbiological soil quality in the Netherlands. Seventy percentof the Dutch soil area is in agricultural use. UniversidadNacional de Heredia(U.N.A), Apartado postal Therefore,180 locations are situated on different types 86-3000Heredia, Costa Rica and Instituto Nacional de offarms, with40 sampled per year on a cycleof Biodiversidad(INBio), Apartado postal 22-3100, Santo Domingode Heredia,Costa Rica 5years.After thebiodiversity conference of Rio de [email protected];[email protected] Janeiro,the Dutch government ratiŽ ed the treaty, and concludedthat functional diversity aspects of non-natural TheInstituto Nacional de Biodiversidad (INBio) isa soilsneed more attention and quality standards. In 1999, non-proŽt scientiŽc institutionwith social orientation, thebiologicalsoil monitoring was extendedto a foodweb- createdin 1989, whose main mission is to promote basedprogramme. The nematode fauna is an important anewawareness of the value of biodiversity and, componentdue totheecological(feeding) groups that can thereby,achieve its conservation in Costa Rica. The bedistinguished. In the second round of the monitoring nematodesubprogram was initiatedofŽ cially in 1998 programme,100 biological farms were addedto serve as inclose cooperation with the Universidad Nacional of areferencefor regularfarming systems. A methodfor Herediaand Ž nancedby the Netherlands government. comparisonand quality assessment expressed in a Soil Weconducteda nematodeinventory in Žveconservation QualityIndex is under development. areasof Costa Rica, chosen because they contain themost representative and diverse ecosystems of the 076Nematode component of the FaunaEuropaea country.Several hundreds of samples were collected project andreferenced with a GPS navigator.All contrasting morphotypesfrom eachsample were representedand 1; 2 Tom BONGERS ¤ and Wouter LOS storedpermanently on Cobb’ s slides.The reference nematodecollection of INBiocontains 17 757specimens, 1Laboratoryof Nematology, W ageningenUniversity, P .O.Box distributedas follows: Dorylamida (39.3%), Rhabditida 8123NL-6700 ES Wageningen,The Netherlands 2 (18.7%),Araeolaimida (10.1%), T ylenchida(9.3%), ZoologicalMuseum Amsterdam, P .O.Box 94766 NL-1090 GT Amsterdam,The Netherlands Enoplida(9.1%), Mononchida (8.2%), Chromadorida [email protected] (2.1%),Aphelenchida (1.9%), Monhysterida (1.0%) and ¤ Mermithida(0.2%). Most of thecollection is identiŽ ed at TheCommunity Biodiversity Strategy of the European genuslevel; only 7% isidentiŽ ed atspecies level of which Commissionprovides a framework for thedevelopment

148 Nematology Symposiumabstracts ofCommunity policies and instruments to comply with enrichment,stress and succession) is critiqued. Applica- theConvention of Biological Diversity. Fauna Europaea bilityof indicesto assessmentof soilquality and associ- contributesby identifying and cataloging components atedsampling methods will be discussed relative to ge- ofEuropean biodiversity to serve as a basictool for ographicalscale, soil and vegetation type. Fundamental scienceand conservation policies. Fauna Europaea will constraintsand research priorities are identiŽ ed: i) clar- assemblea databaseof scientiŽ c namesand distribution ifyingtrophic afŽ nity of some species; ii)integrating ofall living multicellular European land and fresh-water morphological,ecological and molecular approaches; and animals,including nematodes. Marine nematodes are iii)intensifyinglong-term studies in natural and disturbed includedin a ‘sister’project, the European Register of environments. MarineSpecies (ERMS). In anticipation of a Key to the EuropeanNematodes ,thebasal species list ( Nematoden 078Nematode faunal analyses to assess food web vanNederland )was extendedto include the whole of enrichmentand connectance Europe.Currently, the nematode database includes a 1; 2 totalof 2500 species, including the full name, author, Howard FERRIS ¤, Tom BONGERS and date,original genus, main synonyms and geographic Ron DEGOEDE 3 distribution.ClassiŽ cation is based on the most recent, 1Departmentof Nematology, University of California, Davis, generallyaccepted standards. Published records or CA95616,USA personalcommunications for materialdeposited and 2 Laboratoryof Nematology, W ageningenUniversity, P .O.Box availablein a publicnematode collection must support 8123,6700 ES Wageningen,The Netherlands databaseentries. Geographic information will be provided 3 Sub-departmentof SoilQuality, W ageningenUniversity, P .O. for eachspecies according to ISO-TDWG standards; Box8055, 6700 EC Wageningen,The Netherlands thereporting region will include Europe, Macaronesian ¤[email protected]. islands,Cyprus, Franz Josef Land and Novaya Zemlya. Thecp classiŽ cation recognises that nematode taxa with Speciesrecorded from non-Želd locations including anatomicaland physiological commonalities are probably glasshousesand aquariums will not be included. similarlyadapted to speciŽ c environmentalconditions. Functionalguild analysis, which integrates cp-scaling 077Past and present status ofnematodecommunity withfood sources, reveals that some guilds respond indicators opportunisticallyto enrichment while others represent thepresence of higher trophic connectance in the food Ewa DMOWSKA¤ andKrassimira I LIEVA-MAKULEC web.This dichotomy was portrayedgraphically in cp Instituteof Ecology,P olishAcademy of Sciences,Dziekanow trianglesas theproportional representation of enrichment Lesny,05-092 Lomianki, P oland opportunists(cp 1), general opportunists (cp 2) andtaxa ¤[email protected] indicatinghigher connectance (cp 3-5). Confounded in thecalculation of cp triangles is interdependence of Differencesin sensitivity of nematodes towards some theaxes; proportionality of the three groupings to the chemicalshave been recognised for nearlyfour decades. wholenematode fauna requires that increase in food Initially,single species or taxonomic groups were con- webstructure (cp 3-5 taxa) concomitantly decreases sidered,followed by parametersof wholecommunities to theenrichment indicator (cp 1 taxa).The categorical measuredifferent properties of naturaland disturbed ter- separationof nematode taxa into Ž vecp classes does restrialecosystems. A shortoverview of nematodecom- notimply unit increments in r orK characteristics.W e munityindices will be presented,progressing from those usebody size and growth rates to weightthe importance consideredtraditional, relatively simple, and widely ap- ofenrichment indicators and estimates of corresponding pliedin environmental studies ( e.g.,abundance,richness, foodweb connectance to weight the importance of diversity,similarity and constancy) to those considered structureindicators. W econsidercp 2 taxabasal to both moresophisticated ( e.g.,Maturityindex and its modiŽ ca- trajectoriesand calculate position along the enrichment tions)and speciŽ c tonematodes ( e.g.,proportionsamong andstructure axes of afaunaldiagram independently as trophicgroups). Emphasis will be placedon indicesused theweighted ratios of theindicator and basal taxa. mostoften. Their contribution to understandingthe func- tioningof soilecosystems ( e.g.,natureof decomposition pathways,N-mineralisation) in different situations ( e.g., 079Selection of sentineltaxa and biomarkers

Vol.4(2), 2002 149 Diversityand indicators (73-80) and Morphology and development (81-84)

1; 2 Deborah A. NEHER ¤, Daniel A. FISCUS Traditionally,assessments of ecologicalrisks of soilpol- and Fafeng LI 1 lutionare based on chemical analyses compared to a set ofstandards.Recently, in the Netherlands, investigations 1Universityof T oledo,Department of Earth, Ecological and were initiatedto explore the possibility of including bi- EnvironmentalStudies, Mailstop 604, 2801 W .BancroftSt, ologicalindicators in ecological risk assessment proce- Toledo,OH 43606,USA dures.Nematodes are among the soil fauna groups that 2 Universityof Maryland Center for Environmental Science havepromising characteristics as indicatorsfor soilqual- AppalachianLaboratory, 301 Braddock Road, Frostburg, ity (i.e.,ubiquitous,high species diversity, sensitive to var- MD21532,USA iouskinds of soil disturbances, speciŽ c nematode-based [email protected] ¤ indicators).However, interpretation depends on compari- Nematodecommunity indices would be morefeasible for sonwith undisturbed controls or references. As localref- usein environmental monitoring programmes by reducing erencesites or dataare often unavailable, there is demand thenumber of genera that need to be enumerated and for anation-widereference system. In the present project, identiŽed. This could be achievedby narrowingindices to we builta database(DA WACO Nematodes)of 1600ne- includeonly sensitive or tolerantgenera or specieswhile matodefauna samples from whichall taxa were identi- eliminatingambiguous ones. IdentiŽ cation of sentinel Žed.The samples were collectedacross the country, rep- taxacan be achieved by employing a combinationof resentinga varietyof soil-,vegetation- and land-use types tools.For example,multivariate statistics can help identify between1985 and 2001. Samplesfrom undisturbedor ref- taxathat demonstrate relative tolerance or sensitivity to erencelocations were analysedwith multivariate analyses physicaland/ orchemical/ nutrienttypes of disturbance. techniquesto select clusters of samples (14 from terres- Lifehistory characteristics of taxameeting these criteria trialsoils, six from aquaticsediments) with similar nema- canbe veriŽ ed in empirical studies to evaluate and todefauna and habitat characteristics. Based on the sta- recommendreŽ nement in coloniser-persister weights tisticalcharacterisation of the composition of taxa from employedin maturity indices. V eriŽcation of coloniser- areferencecluster, the probability of membershipof any persistervalues can be achieved using independent samplerequiring assessment can now be calculated and molecularbiomarkers. Once sentinel taxa are identiŽ ed, evaluated. moleculardiagnostic tools are possible. A vailabilityof commercialkits then becomes approachable to non- 081Spermatogenesis and nematode phylogeny specialistsand cost-effective (time and labour) for 1; 2 implementingnematode bioindicators within large-scale VladimirV .Y USHIN ¤ andVladimir V .M ALAKHOV environmentalmonitoring programmes. 1Instituteof Marine Biology FEB RAS, Vladivostok690041, Russia 2 080A nematodereference database as Departmentof Biology, Moscow State University, Moscow aninstrument forbiological soil assessment: 119899,Russia [email protected] acasestudy fromthe Netherlands ¤ Dataon sperm structure and spermatogenesis are 1; 1 R.G.M. DE GOEDE ¤, T. BONGERS , W.A.M. frequentlydiscussed in the analyses of the phylogenetic 1 2 3 4 DIDDEN , P. DOELMAN , H. KEIDEL , F. KERKUM , relationshipsinside many classes of the metazoan 5 6 7 R. KNOBEN , T. SCHOUTEN , J. VAN DER WAARDEN animalswith the exception of the class Nematoda. 7 and M. WAGELMANS Oneof the reasons for thissituation is a lackof studieson the relatively primitive aquatic nematodes 1 Sub-departmentof SoilQuality, W ageningenUniversity, P .O. from severalimportant orders. Our latestobservations Box8005, 6700 EC Wageningen,The Netherlands onsperm development in free-living nematodes from 2 DoelmanAdvisory 3 Blgg theorders Enoplida (suborders Enoplina, Oncholaimina, 4 RIZA Tripyloidina),Chromadorida (suborders Chromadorina 5 RoyalHaskoning andCyatholaimina), Desmodorida, Araeolaimida and 6 RIVM Monhysterida,together with the data available on many 7 Bioclear parasiticspecies, allow deŽ nition of fourmain patterns of ¤[email protected] nematodespermatogenesis. Each pattern is distinguished

150 Nematology Symposiumabstracts clearlyby special way of development of aberrant approacheshave to be be taken into account and the organelles– membranousorganelles (MO) andŽ brous numberof analysednematode species must be increased. bodies(FB): i)MOandFB developas the MO- FBcomplexes(most Rhabditia (Rhabditida, Strogylida, 083Nematode embryology and its importancefor Ascaridida,Rhigonematida, Spirurida) and some of the classiŽcation of highernematode taxa Chromadoria(Araeolaimida (including Comesomatidae) andMonhysterida); ii)MOandFB occurbut their VladimirV .M ALAKHOV developmentis asynchronous and independent (some Enoplia:Enoplida ( Enoplus,P ontonema );Dorylaimida Departmentof Biology,Moscow State University, Moscow (Xiphinema)); iii)MOdonot appear, only FB are 119899,Russia [email protected] developed(T ylenchidafrom Rhabditia,Chromadorida andDesmodorida from Chromadoria); iv) FB do not Investigationson the embryology of nematodes of dif- appear,only MO aredeveloped (Enoplia: Enoplida ferenttaxa revealed the great variety of thedevelopment (Anticoma),Mermithida,Trichurida, Dioctophymida). withinthe class. There are three principal patterns of ne- Patternof sperm development may be proposed as a matodedevelopment. The Ž rst oneischaracteristic for ne- newdistinct cytological character for theanalysis of the matodesof theorder Enoplida. The cleavage of theeno- nematoderelationships. plideggs is variable. The bilateral symmetry forms rela- tivelylate, usually after the closing of theblastopore. The fateof theblastomeres is not determinedstrictly as ischar- 082Phylogeny and ontogeny of free-livingand acteristicfor othergroups of nematodes. The second pat- parasiticnematodes ternis characteristicfor nematodesof theEnoplia line of evolutionwith the exception of theEnoplida. This line in- Einhard SCHIERENBERG ¤, Vera LAHL, Magdalena LAUGSCH and Thomas GÖDDE cludesthe orders Monochida, Dorylaimida, Mermithida, Dioctophymidaand Trichurida. The cleavage of the ne- ZoologicalInstitute, University of Cologne,50933 Kö ln, matodesof the above-mentioned orders is bilateral with Germany earlydetermination of the fate of blastomeres. The key ¤[email protected] featureof the second pattern is the origin of the endo- dermfrom theanterior blastomere of thetwo-cell stage. Phylogeneticrelationships between nematodes have Thethird pattern is characteristicfor theChromadoria C alwaysbeen discussed controversially. In recent years Secernentealine. In this line, the egg cleavage is alsobi- analysisof molecular data, particularly sequence data lateraland strictly determined, but the endoderm comes from genescoding for ribosomalRNA, havechallenged from theanterior blastomere of thetwo-cell stage. Three someof the traditional taxonomic proposals based on principalpatterns of nematode development correspond morphologicalcriteria. As athirdmeans to identify tothemain trend of nematodeevolution and could be the charactersof potential phylogenetic value, we are basefor anewnematode classiŽ cation. studyingselected developmental processes during early embryogenesis.These include establishment of axial 084Division and fate ofvulvalprecursor cells: is polarity,separation of soma and germline, assignment polymorphisman imageof evolutionarydivergence? ofcell fates and the pattern of gastrulation. Some of theexisting differences can be readily identiŽ ed by Laure BONNAUD¤, Marie DELATTRE and comparativeanalysis while others require experimental Marie-AnneF ELIX interference.A varietyof free-living and also several parasiticspecies have been studied. Our resultssupport InstitutJacques Monod, CNRS-Université s Paris6 et7,T our 43,2 placeJussieu, 75251 P ariscedex 05, France thenotion that the classical separation into Secernentia [email protected] andAdenophorea is artiŽ cial, and that at least three ¤ supertaxashould be deŽned. In some cases we identiŽed Inorder to approach evolutionary processes of develop- majordifferences between species considered as close mentalmechanisms, we arestudying development of dif- relatives,questioning their present position in the ferentnematodes at the cellular level. The cell lineage phylogenetictree. T oreacha morelucent picture of ofnematodesis mostlyinvariant for agivenspecies, but nematodephylogenetic relationships, data from different variesbetween species. W ehaveperformed a microevo-

Vol.4(2), 2002 151 Marketablebiological control agents for plant-parasitic nematodes (85-88) lutionaryapproach with the nematode vulva as a model 086The commercialisation of Paecilomyceslilacinus systemto deŽ ne how a celllineage varies during evolu- asanagent for the controlof plant-parasitic tion.The vulva is formed by precursors in the ventral ep- nematodes ithelium,called Pn.p cells. Each Pn.p cell has a speciŽc fate,which is speciŽed bycellinteractions that are differ- Diane NEETHLING ent in Caenorhabditiselegans and Oscheius sp. 1 CEW1. BiologicalControl Products SA (Pty)Ltd, P .O.Box1561, Twodistinct networks of cell interactions can therefore Pinetown3600, South Africa leadto an identical pattern that forms asimilarmorpholo- gicalstructure. Moreover, the same vulva lineage charac- Paecilomyceslilacinus isone of anumberof fungiwith tersthat diverge between closely related species are poly- nematophagousproperties that have been examined over morphicwithin a species(between laboratory strains). In thepast two decades for potentialuse as agents for the orderto understandevolutionary mechanisms of celllin- controlof plant pathogenic nematodes in agriculture. eagevariation, we performeda geneticanalysis of P3.p Whileshowing promise, their commercialisation has in divisionbetween strains of C. elegans.Recombinantin- thepast been constrained by Želdresults which have not bredlines show intermediate levels for thelineage char- deliveredsufŽ cient promise to attract commercial atten- acter.Polymorphismbetween two strains is therefore due tion,as well as a certainlack of appreciation by farm- toseveral loci. ingcommunities at large of theeconomic signiŽ cance of nematodeinfestation. The South African company, Bio- 085Biocontrol – arouteto market logicalControl Products (BCP) was establishedin 1995 withfunding secured from privateinvestors together with Dave CRUMP asubstantialgrant from theSouth African government. BCP’s primeobjective was toconduct the necessary ap- BioNemLtd, 46, Chaul End Road, Caddington, Bedfordshire, pliedresearch in areas of product formulation, Ž eldap- LU1 4AS, UK plicationand solid substrate manufacture which would [email protected] leadto acceptance by theagricultural community and reg- ulatoryauthorities of Paecilomyceslilacinus as an eco- Withnematicides declining in popularity for environmen- nomicallyjustiŽ able addition to strategies for theman- talreasons, and resistance not being a long-termsolu- agementof plant-parasiticnematodes. While the develop- tiondue to the ability of nematode populations to over- mentof the manufacturing process and product formu- comeit, there is a needfor anenvironmentallysafe con- lationwas largelyconducted in-house, the research in- trolmeasure that can give effective, long-term control of stitutesattached to the Department of Agriculture were nematodes.Biocontrol has been studied for manyyears, commissionedto assistwith efŽ cacy trials. BCP nowhas withsome encouraging results, and there is now mount- registrationfor theuse of this biological agent sold un- ingpressure to develop it towards practical application. I derthe brand name Pl Plus, for thecontrol of nematodes willbe presentingthe route I amtakingto tryand achieve inbananas, papinos, tomatoes, tobacco and citrus. Fur- this.It is notoriously difŽ cult with biocontrol to obtain thertrials are in progress on othercrops. A solidsubstrate good,consistent data from small-scaleexperiments. Such manufacturingplant capable of processingup to50 tper datais bestobtained from large,long-term trials, but to do yearhas been in operation since 1997 and local farmers thisit isnecessaryto havelarge-scale production systems, currentlyuse Pl Plus in integrated crop protection pro- preferablywith industrial support. Most production com- grammes. panies,however, need to seegood efŽ cacy data from Želd trialsbefore they will invest in new products. BioNem Ltd 087 In vitro culture of Pasteuriapenetrans attemptsto Ž llthis gap between research and commer- cialisationby selectingeffective agents (fungal parasites), 1; 2 Thomas E. HEWLETT ¤, John F. GERBER , developingsuitable production systems, and conducting Kelly S. SMITH 1 andJames H. W HITE 1 realisticŽ eldtests. The two fungi I amconcentratingon are Paecilomyceslilacinus and Verticilliumchlamydospo- 1EntomosLLC, 4445SW 35 th Terr.,Suite310, Gainesville, rium. FL32608,USA 2 CDG Laboratories,Inc., Gainesville, FL, USA ¤[email protected]

152 Nematology Symposiumabstracts

Pasteuriapenetrans hasbeen acknowledged worldwide cooperativeresearch resulted in a commercialproduct, asan effective biological control agent of plantparasitic BioYield™,for useon transplantedcrops. nematodes.The inability to mass-produce this fastidious bacterium in vitro hasbeen the barrier to making P. 089Nematode movement through soil: simple penetrans amarketableproduct. Enterobactercloacae , responsesin a complexenvironment aubiquitoussoil bacterium, has been found in close associationwith the cuticle of Meloidogynearenaria A. Forest ROBINSON femalesextracted from tomatoroots. Co-cultures of E. cloacae and P.penetrans insimple nutrient broth have USDA-ARS, 2765F&B Rd,College Station, TX 77845,USA producedviable P.penetrans endospores.Filtrates from Intheory, similar responses to simple cues can guide eco- E. cloacae grownon simple nutrient broth are suitable logicallydifferent nematodes through the soil in com- culturemedia for bothvegetative growth and sporulation pletelydifferent ways, when the unique characteristics of of P.penetrans ,producingviable, infective endospores. gasand solute diffusion, water movement, and tempera- In vitro cultureand therefore mass productionof P. ture uctuationin soil are considered in relation to sub- penetrans isnowpossible. tledifferences in response thresholds, rates of locomo- tion,and sensory adaptation. V alidationof modelsof ne- 088Development of multi-componenttransplant matodemovement within the diurnally  uctuating,verti- mixesfor plant growth-promotion and disease caltemperature gradients of soilconŽ rmed that two root- suppression parasiticspecies that respond similarly under static con- ditionson agarmove in oppositedirections in thermody- Nancy KOKALIS-BURELLE namicallynatural soil. Experiments on attractionto CO 2 insoilshowed that gradients inducing maximal nematode USDA, ARS, U.S.Horticultural Research Lab, F ortPierce, accumulationat a pointsource are achieved at optimal FL34945,USA [email protected] soilmoisture for nematodemovement, are not generated insaturated soil, and can be sustained for 24h insoil Researchwas undertakento develop a biologically-based witha totalgas volume that would not induce attraction productthat, when added to transplant mixes, would en- onagarin mostassays. Evidence from entomopathogenic hanceplant growth, increase yield, and provide protec- andnematophagous nematodes suggests additional chem- tionagainst pathogens. Studies at Auburn University in icalstimuli modulate responses to CO 2 asobserved for cooperationwith Gustafson LLC testedorganic compo- insects.Future progress will be acceleratedby integration nentsand plant growth-promoting rhizobacteria (PGPR) ofmolecular,anatomical, and physiological data from di- for effectson tomatogrowth and nematode viability. Or- versenematodes, direct comparisons within soil, better ganiccomponents and PGPR were combinedand eval- conceptualpreparation, and improved techniques to mea- uated.Gustafson LLC andUSDA, ARSthenentered sureand simulate the conditions and stimulusgradients of intoa CooperativeResearch and DevelopmentAgreement soil. (CRADA) for Želd-testingin Florida, where combina- tionsof amendedtransplants and methyl bromide alterna- 090Sensory responses of plant-parasiticnematodes tivesoil treatments were evaluated.SigniŽ cant increases tosemiochemicals intomato and pepper growth, vigor, and transplant sur- vivaloccurred with formulations of PGPR. Oneformula- Ekaterini RIGA tionreduced root-knot nematode galling and several im- provedpepper root condition. Four PGPR treatmentsre- WashingtonState University, IAREC, 24106N. BunnRd., ducedangular leaf spot lesions caused by Pseudomonas Prosser,WA 99350,USA [email protected] syringae pv.lachrymans, and gummy stem blight, caused by Didymellabryoniae, onwatermelon.One PGPR treat- Theunderstanding of nematodesemiochemicals has im- mentreduced root-knot nematode disease severity on provedin recentyears due to incorporation of noveltech- muskmelon.The effects of bare root, plug, and PGPR niquesto study nematode behaviour. Extracellular elec- amendedplug transplants on growth and yield of straw- trophysiologicalrecordings from thecephalic region of berryshowed that PGPR amendedplugs had highest plant-parasiticnematodes in conjunctionwith behavioural yields,and bare root transplants had lowest yields. This bioassayshave revealed that chemoreception is essential

Vol.4(2), 2002 153 Sensoryresponse and behaviour (89-92) and V ertebrateparasitic nematodes (93-96) for nematodesurvival. The function of the amphids as individual  p-geneshave restricted expression patterns amainchemoreception organ has been afŽ rmed in all indicatingspeciŽ c rolesfor encodedpeptides. Sequence nematodebehavioural studies. Principal semiochemicals datareveal that a signiŽcant number of FaRPs are thatelicit nematode responses are plant host exudates, structurallyhomologous across a broadrange of nematode foodstimulants, food deterrents, and homospeciŽ c sex species;a similarpattern of functionalhomology remains pheromones.Several amino acids and plantroot diffusates tobe established. While information on FaRP receptors elicitedpositive responses from plantnematodes. In addi- islimited, physiological data suggest that the diversity tion,sensory adaptation and chemical isomer speciŽ city inFaRP structure is not re ected by endogenous FaRP havebeen demonstrated. Recordings from thecephalic receptors.The FaRP-signalling system likely harbours regionof apotatocyst nematode, Globoderarostochien- numerouspotential targets for novelcontrol measures. sis,veriŽed the species-speciŽc natureof sexpheromones. However,the selection and exploitation of these is Biologicalactivity associated with G.rostochiensis and madedifŽ cult by the complexity inherent within the Heteroderaschachtii sexpheromones has been linked to FaRPsignalling system and the potential for functional aspeciŽc HPLC-derivedfraction, and although the sex redundancy. pheromoneshave not yet been identiŽ ed, their physical propertieshave been reported. Sensory physiology is a 092Chemoreceptor genes: what canwe learnfrom newand promising Ž eldin nematology that will lead us C. elegans andhow canwe applythis informationto toa betterunderstanding of nematode biology and will studies onothernematodes? providea meansto control nematodes by interferingwith theirsensory perception. Ann M. BURNELL¤ andDamien M. O’H ALLORAN Instituteof Bioengineeringand Agroecology and Department of 091Neuropeptide signalling systems in parasitic Biology,National University of IrelandMaynooth, Maynooth, nematodesand their potential as novelcontrol targets Co.Kildare, Ireland ¤[email protected] Aaron G. MAULE1; , Nikki J. MARKS 1, Angela ¤ Soildwelling nematodes encounter many types of volatile MOUSLEY 1, Michael KIMBER 1, Colin F. FLEMING 2;3, andwater-soluble molecules in their environment. For David W. HALTON 1,TimothyG. G EARY 4 and free-livingnematodes like C. elegans,successfulforaging David P. THOMPSON 4 dependson the ability to detect a gradientin one 1ParasitologyResearch Group, School of Biology and odorantwhile ignoring extraneous odours. The infectious Biochemistry,Belfast BT9 7BL, UK stagesof plant and animal parasitic nematodes also 2 AppliedPlant Science, Queen’ s UniversityBelfast, Belfast relyon chemoreception as their primary host Ž nding BT9 7BL, UK cue.Using a combinationof genetic, molecular and 3 Departmentof Agriculture and Rural Development for bioinformaticapproaches chemoreceptor genes have been NorthernIreland, Newforge Lane, Belfast, BT9 5PX, UK identiŽed in C. elegans. These C. elegans chemoreceptor 4 PharmaciaAnimal Health, Kalamazoo, MI 49001,USA [email protected] genesencode seven-transmembrane G-protein coupled ¤ receptors(GPCRs) andcomprise the largest gene family Thediversity of neuropeptide signalling molecules inthis nematode. GPCRs are also involved in olfactory innematodes is as great as that known for any signaltransduction across a broadspectrum of animals organism.By far thelargest and most multifaceted includinginsects, crustaceans, Ž shand mammals, but the nematodeneuropeptide family is the FMRFamide-related C. elegans(and Drosophila)chemoreceptorgenes have no peptides(FaRPs). In Caenorhabditiselegans ,thisfamily sequencehomology to vertebrate GPCR odour receptor comprises60 distinct peptides that are encoded on genesand they also differ from vertebrateodour receptor 22genes (designated  p-genes)and available evidence genesin their genomic structure. In this presentation indicatesthat this complexity is mirrored in plant and we willprovide overview of the genomic structure and animalparasitic nematodes. Neuropeptides are known diversityof odorant and chemoreceptor gene families tobe expressed in all neuronal subtypes and to have invertebrates and invertebrates and we describeour potentand wide-ranging effects on motility,alimentation, attempts,using homology-based approaches, to isolate reproductivefunction and sensory perception. Although chemoreceptorgenes in the entomopathogeni cnematode theeffects of FaRPs are widespread in nematodes, Heterorhabditisbacteriophora .

154 Nematology Symposiumabstracts

093Ecological aspects of smallmammal nematode (Ancylostomamartinezi in Genettagenetta (common infections genet); Vigisospirurapotekhina hugoti in Meles meles (badger); ii)thehigh richness in nematodes species in S. MAS-COMA Vulpesvulpes (fox) (16species), Martesfoina (stone Departamentode Parasitología, Facultadde Farmacia, marten)(14 species) and Genettagenetta (12species), Universidadde Valencia,Av. V icentAndré s Estellés s/n,46100 probablyas a theresult of a widerange of distribution Burjassot-Valencia,Spain ofthese hosts as habitat generalists. From anecological [email protected] pointof view: i)thereis an high ethologic incidence of thehost to the nematode fauna; this is specially notable Nematodeshave many adaptation and colonisation inthe case of Mustelidae, which include hosts with capacities.This explains how they were successful aquatic,amphibian or terrestrialbehaviours; the lastgroup incolonising animal hosts from evolutionarylines of presentsthe highest number of nematodespecies; ii) the free-livingnematode groups. Recent results on 18S dietof thehosts is another important determining factor; rRNA genesequencing and corresponding phylogenetic moreverfrom indirectlife cycle nematodes, it is possible analysesshow how the adaptation of a parasiticway toknowwith high precision which taxa are prey of these oflife was acquiredby different free-living nematode hosts.The speciŽ city of reported nematodes in Iberian groupsindependently. These analyses also suggest that Carnivoraalso gives information about the phylogeny of phylogenyinferred from DNA sequencesdoes not always the host. agreewith the present systematic groupings within the Secernentea.Moreover, studies carried out on nematode parasitesof small mammals (rodents and insectivores) 095Nematode parasitic fauna in CanaryIsland onislands in the last 20 years show that the resulting vertebrates evidencedoes not support mathematical models which Basilio VALLADARES provethat nematode species following a one-hostlife cycleare more capable to colonise new environments Departamentode P arasitología, F acultadde F armacia, andexpand. Small mammal nematodes are excellent Universidadde La Laguna, Spain modelsfor ecologicaland evolutionary studies because [email protected] theyinclude many nematode groups presenting very Inthe Department of Parasitology in La Laguna differenttypes of life cycle patterns, from monoxenous University,many faunistic, ecological and biological toheteroxenous life cycles. Monoxenous nematode studieson parasitic helminths in vertebrates have been speciesinfecting rodents and insectivores comprise undertakenover several years. Despite this, nematode ageohelminths,pseudogeohelmin thsand geohelminths. studieshave only been carried out on Canisfamiliaris Heteroxenousnematodes of small mammals include andin the species which can cause zoonosis in man, speciestransmitted by intermediate hosts pertaining to Toxocaracanis and DiroŽlaria immitis .Thein uence differentinvertebrate and vertebrate groups. This wide ofthe Canary Island microclimate on their distribution biologicalspectrum becomes a veryuseful tool for andthe insularity effect were alsostudied. Since 1999, ecologicaland short-term evolutionary studies. parasitichelminths in Canary Island birds have been studiedand the nematofauna is the best known group 094Studies onnematodesof wildcarnivores atthe moment. The bird species (105) analysed include AlectorisBarbara , Laruscachinnans , Columbalivia , Carlos FELIU, Alexis RIBAS Fulica atra and Chalmydotisundulata .Thenematodes andJuan Carlos C ASANOVA obtainedwere studiedunder optical microscopy by clearingwith Amman lactophenol. In order to study the Laboratoride P arasitologia,F acultatde F armàcia, Universitat samplesusing scanning electronic microscopy (SEM), deBarcelona, Avda Diagonal s/ n.08028 Barcelona Mercerand Birbeck (1979), Ba and Marchand (1994) TheIberian carnivores constitute a groupof hostswhere andMiquel and Marchand (1998) techniques were used. thediversity of its nematodes is not very high (45 Thehelminth species identiŽ ed in the different birds species)compared with other mammal groups. From understudy were asfollows: in AlectorisBarbara : afaunisticpoint of view it is signiŽ cant that: i) two Aonchotecacaudin ata , Ascaridiagalli (Žrst reportin endemicspecies are restricted to the Iberian Peninsula this host), Baruscapillariaobsignata , Eucoleusannulatus

Vol.4(2), 2002 155 New technologiesand chemicals for control of plant pathogenic nematodes (97-100) and Heterakisgallinarum : in Columbalivia : Aonchoteca Thereare many time consuming and costly processes sp., TetrameresŽ ssipina , Synhimantus (Dispharynx) involvedwith taking a newpesticide to market. spiralis and Ascaridiacolumbae , and in Fulica atra, Whilemany researchers are searching for drop-in Capillaria sp.were foundand in Laruscachinnans, replacementsfor methylbromide (MeBr), developing a Cosmocephalus sp.The effect of insularity on the newformulation or improving on an old one is just the parasitesis studied. beginningof the process. EfŽ cacy data and Ž eldtesting underone’ s ownresearch programme might indicate promisingresults but the developer needs to be ready 096Some problems related to the controlof for a3-4year process which can cost in excess of nematodeinfections in domestic ruminants US$12m. It is important that no steps or agencies are FranciscoA. R OJO-VÁZQUEZ 1, overlookedin the long road to a marketableproduct. Antonio R. MARTÍNEZ-FERNÁNDEZ 2, Thereare many considerations, such as venture capital, M.A. ÁLVAREZ-SÁNCHEZ 1, J. PÉREZ-GARCÍA 1 thepatenting process for bothUS andforeign proprietary and R.C. MAINAR-JAIME 1 rights,support for minoruse pesticide Ž eldtesting and addressingall steps required by the US Environmental 1 Dptode SanidadAnimal, Universidad de Leó n, Spain ProtectionAgency. This presentation will cover what it 2 Dptode Parasitología, UCM, Madrid,Spain takesto get a MeBralternative to market in the US and beyond. Gastrointestinalnematode infections in small ruminants areof greatimportance in animal husbandry worldwide. 098Soil fumigation: new uses forold chemicals and Thecontrol is basedon theknowledge of theepidemiol- new compounds ogy,the detection of nematode eggs in faeces on a rep- resentativenumber of animalsfrom the ock,the use of R. RODRIGUEZ-KABANA anthelminticdrugs and management practices. However, faecalegg counts are highly variable and must be doneon Departmentof Entomology & PlantP athology,Auburn alargenumber of animals in order to get accurate esti- University,Auburn, AL 36849,USA mationsof theparasite burdens within the  ock.On the [email protected] otherhand, the routine and inadequate application of an- thelminticsis anotherproblem, due to thedevelopment of Theimminent removal of methyl bromide (MBR) as a anthelminticresistance (AR). InSpain, some recent re- soilfumigant has stimulated a world-widesearch for al- portson ARinsmall ruminants shows a ockprevalence ternativetreatments. This decade-long effort identiŽ ed the of21%.Although benzimidazoles (BZ) hasbeenthe most oldfumigants: 1,3-dichloroprope ne(1,3-D), chloropicrin, usedanthelmintic during the last decade, BZ-resistance is methylisothiocyanate (MITC), andcompounds that gen- stilllow. However, there is ahighlevel of ARagainstim- erateMITC, as the only materials available at present idazothiazoles.T wofactors were signiŽcantly associated for developmentof preparations capable of substituting withresistance or suspicion of resistance: breed and an- MBRinsoil fumigation. New formulationsand applica- thelminticefŽ cacy according to the farmer. Foreignand tiontechnologies have been devised to increase the efŽ - crossbreed ocksseemed to be moreprone to present AR. cacyand practicality of thesevery old chemicals as MBR Higherprobabilities of ARseemedto be relatedto ocks alternatives.Other compounds showing promise for future thatused mostly private pastures or to those that bought developmentas MBR substitutesare: inorganic azides, replacementanimals. propyleneoxide, methyl iodide and otheriodinated hydro- carbons.Several naturally occurring ‘ biofumigant’com- pounds (e.g.,citral,fufural, various mustard oils such as 097The cost of licensingand labeling a methyl allylisothiocyanate) are also being explored as MBRal- bromidealternative ternatives.The main difŽ culty in Ž ndingMBR substitutes Robin N. HUETTEL for soilfumigation resides in theneedto develop econom- icalsoil treatments that are safe, environmentally accept- 105Comer Hall, College of Agriculture,Auburn University, ableand effective, not only against plant-pathogenic ne- Auburn,AL 36849,USA matodesbut also weeds, and other soil-borne pathogens [email protected] and pests.

156 Nematology Symposiumabstracts

099New technologiesfor challenging situations – typeof phytoalexins,hitherto undescribed. The pathogen perennialand nursery crops in California resistantbanana hybrid SH-3481 (from FHIA), produces largeamounts of these phytoalexins and phytoanticipin s Sally SCHNEIDER (constitutivenatural antibiotics), suggesting that this USDA ARS SJVASC,9611 S. RiverbendAve, P arlier,CA93648, newtype of phytoalexins plays an important role in USA theresistance mechanisms of banana plants against [email protected] fungaland nematode diseases. The signiŽ cance of theseŽ ndingswill be discussed as they may open Nurserycrops that are grown in Californiafor commercial theway to development of new sustainable agricultural plantingsare required to be free ofplant-parasitic methodologiesto manage plant diseases. nematodes.Nursery certiŽ cation can be accomplished byextensive sampling or by use of an approved soil treatment.Approved treatments (materials and rates) 101Quest forresistance to the peanut podnematode varywith soil type, moisture, temperature, and cropping A.H. MCDONALD, H. FOURIE and S. STEENKAMP historyof the Ž eldand show efŽ cacy to a depth ¤ of150 cm. Historically, the most commonly used materialwas methylbromide. Prior toplanting the ARC-GrainCrops Institute, Private Bag X1251, P otchefstroom certiŽed trees and vines, growers often fumigated with 2520,Republic of South Africa [email protected] methylbromide to prevent replant disorder in their ¤ orchardsand vineyards. Combinations of new and existingtechnologies, novel application methods, and Sinceits discovery in 1987, the peanut pod nematode registeredand experimental materials are being tested for Ditylenchusafricanus isregarded as oneof theeconom- usein nursery and perennial replant situations. Global icallymost important pests of peanut in South Africa. positioningsatellite systems can be used to implement Thenematode is omnipresentin peanut production areas managementstrategies into areas selected for optimal inthis country and its most signiŽ cant impact is on ker- performancebased on the spatial variability of soil neland seed downgrading, which causes serious losses conditions.Drip irrigation technologies can be used inincome for theproducers. This, in turn, has a detri- todeliver emulsiŽ able formulations throughout the soil mentaleffect on net national peanut production. Although proŽle to the depth needed – especiallyfor materialsthat asigniŽcant number of scientiŽc publicationshave been donot move as readily as methyl bromide. Currently publishedover the past few years,the occurrence of the registeredand experimental materials can be re-evaluated peanutpod nematode seems to be restricted to South inlight of thesenew technologies. Rescue strategies are Africa. Screeningof local peanut germplasm, selected necessaryfor situationswhere the management strategy elitebreeding lines and hybrids provided at best a par- has failed. tiallevel of resistanceto D.africanus .Thispartial resis- tance,however, is overcomeat high nematode infestation levels,which is common in peanut Ž elds,given the nema- 100Basic strategies for defensive response of plants todehigh reproductive potential. The local genetic vari- againstnematodes ationin peanut breeding material is small. Most popular J.G. LUIS hybridsare Spanish or Virginia‘ bunch’types, with a few ‘runners’still produced in remote areas. The uniqueness InstitutoUniversitario de Bio-Orgá nica ‘ AntonioGonzá lez’ , ofthis nematode problem and the narrow genetic back- Universidadde LaLaguna, Avda, Francisco Sá nchez, 2, 38206 groundof availablepeanut germplasm present a challenge LaLaguna, T enerife,Canary Islands, Spain for theselectionand applicationof resistanceas a sustain- Inthe last few years,our Ecological Chemistry research ablestrategy to manage D.africanus in peanut. grouphas been engaged in molecular level studies on the host-plantinteractions between banana cultivars and the 102Resistance to root-lesion nematodes in Coffea pathogens: Fusariumoxysporum , MycosphaerellaŽ jensis canephora and Radopholussimilis .Severalsusceptible banana 1; 2 cultivarselicit a defenceresponse to attack by the Luc VILLAIN ¤,FranciscoA NZUETO and pathogenscharacterised by de novo productionof anew Jean-LouisS ARAH 3

Vol.4(2), 2002 157 Resistancebreeding against non sedentary nematodes (101-103) and F oodwebs (104-106)

1CIRAD TA 80/PS3Avenue Agropolis, 34398 Montpellier Therefore,early and rapid screening of plant material Cedex5 France for detectionof nematode resistance is desirable. It 2 ANACAFE, 5aCalle 00-50, Zona 14, Guatemala Ciudad, allowsa rapidselection at the very early plant stage of Guatemala potentiallyinteresting candidates. An in vitro screening 3 CIRAD TA 40/02Avenue Agropolis, 34398 Montpellier Cedex methodallows the elimination of susceptiblegermplasm, 5 France evenearlier compared to early screening in climatic [email protected] ¤ chambersor glass-/ screenhouses.However, the Ž nal Root-lesionnematodes ( Pratylenchus spp.)are widely selectionmust still be performedin theŽ eld,but massive distributedin coffee plantations worldwide. In Central eliminationof clearlynon-resistant plant material would America,many of them are highly pathogenic on considerablyreduce the size of such Ž eldscreening. Coffeaarabica causingimportant economic damage. Todate,the use of in vitro culturingtechniques has Thepoor efŽ ciency of nematicide treatments to control beenwell demonstrated as a rapidand reliable tool for Pratylenchus spp.was demonstratedin Guatemala. In determiningreproductive capabilities of plant-parasitic thiscountry, C.canephora was usedas a rootstock nematodeson known hosts, new hosts and resistant for 30years empirically. Analysis pointed out the varieties. In vitro screeningof varieties for resistanceto lowlevel of root-lesion nematodes in the Ž eldswhere plant-parasiticnematodes was Žrst demonstratedfor root- thisgrafting technique was used.Comparative studies knotnematodes, Meloidogyne spp.,using excised tomato ofpenetration dynamics and reproductive Ž tnesson roots.Nowadays, root-explant cultures of manydifferent coffeeseedlings as well as Ž eldtrials demonstrated the plantshave been used for evaluatingresistance to both existenceof pre- andpost infection factors of partial sedentaryand migratory endoparasitic nematodes. In vitro resistanceto Pratylenchus spp. in some C.canephora screeningprocedures were successfullydeveloped for genotypes.Complementary histological studies of roots Musa,allowingselection of constitutiveresistance against showedno noticeable structural differences between theroot-lesion nematode, Radopholussimilis . C. arabica and C.canephora .Onthe other hand, thepresence of high amounts of polyphenols was 104Structure andfunction offoodwebs observedin the roots of a C.canephora resistant genotypeeven before nematode penetration, suggesting 1; 2 Bryan S. GRIFFITHS ¤, Wim VAN DER PUTTEN and theexistence of constitutivefactors of resistance.Sources Peter C. DE RUITER 3 andmechanisms of resistancemust be investigatedmore exhaustivelyamong C.canephora germplasm.Grafting 1ScottishCrop Research Institute, Invergowrie, Dundee onto C.canephora constitutestoday the best alternative DD2 5DA, UK 2 tonematicide treatments for controllingphytoparasitic NetherlandsInstitute of Ecology,Centre for T errestrial Ecology,P .O.Box40, NL-6666 ZG Heteren,NL nematodecommunities in C. arabica coffeecrop, as some 3 Departmentof Environmental Sciences, University Utrecht, C.canephora genotypesalso show resistance to highly P.O.Box80115, 3508 TC Utrecht,NL pathogenicroot-knot nematodes ( Meloidogyne spp.). ¤[email protected] Severalsoil food webs have been thoroughly described 103 In vitro screeningas anearlyrapid and reliable andthe importance of nematodes is evident from their toolto detect resistanceagainst migratory representationat differenttrophic links – asplant-feeders, endoparasiticnematodes secondarydetritivores, predators and omnivores. Car- Annemie ELSEN bon owmodels have shown the importance of several pathways,including those through bacterial- and fungal- Laboratoryof Tropical Crop Improvement, K.U.Leuven, feedersto predaceous nematodes, and from plantroots KasteelparkArenberg 13, 3001, Leuven, Belgium throughplant-feeders. The herbivory pathway has been [email protected] observedexperimentally by followinga pulseof 13C, ap- Growingnematode-resistant crop varieties is considered pliedabove-ground, through the soil community. Novel asan effective and sustainable method for nematode observationsare also emerging on the effects of root- management.IdentiŽ cation and selection of resistant feedingfauna on above-ground food webs. Disturbance varietiesoften occurs in the Ž eld,but Ž eldtesting offoodweb groups causes a lossin stability,reduced bio- istime consuming, especially with large-sized plants. diversityand changes in soilprocesses. Consideration of

158 Nematology Symposiumabstracts biodiversityin food web models will allow exploration functionalredundancy among bacterivorous nematode ofbiodiversity-product ivityand biodiversity-stabilit yre- species.T orelatelaboratory-derived feeding rates to lationships.Modelling interactions according to func- populationgrowth rates under natural conditions, we tionrather than species reveal interesting interactions in addressthe effects of external in uences on nematode above-groundfood webs. Current soil food web models activityin soils, including texture, aeration, temperature grouporganisms by function and taxa, but for improved andmoisture. understandingof thesystem information on broaderfunc- tionalgroups may be moreappropriate. Current initiatives 106Decomposition pathways and successional anddevelopments in foodweb research and theory high- changes lightthe importance of soilfood webs in biosphere func- 1; 2 tionand are providing insights into the preservation, con- Liliane RUESS ¤ and Howard FERRIS servationand management of soilsystems. 1Instituteof Zoology,TU Darmstadt,Schnittspahnstr .3,64287 Darmstadt,Germany 105Carbon and energy sources for nematodes 2 Departmentof Nematology, University of California,Davis, CA95616,USA 1; 2 3 Tom MOENS ¤, Gregor W. YEATES and Paul DE LEY ¤[email protected] 1BiologyDepartment, Ghent University, K.L. Ledeganckstraat Insoils, energy and nutrient pathways are primarily me- 35,B-9000 Gent, Belgium diatedby bacteria or fungi. Bacteria-dominated systems 2 LandcareResearch, Private Bag 11-052, P almerstonNorth, rapidlytransfer nutrients, directly and via consumers,to New Zealand plants.In contrast, fungal-based decomposition channels 3 Departmentof Nematology,University of California, areslower; they are driven by morecomplex organic re- Riverside,California, USA sources.There are strong linkages between nematodes ¤[email protected] andtheir fungal or bacterialfood sources. On one hand, Nematodespecies diversity, even at small spatial scales, consumerorganisms affect rates of energy and nutrient providesan intriguing system for studieson resourcepar- releasefrom theirprey; on theother hand, they may regu- titioningand on links between biodiversity and ecosys- latethe prey biomass. The nature and abundance of avail- temfunction. The ubiquity and abundance of nema- ableresources can be monitored by faunal analysis of todesin soils and aquatic sediments interest ecologists fungal-and bacterial-feeding nematodes (f/ bratio,chan- seekingmodels for uxesof carbon, nitrogen, phos- nelindex). The resourceschange constitutively with time. phorus etc.However,the complete spectrum of roles Readilydecomposable portions are rapidly consumed by andthe quantitative importance of nematodes in soils bacteriaand their predators so that the recalcitrant frac- andaquatic sediments remain poorly documented. Food tionbecomes proportionally greater. That change is mir- sourcesof nematodes are usually inferred from buccal roredby correspondingincrease in fungaldecomposition morphologybut there are many unresolved questions andre ected in the nematode fauna. W ediscussthe re- onfood sources and feeding rates. Current functional lationshipof nematode trophic structure with the nature guildclassiŽ cations are useful but in some cases, in- oftheincoming organic material and the prevailing state cludingnematodes with minute buccal cavities and ty- ofthephysical environment. For example,decomposition lenchidswith delicate spears, food sources are uncer- pathwaysof naturalforests are predominantly fungal and tain.W ediscusswhether, and how ,alimiteddiversity thoseof agriculturalsystems are bacterial. W ediscussthe offunctional types can be reconciled with great species signiŽcance of the pathways in relation to the structure diversityand present a laboratoryapproach for studying andfunctions of theentire soil food web.

Vol.4(2), 2002 159 Detectionand diagnostics (107-122)

107Immuno-magnetic capture (IMC): anovel 2Istitutoper la Protezione delle Piante, Consiglio Nazionale approachto nematode diagnostics delleRicerche, V iaAmendola 165/ A,70126Bari, Italy ¤[email protected] 1; 2 3 Qing CHEN ¤, Lee ROBERTSON , Rosane H. CURTIS , APCRbasedmethod was appliedto the detection of 1 1 John T. JONES andDerek J.F .B ROWN grapevinefanleaf virus (GFL V). Thevirus was ampliŽed from specimensof itsvector Xiphinemaindex , collected 1Plant-PathogenInteractions Programme, Scottish Crop ina grapevineorchard at Palagiano, Italy. The detection ResearchInstitute, Invergowrie, Dundee, DD2 5DA,Scotland, UK was carriedout with a real-time uorescentRT -PCR 2 Universidadde Castilla-LaMancha, F acultadde Cienciasdel assay.A setof primerswas designedfor theGFL VRNA- MedioAmbiente, EdiŽ cio Sabatini, Laboratorio 0.8, E-45071 2ampliŽcation. A 2500bp fragment was ampliŽed by Toledo,Spain RT-PCR,from X. index adultsand juveniles, cloned and 3 NematodeInteractions Unit, IACR Rothamsted,Harpenden, sequenced.W ecomparedthe obtained GFL Vsequences HertsAL5 2JQ,UK withthose available in GenBank and two regions were ¤[email protected] selectedfor virusdetection and/ orstrain identiŽ cation. AuorescentScorpion probe was designedto amplify Simple,rapid and reliable diagnostic techniques are a100bp fragment within the conserved region of the importantfor identiŽcation and quantiŽ cation of plant- cpgene. A 21bp conserved motif at position 2855 parasiticnematodes. Current diagnostic methods have was usedas probe target. An additional region with limitationsfor Želdsample detection. ELISA diagnosis strain-speciŽc nucleotidevariations at position 2502 isless sensitive than PCR andthe sensitivity of PCR was usedto discriminate between the Palagiano and basedmethods is strongly affected by inhibitors from otherGFL Vstrains.SpeciŽ c oligos,constructed on the soil.IMC offers severaladvantages which overcome basisof GenBank sequences AF304015 and X16907, thesedrawbacks. In IMC anantibody which recognises were usedas controls. Furthermore, a setof three thesurface of target nematodes is incubated with a strain-speciŽc molecularbeacons was designedon this nematodesuspension extracted from aŽeldsample. Then, regionand used with the ampliŽ ed DNA oroligos. The secondary-antibodycoated magnetic beads are added and successfuldetection of the probe targets was shown amagnetis used to capture target nematodes while by uorescentsignals emitted during ampliŽ cation or othernematodes are discarded. W ehaveused IMC to underUV excitation.The Scorpion probe proved to detect Meloidogynearenaria andavirus-vectornematode beuseful in virus detection. Due to their single-base Xiphenemaamericanum. Eightyand 60% of target mismatchsensitivity, the molecular beacons appeared nematodes,respectively, were recoveredfrom mixedsoil suitablefor speciŽc strainrecognition. The potential of samples.These results show that IMC isan effective thesetechnologies in thestudy of transmission and vector methodof detecting speciŽ c nematodesin mixed soil epidemiologyare brie y discussed. samples.IMC convertsŽ eldnematode samples to lab- friendlysamples, containing highly concentrated target nematodessuitable for furtherserological or molecular 109Advances in applied nematode research in techniques,such as ELISA, PCRor Taqman, which South Africaafter introduction of the SCAR-PCR canbe used to detect and quantify speciŽ c nematodes technique fornematode identiŽ cation orviruses. W earecurrently examining the potential Hendrika FOURIE1; , Caroline ZIJLSTRA 2, Alexander commercialapplications of combining IMC withPCR- ¤ Henrique MCDONALD 1 andGertruida Anna V ENTER 1 baseddiagnostics for routinedetection of virus vector nematodes. 1ARC-GrainCrops Institute, Private Bag X1251, Potchefstroom,2520, South Africa 2 PlantResearch International, P .O.Box 16, 6700 AA 108Detection of GFLVin Xiphinemaindex with Wageningen,The Netherlands DNA-based uorescentprobes ¤[email protected] 1 2; Mariella FINETTI-SIALER andAurelio C IANCIO ¤ Theavailability of well-deŽned nematode populations is essentialfor hostplant resistance screening, breeding and 1 Dipartimentodi Protezionedelle Piante, Università degli croprotation purposes. IdentiŽ cation of monocultureroot- Studi,Bari, Italy knotnematode species occurring in South African soils

160 Nematology Postersessions anddifferentiation between species in mixedpopulations andto chemical treatments for eradicationof the nema- were doneby means of the sequence characterised todes,which was successfulin all but a few accessions. ampliŽed region – polymerasechain reaction (SCAR- Withthese phytosanitary methods the Laboratoryof Plant PCR)technique. Meloidogynefallax , M.chitwoodi , M. Quarantineof EmbrapaGenetic Resources and Biotech- javanica, M.incognita , M. arenaria and M. hapla nologycollaborates actively to reduce the riskof introduc- were identiŽed from variouscrops. Multiplex internal tionof newplant-parasitic nematodes species in Brazil. transcribedspacer (ITS)-PCR ampliŽed a fragmentin anunknown root-knot nematode species for whichno 111Professional automated extracting apparatus SCAR-marker ispresently available. This technique forfree-living nematodes enabledthe identiŽ cation of M. fallax,aquarantine organismin Europe, as a newrecord for South Gerard HENDRICKX Africa wherepresence at two localities was established. Resultingfrom thisstudy, the geographical distribution CropProtection Department, Agricultural Research Centre, of M.chitwoodi was expandedfrom twoto Ž veknown Burg.V anGansberghelaan 96, 9820 Merelbeke, Belgium [email protected] localitiesin this country. Root-knot nematode species compositionof Ž eldand glasshouse isolates can be Nematodeextraction by elutriation and subsequent monitoredfrequently using the SCAR-PCR technique, separationon acottonwool Ž lteris labour-intensive, time whichallows for routineanalyses of root-knotnematode consuming,and inefŽ cient. Full automation is difŽ cult. speciesused in research programmes. Extractionbased on centrifugal  otationin swinging bucketsis faster and more efŽ cient but does not allow 110Detection and eradication of plant-parasitic for extractionof large volumes and is still difŽ cult to nematodesin importedgermplasm in Brazil automate.A prototypeof a fullyautomated extraction apparatuswas builtusing a zonalcentrifuge. With this 1; 1 2 3 V. GONZAGA ¤, R.C.V. TENENTE and J.K.A. MATOS prototypethe sample volume was limitedto 100 cm soil (10 cm3 extracted);subsequently, a versionprocessing 1 EMBRAPA,GeneticResources and Biotechnology, P .O.Box 200 cm3 (100 cm3 extracted)was constructed.Besides 2372(70849-970) Brasilia, DF ,Brazil previousmodiŽ cations, changes were madeaiming at an 2 Universidadede Brasília, P .O.Box 4508 (70910-970) Brasilia,DF ,Brazil easierand safer operationby using a conveyor-belt,a pneumaticdrive, and a fool-proofdesign. The apparatus ¤[email protected] iscommercial and can be used to extract other plant TheLaboratory of Plant Quarantine of Embrapa Ge- pathogensfrom soil. neticResources and Biotechnology makes phytosanitary analysesof most of the plant germplasm introduced in Brazil.During 2001, plant-parasitic nematodes were de- 112Magnetic separation as atoolin asample tectedassociated with 1738 germplasm accessions com- preparationprocedure for direct detection and ingfrom severalcountries. The material infected with quantiŽcation of trichodoridnematodes nematodeswere corn,wheat, rice, soybean, oats, melon, RodanthiC. H OLEVA1; , Qing CHEN 1, sesame,cocoa, coffee, orchid, neen, clover and lespedeza. ¤ Rosane CURTIS 2, Roy NEILSON 1, John JONES 1, Thosematerials were introducedfrom thefollowing coun- Vivian BLOK 1 andDerek J.F .B ROWN 1 tries:France, Mexico, USA, Colombia,Japan, Argentina, Venezuela,Trinidad-T obago,Guatemala, Dominican Re- 1ScottishCrop Research Institute, Invergowrie, Dundee, DD2 public,Philippines, Spain and Holland. The techniques 5DA,Scotland, UK 2 usedfor extractionand detection of thenematodes were IACR Rothamsted,Harpenden, Herts AL5 2JQ,England, UK Baermannfunnel, tray technique, sieving and blending ¤[email protected] andcyst detection by Fenwickcan. The plant-parasitic ne- Thedetection of trichodorid nematodes in soil samples matodesdetected were Ditylenchusdipsaci , Ditylenchus isdifŽ cult as conventional methods are laborious, parvus, Ditylenchus sp., Aphelenchoidesbesseyi , Aphe- time-consumingand rely on specialist knowledge of lenchoides sp., Aphelenchusavenae , Aphelenchus sp., morphologicalcharacteristics of thespecies.T oovercome Helicotylenchus sp.and Hoplolaimidae. The accessions thesedifŽ culties the potential of magnetic separation were submittedto dry and humid thermal treatments for recoveringtarget nematodes from soilpopulations

Vol.4(2), 2002 161 Detectionand diagnostics (107-122) ofnematodes was investigated.Lectins and polyclonal standarddilutions. Currently, simultaneous detection and antiserumthat bound speciŽ cally and reproducibly to quantiŽcation of the above trichodorid species is being theoverall surface of Paratrichodorusanemones were evaluated.Our dataconŽ rm thisto be a rapid,accurate identiŽed and bound to monodispersesuperparamagnetic andsensitive molecular diagnostic. particles(Dynabeads) to capture target nematodes from testsuspensions. In recovery experiments, while both 114A methodfor estimating the ratioof typesof probe isolated nematodes from suspension, Meloidogyneincognita to M. arenaria in mixed antibody-coatedbeads recovered them more efŽ ciently populationsusing PCR-RFLP thanbeads coated with lectins. The results obtained suggestthat the immuno-magnetic separation (IMS) Hideaki IWAHORI¤ andZen-ichi S ANO techniquehas potential to provide a serological-based targetnematode enrichment process. Furthermore, it NationalAgricultural Research Center for Kyushu Okinawa representsa signiŽcant advance over current methods for Region,Nishigoushi, Kumamoto 861-1192, Japan [email protected] recoveringnematodes as itfacilitates high throughput to ¤ screenlarge soil populations quickly and economically Themost important root-knot nematodes in southwestern and,being technically simple, requires minimal labour Japan are Meloidogyneincognita and M. arenaria. These input. twospecies are sometimes present in the same Ž eld.W e deviseda methodto estimate the ratio of M.incognita 113Development and evaluation of auorogenic to M. arenaria inmixed populations using PCR-RFLP ™ andimage analysis. V ariousratios of thetwo nematodes 50nucleasePCR assay(T aqMan )forthe detection andquantiŽ cation of virus-vectortrichodorid species were prepared,then PCR-RFLP was performedusing eachratio. Brightness of species-speciŽc RFLP bandsfor

RodanthiC. H OLEVA¤, Vanessa YOUNG,Konstantina M.incognita and M. arenaria was quantiŽed withimage BOUTSIKA, Mark PHILLIPS,DerekJ.F .B ROWN, analysisusing a computerprogram. The brightness of the Roy NEILSON and Vivian BLOK M.incognita -speciŽc bandto that of the M. arenaria band for eachRFLP patternwas usedas anestimate for theratio ScottishCrop Research Institute, Invergowrie, Dundee, DD2 ofthe two nematodes. A regressionline was calculated 5DA,Scotland, UK from thesevalues to estimate actual ratios. W eapplied [email protected] ¤ thismethod successfully to Želdsamples. Attempts have Moleculardetection of pests and pathogens relies on alsobeen made to apply this method to other nematode rapidand dependable methods for theidentiŽ cation and combinations,such as M.incognita and M. hapla or two quantiŽcation of theseorganisms. Here, thedevelopment differentpopulations of Pratylenchuscoffeae . ofa qualitativeand quantitative diagnostic method for trichodoridnematodes based on T aqMan ™ chemistry isreported. T woindependent primer/ probesets were 115Detection of the reniformnematode in cotton designedtargeting the 18S gene of theribosomal cistron usinghyperspectral imagery andproducing an amplicon of 83bp for thevirus-vector A.T. KELLEY 1, G.W. LAWRENCE 1, K.S. MCLEAN 2 species, Paratrichodoruspachydermus and Trichodorus and H.K. LEE 1 similis.Theassay was appliedto puriŽ ed plasmid DNA containingclones of the18S region from bothnematode 1 Departmentof Entomology and Plant P athology,Mississippi species,as well as to genomic DNA extractedfrom StateUniversity, Mississippi State, MS 39762,USA 2 individualnematodes. Both primer/ probesets displayed Departmentof Entomologyand Plant P athology,Auburn highspeciŽ city as no cross-reaction was observedwhen University,Auburn University, USA testedwith samples of two other trichodorid species. In [email protected] experimentswhere dilutions of puriŽed plasmidstandards Thereniform nematode ( Rotylenchulusreniformis Lin- were usedto test the analytic sensitivity, the T aqMan ™ fordand Oliveira, 1940) is one of the two most preva- assaydetected nematode DNA tothe femtogram level. lentnematodes on cotton ( Gossypiumhirsutum L.) and QuantiŽcation of thetarget present in unknown samples isquickly spreading throughout the southeastern United was performedby comparison of the  uorescence States.This nematode inhibits cotton plant development signalsof the samples to those obtained from plasmid resultingin reduced plant growth and sometimes plant

162 Nematology Postersessions death;cotton crop yield loss up to 40-60% has been due Sevenwheat varieties and 13 doubled haploid wheat toreniform nematode infestations. A productionŽ eldnat- lineswere inoculatedwith P.neglectus or P. thornei and urallyinfested with the reniform nematode and controlled grownin a glasshouseexperiment for 8weeks.Root Želdmicroplots are currently being used to study the cor- systemswere washedand nematodes were quantiŽed relationbetween reniform nematode population thresh- usingeither a microscopeor PCR.For assessmentusing oldsand re ectance properties exhibited by infectedcot- themicroscope, root systems were washedto remove tonplants. Re ectance properties, measured by a hand- soil,cut and misted for 4daysand the nematode heldhyperspectral spectroradiometer, will be used to suspensioncounted using a dissectionmicroscope. For develophyperspectral images. These hyperspectral im- assessmentusing PCR, plant roots were washedand agesalong with nematode population threshold data will nematodeDNA extracted.PCR (speciŽc toeither P. beused to determine relationships between cotton plant neglectus or P. thornei)was usedto quantify DNA stressand nematode population thresholds. Hyperspectral levels.There was highervariation between replicates with imagerymay be ausefulremote sensing tool enablingcot- microscope(c.v. 63%)compared with PCR assessment D tonproducers to test for nematodepresence while avoid- (c.v. 22%).This was attributedboth to errors associated D ingthe time consuming and sometimes costly soil sam- withextraction using the misting chamber and in sub- plingprocess presently used today. samplingand counting the nematode suspension using themicroscope. For both P.neglectus and P. thornei, signiŽcant positive linear correlations were obtained 116Possibilities of autouorescence in betweenthe microscope and PCR methods ( P.neglectus anematologicaldiagnostic laboratory r 2 0:80: P < 0:001; P. thornei r2 0:512: P < 0:05). D D ThePCR technique for quantifying P.neglectus or P. Loes den NIJS¤ and Gerrit KARSSEN thornei iscurrently being used for screeningcereals in PlantProtection Service, P .O.Box9102, 6700 HC Wageningen, southernAustralia. TheNetherlands ¤[email protected] 118Use ofthe mistiŽer forextraction of rootlesion Autouorescence is a phenomenonthat scientists often nematodes (Pratylenchus spp.) fromsoil tryto avoid because it interferes with the induced  uo- 1; 1;3 rescencethey want to measure.W estudiedgreen auto u- Vivien A. VANSTONE ¤,CarolineA. V ERSTEEG , orescencein free-living and plant-parasitic nematodes at MichelleH. R USS 1;2 andSharyn P .T AYLOR 2 lowmagniŽ cation (12.5-100 ).Firstobservations indi- £ 1 cateit isuseful for thedistinction between plant-parasitic Universityof Adelaide,Department of PlantScience, pmb 1, GlenOsmond, 5064, South Australia, Australia andnon-plant-parasiti cnematodes,endo-parasitic nema- 2 SouthAustralian Research and Development Institute, GPO todedetection in roots, nematode viability tests such as Box397, Adelaide, 5001, South Australia, Australia liveand dead counting of juvenilesof potatocysts nema- 3 Departmentof Primary Industries, Centre for W etTropics todesand for detectionof other speciŽ c itemslike Pas- Agriculture,South Johnstone, 4859, Queensland, Australia teuria endospores.The Leica MZ FLIII dissectionmi- ¤[email protected] croscopehas been tested on a varietyof nematological objects.The advantages and disadvantages will be high- Since1995, we haveused the mistiŽ er for extractionof Pratylenchus from Želdsamples of soil roots. This lighted. C requireda Žlterthat retained soil, allowed passage of nematodes,and remained intact through 96 hofmisting. 117Comparison of quantitativePCR andmister CoffeeŽ lters(Autocup ®,Thomasand Green Ltd, UK extractionfor the assessmentof Pratylenchus or Altra® FiltersInc., USA) were themost suitable neglectus or P. thornei material.Four Ž ltersare used per 50 gsample,providing cleanextract without hindering passage of nematodes. Sharyn TAYLOR , Alan MCKAY, ¤ Extractionand sample variation were comparedwith Kathy OPHEL-KELLER, Danuta SZOT and HERDINA thetray method of Whitehead and Hemming using SouthAustralian Research and Development Institute, GPO threenaturally infested Ž eldsoils and pasteurised soil Box397, Adelaide, South Australia 5001, Australia inoculatedwith cultured P.neglectus .Eightypercent of ¤[email protected] nematodesfrom theŽ eldsoils were Pratylenchus . Forty

Vol.4(2), 2002 163 Detectionand diagnostics (107-122) eightpercent more nematodes were extractedby mister 2 INRA-USVE, Nematologygroup, B.P .2078,06606 Antibes, thanwith the tray. Sample variation was similarfor mister France (16-23%)and tray (12-20%). Numbers recovered (64%) ¤[email protected] from inoculatedsoil did not differ between methods. Xiphinemaindex (XI), X.diversicaudatum (XD), X. Nematodesin natural soil could have been attached to vuittenezi (XV) and X. italiae (XIT) areestablished or soilparticles or organicmatter, contained within roots or putativevectors of several nepoviruses of grapevine. anhydrobiotic.These nematodes were thereforeextracted Allfour species are closely related, which makes them moreefŽ ciently by themister than from thetray, probably difŽcult to identify reliably when only single or few dueto the greater aeration and water  owon themistiŽ er . individualsare available. With the aim of Ž ndinga For Želdsoils, extraction by mistiŽ er resultedin higher solution,a simplediagnostic method was developed. Pratylenchus recovery. TheITS1 regionspanning the 18S and 5.8S ribosomal geneswas sequencedin one population of each species, 119PCR-based techniques fornematode diagnosis usingtwo conserved primers from theserespective genes. derivedfrom the study ofgeneticdiversity Sequencecomparisons of ITS1, sized 1132 (XV), 1153 (XI), 1175(XD) and1190 (XIT) bp,suggested a genetic L. WAEYENBERGE¤, M. MADANI, H. YU, proximitybetween the two established vector species X. S.A. SUBBOTIN and M. MOENS index and X.diversicaudatum onthe one hand and X. CropProtection Department, Agricultural Research Centre, vuittenezi and X. italiae onthe other hand. The sequence Burg.V anGansberghelaan96, 9820 Merelbeke, Belgium variabilityof ITS1 allowedthe design of internal sense ¤[email protected] primers,speciŽ c for eachspecies, which ampliŽ ed, in combinationwith the same antisense ITS1 primer,a Oneof the main activities in the Department of Crop singlesignature fragment (340 bp for XI, 414bp for Protection(Agricultural Research Centre), is the devel- XIT, 591bp for XVand813 bp for XD). Theprimers opmentof identiŽ cation and detection protocols. Identi- were successfullyused in amultiplextest for thereliable Žcationof nematodesbased on morphologyand morpho- detectionof two to four mixed species each represented metricsis time-consumingand difŽcult due tooverlapping byasingleindividual. ofmany characteristics. Recently molecular identiŽ cation techniquesbased on PCR have been developed and suc- cessfullyapplied for nematodediagnostics. These tech- 121PCR basedmethods forthe analysisof Dutch niquesallow identifying genetic markers speciŽ c for cer- soilsamples for the presenceof root-knotnematodes tainnematode species or groupsof populations.Genetic markersare identiŽ ed bytheanalysisof PCRproducts us- Carolien ZIJLSTRA¤, Dorine DONKERS-VENNE, ingRFLP ,directsequencing and random PCR approaches Richard VAN HOOF, Linda KOX and Gerrit KARSSEN likeRAPD andAFLP .Markersare used to develop PCR PlantResearch International (PRI), P .O.Box 16, 6700 AA withspecies-speciŽ c primersfor rapidnematode detec- Wageningen,The Netherlands; Plant Protection Service (PD), tionand Real Time PCR for nematodequantiŽ cation in P.O.Box9102, 6700 HC Wageningen,The Netherlands samples.The poster presents several PCR-applications ¤[email protected] usedfor nematodeidentiŽ cation in CLO-DGB. Theroot-knot nematodes Meloidogynechitwoodi , M. fallax, M. hapla and M. naasi areregularly found as 120PCR multiplexidentiŽ cation of single plant-parasiticnematodes in Dutch agriculture, either individualsof the Longidoridnematodes, Xiphinema presentalone or as mixtures. The former twohave index, X.diversicaudatum , X.vuittenezi and X. italiae acquiredimportance as quarantine pests while M. usingspeciŽ c primersfrom ribosomal genes hapla hasalso increased importance. All three species causeconsiderable damage on most agricultural crops. Xirong WANG 1, Nathalie BOSSELUT 2, Meloidogynenaasi isalso found more frequently in Chantal CASTAGNONE 2, Roger VOISIN 2, Pierre ABAD 2 agriculturalplots, but its relevance is limited to grasses. and Daniel ESMENJAUD2; ¤ Inspectionservices identify root-knot nematode species 1 NematologyLaboratory, South China Agricultural University, generallybased on morphological characteristics. In GuangZhou,China acollaborationbetween PRI andthe PD, analyses

164 Nematology Postersessions ofnematode samples have been compared. Root-knot toensure safe andsustainable agriculture. However, the nematodecomposition of samples of approximately multiplicityof detection assays available for aspeciŽc 100nematodes was Žrst determinedmorphologically; pathogenleads to a lackof consistency among the subsequently,the samples were dividedbetween PRI and varioustesting agencies and hampers standardisation. PDandanalysed using PCR assays for thedetection Micro-arraytechnology, in which thousands of different of M.chitwoodi , M. fallax, M. hapla and M. naasi. oligosor proteins can be spotted on little more than For theformer threespecies, species-speciŽ c PCR 1 cm2,enablesthe detection of many different target primersalready existed; for M. naasi anewspecies- moleculesin the same sample with increased speciŽ city. speciŽc primerwas developed.The results between Therefore,micro-arrays can meet the demands for fast, thethree analyses were comparable.With respect to speciŽc, efŽ cient, cost-effective, user-friendly and reliable routinediagnostic assays, PCR based assays seem to multiplexdetection methods for differentorganisms. offer attractivealternatives: they are reliable, sensitive, Todevelopthe micro-array technology for diagnostic relativelyfast, easy to perform, and do not require purposes,generic extraction methods for DNA andRNA nematologicalexpertise. haveto be developed. Secondly, the sensitivity must beimproved in order to detect low concentrations of extractednucleic acids, with generic pre-ampliŽ cation 122Multiplex detection of nematodesand other soil methods.Recently a revolutionaryporous capillary solid relatedorganisms using three dimensional phasemicro-array has been developed. The capacity micro-arrays ofthis three-dimensional array to bind oligonucleotide s ishigher than that of a two-dimensionalglass array Carolien ZIJLSTRA¤, Arjen SPEKSNIJDER, Peter BONANTS and Cor SCHOEN resultingin a highersensitivity. Moreover, the porous solidphase allows  owthrough measurements, resulting PlantResearch International (PRI), P .O.Box16, 6700 AA infasthybridisation times of only15 mininstead of 18h Wageningen,The Netherlands asonglass. Recent data to detectplant pathogens in this [email protected] ¤ multiplexsetting look promising. Detectingthe presence of nematodesand other soil related plantpathogens or beneŽ cials in soil samples is helpful

Vol.4(2), 2002 165 Evolution,phylogeny and classiŽ cation (123-175)

123Comments onintraspeciŽc variabilityin maticmachinery necessary for homologousrecombina- the genus Chiloplacus Thorne,1937 tionis present in animal mitochondria, along with phy- logeneticevidence that recombination has been a feature Joaquín ABOLAFIA, Gracia LIÉBANAS, ofthe evolution of primate mtDNA, haschallenged the Pablo GUERRERO and Reyes PEÑA-SANTIAGO¤ receivedorthodoxy that animal mtDNA isinherited clon- Departamentode Biología Animal,V egetaly Ecología, ally.The existence of a persistentheteroplasmic state as Universidadde Jaé n, Campus‘ LasLagunillas’ s/ n, found in G. pallida wouldseem likely to provide am- 23071-Jaén, Spain pleopportunityfor recombination.W enowhave evidence ¤[email protected] ofmtDNA recombinationboth between distinct G. pal- The genus Chiloplacus Thorne,1937 is one of themost lida mtDNA lineages,and between sub-types of single widelydistributed taxa among cephalobid nematodes, and mtDNA lineages.Recombination is demonstrated using someof its species are considered to be cosmopolitan. thelatest methods from statisticalphylogenetics, includ- Thegenus currently includes 30 valid species, plus ten inghidden Markov models and Bayesian (Markov chain incertaesedis ,whichshow a remarkableinterspeciŽ c MonteCarlo) approaches. variabilitywhen they are studied under both optical microscopyand SEM. In Andalucí a Oriental(SE Iberian 125Conversion of AFLPmarkers to PCR-based Peninsula)the genus is represented by six species: C. STSmarkersto identify Heteroderaglycines demani (Thorne,1925) Thorne, 1937; C. magnus Rashid populations &Heyns,1990; C. minimus (Thorne,1925) Andrá ssy, 1959; C. tenuis Rashid & Heyns,1990; C.trilineatus N. ATIBALENTJA¤, G.R. NOEL and L. DOMIER Steiner,1940 and Chiloplacus sp.In addition to the usual variabilityaffecting the body length, number of lateral Departmentof CropSciences, University of Illinois,Urbana, Želdincisures, postvulval sac length, tail morphology, IL61801,USA; USDA-ARS, Urbana,IL 61801,USA [email protected] etc.,presentstudy has revealedan interestingvariability in ¤ severalfeatures of thelipregion: separated or moreor less NineAFLP markerswere identiŽed that provide varying amalgamatedlips, labial probolae differing in the shape degreesof speciŽ city to some of the 48 selected (biacuteor bifurcated) and the length of their branches, populationsinvolved in apreviousstudy that investigated cephalicprobolae also differing in shape and size, more theparasitic ability of the soybean cyst nematode, H. orless distinct primary and secondary axils, and cephalic glycines.The48 populationswere obtainedby subjecting marginsmooth, serrated or bearingtines. The taxonomic Želdpopulations of races1 through6, 9,and14 toseveral interestof these features is discussedbrie y. cyclesof selectionon eachof sixsources of resistanceto H. glycines:Peking,Cloud, PI88788, PI89772, PI209332 124Evidence for heteroplasmy and recombination andPI90763. The nine AFLP markers were cloned innematode mitochondrial DNA andtheir DNA nucleotidesequence determined before theywere usedto design oligonucleotide primers for 1; 1 Miles ARMSTRONG ¤, Mark PHILLIPS , PCR-ampliŽcation of genomic DNA from theoriginal Dirk HUSMEIER 2, Frank WRIGHT 2 and Vivian BLOK 1 unselectedŽ eldpopulations. Following this procedure, we 1ScottishCrop Research Institute, Dundee, DD2 5DA, havegenerated STS (sequencetag site) primers that are Scotland,UK speciŽc for races1 through6, 9, and 14 of H. glycines. 2 Bioinfomaticsand Statistics Scotland, Scottish Crop Research TheseSTS markers shouldbe very useful as diagnostic Institute,Dundee, DD2 5DA,Scotland, UK probesfor theidentiŽ cation of Ž eldpopulations of H. ¤[email protected] glycines. Wepreviouslyreported the unusual multipartite arrange- mentof the mtDNA ofthe potato cyst nematode Glo- 126Phylogenetic relationships between boderapallida. Its mtDNA consistsof a populationof Paratrichodorus and Trichodorus species(Nematoda: small,genetically distinct mtDNAs which,presumably, Trichodoridae) existin as yet unknown combinations in order to en- codeall the proteins necessary for mitochondrialfunction. KonstantinaB OUTSIKA¤, Mark S. PHILLIPS, Recentlypublished biochemical evidence that the enzy- Derek J.F. BROWN andVivian C. B LOK

166 Nematology Postersessions

ScottishCrop Research Institute, Invergowrie, Dundee DD2 for P. thornei.Thisfragment was notampliŽ ed when to- 5DA,Scotland, UK talgenomic DNA from other Pratylenchus spp., Meloido- ¤[email protected] gyne spp., Heteroderamediterranea , Zygotylenchusgue- Paratrichodorus and Trichodorus species(trichodorids) varai, Ditylenchusdipsaci and Radopholussimilis was aresoil-inhabiting ectoparasitic nematodes, several of usedas template in PCR reactionsusing the newly de- whichare natural vectors of T obraviruses.Trichodorids signedprimer pair. usedin this study were obtainedfrom colleaguesin Europe,North and South America, Asia, and from British 128Studies onthe taxonomyof importantgenera soilsamples. Phylogenetic analysis of the18S gene from andspecies of Criconematoideafrom China ribosomalDNA sequencedata obtained from 17different trichodoridspecies revealed a majordifferentiation Lijie CHEN¤ and Weizhi LIU Paratrichodorus correspondingto the two genera and Departmentof PlantProtection, Shenyang Agricultural Trichodorus , but P. minor and P. porosus were only University,Shenyang 110161, P .R.China distantlyrelated to the other members of their genus. ¤[email protected] Analysisof thealigned sequences indicated the presence of‘ mosaic’sequences in the 18S gene of P. minor Morethan 895 samples from 67families, 189 genera, and P. porosus populationsand suggested recombination 246species of plants in 73 cities and counties of 17 events.The cuticle is one of the main characters used provincesand autonomous region were examinedin todistinguish between the two genera and, recently, Ž ne China.The important genera, Criconema, Ogma, Dis- structureof the body cuticle in several trichodorids was cocriconemella , Hemicycliophora , Hemicriconemoides , foundnot to be genusspeciŽ c. This observation, together Criconemella , Paratylenchus and Gracilacus,ofCricone- withour resultswith P. minor and P. porosus, suggeststhat matoideafrom Chinawere systematicallyobserved and thecurrent divisions in the Family Trichodoridae require identiŽed. Forty species from eightgenera of Cricone- areassessment. matoideawere identiŽed, including one species of Criconema,onespecies of Ogma,onenew species of Discocriconemella ,onespecies of Hemicycliophora , Ž ve 127Molecular diagnosis of Pratylenchusthornei species of Hemicriconemoides ,tenspecies of Cricone- 1; mella,13species of Paratylenchus andeight species of Susana CARRASCO-BALLESTEROS ¤, Gracilacus.Of the40 species,there were Žvenewones: Pablo CASTILLO 1,Encarnación P ÉREZ-ARTÉS 1 Discocriconemellasinensis n. sp., Hemicriconemoides andRafael M. J IMÉNEZ-DÍAZ 1;2 parasinensis n. sp., Paratylenchushandansis n. sp., 1Departamentode Protecció n deCultivos,Instituto de bayansis n. sp., and Gracilacuspopulus n.sp., and there AgriculturaSostenible (IAS), Consejo Superior de were 19newrecorded species in China. InvestigacionesCientí Ž cas(CSIC), Apartado 4084, 14080 Córdoba, Spain 2 EscuelaTé cnica Superior de IngenierosAgró nomos y Montes, 129The nematode collection at the Museum of Universidadde Córdoba, Apartado 3048, 14080 Có rdoba, ComparativeZoology, Harvard University Spain ¤[email protected] DavidBruce C ONN Root-lesionnematodes are economically important patho- Schoolof Mathematical and Natural Sciences, Berry College, gensof awiderange of crops.This study describes the de- MountBerry, Georgia 30165 USA andMuseum of Comparative velopmentof aspecies-speciŽc primerpair designed from Zoology,Dept. of InvertebrateZoology, Harvard University, aRAPD fragmentassociated with Pratylenchusthornei , Cambridge,Massachusetts 02138 USA [email protected] thecereal and legume root-lesion nematode. The RAPD fragmentwas clonedand sequenced and primers for spe- TheMuseum of ComparativeZoology (MCZ) of Harvard ciŽc PCRwere developedfrom sequencedata. SpeciŽ c Universitymaintains several globally important research PCRanalyses using the new primer pair and total ge- collections.The MCZ’ sDepartmentof Invertebrate nomicDNA extractedfrom amixturecontaining all life Zoologycurates nearly one million specimens, 10 000 stagesof thenematode (eggs, juveniles and adult females) ofwhich are type specimens; additionally, entomology rendereda 1050bp DNA fragmentwhich proved speciŽ c andmalacology collections are curated by separate

Vol.4(2), 2002 167 Evolution,phylogeny and classiŽ cation (123-175) departments.The nematode collection is growing, and 131T estinga molecularbarcode system against the includessigniŽ cant holdings obtained from several biologicalspecies concept in cultureof Panagrolaimus expeditionsand studies conducted around the world. populations Mostspecimens are parasitic species from animalhosts, butsome free-living and plant-parasitic species are Abebe EYUALEM and Mark BLAXTER included.Most were collectedwithin the past 100 years, Instituteof Cell, Animal and P opulationBiology, Ashworth butsome date closer to MCZ’ sfoundingin 1859. In Laboratories,King’ s Buildings,W estMains Road, The 1980,MCZ transferredmany nematode type specimens Universityof Edinburgh,Edinburgh EH9 3JT,Scotland,UK tothe United States National Parasite Collection, but retainedall other nematode specimens. Current collection Previouslywe developeda molecularbarcode system activitiesfocus primarily on voucherspecimens for biotic usingthe 18S of rDNA for deŽning a molecular surveysand ecological and experimental studies. Use operationaltaxonomic unit (MOTU) for theidentiŽ cation ofand contribution to the collections by nematologists ofnematodesfrom anexperimental Ž eldsite in the UK. outsideHarvard for scientiŽc andeducational purposes is Here we presentthe result of our attempt to identify encouraged,subject to approval by MCZ ona case-by- Ž ve Panagrolaimus populationsfrom thissite, which case basis. we maintainedin culture, morphologically, molecularly andbiologically, i.e.,throughbreeding each population againstthe other. The results showed that biologically 130Protein variation in potatocyst nematodes theybelonged to two reproductively isolated species. revealedby capillary electrophoresis Theavailable morphological criteria, including SEM, 1; were insufŽcient to differentiatebetween them; thus they MariaJosé M. DA CUNHA ¤,IsabelLuci P .M. DA CONCEIÇÃO 2, Lúcia M. PIRES 1, Isabel M. DE O. probablybelong to one species. The variation of the ABRANTES 2 andM. SusanaN. DE A. SANTOS 2 morphometricaldata rendered its use subjective, more soit did not discern between the two biological species. 1 EscolaSuperior Agrá ria de Coimbra, Bencanta Themolecular operational taxonomic unit resulted from 2 Institutodo Ambientee Vida,Departamento de Zoologia, ourbarcode system, perfectly agreeing with the breeding Universidadede Coimbra, 3004-517 Coimbra, P ortugal results,clearly separated the populations into two [email protected] ¤ groups.While understanding that the work was limited Capillaryelectrophoresis was usedto characterise 49 toonly Ž vepopulations of one genus, we maintain Portuguesepopulations of potato cyst nematodes, 46 of thatthis congruence of the MOTU conceptwith the Globoderarostochiensis and three of G. pallida, and biologicalspecies concept in the taxonomically difŽ cult eightrepresentative populations of pathotypes deŽ ned genus, Panagrolaimus ,showsthat the method could be for thesetwo species (Ro1, Ro2, Ro3 and Ro4 for G. promisingas it is simple, comparable and transferable, rostochiensis andPa1, Pa2 and Pa3for G. pallida). Protein thusuniversal. extractsof 50 cysts per sample were separatedusing BeckmaneCAP SDS 14-200kit, in a capillarywith 57 132Large-scale molecular barcode survey of soil cmlength and 100 ¹minternaldiameter and a runtime nematodes of40 min. Reproducible protein proŽ les were obtained andthe populations were comparedtaking account the Robin M. FLOYD¤ and Mark L. BLAXTER relativemigration time and area of each peak recorded by ICAPB, Universityof Edinburgh,King’ s Buildings,W estMains theGold Software DataSystem. Similarity indices (F) and Road,Edinburgh EH9 3JT,Scotland,UK geneticdistances (D 1-F) betweenpopulations were [email protected] calculatedusing peak Darea data and a dendrogramwas ¤ constructedaccording to the UPGMA method.Protein Wehavedeveloped a molecularbarcoding system for proŽles of eachpathotype were obtainedand some of the identiŽcation of soil nematodes by DNA sequencing, proteinswere characteristicof eachpathotype. However, andhere report on the results of our Ž rst major Ro1and Ro4 were notplaced together, and Pa1 was not survey.PCR was carriedout on individual nematodes, placedtogether with Pa2. Portuguese populations revealed and the 50 segmentof the small subunit ribosomal agreatvariability and it seemsthat protein composition is RNA (SSU) geneampliŽ ed and sequenced. Resulting notrelated to geographic origin of populations. sequences,typically 450-500 bases, were alignedand

168 Nematology Postersessions clusteredusing a neighbour-joiningalgorithm.Groups InvertebrateIdentiŽ cation T eam,Plant Health Group, Central ofsimilar or identical sequences were designatedas ScienceLaboratory, Sand Hutton, Y ork,YO41 1LZ, UK molecularoperational taxonomic units (MOTU). Two [email protected] large-scalesurveys were carriedout (in July and October The genus Aphelenchoides consistsof over140 nominal 2001)on a Scottishupland Agrostis-Festuca grassland species,some of which are pathogenic to plants. In soil(the Sourhope Soil Biodiversity Programme study thisstudy, a revisedlist of species was produced, site),using both molecular and morphological methods deletingdescriptions considered too poor for subsequent inparallel, in collaboration with Dr EyualemAbebe. recognition.Key diagnostic characters were identiŽed Resultsindicate a generalcongruence between the andpromising features for futurediagnostic use were twomethods, afŽ rming the reliability of the molecular investigatedusing 14 populations. The primary key barcodeapproach. Chemically treated plots (nitrogen, characterswere identiŽed asthe length of thepost-vulval lime,nitrogen limeand biocide, along with untreated C sac(pvs) as a percentageof the distance between the controls)were includedin this study, and the effect of vulvaand the anus, the shape of the tail terminus and thesetreatments on nematode diversity and distribution tail,body length, and the ratios ‘ a’and ‘ c’. Promising was analysed.This study establishes molecular barcoding keycharacters for thefuture are c ,thedistance from asa rapid,time-efŽ cient and robust method for largescale 0 thevulva to the anus, vulval body width, pvs length as surveysof nematodediversity. ameasureof the vulval body width, head width and allmeasurements associated with the median bulb. A 133A phylogeneticview of longidorids polytomouskey was produced,and can be adapted for computeruse. Diagnosticians and nominators of new 1; 1 2 Yu HE ¤, Tatjana RUBTSOVA ,DerekJ.F .B ROWN , Aphelenchoides speciesare invited to use the key and 3 1 Franco LAMBERTI andMaurice M OENS itsassociated recording form tocollect more details of morphologicaland morphometriccharacters in thisgenus. 1CropProtection Department, Agricultural Research Centre, Burg.V anGansberghelaan96, 9820 Merelbeke, Belgium 135An interactiveelectronic key to nematode 2 ScottishCrop Research Institute, Invergowrie, Dundee genera DD2 5DA,Scotland, UK 3 Istitutodi NematologiaAgraria, CNR, 70126Bari, Italy Mike HODDA ¤[email protected] CSIRO Entomology,GPO Box1700, Canberra ACT 2601, Themolecular phylogeny for 77populations including Australia 62species of the Longidoridae (24 Longidorus spp., [email protected] two Paralongidorus spp., 34 Xiphinema spp., and Usingthe DEL TA ® system,a databaseof the character- two Xiphidorus spp.)was constructedusing Paup 4.0. isticsof genera of Australian freshwater aquaticnema- Sequencesobtained from theD2D3 region of the 26S todesand Dorylaimida has been constructed. This data- subunitgene located in ribosomal RNA geneclusterswere baselists over 700 characteristics for over300 genera. alignedand processed for constructingthe phylogram Thecomputer program INTKEY ® canbe used to inter- usingthe maximum likelihood criterion. The species rogatethis database to key out any genuseasily. The data- areclustered in two main groups: Longidorus and basehas been constructed so that the key will serve a Xiphinema. Paralongidorus speciesare grouped with rangeof users, from noviceto expert, and a rangeof Longidorusafricanus . Xiphidorus speciesare grouped situations,from abundantgood material to scarce dam- with Xiphinemakrugi andthe subgroup Xiphinema agedspecimens. Characters from anyparticular part of americanum.Mostof thebranches obtained good support thebody may be omitted or used exclusively. Charac- from non-parametricbootstrap analysis. tersvisible with high power microscopy may be usedor thosevisible at relatively low magniŽ cations only may 134A proposedpolytomous key for the genus beused (although these do not always lead to identiŽ - Aphelenchoides cationof a singletaxon). All characters may be used, includingthose speciŽ c tolocal populations, or strictly Sue HOCKLAND taxonomiccharacters only may be used,leading to iden-

Vol.4(2), 2002 169 Evolution,phylogeny and classiŽ cation (123-175) tiŽcations which are by deŽ nition robust for areasout- americanum groupof species, X.pachtaicum , and the sidethe area speciŽ cally covered. Characters suitable for other Xiphinema specieswith which the genus Xiphidorus noviceand experienced users can also be selected. The clustered.The genus Xiphinema was mostclosely related keyis illustrated, and is available now online free at to L.camelliae ,followedby L. litchi, and then to http://www.ento.csiro.au/science/nematodes. aclusterthat included several Longidorus spp., and two Paralongidorus spp.The remaining clusters of Longidorusspp.revealed strong evidence of geographical 136A phylogenyof selectedLongidoridae based on originof thespecies. 18srDNA genesequences

1; 2 J. HÜBSCHEN ¤, M. DE OLIVEIRA , 137Phylogenetic relationships based on 18S 3 4 5 A. AUWERKERKEN , L. BARSI , L. FERRAZ , sequencesamong some Xiphinema specieswith three U. IPACH 1, S. LAZAROVA 6, M. LISKOVA 7, andfour juvenile development stages V. PENEVA 6, R. ROBBINS 8, A. SUSULOUSKY9, 10 8 11 1; 2 1 M. TZORTZAKAIS , W. YE , J. ZHENG , Judith HÜBSCHEN ¤, Roy NEILSON , Ulrike IPACH R. NEILSON12 and D. BROWN12 andDerek J.F .B ROWN 2

1Staatl.Lehr-und F orschungsanstaltfur Landwirtschaft, 1SLFA,Neustadt/Weinstrasse,Germany Weinbauund Gartenbau, F achbereichPhytomedizin, 2 SCRI,Dundee, Scotland, UK Breitenweg71, D-67435 Neustadt/ W,Germany ¤jhübschen.slfa-nw@agrarinfo. rlp.de 2 InstitutoBiologico, Rodovia Heitor P enteadokm3, Campinas, SP, Brazil Xiphinemaamericanum -groupnematodes, comprising of 3 Instituteof TropicalAgriculture, High Rainfall Station, P ort approximately50 putative species, have a cosmopolitan Harcourt,Nigeria distributionand are known to vector four economically 4 Instituteof Biology, Novi Sad, FR Yugoslavia damagingviruses; consequently these nematodes are an 5 DepartamentoZoologia, Universidade de Sã o Paulo, importantglobal agricultural pest. Previous studies have Piracicaba,SP ,Brazil concludedthat some species within the X.americanum - 6 Laboratoryof GeneralEcology, SoŽ a, Bulgaria grouphave three juvenile stages with other species having 7 ParasitologicalInstitute SAS, Koš ice, Slovak Republic 8 NematologyLaboratory, University of Arkansas, F ayetteville fourjuvenile stages. Morphological characterisation of AR, USA thisgroup is problematical due to similarities of 9Museumof Natural History, L’ viv, Ukraine manyof the taxonomic characters across the entire 10PlantProtection Institute, Agricultural Research F oundation, group.This lack of taxonomic resolution has crucial Crete,Greece concomitantimplications when trying to assess virus- 11Collegeof Agriculture, Zhejiang University, Hangzhou, vectorrelationships, i.e.,whichspecies vector a particular China virus.Analyses of 18S gene sequence have previously 12ScottishCrop Research Institute, Dundee, Scotland, UK beenreported to discriminate taxonomic units in many ¤jhuebschen.s[email protected] organismsincluding nematodes. 18S data showed that The18s rDNA geneis highly conserved within taxa, allthe Asiatic (Chinese) populations of X.americanum , whichmakes it particularly useful for phylogenetic includingthose with three or four juvenile stages, were studies.Phylogenies of several groups of plant-parasitic geneticallyidentical inferring a singlespecies. In contrast, nematodeshave been produced by various authors, but speciationwas evidentin the studied populations from theserefer onlyto endo-or semi-endoparasitic nematodes bothNorth America and Europe. Furthermore, our data andhave not involved 18s sequence data. The family suggestedthat a greatergenetic diversity existed in the Longidoridaeis comprised of Ž vegenera, with two of studiedEuropean populations that originated from a these, Longidorus and Xiphinema, comprisedof over smallergeographic area than the studied populations that 100and 200 species, respectively. Individual specimens originatedin eitherNorth America or Asia. from representativepopulations of longidorids from aroundthe world had their 18s gene fully sequenced. 138Molecular diagnostics for virus-transmitting Analysesof thesequence data revealed a clearseparation nematodesin German viticulture between Xiphinema and Longidorus;threemajor clusters 1; 1 ofspecies were apparentin the genus Xiphinema: X. Judith HÜBSCHEN ¤, Ulrike IPACH ,

170 Nematology Postersessions

Volker ZINKERNAGEL 2, Daniel ESMENJAUD 3, Meloidogynemayaguensis Rammah& Hirschmann,1988 Derek J.F. BROWN 4 and Roy NEILSON 4 hasbeen recently identiŽ ed in the French W estIndies from guava (Psidiumguayava )orchards.This very 1StaatlicheLehr- und F orschungsanstaltfü r Landwirtschaft, aggressiveroot-knot nematode seems already widely Weinbauund Gartenbau, F achbereichPhytomedizin, distributedin the Caribbean (Cuba, Dominican Republic, Breitenweg71, D-67435 Neustadt/ W.,Germany 2 PuertoRico, Guadeloupe, Martinique). The objective TechnischeUniversitä tMü nchen, Lehrstuhl fü r ofthis study was toevaluate the genetic variability of Phytopathologie,Am Hochanger2, D-85350 M.mayaguensis isolatesfrom severallocations in the Freising-Weihenstephan,Germany 3 InstitutNational de laRecherche Agronomique, F-06600 Caribbean,in comparison with African isolates from Antibes,France Burundiand Congo. For thatpurpose, nematodes were 4 ScottishCrop Research Institute, Invergowrie, Dundee DD2 characterisedusing both isozyme electrophoresis and 5DA,Scotland, UK RAPD-PCR. At thebiochemical level, for thetwo ¤[email protected] enzymaticsystems used (EST andMDH), allthe isolates sharedthe same electrophoretic proŽ les. At themolecular Virus-free grapevineplants required for viticulturehave to level,except one isolate from Martinique,RAPD analysis begrownin soilfree from virus-transmittingnematodes. revealeda highlevel of genetic similarity ( >78%) Currentsoil testing methods are laborious and require betweenall isolates. The UPGMA dendrogramdeduced highlyexperienced staff asthey rely on morphological from theRAPD patternsshowed that clustering of isolates examinationof the nematodes. Molecular diagnostics neitherre ected their geographic origin nor indicated providea simpleand reliable alternative, but are not howthey might have been spread. Despite having been availablefor virus-transmittingnematodes. T odevelop sampledfrom verydistant locations, such a relativelack thismethod for applicationwith samples from German ofgenetic polymorphism suggests a commonorigin for vineyards,we investigatedspecies-speciŽ c primersfor all the M.mayaguensis isolatestested, and indicates that themost important nematodes in German viticulture viz., theymay have been dispersed from asinglesource instead Xiphinemaindex, X. diversicaudatum and X.vuittenezi . ofrepresentinglocal indigenous populations. The18S gene of rDNA, whichis highly conserved in taxa,was sequencedusing three primer pairs for ten, 140Four new speciesof soilnematodes (Nematoda: eightand four populations of these three species to Dorylaimida)from Japan conŽrm thehomogeneity of each species. The genetic variationbetween the species was low.Consequently,in Zakaullah KHAN¤ andMasaaki A RAKI theŽ rst instance,species-speciŽ c primersidentiŽ ed from thegenetically variable ITS-1 regionof rDNA for eachof NationalInstitute for Agro-Environmental Sciences, Tsukuba, thesespecies, developed by INRA France,were usedin Ibaraki305-8604, Japan [email protected] interactiontests in PCR toconŽ rm theirspeciŽ city and ¤ robustnesswhen used for detectingthe species occurring Duringthe course of studies on biodiversity of soil insoil samples from German vineyards.Primer setsare nematodesin Japan, four new species of soil nematodes alsobeing developed for Longidorus and Paralongidorus belongingto theorder Dorylaimida were isolatedfrom the virus-vectorspecies. The preliminary results from these soilsamples collected from anon-tillageŽ eldof National testsare presented and discussed. Institutefor Agro-EnvironmentalSciences, Tsukuba, Japan.Specimens killed in hot water were Žxedin T AF, processedto anhydrous glycerin and mounted on slides. 139Genetic variability of Meloidogynemayaguensis Specimenswere observedunder optical microscope and isolatesfrom the Caribbean measuredwith the help of drawing tube attachment. A. KERMARREC 1, S. PANOMA 1, P. QUÉNÉHERVÉ 2, Measurementsand characteristicsof thesenew species are 1 3 as follows: Egtitus n.sp.is 1.7-2.0mm long,a 41-46, H. MAULEON and P. CASTAGNONE-SERENO D b 3.9-4.3, c 15-19, V 50-53,odontostyle 21- D D D D 1 INRA-URPV,DomaineDuclos, 97270 P etitBourg, 23 ¹m,andis characterisedby havingsinuate odontostyle Guadeloupe andsmaller prerectum. Axonchium n.sp. having 1.5-1.6 2 IRD, BP8007,97259 F ort-de-FranceCedex, Martinique mmlongbody, a 39-43,b 2.5-2.6,c 48-54, V 50- D D D D 3 INRA-IPMSV,BP2078;06606 Antibes Cedex, France 52,odontostyle 9-11 ¹m,and is distinguishable by its D Vol.4(2), 2002 171 Evolution,phylogeny and classiŽ cation (123-175) largerbody and absence of post-uterinesac. Chitwoodius 1Laboratoriode Nematologí a, Centrode Zoología Aplicada, n.sp.measuring 1.0-1.2 mm long,a 26-31, b 3.4-4.1, UniversidadNacional de Có rdoba, C.C. 122,5000. Có rdoba, D D c 47-55, V 61-64,odontostyle 30-33 ¹m, spicules Argentina D D D 2 48-55 ¹mandis characterised by having posteriorly Unidadde ProcesamientoElectró nico de Datos,F acultadde D locatedguiding ring and larger prerectum. Labronema n. CienciasAgropecuarias, Universidad Nacional de Córdoba, C.C. 509,5000, Có rdoba, Argentina sp.is 1.0-1.2mm long,a 22-25, b 3.6-4.0, c 47-50, 3 AgromilloraCatalana S.A. El Rebatos/ n,08737Subirats, V 50-52,odontostyle D 20-21 ¹m,D spicules D55 ¹m D D D Barcelona,Spain andis characterisedby smaller body and sub-digitate tail. †deceased ¤[email protected] 141Four new speciesof soilnematodes (Nematoda: Dorylaimida)from Japan Datacorresponding to morphometrical characters of malesand females belonging to populations of Praty- Zakaullah KHAN andMasaaki A RAKI ¤ lenchusvulnus from differentgeographic origin and under differentdevelopmental conditions were considered.An NationalInstitute for Agro-Environmental Sciences, Tsukuba, interand intra-population variance was estimatedfor each Ibaraki305-8604, Japan characteras well as the percentage of inter-population ¤[email protected] variance.Characters were groupedinto three categories (low,mediumand high variability). Considering those Duringthe course of a studyon biodiversity of soil percentages,it was observedthat the number of characters nematodesin Japan, four new species of soilnematodes withlow variability was higherin females than in males, belongingto the order Dorylaimida were isolatedfrom beinginverse in thecase of characterswith high variabil- thesoil samples collected from anon-tillageŽ eldof ity.Depending on thepercentage of inter-populationvari- theNational Institute for Agro-EnvironmentalSciences, abilityfor eachcharacter, it was inferredthat those char- Tsukuba.Specimens killed in hotwater were Žxedin T AF, actersincluded in the low variability category would be processedto anhydrous glycerin and mounted on slides. barelyin uenced by environmental conditions, whereas Specimenswere observedunder optical microscope thosecharacters related to high values would be consider- andmeasured with drawing tube. Measurements and ablyin uenced. Therefore, not all characters would have characteristicsof these new species are as follows: thesame signiŽ cance. Some of them appear to be more Axonchium n.sp. is 1.5-1.6 mm long,a 39-43, b signiŽcant than others for thespecies identiŽ cation (V ra- D 2.5-2.6, c 48-54, V 50-52,odontostyle 9- tioand spicule length for malesand females, respectively, D D D D 11 ¹m,and is distinguishable by its larger body and ratiosO, MB,c 0,pharynxlength) whereas other group of absenceof post-uterinesac. Egtitus n.sp.is 1.7-2.0 mm charactersshould be preferablyconsidered when evaluat- long, a 41-46, b 3.9-4.3, c 15-19, V 50-53, ingthe existence of possibledifferences between popula- D D D D odontostyle 21-23 ¹m,andis characterised by having tions. sinuateodontostyle D and smaller prerectum. Labronema n. sp.is 1.0-1.2 mm long,a 22-25, b 3.6-4.0, c D D D 143Genetic variability in two populationsof 47-50, V 50-52,odontostyle 20-21 ¹m, spicules D D Heteroderaglycines Ichinohe,1952 from Argentina 55 ¹mandis characterised by smaller body and sub- D estimatedwith RAPD-PCR markers digitatetail. Prodorylaimus n.sp.measuring 1.3-1.5 mm long, a 37-40, b 3.8-4.0, c 5.1-6.0, V 50-53, Paola LAX1; , Juan C. RONDAN DUEÑAS 2, D D D D ¤ odontostyle 21-24 ¹m,andis characterised by having CristinaN. G ARDENAL 2 andMarcelo E. D OUCET 1 D roundedlip region and gradually tapering to long Ž liform tail. 1Laboratoriode Nematologí a, Centrode Zoología Aplicada, Facultadde Ciencias Exactas, Fí sicas y Naturales,Universidad 142Inter-population variability in Pratylenchus Nacionalde Có rdoba, C.C. 122,5000, Có rdoba, Argentina vulnus Allen& Jensen,1951 (Nematoda: Tylenchida) 2 Cátedra de Bioquímica y Biología Molecular,Facultadde CienciasMé dicas, Universidad Nacional de Córdoba, C.C. 35, 1; 1 Paola LAX ¤,MarceloE. D OUCET , Julio A. DI Sucursal16, 5016, Có rdoba, Argentina RIENZO 2, Pierre BAUJARD† and Jorge PINOCHET 3 ¤[email protected]

172 Nematology Postersessions

Thegenetic variability of two populations of H. glycines numberof ampliŽed polymorphic fragments of 1267from was analysedusing Random AmpliŽ ed Polymorphic tenprimer combinations were analysedwith NTSYS-pc DNA (RAPD-PCR) asmarkers. Thirty females of a package.Populations from eachspecies form asingle populationbelonging to race 1 from LagunaLarga cladeand the phylogenic relationship matches very well (Córdoba Province) and 31 femalesfrom apopulationof withprevious phylogeny. Meloidogynearenaria and M. race3 from Tortugas(Santa Fe Province)were studied javanica areclosely related and form abranchthat is individually.T wentyrandom primers from Biodynamics monophyleticto M.incognita.Meloidogyne panyuensis is (BuenosAires, Argentina)were tested.Five of them distantlyrelated to theother three species. W econcluded were selectedbecause they originated reproducible thatAFLP ismore reliable than rDNA-ITS sequences polymorphicbands. Thirty nine consistent fragments for nematodephylogenic study since AFLP coversthe were consideredfor theanalysis. Allele frequencies were wholegenome whereas rDNA-ITS sequencescome from estimatedassuming RAPD segregateas dominant and onlyone locus. Furthermore, AFLP ismuch simpler than thatthe populationswere inHardy-Weinbergequilibrium. rDNA-ITS sequencing. Proportionof polymorphic loci was 100%in both populations;mean expected heterozygosity (He) among 145Enzymatic characterisation of Meloidogyne spp. individualswas 0.39and 0.42 for femalesfrom Laguna associatedwith ornamentalsand agronomic crops in Largaand T ortugas,respectively. F values were ST Florida,USA signiŽcant ( P < 0:05)for 67%of theloci. The AMOV A testshowed a variationof 81.2% within specimens of 1; 2 3 R.D. LIMA ¤, J.A. BRITO , D.W. DICKSON , apopulation,while the remaining 18.8% accounted for W.T. CROW 3, C.A. ZAMORA 2 and M.L. MENDES 3 differencesbetween populations. None of thepopulations presentedexclusive bands, although many fragments showedsigniŽ cant differences in their frequencies. This 1Deptode Fitopatologia, Universidade F ederalde V içosa, studyrevealed a highintra-population variability and 36571-000V içosa, MG, Brazil 2 animportant degree of genetic differentiation between Divisionof PlantIndustry, Gainesville, FL, 32618-7100,USA 3 Entomologyand Nematology Dept., University of Florida, populationsof races1 and3. Gainesville,FL 32611-0620,USA ¤[email protected] 144Phylogeny of Meloidogyne spp. basedon rDNA-ITS sequenceand  uorescentAFLP Theuseof enzymephenotypes has madethe identiŽcation 1;2; 2 1 Meloidogyne Jinling LIAO ¤, Zhixin FENG and Weicai YANG ofspecies of lesssubjective and more ac- curate.Biochemical analysis based on phenotypes of es- 1 Instituteof MolecularAgrobiology, Research Link 1, National terase(EST), malatedehydrogenase (MDH), superoxide Universityof Singapore,Singapore 117604 dismutase(SOD), andglutamate-oxaloace tatetransami- 2PlantNematology Laboratory, South China Agricultural nase(GOT) was usedto characterise 52 populations of University,Guangzhou, PR China510642 [email protected] Meloidogyne spp.collected from ornamentalsand agro- ¤ nomiccrops in Florida. Meloidogynejavanica, J3 phe- Aphylogeneticstudy of 12populations of M.incognita , notype,was themost common species (38.4%), followed twopopulations of M.arenaria, twopopulations of by M. arenaria phenotypeA2 (25.0%), M.incognita phe- M. javanica andnewly-described M.panyuensis n. sp. notypesI1 (15.4%) and I2 (7.6%), M. graminis (9.6%), was performedon rDNA-ITS sequencesand  uorescent and Meloidogyne sp. (3.8%). Meloidogynegraminis had AFLP.For therDNA-ITS sequenceanalysis, genomic onlyone slow moving activity EST bandwhich was not sequencescontaining the partial 18S, complete ITS presentin any of theother phenotypes, three SOD bands andpartial 26S regions were PCRampliŽed, cloned (twointensely stained and a thirdless intensely stained andsequenced. Sequence analysis using the neighbour- butfaster moving band), and one intensely stained MDH joiningcluster method showed that it is difŽ cult to bandidentical to that of M.mayaguensis .Theoccurrence establisha reasonablephylogenic relationship among the of M. javanicainpeanutis being reported for theŽrst time fourspecies since the tree topology does not match inFlorida,USA. withthe established taxonomy. For AFLP analysis,42 primercombinations were screenedand AFLP witha total

Vol.4(2), 2002 173 Evolution,phylogeny and classiŽ cation (123-175)

146V ariabilityamong isolates of Pratylenchus severalagriculturally important cyst nematode species penetrans foran interactionwith Verticilliumdahliae andseparation from theirsibling species. RFLP proŽles andthe potatoearly dying disease for someEuropean populations of Globoderapallida , G. rostochiensis,Heterodera carotae, H. Žci,H. Žlipjevi,H. A. MACGUIDWIN¤, A. REID-RICE, R. ROWE goettingiana,H. hordecalis,H. humuli,H. mediterranea, and M. OMAR H. riparia and H.schachtii aregiven. The species- Universityof Wisconsin,Madison, WI; OARDC, OhioState speciŽc primerdeveloped by Bulman and Marshall University,W ooster,OH, USA clearlydiscriminated G. pallida from G.rostochiensis. ¤[email protected] Bsh1236I digestionof the PCR productseparated both Globodera speciesfrom eachother and from G. tabacum. Asynergisticinteraction of Pratylenchuspenetrans and Verticilliumdahliae for thepotato early dying disease has Comparisonof obtained sequences with those deposited inGenBank showed high similarities (99.8-100%). beendemonstrated using a varietyof potatocultivars and Relativehigh level of sequence divergence between soiltypes. The purpose of our study was todetermine populationsof H.hordecalis (1.5%)suggests the presence ifgeographic isolates of P.penetrans varyfor this ofseveralbiological species presently grouped under this interaction.W etestedeight P.penetrans isolatesfrom Wisconsinand one eachfrom Washington,Minnesota and taxon. Ohioin a seriesof growthchamber experiments. Three- week-oldpotato cv. Russet Burbank grown from tissue 148Intraspecies DNA polymorphismin the tobacco culturewas inoculatedwith nematodes and 1 weeklater cystnematode complex ( Globoderatabacum ) using withconidia of V. dahliae.Controlplants were inoculated AFLP with only V. dahliaeormock inoculated with water and no pathogens.The isolates were rankedfor thetime required Laurent MARCHɤ, Eric GRENIER, Sylvie VALETTE and for threeleaves to become symptomatic and area under Didier MUGNIÉRY thedisease progress curve, as well as for population INRA UMR Biologiedes Organismes et des P opulations attributessuch as female:male ratio and the rate of root appliquée àlaProtectiondes Plantes, Domaine de la Motte, egress.Three isolates from Wisconsinrepresented the B.P.35657,35653 Le Rheucedex, France rangeof variabilityfor symptomexpression; two ranked ¤[email protected] inthe top third and one ranked in the bottom third consistently,but all isolates were ableto cause disease in AFLP was usedto obtain information on the within- combinationwith V. dahliae. speciesgenetic variability of thetobacco cyst nematodes (TCN) complex.AFLP was foundto be well suited for thistype of study.The current classiŽ cation of TCNwas 147IdentiŽ cation of somecyst-forming nematodes conŽrmed. Results indicate that the Globoderatabacum basedon PCR-RFLP,sequenceof rDNA andspeciŽ c solanacearum group,believed to be restrictedto the USA, primers alsooccurs in Mexico. The within-species variability of 1; 2 theTCN isconsiderable. Populations from Mexicomay Mehrdad MADANI ¤, Nicola VOVLAS , form anewsub-group. AFLP group-speciŽc markers were Sergei A. SUBBOTIN 3 andMaurice M OENS 1 identiŽed for twoTCN subgroups: Globoderatabacum 1 Departmentof Crop Protection, Agricultural Research Centre, tabacum and Globoderatabacum solanacearum . Burg.V anGansberghelaan96, 9820 Merelbeke, Belgium 2 Istitutodi NematologiaAgraria del C.N.R., 70126Bari, Italy 3 Instituteof Parasitologyof theRussian Academy of Sciences, 149Reconstructing a phylogenyof Dorylaimida Leninskiiprospect 33, Moscow ,117071,Russia (Nematoda)with partial18s sequence data using ¤[email protected] Bayesianinference Morethan 50 populations of cyst-forming nematodes Peter G. MULLIN¤,TimothyS. H ARRIS and collectedin Italy and Belgium were analysedusing PCR- Thomas O. POWERS RFLP,ITS-rDNA sequences,and PCR withspecies- speciŽc primers.Restriction of PCR products by eight Departmentof Plant P athology,406 Plant Sciences Hall, restrictionenzymes AluI, AvaI, Bsh1236I, CfoI, HaeIII, Universityof Nebraska-Lincoln,Lincoln, NE 68583-0722USA MvaI, PstI and RsaI allowedthe identiŽ cation of ¤[email protected]

174 Nematology Postersessions

Evolutionaryrelationships among members of Dorylaim- and b) mean FST valuesfrom ananalysis of molecular idaare investigated using approximately 600 bp of the variance,separated the eight G. pallida populations 18sribosomal DNA repeat.Sequence data obtained from intofour similar putative groupings. As withprevious selectedtaxa within Dorylaimida and from severalre- studies,P5A was distinctfrom allEuropean populations latedgroups form thebasis for theanalyses. Results ob- andLuffness (and Lindley) were disparatefrom the tainedunder a rangeof optimalitycriteria and using sev- otherUK populations.The remaining populations formed eraldifferent methods, including parsimony, distance, and twogroupings suggesting further genetic differentiation likelihoodapproaches, are presented and compared. The betweenUK populations. useof Bayesianinference in phylogeny reconstruction al- lowsthe combination of detailed likelihood models of 151V ariabilityof Meloidogyneexigua on coffee DNA evolutionwith an assessment of the reliability of cropsin the Zonada Mata of MinasGerais State, theresults by summarising the posterior probabilities of Brazil anextensive sample of treesgenerated under a givenset ofmodel parameters. The resulting consensus tree, with DagobertoSaunders O LIVEIRA,RosangelaD’ Arc L IMA summarybranch lengths and clade credibility values, is andRodrigo Vieira S ILVA analogousto a maximumlikelihood bootstrap tree, but canbe generated in a fractionof the time. W epresenta Deptode Fitopatologia,Universidade F ederalde Viçosa, phylogenetichypothesis for Dorylaimidaand relatedtaxa, 36571-000,V içosa, MG, Brazil alongwith discussion of severalpoints of departurefrom [email protected] recentclassiŽ cation schemes of this important group of terrestrialnematodes. MinasGerais Stateis the most important producer of cof- feein Brazil and 28% of its production occurs in the 150Genetic differentiation of individualsand Zonada Mata region. Four major species of root-knot populationsof the endoparasiticnematode, Globodera nematodesattacking coffee plants have been reported in pallida,usingmicrosatellites Brazil,and some of themcan cause plant death. The cor- rectidentiŽ cation of species and/ orrace(s) of Meloido- Roy NEILSON1; , Vivian C. BLOK 1, Jim W. MCNICOL 2 ¤ gyne presentin roots of coffeeis extremelyimportant in and Mark S. PHILLIPS 1 decidingwhich measures are more appropriate for con- 1ScottishCrop Research Institute, Dundee, DD2 5DA, trollingthe pathogens. In order to determine the occur- Scotland,UK renceand variability of Meloidogyne spp.in the region, 2 Bioinfomaticsand Statistics Scotland, Scottish Crop Research 57populationsfrom 16differentlocations were evaluated Institute,Dundee, DD2 5DA,Scotland, UK basedon morphologic, enzymatic and physiologic traits. ¤[email protected] Allof the57 populations were identiŽed as Meloidogyne Thegenetic differentiation of eight populations of the exigua basedon theirperineal patterns. This characterisa- endoparasiticnematode Globoderapallida with known tionwas conŽrmed by phenotypesof esterase,malate de- virulencecharacteristics was assessedat the individual hydrogenase,sulphoxide dismutase and glutamate oxalo- nematodelevel using six microsatellite loci. This provided acetatetransaminase. Thirteen populations presented the anopportunity to assess levels of genetic diversity typicalesterase phenotype showed by one band (VF1), betweenand within populations at a resolutionthat whilemost of the populations (77.2%) exhibit a pheno- hithertohas not been possible. Intra-population genetic typeshowed by two bands. No intraspeciŽ c variability variabilitywas evident,with four populations, Luffness, was observedin thepopulations studied, and all of them P5A,Gourdie and Halton being least genetically variable. were ableto reproduce on tomato, pepper, cocoa, onion, Although Globoderapallida isa sedentaryendoparasite beanand soybean. andsexually reproducing species, F ST values and heterozygositydata suggested that it hadcharacteristicsof 152Molecular characterisation of somespecies of anin-breeding species and supports published hypotheses the genus Meloidogyne from China postulatingthat many of the European G. pallida 1; 2 populationsarose via multiplefounder effects. Principal Deliang PENG ¤, Sergei A. SUBBOTIN and coordinateanalyses derived from a) dissimilarityvalues Maurice MOENS 3

Vol.4(2), 2002 175 Evolution,phylogeny and classiŽ cation (123-175)

1Instituteof PlantProtection of Chinese Academy of accordingto Subbotin et al. (2000).Digestion by AluI AgriculturalSciences, Beijing, 100094, China alsoshowed heterogenecity in ITS regionsof Morocco 2 Instituteof Parasitologyof RussianAcademy of Sciences, population,and two additional bands were obtained; Leninskiiprospect 33, Moscow ,117071,Russia thesum of the three fragments was approximately 3 AgricultureResearch Center ,CropProtection Department, 2120 bp. HinfIdigestedPCR ampliŽed ITS products Burg.V anGansberghelaan96, 9820 Merelbeke, Belgium ofChinesepopulations obtained two fragments (850 bp, [email protected] ¤ 200bp), but obtained three fragments (510 bp, 340 bp, rDNA-ITS RFLP andsequence analyses were usedfor 200bp) from Moroccopopulation. The results showed identiŽcation and characterisation of ten Meloidogyne thatChinese populations may be distinctlydifferent from populationsfrom China.The ITS regionswere ampliŽed Moroccopopulation. Seven enzymes CfoI, Bsh1236I, usingF195 and 5367 primers. PCR resultedin a single MsrFI, ScrFI, HaeIII, MvaI, MspIproducedrestriction fragmentof about 700 bp for allsamples. Based on proŽles identical for allCCN populations. HindIII and morphologicaland morphometrical characters, and ITS AvaIdidnot digest the ITS productsof Chinese and sequences,seven populations were identiŽed as M. MoroccoCCN populations. hapla, M.javanica,M. arenaria , M.incognita and M. enterolobii ,whilethree populations belonged to two 154Genetic variability of Heteroderaschachtii undescribedspecies. T welverestriction enzymes were populationsin the north ofFranceinferred by usedfor digestionof PCR products. The RFLP patterns microsatelliteloci generatedby HinfI RsaI separatedall populations C M. intothree groups. None of theenzymes differentiated Olivier PLANTARD¤,CatherineP ORTE and incognita, M. arenaria and M. javanica from eachother. Eric GRENIER TheRFLP andsequence analyses of the D2-D3 regions ofthe 28S gene permitted separation of severalroot-knot INRA UMR Biologiedes Organismes et des P opulations nematodespecies. appliquée àlaProtectiondes Plantes, Domaine de la Motte, B.P.35657,35653 Le Rheucedex, France ¤[email protected] 153rDNA restrictionfragment length Abetterknowledge of thegenetic structure of phytopar- polymorphismof Heteroderaavenae in China asiticnematode populations is essential to control these 1; 2 organisms,notably in the context of the use of resistant Deliang PENG ¤, Sergei A. SUBBOTIN and Maurice MOENS 3 varietiesleading to the selection of virulent individuals ableto overcome resistance genes. With that aim, we Žrst 1Instituteof PlantProtection of Chinese Academy of studiedthe genetic variability of nematode populations AgriculturalSciences, Beijing, 100094, China atvarious spatial scales using neutral genetic markers in 2 Instituteof Parasitologyof RussianAcademy of Sciences, Želdswhere no resistant varieties have been grown until Leninskiiprospect 33, Moscow ,117071,Russia now.Our studyfocused on the sugar beet cyst nematode 3 AgricultureResearch Center ,CropProtection Department, Heteroderaschachtii Burg.V anGansberghelaan96, 9820 Merelbeke, Belgium ( )inthe north of France.Using Ž ve microsatelliteloci on individualssampled according to a ¤[email protected] hierarchicaldesign, an Analysis of MOlecular V Ariance TheampliŽ cation of therDNA-ITS regionof Heterodera (AMOVA)indicatesthat almost all the genetic variability avenae (CCN) from Chinaand Morocco with the current isobservedat theŽ eldscale (94% of thetotal genetic vari- primersAB28 and TW81 yielded one fragment of ability),with only 4% variabilityobserved among Ž eldsof approximately1060 bp. A totalof 27 scored fragments thesameregion and 2%amongregions. This suggests that were obtainedwith 12 restriction enzymes. IntraspeciŽ c signiŽcant gene owsoccur among Ž eldsand regionspre- polymorphismwas revealedwithin H. avenae by AluI, ventinggenetic differentiation. In this diploid amphimic- RsaI and Hinf Idigestion. AluI and RsaIdigestionof ITS ticspecies, Hardy-W einbergequilibrium is observed only productsof sevenChinese CCN populationsyielded two atthesmallerspatial scale ( i.e.,theindividualplant), prob- fragmentsrespectively (560 bp, 500 bp; 720 bp, 320 bp); ablyas a resultof limited dispersal of individualsin the but neither AluI nor RsaIcoulddigest ITS productof soilbetween hatching and mating. Morocco.Those RFLP proŽles revealed by AluI and RsaI classiŽed the ITS ofChinese populations as ‘ TypeB’

176 Nematology Postersessions

155The differential host test, mtDNA andrDNA 9.7%of the total genomic DNA. Twentyindependent PCR todistinguish Meloidogynearenaria, monomershave been cloned and sequenced. They are M.incognita and M. javanica fromvineyards veryhomogeneous in sequence, with 2.4% average divergencefrom thededuced consensus. This result Motiul QUADER¤ and Ian T. RILEY stronglysuggests that this repeated family originated CooperativeResearch Centre for V iticulture,P .O.Box154, and from arecentampliŽ cation event. Dot-blot experiments Appliedand Molecular Ecology, Adelaide University, Glen demonstratedthat the satelliteDNA distributionis limited Osmond,SA 5064,Australia to M. exigua isolatesonly. In squash-blot assays, it ¤[email protected] was possibleto detect and positively identify single nematodes,independently of their developmental stage. TheNorth Carolina (NC) differentialhost test and Therefore,because of itshigh reiteration in the genome, mtDNA were usedto distinguish a collectionof root- andits species-speciŽ city, this sequence could be usedas knotnematodes from SouthAustralian vineyards. PCR aprobefor themolecular diagnosis of thenematode. ampliŽcations of D3expansion region of 28SrRNA gene andintergenic sequences of ribosomalDNA (IGS-rDNA) were alsomade to distinguish Meloidogynearenaria, 157Genetic diversity in Želdpopulations of tobacco M.incognita and M. javanica.TheNC differentialhost cystnematode in North Carolina Meloidogyneincognita M. testdifferentiated but not 1; 2 arenaria race 2 from M. javanica.Thecombination Uma RAO ¤ and D. MCK.BIRD ofthe NC hosttest and mtDNA analysisdifferentiated 1Divisionof Nematology,Indian Agricultural Research among M.arenaria,M. incognita and M. javanica. The Institute,New Delhi110012, India differentiationof thesespecies with D3 expansion region 2 Departmentof PlantP athology,North Carolina State of28S rRNA genewas notpossible. The sequences of University,Raleigh, USA thisregion are highly conserved among the species. The ¤[email protected] PCRampliŽ cation of IGS-rDNA from singlefemales ofeach species produced distinct banding patterns that Tobaccocyst nematode (TCN) isa fastspreading differentiatedthe species from eachother. These species- nematodeproblem in North Carolina requiring an efŽcient speciŽc bandingpatterns were reproducibleacross a methodof management. Resistant varieties are a good rangeof individual nematodes of each species collected alternativeto chemical nematicides. QuantiŽ cation of from differentgeographical locations of Australia. This geneticdiversity is essential to decide the type of methodalso produced DNA Žngerprintvariability within resistanceto be incorporated. Cysts collected from someindividuals of each species. This variability could infestedŽ eldsof tobacco in V ance,W arrenand Surry beapplied to the examination of intraspeciŽ c variation countieswere usedfor RAPD-PCR analysis.Fourteen andpotentially development of race speciŽ c diagnostic Želdsamples for inter-populationdiversity and four marker(s). populations,one from eachcounty, for intra-population diversitywere used.The size of the PCR ampliŽed productsvaried from 220to 2500 bp and the number 156A new species-speciŽc satelliteDNA familyin ofproducts varied from 11to 22. UPGMA clustering the genomeof the coffeeroot-knot nematode, grouped13 populations into two major clusters at nodal Meloidogyneexigua interjunctionsof 47 and 58%. One population was heterogeneousenough to fall out of these two clusters. Onivaldo RANDIG, Michel BONGIOVANNI and For withinpopulation, percentage similarity ranged from Philippe CASTAGNONE-SERENO ¤ 36to 56 indicating considerable amount of genetic INRA UnitéIPMSV ,BP2078,06606 Antibes cé dex, France diversity.The heterogeneity observed in TCN suggests ¤[email protected] thatbreeding for resistanceshould be horizontalor broad enoughto managethe damage at threshold levels. Anewsatellite DNA familyhas been cloned in the coffee root-knotnematode, Meloidogyneexigua. Itis represented astandemly repeated sequences with a monomericunit 158The development of aquerysystem for of277 bp. The repeats are present at approximately the Braziliannematological bibliography references 17900copies per haploid genome, and represent about throughthe Internet

Vol.4(2), 2002 177 Evolution,phylogeny and classiŽ cation (123-175)

1; 2 2 V. RISSOLI ¤, R. TENENTE , L. PALHARES possessesa rare nucleotidesubstitution within the region and M. TENENTE 2 ofhairpin35 oftheSSU rRNA secondarystructure. The 1UniversidadeCató lica Brasí lia, QS 7-Lote1 (72022-900) samesubstitution is common for all17 Enoplida species TaquatingaSul, DF ,Brazil studiedto date, including Trefusia and Xenella (Trefusi- 2 EMBRAPARecursosGené ticos e Biotecnologia,P .O.Box idae),but is absent from othernematode lineages includ- 2372(70849-970) Brasí lia, DF ,Brazil ingTriplonchida and Dorylaimia. Thus, the substitution ¤[email protected] canbe considered a molecularsynapomorphy of Enopl- Severalbibliographical sources regarding nematode idaandTrefusiidae.Apart from thissubstitution, Enoplida themesare organised in a bankand this will be put arefound to share a rare transversionin theloopofhairpin ina databasewhich the Nematode Query System will 48.This substitution is notstrictly speciŽ c for Enoplida, access,for use via theInternet. When one wants to asit is also known from Mesorhabditoideaand selected accessnematode data, the search for suchdata becomes non-nematodetaxa. These signatures may be used in de- anarduous task for professionals.A standardform of signingspeciŽ c primersand be consideredas markersof systematicclassiŽ cation of such bibliographical sources thedivergent sequences of marineEnoplida. isvery important. In this context, the construction of adatabasein computational environment allows the 160SSU rRNA geneof Soboliphymebaturini datato be stored in a systematicand standard way, Petrow,1930(Nematoda: Dioctophymida) and allowingefŽ cient data recovery. Recently, EMBRAP A areappraisalof molecularbounds ofDorylaimia GeneticResources and Biotechnology and Brasilia 1; 1 2 CatholicUniversity have been constructing a database L. ROUSSINE ¤, V.V. ALESHIN , A.V. TCHESUNOV ofthese references which include several species of and G.I. ATRASHKEVICH 3 nematodesand plants belonging to different botanical 1BelozerskyInstitute of Physico-ChemicalBiology, Department families.This database allows a searchof Brazilian ofEvolutionaryBiochemistry publicationsincluding the distribution in Brazil and 2 Facultyof Biology,Moscow State University, Department of typesof control. Currently, this database has 1850 InvertebrateZoology, 119899 Moscow ,Russia bibliographicalreferences recovered from 1981to 2001. 3 Instituteof the Biological Problems of the North, F ar-Eastern Thenematode query system, which is asystemof access Branchof the Russian Academy of Sciences,685000 Magadan, tobibliographical references databases, is now under Russia constructionand will be accessibleat EMBRAP ACenter. ¤[email protected] Asearchof the nematode query system can be done by Molecularphylogenies based on SSU rDNA sequence usingthe scientiŽ c nameof either the nematode or the datasuggest that Dorylaimia, along with Enoplia host plant. andChromadoria, constitutes the third mainstream innematode evolution. Its contentsare represented 159A simplemolecular diagnostic of marine asfollows: (Trichinellida (Dorylaimida (Mononchida: Enoplida Mermithida))).However, high rates of sequence evolu- tioninherent in the trichinellid lineage render this topol- 1; 2 1 L. ROUSSINE ¤, P. DE LEY , V.V. ALESHIN ogyunreliable. Novel molecular data on the SSU rRNA 1 and N.B. PETROV genestructure of Soboliphymebaturini (Dioctophymida: 1BelozerskyInstitute of Physico-Chemical Biology, Department Soboliphymidae)revealed the Ž fthlineage within Dory- ofEvolutionary Biochemistry, Moscow State University, laimiaand provided new information for establishingits 119899Moscow ,Russia internaltopology. Maximum parsimony, maximum like- 2 Departmentof Nematology,University of California, lihoodand neighbour-joining algorithms with bootstrap Riverside,CA 92521,USA replicatesconsistently inferred S. baturini asa sistertaxon ¤[email protected] toTrichinellida.Inclusion of S. baturini hada stabilising MarineEnoplida constitute a groupof free-living ma- affecton the integrity of the Dorylaimia clade through rinenematodes, which do not share any known morpho- breakingthe long branch leading to Trichinellida. Puta- logicalsynapomorphy. On SSU rRNA phylogeniesthe tivemolecular synapomorphies reconstructed by parsi- groupalso lacks solid statistical support. In recent studies monyfor eachnode of thetree were analysedon alarge itwas foundthat an enoplidnematode Trefusiazostericola datasetof metazoan SSU rDNA sequencesto sort out

178 Nematology Postersessions homoplasiouscharacters that may potentially bias par- highlyinconsistently with this problematic dataset and simonyestimations. Strong synapomorphies located in thatbootstrap analyses (both NJ andMP) greatlyunder- highlyconserved parts of the gene support monophyly estimaterepeatability of inferringeven basal nodes of the ofall Dorylaimia and suggest the following internal bi- nematodetree. Instead, comparing equally parsimonious furcationpattern: ((Trichinellida: Dioctophymida): (Do- topologiesobtained in a seriesof MP runswith a num- rylaimida(Mononchida: Mermithida))). Molecular phy- berof statistical tests under the ML optimalitycriterion logenyof Dorylaimiajuxtaposed with morphological evi- allowedthe reconstruction of the major lineages within dencemay alter our perceptionof morphologicaland eco- Enoplia. logicalchange in nematodeevolution. 162Are datafrom the beta-tubulin genefamily 161Phylogeny of Enoplia:parsimony estimations useful forcyst nematode phylogeny? with SSU rDNA sequencedata Aniko SABO¤ and V.R. FERRIS 1; 2 1 L. ROUSSINE ¤, P. DE LEY , V.V. ALESHIN , 3 4 Departmentof Entomology,Purdue University, W estLafayette, J.R. VANFLETEREN , D. STURHAN , IN47907-1158,USA A. VIERSTRAETE 5 and M. BLAXTER 6 ¤[email protected] 1BelozerskyInstitute of Physico-Chemical Biology, Department Manyeukaryotic genes have been found to be members ofEvolutionary Biochemistry, Moscow State University, ofgenefamilies. When similar, but different members of 119899Moscow ,Russia agenefamily are recovered with the same PCR primers, 2 Departmentof Nematology,University of California, itcan be difŽ cult to use such data for phylogenetic Riverside,CA 92521,USA inference.T odoso, we Žrst inferredevolutionary 3 Universityof Ghent, Department of Biology, B-9000 Gent, relationshipsamong 24 species of cyst nematodes from Belgium 4 BiologischeBundesanstalt, Institut fuer Nematologie und ribosomalDNA (rDNA) sequencedata. Separately, we Wirbeltierkunde,48161 Muenster ,Germany usedbeta-tubulin data, now known to be a genefamily 5 Universityof Ghent, Department of Biology, B-9000 Gent, ofsix members in Caenorhabditiselegans .Topologies Belgium from thetwo trees were incongruent.Following restriction 6 Instituteof Cell, Animal and P opulationBiology, University of andhybridisationanalysis, we concludedthat beta-tubulin Edinburgh,Edinburgh, EH9 3JT,UK isa smallgene family in cyst nematodes as well. W e ¤[email protected] attributedsix monophyletic groups on our beta-tubulin treeto putative gene family members. Some taxa appeared Molecularanalyses suggest that Enoplia is the mostdiver- inmore than one group, indicating the presence of gentmajor clade within the phylum Nematoda. Dispari- multiplefamily members. The evolution of the gene tiesin ratesof sequenceevolution between lineages, lack familywas assessedby tracing putative beta-tubulin ofintermediatetaxa to break possible long branches and geneson the rDNA tree,which we hypothesisedto be thelimited amount of phylogenetic information shared atruespecies tree. A reasonableevolutionary scenario bysampled taxa make phylogenetic inference for Eno- couldbe devised involving gene duplication, gene loss, pliaa difŽcult task. The dataset was carefullyselected anddeep coalescence. Each of three different groups tomaximally encompass enoplian biodiversity and utilise inHeteroderidae, monophyletic on our species tree, had basalrepresentatives of other clades, thus breaking long differentputative beta-tubulin gene members, and thedata branchesat all levels of the tree. It includes several ge- thereforecorroborated the rDNA speciestree. nerafor whichmolecular evidence has not been previ- ouslypublished: Campydora, Leptosomatum,Thoracos- tomopsis,Bathylaimus, Tripyloides, Calyptronema, V is- 163 RAPD of Pratylenchus populationsfrom coffee, cosia,Halalaimus, Ironus, Alaimus, Xenella, Rhabdode- banana,ornamental plant and citrus in Brazil mania,T obrilus,Aporcelaimellus and Nygolaimus. The SilviaRenata S ICILIANO-WILCKEN1; , datawas examinedfor presenceof phylogeneticsignal us- ¤ Mário M. INOMOTO 2,LuizCarlos C.B. F ERRAZ 2, ingapproaches based on sequentialremoval of categories Cláudio M.G. O LIVEIRA 3 andEdson S. M ORI 1 ofvariable sites, with phylogeny reconstruction at each step.This case study proves that the NJ methodperforms 1FCA/UNESP,P.O.Box 237, 18603-970, Botucatu, SP ,Brazil

Vol.4(2), 2002 179 Evolution,phylogeny and classiŽ cation (123-175)

2 ESALQ/USP,Piracicaba,SP ,Brazil betweennematodes and other organisms. W eperformed 3 InstitutoBioló gico, Campinas, SP ,Brazil anotherstudy to detect the presence of hsp90 polymor- ¤[email protected] phismsthat may be useful for discriminationof nema- todesat the sub-species level. Comparison of hsp90 ge- RAPD analysesof seven Pratylenchus populationsfrom nomicsequences from severalhighly inbred strains of differenthosts and regions of Brazil identiŽ ed as thesoybeancyst nematode, Heteroderaglycines, revealed Pratylenchuscoffeae were studied.Three populations uniquecombinations of intron polymorphisms in some were extractedfrom rootsof bananaplant (Minas Gerais strains,thus demonstrating the potential of thisgene for State),two from citrus(Sã o PauloState), one from coffee discriminationof soybeancyst nematode genotypes. (São PauloState) and one from Aglaonema (Rio de JaneiroState). Pratylenchuspenetrans from oilpalm was usedas outgroup. They had beenculturedon alfalfacallus 165Functional analysis of heatshock protein, tissues.Baermann extracted specimens of thepopulations HSP90,from the soybeancyst nematode, Heterodera from calluswere smashedindividually into a 200 ¹l glycines thermocyclertube with 5 ¹loflysesbuffer and frozen at Andrea M. SKANTAR , Keli AGAMA and 20±C.After freezingfor atleast 1 week,the individuals ¤ of¡ each population were pooled,and 5 ¹l of the bulk Lynn K. CARTA ofthepopulation was usedfor PCRampliŽ cation. They were thenrun in 3% agarose gel for 3hat80 V . USDA-ARS, PlantSciences Institute, Nematology Laboratory, Thegel was submittedin an ethidium bromite 0.25% 10300Baltimore Blvd, Bldg 011A BARC West,Rm 165B, Beltsville,MD 20705,USA andphotographed. Polimorphic bands were analysedin [email protected] Popgenesoftware which grouped them in four clusters ¤ accordingto thehost plants. Developmentallyarrested second-stage juveniles of many plantparasites are functionally similar to Caenorhabditis 164Utility of the heatshock protein gene, hsp90, for elegans dauers,and likely use molecules similar to those phylogeneticanalysis and diagnostics of nematodesat deŽned by the C. elegans dauerpathway to regulate severaltaxonomic levels developmentalarrest, lifespan, and chemosensation. One C. elegans dauerpathway gene, daf-21, encodes an Andrea M. SKANTAR¤ and Lynn K. CARTA HSP90molecular chaperone. The precise function of daf- 21 isunclear; however, HSP90 chaperones are known to USDA-ARS, PlantSciences Institute, Nematology Laboratory, 10300Baltimore Blvd, Bldg 011A BARC West,Rm 165B, refolddenatured or misfolded proteins, especially under Beltsville,MD 20705,USA conditionsof stress. In many systems, HSP90 guides ¤[email protected] theproper folding of speciŽ c targetproteins, including nuclearhormone receptors and protein kinases. W eare Novelmolecular traits are needed for nematodephy- currentlyexamining the effect of an HSP90 inhibitor, logenyand diagnostics, particularly when source speci- geldanamycin,on C. elegans development.Disruption of mensare limited or when currently used molecules do normaldevelopment by this compound would implicate notprovide enough molecular discrimination to distin- arolefor HSP90in this process, and form thebasis guishgenera, species, or isolates.W ehaveexamined the for similarstudies in plant-parasiticnematodes. W ehave hsp90 geneas asingle-copydiagnostic molecule for ne- alsoinitiated a yeasttwo hybrid screen using Heterodera matodesat several taxonomic levels. W euseddegener- glycines HSP90as bait, to identify co-chaperones or ateprimer PCR to amplify partial hsp90 genomic se- targetproteins, some of whichmay include dauer pathway quencesfrom abroadrange of bacterial-feeding and homologsthat are critical to plant-parasitic nematode plant-parasiticnematode genera. Genomic DNA align- development.In the absence of robust transformation or mentsshowed that hsp90 intronshad species-speciŽ c RNAi strategiesfor plant-parasiticnematodes, the yeast arrangementsand genus-speciŽc sizes.HSP90 amino acid twohybrid approach should provide a tractablesystem for phylogenyrevealed several nematode-speciŽ c residues, analysinggene function and identifying critical protein- andplaced nematodes paraphyletic to insects. These re- proteininteractions. sultsdemonstrate that hsp90 isinformative and useful for evaluatingthe evolutionary relationships within and

180 Nematology Postersessions

166Phylogenetic and sequence analysis of ITS1, 2 BiologischeBundesanstalt, Institut fü r Nematologieund 5.8Sgene, and ITS2 regions of the rDNA of Wirbeltierkunde,T oppheideweg88, 48161, Mü nster ,Germany Steinernema :comparisonof populationsand species 3 CropProtection Department, Agricultural Research Centre, Burg.V anGansberghelaan 96, 9820 Merelbeke, Belgium 1; 2 2 S.E. SPIRIDONOV ¤, A.P. REID , K. PODRUCKA , ¤[email protected] 1 3 S.A. SUBBOTIN and M. MOENS Oneof the exciting developments in nematology during 1Instituteof Parasitologyof RAS, Leninskiiprospect 33, thelast years has been the application of nucleic acid Moscow,117071,Russia dataanalysis to problemsof systematicsof certaingroups. 2 CABI-Bioscience,Egham, TW20 9TY ,UK Inmany cases, molecular data conŽ rm themonophyly 3 CropProtection Department, Agricultural Research Centre, ofspecies groups recognised by morphological studies. Burg.V anGansberghelaan96, 9820 Merelbeke, Belgium Inother cases, the molecular data reveal taxa of which ¤[email protected] thephylogenetic relationship based on the morphology Ninetynew and 16 known complete ITS1 5.8S ITS2 seemsto be problematic. Molecular data put more light C C ofrDNA sequencesbelonging to various popula- onproblems of species boundaries and co-evolution of tionsof 22 nominal and several unidentiŽ ed Stei- nematodeswith their hosts. The early presumption that nernema specieshave been analysed using maximum phylogenetictrees based on molecular data would more parsimonyand maximum likelihood methods. Analy- preciselyre ect the true phylogeny than morphological sesshowed that more than 90% of the studied se- data,seems to be wrong. Analyses of in uence of quencesformed four main moderately or highly sup- alignmentor tree building methods on phylogenies portedclades ‘ intermedium -afŽ ne’, ‘carpocapsae-tami- showedthat molecular data could be subjected to many siamkayai-scapterisci ’, ‘feltiae-kraussei-oregonense ’, problems.Congruence and incongruence of results of ‘glaseri-cubanum -arenarium-karii-longicaudum ’) and analysesof molecular and morphological data sets oneweakly supported clade (‘ bicornutum-ceratophorum- andthe possible reasons for conicts are discussed riobrave’).Thisgrouping was obtainedin most of the withexamples of plant-parasitic nematodes from the phylogenetictrees obtained from differentITS alignments familyHeteroderidae, Anguinidae, Longidoridae, and andphylogenetic procedures. A shortmorphological char- entomoparasiticnematodes of the T ylenchidaand the acterisationof theseclades based on infectivejuvenile (IJ) familiesSteinernematidae and Heterorhabditidae. bodylength, amoeboid cell and lateral Ž eldstructure is given.Groupings of species in the ITS basedtrees are 168Molecular phylogenetics of the cyst-forming largelycongruent with previous D2-D3 basedtrees. Mole- nematodes(T ylenchida:Heteroderidae) cularanalysis revealed at least ten new species among 1; 2 studied Steinernema isolatesand proved the identity for Sergei A. SUBBOTIN ¤, Dieter STURHAN twoundescribed new species from Germany(types ‘ B’ andMaurice M OENS 3 and‘ F’)withBritish undescribed cultures (types ‘ B3’ 1Instituteof P arasitologyof RAS, Leninskiiprospect 33, and‘ D1’), respectively.Sequence differences between Moscow,117071,Russia morethan ten isolates of S. feltiae and S. kraussei were 2 BiologischeBundesanstalt, Institut fü r Nematologieund estimated.Several isolates from Switzerland,Sri-Lanka, Wirbeltierkunde,T oppheideweg88, 48161, Mü nster ,Germany Venezuela,V ietnamand USA withlong and medium- 3 CropProtection Department, Agricultural Research Centre, sizedIJ were foundto beindependent from relatedknown Burg.V anGansberghelaan 96, 9820 Merelbeke, Belgium species. ¤[email protected] Sequencesof the complete ITS andthe D2-D3 expan- 167Molecular phylogenies of plantand sionsegment of the 28S gene of rDNA obtainedfrom entomoparasiticnematodes: congruence and 54valid and seven undescribed species of cyst forming incongruencewith morphologicaland biological data nematodesfrom thegenera Afenestrata,Betulodera, Cac- todera, Dolichodera,Globodera, Heterodera and Punc- 1; 2 Sergei SUBBOTIN ¤, Dieter STURHAN and todera havebeen analysed using maximum parsimony. 3 Maurice MOENS Onthe phylogenetic trees the species clustered in sev- 1Instituteof Parasitologyof RAS, Leninskiiprospect 33, eralmajor clades corresponding to the morphological Moscow,117071,Russia groupingand present taxonomic division. The analy-

Vol.4(2), 2002 181 Evolution,phylogeny and classiŽ cation (123-175) sisyielded six main morphological groups within the withthe anguinidgroups supported by theITS phylogeny. genus Heterodera:Avenae,Sacchari, Humuli, Schachtii, Thetest of topologiesconducted by maximumlikelihood Cyperiand Goettingiana. Phylogenetic relationships be- analysesshows that the monophyletic origin of anguinids tweensome of the Heterodera groupswere notwell re- parasitisinggrasses and sedges could not be rejected. solved.The Goettingiana group occupied a basalposi- Themain anguinid groups are generally associated with tionwithin Heteroderinae. Molecular data strongly sup- planthosts belonging to the same or related systematic portedmonophyly of thePunctoderinae containing the ge- groups. nera Cactodera,Globodera, Punctodera , Betulodera and Dolichodera.Punctodera and Dolichodera havesister re- 170A new anguinidfrom dune grassin South lationships.Analysis of the D2-D3 expansionsegment Africa ofthe28S gene only partly resolved relationships within basalgroups of Heteroderidae. 1; 2 3 A. SWART ¤, S.A. SUBBOTIN , L.R. TIEDT and I.T. RILEY 4 169Phylogenetic relationships within the stem and 1BiosystematicsDivision, Plant Protection Research Institute, gallforming plant-parasitic nematodes (T ylenchida: PrivateBag X134, Pretoria 0001, Republic of South Africa Anguinidae)as inferredfrom analyses of the 2 Institutefor P arasitology,Russian Academy of Sciences, ITS-rDNA sequences Leninskiiprospect 33, Moscow 117071, Russia 3 Laboratoryfor Electron Microscopy, P otchefstroom 1; 2 Sergei A. SUBBOTIN ¤ , Eino L. KRALL , Ian T. Universityfor CHE, PrivateBag X6001, P otchefstroom,2520, 3 4 5 RILEY ,VladimirN. C HIZHOV , Ariane STAELENS , Republicof South Africa Marc DE LOOSE 5 andMaurice M OENS 6 4 Appliedand Molecular Ecology, Adelaide University, PMB 1, GlenOsmond, SA 5064,Australia 1Instituteof Parasitologyof RAS, Leninskiiprospect 33, ¤[email protected] Moscow,117071,Russia 2 Instituteof PlantProtection, Estonian Agricultural University, InOctober 2000, the senior author received dune grass 51014T artu,Estonia (Ehrhartavillosa var. villosa)withpurple coloured galls 3 Appliedand Molecular Ecology, Adelaide University, PMB1, onthe stems and leaf sheaths. On opening, each gall GlenOsmond, SA 5064,Australia containedfour to six adult anguinid nematodes and 4 ProspectMira 184-1-53, Moscow ,129301,Russia numerouseggs and juveniles. Some galls were Žlledwith 5 Departmentof PlantGenetics and Breeding, Agricultural ayellowsubstance containing bacteria. The bacteria were ResearchCentre, Caritasstraat 21, 9090 Melle, Belgium 6 CropProtection Department, Agricultural Research Centre, isolatedand provisionally identiŽ ed as Clavibacter . The Burg.V anGansberghelaan96, 9820 Merelbeke, Belgium gallsand the nematodes were studiedwith the aid of ¤[email protected] thescanning electron microscope, the results of which aredepicted on the poster .Molecularanalysis of the Internaltranscribed spacer (ITS) sequencesof rDNA from anguinidsshowed that they are close to both Anguina 53populations and species of gall-forming nematodes of australis and A.microlaenia .Boththese species have thesubfamily Anguininae, along with Ž vepopulationsof beenidentiŽ ed in Australia where they infect plants of the Ditylenchusdipsaci speciescomplex were usedfor thePoaceae, tribe Ehrharteae. According to molecular phylogeneticanalyses. The molecular analyses support a analysis,a hosttest and a lightmicroscope study, the conceptof narrowspecialisation for seed-gallnematodes anguinidpopulation from SouthAfrica isnew to science andreveal distinction of atleastnine undescribed species andin the process of beingdescribed. of Anguina inducingseed galls, previously identiŽ ed as A.agrostis, andtwo species within the D. dipsaci 171Phylogenetic analysis of Longidorus and speciescomplex. Both the maximum parsimony and Xiphinema species(Nematoda: Dorylaimida) using maximumlikelihood analyses of the ITS datastrongly ITS1sequences of nuclearribosomal DNA supportmonophyly of the genus Anguina. Also, non- monophylyfor Subanguina inthe broad sense of Weimin YE 1, R.T. ROBBINS 1 and A.L. SZALANSKI 2 Brzeski(1981) and of Mesoanguina and Heteroanguina accordingto Chizhov and Subbotin was indicated. 1 Cralley-WarrenCenter ,2601N. YoungAvenue, F ayetteville, Morphologicaland biological characters are congruent AR72704,USA

182 Nematology Postersessions

2 Departmentof Entomology,University of Arkansas, Theanalysis of the ITS sequencealignment of the four Fayetteville,AR 72704,USA populationsof B.mucronatus andof three of the B. SomeLongidoridae in the genera Longidorus and xylophilus populationsrevealed minimal (one nucleotide) Xiphinema areimportant plant parasites as well as intra-speciŽc divergence.The inter-speciŽ c divergence vectorsof plant nepoviruses. Species discrimination was morethan 100 bp. There were only3 bpdifferences basedentirely on morphology is often questionable withinpopulations of thesame species, and 18 or 19bp andcontroversial because of morphometric overlap differencebetween species in sequencesof theD2 and D3 andthe lack of a phylogeneticframework for this expansionsegments of the28S gene. group.Phylogenetic analysis using DNA sequencing ofnuclear ribosomal DNA ITS1 was conductedto 173Study ofpopulationsof Ditylenchusdipsaci constructan evolutionary tree. Extensive interspecies fromthe Czech Republicby molecular methods sequencevariation and minor intraspecies sequence variationwere observed.The analyses suggest two groups Miloslav ZOUHAR¤ and Pavel RYSANEK withinArkansas Longidoridae species. The Xiphinema Departmentof PlantProtection, Czech University of groupincludes X.americanum , X. bakeri, X.chambersi , Agriculture,Kamycka 129, Prague, 165 21, Czech Republic and Longidorusdiadecturus . Longidorusdiadecturus ¤[email protected] sharessome morphological characters of both Xiphinema Ditylenchusdipsaci isone of the most harmful parasitic and Longidorus. The Longidorus groupincludes L. nematodesin Central Europe. It is able to survive for a breviannulatus , L. crassus, L. fragilis,Žveundescribed longtime in soil without its host plants and that is why species,and two non-Arkansas species ( L.elongatus itbelongsamong organisms with quarantine importance. from Europeand L.africanus from California).The Nothingis known about D. dipsaci distributionin the phylogeneticanalysis largely corresponded to thegenera CzechRepublic. The aim of the study was tocollect andspecies deŽ ned by morphology.Results demonstrated samples of D. dipsaci from theCzech Republic and to thepotential for theuse of rDNA ITS1 and18S gene identifythem by molecular methods. A regionof rDNA toinfer phylogenetic relationships and PCR-RFLP based including 3 endof S18 gene, ITS1, 5,8S gene, ITS2 identiŽcation among longidorids. 0 and 50 endof S26 gene was ampliŽed using general primersdesigned according to the DNA sequenceof 172Inter- andintra-speciŽ c variationin ribosomal Caenorhabditiselegans .Theamplicon (900 bp) was DNA insomeisolates of Bursaphelenchusxylophilus analysedby RFLP andSSCP .Restrictionendonucleases and B.mucronatus EcoR1, HincII andAluI canbe usedfor differentiationof certainbioraces of D. dipsaci.At thesame time, methods 1; 2 Jingwu ZHENG ¤, Sergei A. SUBBOTIN , for DNA extractionfrom plantmaterial and contaminated 3 3 4 Shuishan HE , Jianfeng GU andMaurice M OENS soilwere optimised. 1Departmentof Plant Protection, College of Agriculture and Biotechnology,Zhejiang University, Hangzhu 310029, China 174Study ofpopulationsof Globoderarostochiensis 2 Instituteof Parasitologyof RussianAcademy of Sciences, and G. pallida fromthe Czech Republicby molecular LeninskiiProspect 33, Moscow 117071, Russia 3 NingboEntry-Exit Inspection and Quarantine Bureau, 144 methods LiuTing Street, Ningbo 315012, China 4 CropProtection Department, Agricultural Research Centre, Miloslav ZOUHAR¤ and Pavel RYSANEK Burg.V anGansberghelaan96, 9820 Merelbeke, Belgium Departmentof PlantProtection, Czech University of ¤[email protected] Agriculture,Kamycka 129, Prague, 165 21, Czech Republic [email protected] Thegenetic diversity of Žveisolatesof Bursaphelenchus ¤ xylophilus and four of B.mucronatus from Chinaor Potatocyst nematodes (PCN) areorganisms of quarantine detectedin package woods from Japanand Korea was importance.That is why speciŽ c andsensitive methods studied.PCR-RFLP analysis of theITS regionof rDNA for theirdetermination are needed. According to theinter- showed that AluI, CfoI, HaeIII, HinfI and MspI yield nationalclassiŽ cation, Ž vepathotypes of G.rostochiensis speciŽc patterns.Using these patterns, both species were andthree pathotypes of G. pallida exist.They can be dif- detectedin a samplefrom Ningbo,Zhejiang, China. ferentiatedby biologicaltests using potato varieties resis-

Vol.4(2), 2002 183 Evolution,phylogeny and classiŽ cation (123-175) tantto certain pathotype(s), but this method is time and Theposter introduces a newkey of thedorylaimid genera labourconsuming. For theapplication of molecularmeth- oftheworld, updated to the latest taxonomic information odsfor speciesand pathotype differentiation, we useda availablein spring2002. The key comprises 234 genera, relativelystable region in the genome of nematodes be- whichare differentiated on the basis of nine characters tweengenes S18 and S26 with the aim of Ž ndingpoly- andeightmeasures. The key is implemented as agraphics- morphismsenabling differentiation of PCN populations baseddecision table in electronic form andis targetedly from theCzech Republic. Samples of PCNfrom different designedfor rapidroutine identiŽ cation. Response time localitieswere testedby PCR, RAPD, RFLP andSSCP . isas low as 100 ms, full identiŽ cation (99% certainty) Completepathotype series from Scotlandand Germany isachievedafter eight mouse-clicks on average.Because were usedas standards.Six populations contained G. pal- theutilised inference mechanisms consistently rely lida. Theseresults were identicalto theresults of biologi- onprobabilistic calculations, the key offers innovative caltests and morphometrical observations of cysts. functionality:like other electronic keys it allows for i)afree sequenceof identiŽ cation decisions, ii) the omissionof decisions, and iii)continuousfeedback on 175An actualand modern identiŽ cation key of identiŽcation progress. In addition, the key dynamically the dorylaimidgenera world-wide iv)indicatesthe best decision sequence onward, based on 1; 2 theprevious selections made by the user, v) provides a Aldo ZULLINI ¤ andKlemens E KSCHMITT quantitativecertiŽ cation of identiŽcation quality, and vi) 1 Universitàdi Milano-Bicocca, Dipartimento Biotecnologia e enablesnematode classiŽ cation into feeding types and c-p Bioscienze,Piazza della Scienza 2, I–20126Milano, Italy classesprior to full identiŽ cation, whenever possible. The 2 JustusLiebig University, IFZ –Departmentof Animal posterexplains and illustrates major characteristics of the Ecology,H.-Buff-Ring 26-32, D –35392Giessen, Germany key. ¤[email protected]

184 Nematology Postersessions

176Molecular surveys of marinenematode diversity 13families belonging to six orders were identiŽed. Station2 registerednot only the highest density of Mark BLAXTER, Ingrid IREDALE, Ronan ROCHE, nematodes(658 ind./ 10cm 2)butalso the highest Robin FLOYD and Abebe EYUALEM numberof families. Monhysteridae, Oncholaimidae and Instituteof Cell,Animal and P opulationBiology, University of Desmodoridaepredominated, and theyrepresented 80.8% Edinburgh,Ashworth, Laboratories, Kings Buildings, ofallnematodes in thisstation. Station 1 was represented Edinburgh,EH9 3JT,UK byonly seven families with a densityof 278 ind./ 10 cm2,Desmodoridaebeing the dominant. The differences Usingthe molecular barcoding system developed for ter- foundin density, evenness and richness depend on restrialnematodes, we performeda setof assayson ma- thedifferential behaviour of the ecological factors. In rinesamples to prove the utility of the method in addi- thissense, stability and granulometric composition of tionalhabitats. The method relies on polymerase chain sedimentswere important. ampliŽcation and DNA sequencingof an informative segmentof the small subunit ribosomal RNA (SSUr- RNA) genefrom individualnematodes. The sequence is 178Trophic composition of free-livingmarine thenused to deŽ ne operational taxonomic units (OTU) nematodesin two stationsfrom Diego Perez Key, for thestudy population, and is compared to sequences Cuba from knowntaxa to attach taxonomic and ecological at- E. CALVO and J. GUERRA tributes.W esurveyednematodes from thelow tide zone ¤ ofthreeScottish beaches (Loch Fyne (W estcoast, muds), Dpto.Biologí a Animaly Humana,F acultadde Biología, Orkney(Northern, sandy) and Gullane (East coast, estuar- Universidadde La Habana, Calle 25 No. 455 entre J eI, ine/sandy)).A secondsurvey of agasseep site in theFirth Vedado,Ciudad de LaHabana, Cuba ofForthwas carriedout toassessthe utility of themethod ¤[email protected] for shorterrange surveys. Over 250sequences were de- terminedand these deŽ ne over 30 OTU. Themolecular Free-livingmarine nematodes are ecologically very OTU identiŽed were assignedto ‘morphological’OTU by heterogeneousand occupy different trophic positions in i)sequencingfrom specimensfrom thesites (identiŽ ed thebenthic food web, but in Cuba they are poorly incollaboration with Dr TimFerrero andProf. J.Lamb- studied.Sediment was collectedat the external side of shead,NHM, London)and ii)bycomparisonto our grow- thereef (Station1) and from amongthe roots of red ingdatabase of nematodeSSUrRNA sequences. mangrovein the interior lagoon of the key (Station 2). Diversity(H 0),evenness(J 0)andrichness were calculated. Nematodeswere dividedinto four trophic guilds: deposit 177Free-living marine nematodes in two stations feeders(DF), epistratefeeders (EF), scavengers(S) and fromDiego Perez Key, Cuba predators(P). Trophicbehaviour at guild level re ected highdifferences among stations. The diversity in the E. CALVO¤ and J. GUERRA guildswas higherin Station 2: this behaviour is due to DptoBiologí a Animaly Humana,F acultadde Biología, thedistribution of individuals by family, more than the Universidadde LaHabana, Calle 25 No.455 entre J eI, amountof represented families in the case of EF ,Sand Vedado,Ciudad de LaHabana, Cuba Pguilds.The component that produces a higherdiversity ¤[email protected] inDF guildis the amount of families. In DF guildthe abundanceand richness presented the highest differences Free-livingmarine nematodes have been poorly studied betweenhabitats. In Station 2, thetwo main families are inCuba and, in general, they are named to class includedin two guilds associated with the detritus chain, levelas components of the meiobenthos. The sediment butin Station 1 thetwo dominant families are in the same accumulatedin the external slope of the reef (Station guildrelated to thegrazing chain. 1)and among the roots of the red mangrove in the interiorlagoon of the key (Station 2) was collected. Thetemperature and salinity values were 29 ±C and 179Biology and taxonomic notes on siblingspecies 34%,respectively. Granulometric composition of the of Oncholaimium (Nematoda:Enoplida) from stationswas analysed.Diversity (H 0),evenness(J 0) and Russian FarEastern part of the Seaof Japan richnesswere calculated.One species, 13 genera and

Vol.4(2), 2002 185 Marineand fresh water nematodes (176-186)

N.P. FADEEVA 46nematode species were recorded;the most numerous speciesis widely-distributed Sabatienapulchra, followed FarEast State University, 8 SukhanovaSt., 690600 Vladivostok,Russia by Anoplostomacuticularia , Halanonchusarenarius and [email protected] Tershellingia sp.Nematode communities were identiŽed usingclassiŽ cation and ordination techniques. The identi- Speciesof the marine nematode genus Oncholaimium are Žedcommunitieswere thancharacterised with respect to widelydistributed in thebenthos of theSea of Japan.W e speciescomposition, density, diversity, size structure and determinedthat the Oncholaimium complexfrom Russian feedingtype. Deposit feeders were mostdominant in all Far Easternpart of theSea of Japanwas composedof at communities,followed by omnivore/predators,explained leastthree sibling species: O.olium,O. paraolium and bythemuddy nature of thesediment. The structure of the O. ramosum. Populationsfrom habitatsin threelocalities nematodecommunities from theT atarskyiStrait is com- havebeen compared. These three species are very similar pared with: i)thecommunities from adjacentoxic deep inmorphology. Multivariate analyses were donefor 14 seasediments; ii)anotherarea of continentalshelf water morphometriccharacteristics in males, and 13 infemales. off thecoast of Sakhalin;and iii)shallowreduced envi- Inaddition, a seriesof minor variations in morphology ronmentssuch as sedimentsof eutrophicbottoms in Peter were found(structure of terminal parts of the tubular theGreat Bay. organ,form ofsupplementary organ). The sympatric siblingspecies Oncholaimium complexform aseries onthe successional scale between the most perturbed 181The embryonic cell lineage of the nematode conditionsoccupied by nematode O. ramosum, and less Pellioditismarina shows evolutionaryadaptations impacted,slightly more predictable and stable conditions, towardsfaster development in the Rhabditidae characterisedby sub-littoral nematode O.paraolium and Wouter HOUTHOOFD , Sandra VANGESTEL, Clarinda bylittoralnematode O.paraolium. DetailedŽ eldstudies ¤ MERTENS, August COOMANS and Gaëtan BORGONIE of O. ramosum havefound that it isa dominantmember ofthe epifauna in areas subjected to severe organic UniversityGent, Department of Biology, K.L. Ledeganckstraat disturbanceand heavy metal and petroleum hydrocarbon 35,9000 Gent, Belgium contamination,and hastheability to withstandhigh levels ¤[email protected] of H2S,acombinationfacilitating the primary opportunist Untilnow only the embryonic cell lineage of themodel or- roleof thisspecies of thecomplex. ganism Caenorhabditiselegans hasbeen described. The completeembryonic cell lineage (until muscle contrac- 180Free-living marine nematode communities in tion) of a Pellioditismarina embryowas establishedby the northernpart of TatarskyiStrait in the Seaof meansof 4D-microscopy. Pellioditismarina isa closerel- Japan ative of C. elegans,buthas adapted to amarine,brackish environment.The overall cell lineage resembles strongly N.P. FADEEVA¤ and N.L. DEMCHENKO that of C. elegans,witha few smalldifferences. At the stageof muscle contraction (when most cells are estab- FarEast State University, 8 SukhanovaSt, 690600 Vladivostok, lished), P. marina hasas many cells as C. elegans (571 Russia cells)but less cell deaths (60 and 106, respectively). In ¤[email protected] morederived forms ofdevelopment,like in P. marina and Thenorthwestern part of the Sea of Japan, especially in C. elegans,developmentalrate is hastened by adapting theregion of Amurskyi Liman, is the most productive theirlineage, so that cells originate close to their Ž nal andsensitive to modern negative anthropogenic activity. position.This could explain the ‘ chaotic’cell lineages of Meiofaunawere sampledin shallow (3-20 m) continen- P. marina and C. elegans wheretissues are derived from talshelf at 15 stations on June to August 2001. T otal differentfounder cells. In the slower developing Hali- meiofaunaabundances ranged from threeto 1730 indi- cephalobus sp.,sublineages form identicalcells, which vidualsper 10 cm 2 witha meanof 810 individuals per migrateto theirexact location. Pellioditismarina and C. 10 cm2.Inmeiobenthos we seereinforced nematodes elegans haveadjusted these lineages to avoidthese migra- dominancecomparative with other groups. Nematofauna tionsand hasten their development. abundancesranged from twoto 1350 individuals per 10 cm2 witha meanof 700individualsper 10cm 2. A total of

186 Nematology Postersessions

182Thallassinidean burrow walls: a cradlefor offer thepromise of being able to resolve con icts nematodes inthis Ž eldby measuring relatedness using a single metricand veriŽable, model-driven analysis. Phylogenetic Helmut KOLLER relationshipsof marine nematodes from UKwaters Departmentof Marine Biology, University V ienna,Altanstrasse were inferredfrom partialsmall subunit ribosomal RNA 14,A-1010 Wien genesequences. As partof the experimental design [email protected] anewsequence of techniques was setup to ensure speciesidentiŽ cation prior to DNA extractionof the As apartof a studyof the biology of the intertidal- wholeanimal. Phylogenetic trees derived from these burrowingshrimp Callianassatyrrhena (Thalassinidea) sequencesshowed that, within ‘ Adenophorea’, onlythe themeiofauna from burrowwalls and chambers was Monhysteridaform amonophyleticgroup, and that the comparedwith that of surface sediment and embedded Chromadoridaand Enoplia are both polyphyletic. Even at ambientdebris. Abundance and size spectrum of highertaxonomic levels, different clusterings of species nematodesfrom allsamples were determined.Nematodes areevident in molecular compared to morphological from burrowwall and surface sediment samples were phylogenies.The data support earlier studies using identiŽed to genus level. The highest number of moleculartechniques in that classical views of nematode nematodesespecially juveniles was foundin the burrow systematicshave to berevised. wallsamples. Halaphanolaimus was themost dominant ofthe19 different genera found in this sample category. Insurface sediment samples 12 different genera were 184Nematodes of the order Dorylaimida (Pearse, found, with Chromadorina beingthe most dominant. 1942)in different high-mountainlakes of the Sierra Sevengenera are common to both sample categories. The Nevada(Granada, Spain) denselypacked sediment of theburrow walls apparently 1; 2 A. OCAÑA ¤, R. PEÑA SANTIAGO , hasenough interstitial space for alargemeiofauna 1 1 community,and also represents a retreatfor juvenile J. HERNÁNDEZ RUIZ and R. MORALES BAQUERO nematodes. 1DeptoBiologí a Animaly Ecología, Universidad de Granada, Granada18071, Spain 183Phylogenetics of marinenematodes – 2 DeptoBiologí a Animal,V egetaly Ecología, F acultadde amolecularapproach CienciasExperimentales, Universidad de Jaé n, Jaén 23071, Spain 1;2; 2 Birgit MELDAL ¤, John LAMBSHEAD , Alex ¤[email protected] ROGERS 3, Adam COOK 2, Mark BLAXTER 4, Nicola MITCHELL 2 andMelanie A USTEN 5 Atotalof 20 lakes was sampledin the Sierra Nevada mountains(Granada, Spain) in thesummers of1990and 1Schoolof Oceanand Earth Sciences, University of 1991.At thebeginning of thisperiod in 1991, the lakes Southampton,Southampton Oceanography Centre, European were stillcovered with ice. Despite their altitudes of 2820- Way,Southampton, SO14 3ZH, UK 2 3050m andbeing oligotrophic lakes, the nematofauna NematodeResearch Group, Department of Zoology, The provedabundant. The orders with the greatest total NaturalHistory Museum, London SW7 5BD, UK biomasswere: Dorylaimusasymphidorus Andrássy, 1969 3 BritishAntarctic Survey, High Cross, Madingley Road, Cambridge,CB3 0ET,UK (45.2%of the total of the dorylaimids in 1990 and 4 Instituteof Cell, Animal and P opulationBiology, Ashworth 62%in 1991); Mesodorylaimus cf. litoralis Loof, 1969 Laboratories,University of Edinburgh,King’ s Buildings,W est (35%in 1990 and 7.7% in 1991) and Paractinolaimus MainsRoad, Edinburgh, EH9 3JT,UK macrolaimus Meyl,1957 (4.5% in 1990 and 14.2% in 5 PlymouthMarine Laboratory, Prospect Place, The Hoe, 1991).No correlation was foundbetween the dorylaimids Plymouth,PL1 3DH,UK andthe physico-chemical parameters analysed; however, ¤[email protected] thisnematofauna correlated positively with surface area, depthand catchment area of thelakes. Historically,the classiŽ cation of nematodes has been basedsolely on the interpretation of the evolutionary historyof morphologicalcharacters, resulting in a setof 185Marine nematodes as hosts for mutuallyincompatible systems. Molecular phylogenetics chemoautotrophicsymbionts

Vol.4(2), 2002 187 Marineand fresh water nematodes (176-186)

1; 1 Joerg OTT ¤, Monika BRIGHT , 186 Draconemahoonsooi n. sp. and Draconema Andrea NUSSBAUMER 1, Katrina VANURA 2, youngeouni n.sp. (Adenophorea:Chromadorida: Irmgard EICHINGER 1 and Irma SCHABUSSOVA 1 Draconematidae).The Ž rst recordof free-living marinenematodes from Korea 1Instituteof Ecologyand Conservation Biology, University of Vienna,Althanstrasse 14, A-1090 V ienna,Austria 1; 1 2 Hyun Soo RHO ¤, Joong-Ki PARK , Dept.of Internal Medicine I, Div.of Hematology & 2 1 Hemostaseology,University Hospital/ Vienna,Wä hringer CheonY oungC HANG and Won KIM Gürtel 18-20, A-1090 V ienna,Austria 1Schoolof BiologicalSciences, Seoul National University, ¤[email protected] Seoul151-747, Korea 2 Marinefree-living nematodes (Stilbonematinae, Desmo- Departmentof Biology, College of NaturalSciences, T aegu University,712-714, Korea doridae,Adenophorea) have ectosymbiotic sulphur- [email protected] oxidisingchemoautotrophic bacteria attached to their ¤ cuticlein agenusor speciesspeciŽ c pattern.In thegenus Twofree-living marine nematodes, Draconemahoonsooi Eubostrichus thebacteria are up to 100 ¹m long non- n. sp. and Draconemayoungeouni n.sp. of the family septateŽ lamentscontaining many nucleoids. The worms Draconematidae,are described on the basis of the supplythe bacteria alternately with oxygen and sulphide specimenssampled at depth 30-60 m ofthe Sea of bymigrating through the chemocline in sheltered sandy Japanand Jeju Island of Korea. Draconemahoonsooi sediments.In turn, the bacteria are eaten by the worms n.sp. is recognised by the possession of the following andprobably constitute their only food. The nematodes characteristics:having a cup-shapedamphideal fovea possessconspicuous epidermal glandular sense organs inboth male and female, Ž vepairs of evenly tapered whichare unique to the Stilbonematine. The mucus analsetae (male), 15 pairs (male) and18 pairs (female) producedby these glands is most probably connected to ofposterior sublateral adhesion tube (SlA T), andŽ ve therecognition mechanism between the worms andthe pairsof setae on non-annulated tail region in both bacteria.The latter are shed with each moult and have sexes. Draconemayoungeouni n.sp. can also be easily tobe recruited from theenvironment. Preliminary data distinguishedfrom allknown congeneric species in suggesta lectine/sugarbond. According to the sequence havinga combinationof the following features: the ofthe16S rRNA genethe primary symbionts all belong greaterbody length and slender body shape, the bowl- to the ° -proteobacteria.In addition, the complex bacterial shapedamphideal fovea in both male and female, 13 pairs coatof somegenera harbours a varietyof otherbacteria, (male) and17 pairs (female) ofposterior sublateral including ®- and "-proteobacteriaand cytophaga. The adhesiontube, Ž vepairs of evenly tapered anal setae stilbonematid/bacteriasymbiosis is found in tropical to (male),and the absence of long setae on non-annulated cooltemperate shallow waters. Highest diversity and tailregion in both sexes. The photomicrographs of the abundance,however, is found in tropical calcareous newspecies by scanningelectron microscopy (SEM) and sediments. differentialinterference contrast (DIC) microscopyare presentedwith a detailedmorphological description.

188 Nematology Postersessions

187Molecular evolution among geographic isolates UK, and S. riobrave 335on cabbage leaves were 43,2, of Steinernemacarpocapsae and0%, respectively, 4 hfollowingapplication. Under Želdconditions, foliar applications of S.carpocapsae All Byron J. ADAMS¤ andKhuong B. N GUYEN provided24.9, 19.4 and 14.9% control of A. rapae on Entomologyand Nematology Department, University of Brusselssprout, broccoli, and cauli ower, respectively. Florida,Gainesville, FL 32611-0620,USA ¤bjadams@u.edu 189Discrimination between isolatesof RibosomalDNA (ITS regions1 and2) from tendisparate Heterorhabditisbacteriophora Poinar,1976 geographicisolates of Steinernemacarpocapsae was (Nematoda:Heterorhabditidae) from the provinceof sequencedand analysed in order to better understand Córdoba, Argentina intraspeciŽc variation,speciation, and the molecular María A. BERTOLOTTI1; ,MarceloE. D OUCET 1 and evolutionof this locus. T axaassumed to represent ¤ Julio A. DI RIENZO 2 panmicticpopulations were sampledfrom theUSA, Poland,Russia, France, Sweden, UK andMexico. No 1Laboratoriode Nematologí a, Centrode Zoología Aplicada, phylogeographicpatterns emerged from thedata set. UniversidadNacional de Có rdoba, C.C. 122,5000, Có rdoba, Amongthe populations, transitions were ascommon as Argentina 2 transversions,and a singleinsertion event was observed. Unidadde ProcesamientoElectró nico de Datos,F acultadde Intra-individualpolymorphic characters with states Ž xed CienciasAgropecuarias, (U.N.C.). C.C. 509,5000, Có rdoba, Argentina amongother taxa indicate an intermediate phase in [email protected] concertedevolution and tokogenetic relationships among ¤ thesampled taxa. These data are used to explore the mode Upto the present, eight isolates of Heterorhabditis andtempo of evolutionat this locus, and as anoperational bacteriophora from theprovince of Có rdoba have modelfor comparingapplications of speciesconcepts. beendescribed, and signiŽ cant differences have been foundfor alltheir morphometrical characters. The aimsof this work were: to discriminate these isolates 188EfŽ cacy of steinernematidnematodes against basedon morphometrical characters, to identify the threeinsect pests ofcrucifersin Quebec principalcharacters that permit their separation and toevaluate the possible relationship between isolates. Guy BÉLAIR¤, Yvon FOURNIER and Isolatesfrom sixdepartments corresponding to two Nathalie DAUPHINAIS differentphytogeographi calregions of the province Agricultureand Agri-F oodCanada, 430 Gouin Blvd, were considered.Hermaphroditic females, amphimictic St-Jean-sur-Richelieu(Quebec), J3B 3E6, Canada females,males and infective juveniles (IJ) were analysed. ¤[email protected] Anevident separation of one isolate with respect Steinernemafeltiae , S.carpocapsae and S. riobrave were tothe others was observedin adults. In IJ the evaluatedagainst imported cabbageworm Artogeiarapae , isolateswere separatedin three groups. The most diamondbackmoth Plutellaxylostella ,andcabbage relevantmorphometrical characters for discrimination looper Trichoplusiani underlaboratory and Ž eld were:distance between anterior end and nerve ring, bodydiameter at anus, distance between anterior end conditions.LD 50 valuesranged from 3.6-18.2for A. rapae L2,from 2.3-24.5for P.xylostella L3,and from 4.7- andexcretory pore, ratios a, c, D inadults; distance 10.1 for T. ni L2. S. feltiae UKwas themost pathogenic betweenanterior end and pharyngeal basal bulb, distance strainon all three pests. Maximum larval mortality rates betweenanterior end and excretory pore, ratios a, D, E recorded on A. rapae were inuenced by temperature: inIJ. Similarityrelationship among isolates was different dependingon thestage considered. 100% with S. riobrave 335 at 30±C,95.8%with S. feltiae UK at 25±C,91.7%with S. feltiae 27 at 25±C, and 75.7% with S.carpocapsae All at 30±C.Mortalityof A. 190The use ofthe glycosylase-mediated rapae L2increased with contact time to S.carpocapsae polymorphismdetection for the study ofgene owin All and S. feltiae UKwitha 76and 78% maximum, entomopathogenicnematodes respectively,reached after 12 h contact.At 20 ±C and 70%RH, survivalrates of S.carpocapsae All, S. feltiae Stephen BOYLE and Thomae KAKOULI-DUARTE¤

Vol.4(2), 2002 189 Entomopathogenicnematodes (187-221)

Departmentof Applied Biology and Chemistry, Institute of developmentis completed12-15 days after oviposition at TechnologyCarlow ,KilkennyRoad, Carlow ,Ireland 26±C.Thepre-parasitic juvenile emerges from theegg, ¤[email protected] livingonly 48 h, when it must Ž ndand penetrate the Entomopathogenicnematodesare traded for controlof host.The parasitic juvenile develops in the host, being the difŽcult pests. The greatest limitation on understanding onlyfeeding stage. The post-parasitic juvenile abandons howindigenous populations are affected by newstrains or thehost through cuticle perforation. It moults into adult, escapedgenes is the problem of recognising the genetic maleor female,in the environment. Moulting, copulation structureof wild populations and the levels of gene andoviposition were observedonly when numerous owbetween them. W eareaiming at studying gene individualswere interlaced.The biological observations owin Irishentomopathogen icnematodepopulations by performedagree with what was indicatedfor otherspecies detectingDNA polymorphismsin the intron regions of of the genus Gastromermis . highlyconserved nematode genes. W ehaveused EPIC- PCRto amplify part of the major sperm protein gene 192Phoretic association of Steinernemafeltiae from Steinernemafeltiae and Heterorhabditismegidis (Nematoda)with Eiseniafetida (Annelida) containinga putativeintron. DNA sequenceanalysis 1; 1 2 willfollow to conŽ rm thepresence of the intron. Raquel CAMPOS ¤, Jesús JIMÉNEZ , Dolores TRIGO , AmpliŽcations were achievedusing degenerate primers Marta RAMAJO 2 and Carmen GUTIÉRREZ 1 designedafter aligning the major sperm protein sequences 1 from sevennematode species taken from GenBank.Our DptoAgroecologí a, CCMA CSIC,Serrano 115 dpdo 28006 researchis still ongoing for additionalsuitable genes. Madrid,Spain 2 DptoBiologí a AnimalI, Facultadde Ciencias Bioló gicas, Geneticvariation in the introns will be studied by the UniversidadComplutense de Madrid, Spain useof theglycosylase-mediat edpolymorphism detection ¤[email protected] (GMPD). Intronswill be reampliŽ ed using the four dNTPs includinguracil and having adjusted the dTTP Earthwormsimprove soil characteristics and have a no- todUTP ratio.Products will then be subjected to the tablein uence on edaphic organisms; however, inter- actionof uracilDNA glycosylase,which results in DNA actionsbetween them have not been studied in detail. cleavageat speciŽ c sitesthus generating an allelicproŽ le After protozoa,nematodes are the most abundant and for eachpopulation studied. diversegroup of soil invertebrates and an important partof food webs. Entomopathogen icnematodes are an environmentally-friendlyalternativefor insectpest con- 191Life cycle of Gastromermismassei Doucet & trol.However, their dispersal capability is limited and it is Cagnolo,1997 (Nematoda: Mermithidae) oftendifŽ cult to reachthe expectedlevel of control.Since earthwormsare able to change soil structure and move Susana R. CAGNOLO¤ andMarcelo E. D OUCET largeamounts of soil,some authors think that earthworms Laboratoriode Nematologí a, Centrode Zoología Aplicada, couldbe used as vectors to introduce and disperse ben- UniversidadNacional de Córdoba, C.C. 122,5000 Có rdoba, eŽcial organisms. As partof thestudy about interactions Argentina [email protected] betweenearthworms and nematodes, we showthe infec- ¤ tivityresults of Steinernemafeltiae afterpassage through Thelife cycle of Gastromermismassei was studiedfor 3 the Eiseniafetida gut.Moreover, entomopathogen icne- years.Samples were takenfrom thebedofCosquín River, matodeshave no deleteriouseffect on earthworms. Córdoba, Argentina and individuals in their free-living stagewere collectedwith Surber trap. Samples were 193Combination of Steinernemacarpocapsae processedthrough suspension and 40 ¹mporesieving. (Weiser)and pheromone lure: a promisingstrategy Larvae,pupae and adults of Simulium (Ectemnaspis ) forbiological control of the bananablack weevil wolfhuegeli (Enderlein)were collectedfor thestudy of Cosmopolitessordidus (Germar)on bananasin theparasitic stage. Parasites and immature stages of Martinique hostswere keptunder laboratory conditions. Cycle was completedin 30-40 days. It includes adults, eggs, and C. CHABRIER 1, H. MAULEON 2 and P. QUÉNÉHERVÉ 3 fourlarval stages. Eggs are deposited freely on the substrate,without showing segmentation. The embryonic 1 CIRAD, BP153,97202 F ort-de-FranceCedex, Martinique

190 Nematology Postersessions

2 INRA-URPV,DomaineDuclos, 97170 P etitBourg, duringthe Ž rst halfmonth after which it decreased. The Guadeloupe lipidcontent of IJwas negativelycorrelated to thestorage 3 IRD, BP8007,97259 F ort-de-Francecedex, Martinique temperatureand time. Lipid content and IJ survivalwere positivelycorrelated; lipid content and IJ infectivitywere Cosmopolitessordidus (Germar, 1824)isoneofthemajor notcorrelated. pestof bananasin the Caribbean. Several techniques can beapplied to avoid use of insecticides. On one hand, laboratorystudies have shown that some speciŽ c strains 195The potential of Heterorhabditismarelatus for of Steinernema spp.could contribute efŽ ciently to the controlof Coloradopotato beetle controlof this pest. Unfortunately, Ž eldapplications of Nathan COTTRELL , E. GRAFIUS and formulationsof S.carpocapsae oftengive inconsistent ¤ H. MELAKEBERHAN resultsdue to formulation and conservation constraints. Onthe other hand, mass-trapping using pheromone lure MichiganState University Department of Entomology,243 hasgiven good Ž eldresults, excepted when the C. NaturalScience Building, East Lansing, MI 48824-1115,USA sordidus populationlevels were veryhigh. In this study, ¤[email protected] we usedpheromone traps to contaminate weevils with Coloradopotato beetle (CPB, Leptinotarsadecemlineata ) entomopathogenicnematodesinstead of killing them. isan important pest of potato and has considerable re- Duringa 10month study, we evaluatedthe interest sistanceto manyinsecticides. With current and future re- ofpheromone traps using pheromone lure on which strictionson pesticides,ecologically safe alternativesare we replacedeach week new Galleriamellonella larvae needed.The goal of thisproject is todevelop Heterorhab- S.carpocapsae previouslyinoculated with (Weiser).In ditismarelatus (Hm)asabiologicalcontrol alternative for thisexperiment, promising results were obtainedwith a CPBthroughunderstanding its pathogenicity and adap- reductionof attacked plants from 98.5to 40% and a tationto Ž eldedaphic factors. Hm hadbest survival and reductionof theattack severity (according to Vilardebo’s pathogenicityin sandand sandy loam soil and at 60-100% coefŽcient) from 42to23. moisturecontent under controlled conditions. In naturally CPBinfestedŽ eldplots (322 m 2),treatmentsof 0, 333 194Effects ofstoragetemperature on survival, million,667 million or 1 billion Hm/m2 ofsoil in 2000 infectivityand lipid content ofthe entomopathogenic and2001 showed higher numbers of beetles in control nematode Steinernemafeltiae (Rhabditida: plotscompared with nematode-treated plots. Field cages Steinernematidae) (2 m3)artiŽcially infested with Ž veCPBadults per plant were treatedwith either 0 or1billion Hm/m2 at the emer- Shulong CHEN¤, Xiuying HAN andMaurice M OENS genceand peak of 4 th instar,or atpeak pupation. Time of applicationwas notsigniŽ cant, but more adults emerged AgriculturalResearch Centre, Crop Protection Department, Burg.V anGansberghelaan96, B-9820 Merelbeke, Belgium incontrols than in Hm treatedplots. Overall, the results suggestthat Hm hasa potentialfor managingCPB under ¤[email protected] Želdconditions. TheBelgian Lxm 31 population of Steinernemafeltiae was cultured in vitro or in vivo andstored in water at 5, 196Abundance ofendemicentomopathogenic 10, 15 or 20±C.Its survival,infectivity and lipid content nematodesfollowing application of Steinernema were evaluatedafter 0, 0.5, 1, 2, 3, 4, 5, 6, 12 or 18 riobrave months.Temperature, storage time and culture method signiŽcantly in uenced the survival of the infective L.W. DUNCAN 1, J.H. GRAHAM 1, D.C. DUNN 1, juveniles(IJ). Survivaldecreased with increasing storage J. ZELLERS 1 and K. NGUYEN 2 temperatureand was negativelycorrelated to storage time. 1 After along-termstorage, the percentage of survivingIJ Universityof Florida, Citrus Research and Education Center , was higherwhen cultured in vivo thanwhen produced 700Experiment Station Rd., Lake Alfred, FL 33850,USA 2 Departmentof Entomology and Nematology, Gainesville, FL in vitro.Storagetemperature also in uenced infectivity. 32611, USA Whenkept at low temperatures (5-10 ±C),IJ infectivity decreasedduring the Ž rsthalf month and then increased. Cagedinsect larvae were buriedat monthly intervals in a WhenIJ were storedat 20 ±C,theirinfectivity increased Floridacitrus orchard during 2000-2001 to monitor nat-

Vol.4(2), 2002 191 Entomopathogenicnematodes (187-221) uralcontrol of Diaprepesabbreviatus byentomopatho- Asingletreatment of carbaryl followed by 4 yearsof genicnematodes (EPN). Thesurvey occurred in plots that treatmentwith S. riobrave increasedthe cumulative yield were treatedor nottreated with Steinernemariobrave dur- (1999-2001)of citrus by 16-23%, a 4-11-foldreturn ing1998-2001. Mean monthly insect mortality in control oninvestment, depending on fruit variety and market plotsranged from 13-74%and endemic EPN ( Steiner- destination. nema sp.,Heterorhabditiszealandica, H. indica, and H. bacteriophora )were recoveredfrom 22-81%of the ca- 198Liquid culture production of biocontrol daverseach month. Endemic EPN ineach plot were in- nematodes verselyrelated to S. riobrave anddirectly related to the numbersof adult weevils ( D.abbreviatus and Pachnaeus R.-U. EHLERS¤ and A. PETERS litus)capturedin modiŽ ed T edderstraps. Insect mortality Institutefor Phytopathology, University of Kiel,Department of was higherand cadavers containing endemic EPN were Biotechnology& BiologicalControl, Klausdorfer Str .28-36, morenumerous in controlplots than in S.riobrave- treated 24223Raisdorf, Germany plots,except during months in which S. riobrave was ap- ¤[email protected] plied.In treated plots, recovery of endemicEPN exceeded For mass productionnematodes are produced in liquid S. riobrave that of inall but the treatment months. The mediapre-incubated with the symbiotic bacterium S. riobrave D.abbrevia- netefŽ cacy of treatmentsagainst for 1daybefore the dauer juveniles are inoculated. tus attenuatedover time, possibly due to competitive dis- Nematodeyields and the process duration can vary placementof EPN thatare better adapted than S. riobrave signiŽcantly. Several different species have nowadays topersist in Floridacitrus orchards. beencultured successfully in liquid media. For example, Heterorhabditismegidis dauerjuvenile yields can reach 197ProŽ tability of applicationsof Steinernema 100000/ml;however, the mean of several processes is riobrave,metalaxyland supplemental fertilisation for usuallynot higher than 40 000/ml.The process time managementof Diaprepesabbreviatus and iseither 15 days (one generation) or 24 days (two Phytophthoranicotianae ina Floridacitrus orchard generationprocess). Y ieldsdepend on the nematode speciescultured. Species with smaller dauer juveniles L.W. DUNCAN, J.H. GRAHAM and J. ZELLERS canreach a dauerjuvenile density of >500000/ml.A majorreason for processinstability is a variationof Universityof Florida,Citrus Research and Education Center , thenematode population dynamics. Another in uencing 700Experiment Station Rd, Lake Alfred, FL 33850,USA factoron yields is the bioreactor design. Internal loop Anexperiment was conducted(1998-2001) in a central bioreactorsequipped with a marinepropeller can provide thenecessary oxygen at low shear forces. The system has Floridacitrus orchard infested by Diaprepesabbreviatus beenproven to be superior to airlift and conventionally and Pachnaeuslitus toevaluate the proŽ tability of stirredbioreactors and has been successfully scaled up to recommendedIPM tacticsfor insectcontrol. Main 3000 l. plots were i)nottreated for insectcontrol, ii) treated twiceannually since 1998 with commercially formulated Steinernemariobrave, or iii)treatedtwice annually with 199Factors in uencing dauer juvenile recovery of S. riobrave and15 times annually with liquid fertiliser Heterorhabditisbacteriophora inliquid culture inplace of dry fertiliser (three times/ year)used in the othertreatments. Split plots were untreatedor treated R.-U. EHLERS¤, J. AUMANN and O. STRAUCH twiceannually with metalaxyl. Carbaryl was applied Institutefor Phytopathology, University of Kiel,Department of once(May 1998) to treatments 2 and3. During 4 years, Biotechnology& BiologicalControl, Klausdorfer Str .28-36, numbers of D.abbreviatus inplots were directlyrelated 24223Raisdorf, Germany toabundance of Phytophthoranicotianae andinversely ¤[email protected] relatedto fruit yield. Fertiliser treatment did not affect Recoveryis thestep of thedevelopmentallyarrested infec- yieldor abundanceof anypest. Treatment with S. riobrave tivethird stage juvenile (dauer) to feeding third stage juve- reducednumbers of both insects and abundance of P. niles,which continue growth to reproductive adults. It is a nicotianae byan average of 45-60% annually except in responseto yet unidentiŽed food signals, which re ect the 2001.Metalaxyl did not affect abundance of thefungus. nutritionalconditions in theenvironment. Unlike dauer ju-

192 Nematology Postersessions veniles(DJ) of Caenorhabditiselegans ,whichrecover in 201Thermal adaptation of entomopathogenic thepresence of yeastextract, entomopathogen icnematode nematodes:acclimation, trehalose accumulation, and DJneedmore speciŽ c signals.In liquid culture a deŽned survivalat environmentalextremes numberof hermaphrodites(approximately 5000/ mlin the testedmedium) results in maximum yields. This density, ParwinderS. G REWAL andGanpati B. J AGDALE however,cannot easily be obtainedby justdeŽ ning the DJ Departmentof Entomology, Ohio State University, W ooster, inoculationdensity. In liquid culture DJ respondto food OH44691,USA signalsproduced by theirsymbiotic bacteria, which cause between18 and90% of theDJ torecoverwithin a period Coldacclimation leads to trehalose accumulation in ofseveraldays. This unsynchronised and low recovery in entomopathogenicnematodes. W ehypothesisedthat in vitro culturesis the major reason for processinstability trehaloseaccumulation in nematodes is ageneralstrategy andvariable yields. In order to increase recovery and thus toprepare for survivalat environmental extremes. improvethe management of nematodepopulation density, Therefore,we testedwhether the nematodes will severalprocess parameters were investigatedfor theirin- accumulatetrehalose during acclimation at sub-lethal uenceof DJrecoveryin liquid culture. warm andcold temperatures and whether the accumulated trehalosecorrelates with enhanced desiccation, heat, andfreezing tolerance. Three species, Steinernema 200Effect ofthe Steinernemafeltiae-Xenorhabdus carpocapsae,S. feltiae and S.riobrave, were acclimated bovienii complex on Meloidogynejavanica at 35 and 5±Cfor 1and4 days,respectively, and theirtrehalose contents were measured.Survival of D.J. FALLON 1, H.K. KAYA 2 and B.S. SIPES1; ¤ acclimated-and non-acclimated nematodes at 20 and ¡ 40±Candin 25% glycerol was compared. Steinernema 1 Plantand Environmental Protection Sciences, University of riobrave and S.carpocapsae accumulatedhigh trehalose Hawaii,Honolulu, HI 96822,USA 2 Departmentof Nematology,University of California,Davis, at 35±C, and S. feltiae at 5±C.Heattolerance in CA95616,USA acclimated S.carpocapsae and S. feltiae was high, butunaffected in acclimated S. riobrave. Freezing ¤[email protected] tolerancein acclimated S.carpocapsae and S. riobrave Metabolitesfrom the Steinernemafeltiae – Xenorhabdus was increased,whereas in acclimated S. feltiae it was bovienii complexwere testedfor theirefŽ cacy against unaffected.Heat acclimated S.carpocapsae and cold Meloidogynejavanica penetrationand egg production acclimated S. riobrave showedthe highest desiccation in soybean. In vitro metabolitesof X. bovienii were survivalat 5 ±C.Overall,the accumulated trehalose levels collectedfrom 4-day-oldYS-broth cultures by ethyl appearto enhancedesiccation, freezing and heat tolerance acetateextraction. In vivo metabolitesof the S. feltiae ofentomopathogenicnematodes. – X. bovienii complexwere collectedfrom 4-day-old S. feltiae-infected Galleriamellonella .Dailyapplications of 202Differences in susceptibility of introducedand in vitro exudatesat 1 mg/plant/day,or S. feltiae juveniles nativewhite grubspecies to entomopathogenic at1000 or 10000infectivejuveniles/ plant/day,for upto3 nematodesfrom diverse geographic localities days,were ineffectiveat reducing M. javanica penetration oregg production, compared to water treatment. A P.S. GREWAL 1, S.K. GREWAL 1 and M.G. KLEIN 2 in vivo singleapplication of metabolitesapplied at 250 1 Departmentof Entomology,Ohio State University mg-insect-cadaver/plantreduced M. javanica penetration 2 USDA, AgriculturalResearch Service, Horticultural Insects by42% compared to water treatment, but the effect ResearchLaboratory, 1680 Madison Avenue, W ooster, was lostusing multiple applications. Daily applications OH44691,USA of in vivo metabolitesdid not affect M. javanica egg production.Multiple treatment applications increased M. Wecomparedthe susceptibility of theintroduced Popillia javanica establishmentin soybeans. Steinernemafeltiae – japonica andthe native Cyclocephalaborealis to 16 X. bovienii complexeffects on M. javanica were highly speciesand strains of entomopathogen icnematodes variable. isolatedfrom withinor outside the geographic ranges ofthe two scarabs. W efoundlarge variation in the virulenceof the species/ strainsof nematodes with over

Vol.4(2), 2002 193 Entomopathogenicnematodes (187-221)

50%mortalityof P. japonicaproduced by Heterorhabditis Carmen GUTIÉRREZ¤ zealandica strainX1 and H.bacteriophora strainGPS11 DptoAgroecologí a, CCMA CSIC,Serrano 115 dpdo 28006 and of C. borealis by H.zealandica and H.bacteriophora Madrid,Spain strainsKMD10 and NC1. Heterorhabditisindica and ¤[email protected] H.marelatus causedless than 20% mortality of both scarabspecies. When considered as a group,the nematode Spodopteralittoralis ,aseriouspest in African and speciesand strains from withinand outside the geographic Mediterraneancountries, has highly polyphagous larvae, rangesof either P. japonica or C. borealis didnot differin whichfeed mainly at night and hide below ground virulencetowards either scarab species. Results suggest byday. Pupation takes place just below the surface thatthe introduction of the exotic H.zealandica into ofthe soil, and there are at least seven generations a thefront-line states with respect to the movement of yearin warm climates.Although chemical pesticides P. japonica and A.orientalis shouldbe explored as a and Bt areused for control,resistances have recently tacticto delaytheir establishment and spread. The results beenreported in Ž eldand laboratory. Entomopathogeni c alsosuggest that the manipulation of the indigenous H. nematodesare virulent against many species of insects. bacteriophora populationsmay help in delaying spread Theirability to actively locate and kill an insect host andmitigating losses caused by theinvasive grub species. ina few hoursas well as their high reproductive potentialand capacity for mass productionmake these organismsa keyto establishingenvironmentally-frien dly 203Control of overwinteringblack vine weevil alternativesfor controllinginsect pests. However, their Otiorhynchussulcatus bya cold-active dispersalcapability in soilcan be affectedby thetexture entomopathogenicnematode Steinernemakraussei andbiological factors of the isolation strain. Thus, soil texture/infectivitybioassays can improve on selection of Andrew J. HART andDeena M. W ILLMOTT ¤ amoreaccurate Ž elddose application. The virulence of Departmentof Entomological Sciences, Horticulture Research Steinernemafeltiae (Riojastrain) against S.littoralis in International,W ellesbourne,W arwickCV35 9EF ,UK threesoils with different clay contents was studiedunder ¤[email protected] laboratoryconditions. Replicate samples of variousdoses of S. feltiae juvenileswere appliedto the soil surface. Otiorhyn- Thesusceptibility of black vine weevil larvae Nematodevirulence against S.littoralis was reducedin chussulcatus toa cold-activeentomopathogeni cnema- soilwith 24% clay content. tode, Steinernemakraussei (isolateL137) and a commer- ciallyavailable nematode, Steinernemacarpocapsae (Ex- hibit)was comparedin winterŽ eldtrials. Nematodes were 205The genetic selection for improved desiccation appliedto outdoorpotted strawberry plants at a rangeof toleranceof Heterorhabditismegidis 01 PL dosesin early winter .Parasitismand mortality of vine weevilswere assessedat theendof thewinter experiment. Jolanta KOWALSKA¤ Upto 83% of vine weevils were infectedand killed by Instituteof PlantProtection, Miczurina Str 20, 60-318 P oznan, S. kraussei atthehigh dose of 60000nematodes per pot, Poland whereastreatments with S.carpocapsae causedno signiŽ- ¤[email protected] cantmortality. S. kraussei (L137)was ableto survivewin- Theprocess of selectionwas inducedin a Polishisolate terŽ eldconditions including prolonged exposure to low of Heterorhabditismegidis (01PL). Theobjective of the temperaturesin contrastto S.carpocapsae whichshowed selectionprocedure was toimprove the nematode desic- poorsurvival. These results suggest S. kraussei (L137) has cationtolerance in extreme environmental conditions. A greatpotential as a commercialbiocontrol agent against seriesof basepopulations, 10 000infective juveniles (IJ) overwintering O. sulcatus. each,was exposedto environments with reduced relative humidity(r.h.) levelsin order to establish the threshold 204Effect ofsoiltexture on Steinernemafeltiae of30% survival after 24 h. The experiments were con- (Filipjev)virulence against Spodopteralittoralis ductedin desiccatorswith various saturated salt solutions (Lepidoptera:Noctuidae) at 20 1 C.Theinitial r .h.levelfor theselectionwas setat § ± 60%.After eachround the selected nematodes were repro- Jesús JIMÉNEZ, Raquel CAMPOS and ducedin last instar larvae of Galleriamellonella . Evalua-

194 Nematology Postersessions

1; 1 1 tionof theselection efŽ cacy was basedon the nematode Miklós NÁDASY ¤, Szilvia PEKÁR , Attila LUCSKAI , survivalafter each round. Although the selection is still András FODOR 2, Gyula SÁRINGER 1 and Csaba BUDAI 3 inprogress, overall changes in the population survival at 1Universityof Veszprém, GeorgikonF acultyof Agriculture, lowhumidity levels are observed. This trial was toassess Deák F.u.57, 8360 Keszthely, Hungary thepossibility of permanentenhancement of thenematode 2 EotvosUniversity, Department of Genetics, Pá zmá ny Pé ter desiccationtolerance although the genetic determination sétá ny 1/ C1117Budapest, Hungary ofthis characteristic still remains unknown. It is planned 3 CsongradCountry Plant Health and Soil Conservation tocontinue the research in the future. The survival and Station,Biological Control and Quarantine Development pestcontrol efŽ cacy of theselected nematode population Laboratory,P .O.Boksz 99, 6800 Hó dmezõ v_s_rhely, Hungary whenexposed to open-Ž eld conditions will be tested. ¤[email protected] Entomopathogenicnematode/ bacterium(EPN/ EPB)sym- 206Identifying nematode-inducible genes in bioticcomplexes have been providing a ratherefŽ cient Xenorhabdusnematophila andenvironmentally friendly way of controllingagricul- turalinsect pests. Since the bacterial partner plays the Eric MARTENS and Heidi GOODRICH-BLAIR importantrole of pathogenicity,we havedeveloped sev- eralnew EPN/ EPBcombinationsbetween Heterorhabdi- Universityof Wisconsin-Madison,1550 Linden Drive, tis spp. and Photorhabdus spp.strains at theGenetic De- Madison,WI 53706,USA partmentof theEö tvö s Loránd University,and tested them onseveralagricultural insect pests at the Plant Protection Thebacterium Xenorhabdusnematophila is a mutual- Instituteof theVE GeorgikonFaculty of Agriculture.The istof theentomopathogen icnematode, Steinernemacar- resultsof thepreviousgnotobiologic alanalyses have been pocapsae.Mostof thelife cycle of these two organisms publishedelsewhere. In 1998 we testedsome EPN/ EPB takesplace inside larval-stage insects where the nema- symbioticcomplexes that were appliedin threedoses (1/ 1, todeand bacterium reproduce individually. However, a 1/10,1/ 100infective juveniles (IJ)/ caterpillar)on lastin- non-feedingform ofthenematode, the infective juvenile starwax moth larvae of Galleriamellonella as well as (IJ), residesin thesoil and is thevector for thenematode intwo doses (3000 and 5000 IJ/ whiteinsect) on second germlineand for bacteriabetween insect hosts. Xenorhab- stagewhite grubs of maybeetle( Melolonthamelolontha ) dusnematophila colonisesa specialisedintestinal organ inlaboratoryconditions. It was concludedthat the effec- oftheIJ andcan survive in this location for longerthan tivenessof thenew symbiotic complexes could be com- 5monthsdespite a lackof nutritional uptake by the ne- paredto that of natural EPN strains.In 1999 we tested matode/bacteriacomplex. Factors mediating colonisation theeffectivenessof various Steinernema species (S. anom- ofthe nematode and the metabolic state of thebacterium ali, S. serratum, S. riobrave, S. glaserii, S.carpocapsae duringexistence inside nematodes have not been char- Mex.,T1, T2, T4, S. feltiae Nyíregyhá za., IS6, (Israel), on acterised.W ehavedeveloped a strategyto identify and Germancockroaches ( Blattellagermanica ) and on Peri- characterisebacterial genes that are speciŽ cally expressed planetaamericana .Twotests were applied: i) Ž lter pa- duringcolonisation and growth inside nematodes. The re- permethod:5000 IJ/ 5cockroach/plate; ii)feedingexperi- sultsof thisanalysis will yield insight into those bacter- ment:wax moth killed by nematodeswere givento cock- ialfunctions that are important for establishingand main- roaches.W axmoth larvae killed by H.bacteriophora Bre- taininga mutualisticinteraction with the nematode. W e conand wax moth larvae killed by freezing were used arecurrently examining several nematode-inducib lebac- ascontrols. In conclusion, S.carpocapsae gavepositive terialloci to identify the speciŽ c genesthat are subjected resultsagainst cockroaches and the cockroaches ate the tonematode-speciŽ c expression.Future analysis will in- waxmoth larvae killed by Steinernema butavoided those cludetargeted mutation of nematode-induciblegenesand killed by Heterorhabditis . determinationof theresulting mutant phenotypes with re- gardto nematode-colonisat ion. 208IdentiŽ cation of Steinernema spp. and Heterorhabditis spp. usingscanning electron 207The bioassay of entomopathogenicnematodes microscopyof the labialregion, lateral Ž elds,vulval onagriculturalinsect pests inlaboratory conditions area,spicules and gubernacula

Vol.4(2), 2002 195 Entomopathogenicnematodes (187-221)

Khuong B. NGUYEN¤ andByron J. A DAMS A.aphidimyza populations,but that the presence of A. aphidimyza doesnot interfere with H.bacteriophora host Entomologyand Nematology Department, University of Florida,Gainesville, FL 32611-0620,USA searchingbehaviour. ¤kbn@u.edu Scanningelectron microscopy techniques were used 210The Ž rst reportof geneticdiversity among toidentify morphological characters that are readily Indianpopulations of entomopathogenicnematodes, observedunder light microscopy for theidentiŽ cation of Steinernema spp. and Heterorhabditis spp. species of Steinernema .Headshape and structures in the Uma RAO , K.K. KAUSHAL, R. RENUKA, K. USHA and labialregion of infective juveniles (IJ) canbe used to ¤ A.K. GANGULY dividesteinernematids into four groups: i)headwithout labialpapillae, ii)withsix labial papillae, iii) with two Divisionof Nematology,Indian Agricultural Research Institute, labialhorns, and iv)withswollen head. Formulae based New Delhi110012, India onnumbers of ridges in the lateral Ž eldfrom anterior ¤[email protected] endto tail tip are also informative. Scanning electron Insectpests are one ofthemajor constraints of agricultural microscopyof Heterorhabditis revealsthat the H. megidis productivityin India. T outiliseentomopathogeni c IJhasa membranoustube associated with its dorsal nematodesfor themanagement of regional insects, tooth.The spicules, gubernacula and vulval areas provide Steinernema spp. and Heterorhabditis spp.from different diagnosticcharacters among species of bothgenera. agro-climaticregions of India are being isolated, culturedand conserved in our laboratory. Nineteen EPN 209Non-target mortality by and host searching populationswere assayedfor theirinsect host range and behaviourof Heterorhabiditisbacteriophora in the analysedfor geneticdiversity using RAPD andPCR- presenceof anaphidpredator , Aphidoletesaphidimyza RFLP ofribosomal DNA. Steinernemacarpocapsae, S. glaseri and S.thermophilum –anewspecies recently

Jeff R. POWELL¤ and John M. WEBSTER reportedfrom India– were includedfor comparisonof Steinernema populations.Both and PCR-RFLP of rDNA Departmentof Biological Sciences, Simon Fraser University, couldnot detect any polymorphism among populations of Burnaby,BC, CanadaV5A 1S6 Heterorhabditis spp.Both the proŽ ling systems revealed [email protected] ¤ ahighdegree of polymorphism among Steinernema Aphidoletesaphidimyza occursnaturally in severalNorth populations.RAPD UPGMA analysisclustered the American,European, and Asian Ž eldcrops and is populationsinto two main clusters. PCR-RFLP of rDNA releasedin glasshouses for aphidbiocontrol. Experiments of 13 Steinernema populationsrevealed similarity of only were conductedto determine how it might interact onepopulation to S. glaseri. Theinternal transcriber withan entomopathogeni cnematode, Heterorhabditis spacers1 and2 ofrDNA hasbeen sequenced and aligned. bacteriophora . Fewer A.aphidimyza adultsemerged from Differencesin the sequence alignment and the correlation soilin 60 mmdiameterPetri dishes inoculated with 1000 betweenthe observed genetic diversity revealed by RAPD and 10 000 H.bacteriophora infectivejuveniles (IJ) than andPCR – RFLP analysiswill be discussed. from uninoculatedsoil. No difference in percent adult emergenceof A.aphidimyza was observedwhen IJ were 211Entomopathogenic nematode occurrence in an applied1, 6 or9 daysprior to emergence. Aphidoletes Irishdune system aphidimyza didnot interfere with H.bacteriophora host searchingbehaviour: e.g.,nodifference was observedin Alec ROLSTON 1, Martin DOWNES 2 and Galleriamellonella thenumber of IJ infecting larvae at Christine GRIFFIN 2 thebottom of 276 ml styrofoam cups in the presence 1 orabsence of 100 A.aphidimyza nearthe soil surface. PopulationEcology Laboratory 2 Heterorhabditisbacteriophora was notpreferentially Instituteof Bioengineering and Agroecology, National Universityof Maynooth,Maynooth, Co. Kildare, Ireland attractedto A.aphidimyza ina soil-Žlled 150 mm diameterPetri dish with one side containing one or 30 Entomopathogenicnematodes are obligate parasites of in- A.aphidimyza andthe other containing none. This study sects.The two genera Steinernema and Heterorhabditis suggeststhat H.bacteriophora applicationsmay reduce areubiquitous soil organisms with world-wide distribu-

196 Nematology Postersessions

tions.Insect-killing nematodes are traded globally as bi- NGUYEN 4, Byron J. ADAMS 4,RichardA. H UMBER 5 ologicalcontrol agents. Infective juveniles (IJ) emerge andMichael J. H ALL 6 from theinsect cadaver to search out a newhost. The IJ invadethe host via themouth, anus or spiracles and re- 1USDA-ARS, SEFTRNL, 21Dunbar Rd, Byron, GA 31008,USA 2 leasea symbioticbacteria which kills the insect by septi- Departmentof Entomology, University of Georgia, GrifŽ n, GA caemia.Development into either males or females ( Stei- 30223, USA 3 Departmentof Entomology, Louisiana Agricultural nernema spp.),or ahermaphrodite( Heterorhabditis spp.) ExperimentStation, Louisiana State University Agricultural eventuallyleads to theproduction of furtherinfective ju- Center,BatonRouge, LA 70803,USA veniles.These then emerge from thecadaver into the soil 4 Departmentof Entomologyand Nematology, University of after10-25 days depending on thespecies. Intensive sam- Florida,Gainesville, FL 32611,USA plingof thedune system on thegeologically new Bull Is- 5 USDA-ARS, USPlantand Nutrition Lab, Ithaca, NY 14853, land(Dublin Bay, Republic of Ireland) revealed the co- USA occurrenceof thetwo genera. The genera were intimately 6 LSU AgriculturalCenter ,Shreveport,LA 71135,USA associated,but Heterorhabditis occurredat lowerfrequen- ¤[email protected] cies.Prevalence was lowwith only 4.4% of samples re- vealingnematode presence ( Steinernema 2.5%; Hete- Thepecan weevil, Curculiocaryae (Horn), isa major D rorhabditis 1.9%).Populations were patchyand thedis- pestof pecans in thesoutheastern US. Entomopathogenic tributionof Deachgenus seems to be relatedto stable and nematodesand fungi are potential alternatives to chemi- unstablehabitats. calinsecticides for C. caryae control.Our objectivewas tosurvey pecan orchards in the southeastern US for en- tomopathogenicnematodes and fungi determine the vir- 212Entomopathogenic nematodes in Norwegian ulenceof the new isolates to C. caryae.Soilwas col- forestsoils lectedfrom 105sites in 21 orchardsin Arkansas,Georgia, SolveigHaukeland S ALINAS Louisiana,and Mississippi. Entomopathogen swere iso- latedby exposingsoil to C. caryae andgreater waxmoth larvae, Galleriamellonella L.Weisolatedentomopatho- TheNorwegian Crop Research Institute, Plant Protection genicnematodes from sixof the 21 orchards surveyed, Centre,Hogskoleveien 7, 1430Aas, Norway respectively.Nematodes included Heterorhabditisbacte- [email protected] riophora Poinar, Steinernemacarpocapsae (Weiser), S. glaseri (Steiner),and S. rarum (Doucet).This is theŽ rst Aprojecton the use of entomopathogen icnematodes report of S. rarum inthe US. Inlaboratory assays, vir- (EPN) for controlof the pine weevil ( Hylobiusabietis ) ulenceof 15 nematode isolates to C. caryae larvae was hasbeen initiated. In the Ž rst phasewe areinvestigating testedin small plastic cups containing soil. Results in- thenatural occurrence of EPNinforest soils. Soil samples dicatedpoor susceptibility of the C. caryae larvae to were collectedfrom severalclear cut forest sites in entomopathogenicnematodes. However, high levels of autumn2001. Sampleswere takenaround the rootsof tree nematodevirulence were observedtoward adult wee- stumpsat each site. EPN were isolatedusing the Galleria vils,and several entomopathogeni cfungalisolates caused technique.Preliminary results indicate that several species signiŽcantly higher mortality in C. caryae larvae than in the genus Steinernema arepresent. So far Steinernema otherstrains (including a commercialstrain of Beauveria krauseii hasbeen identiŽ ed which is a newrecord for bassiana). Norway.IdentiŽ cation of thenematodes isolated is being conductedusing morphometric and moleculartechniques. 214Integration of Azadirachtaindica with entomopathogenicnematodes for control of root 213Survey of entomopathogenicnematodes grubs (Holotricaserrata ) endemicto pecan orchards of the southeasternUS andtheir virulence to the pecanweevil (Coleoptera: S. SIVARAMAKRISHNAN , K. MURUGAN and Curculionidae) N. POONKODI

1; 2 David I. SHAPIRO-ILAN ¤, Wayne A. GARDNER , Divisionof Entomology, Department of Zoology, Bharathiar James R. FUXA 3, Bruce W. WOOD 1, Khuong B. University,Coimbatore-641 046, T amilNadu, India

Vol.4(2), 2002 197 Entomopathogenicnematodes (187-221)

Studieswere conductedto explore the integration adverseeffect on free-living nematodes that play a role betweenneem seed kernel extract (NSKE), neemoil innutrientcycling may be abeneŽcial non-target effect of (NO) and Steinernemacarpocapsae (SC) onroot grub entomopathogenicnematodes. Holotricaserrata .Fieldcollected third instar grubs were keptindividually in microcosms Ž lledwith soil. Soil 216Detection of entomopathogenicnematodes and was exposedto various concentrations of NSKE, NO theirpossible use under IPMinPakistan and Steinernemacarpocapsae .Infectivejuveniles were added0 to16 days after application of NSKE and Manzoor H. SOOMRO 1, F. SHAHINA, M.A. MAQBOOL 2 NOrootgrub mortality was notedevery day. After andIftikhar A HMAD 1 combiningand immediately applying the Steinernema carpocapsae, highermortality was obtainedagainst root 1 NationalIPM Programme,National Agricultural Research grub.The survivability and virulence of SCinneem were Centre,Islamabad, P akistan maintainedfor 20days with or without aeration. SC 2 NationalNematological Research Centre, University of survivedwell in the neem concentrations and also the Karachi,Karachi, P akistan virulencewas notaffected by the neem concentrations withaeration and without aeration. Neem enhancesthe InPakistan the studies on biology,taxonomy and efŽ cacy activityof nematodes. Entomophagous nematodes and ofentomopathogeni cnematodes(EPNs) were initiated neemcan represent an eco-friendlystrategy to control root in1996. Since then during extensive surveys, over 1000 grubin agriculture and forest ecosystems. soilsamples were collectedfrom diversehabitats of Pakistanwith emphasis on Sindh Province. Nematodes were collectedusing Galleriamollonella soiltrap method. 215Ecological consequences of inundative Threeisolates of Steinernema and 59 of Heterorhabditis biologicalcontrol: impact of entomopathogenic were detected;of which,22 isolateswere identiŽed as H. nematodeson the nematodecommunity in turfgrass indica. Outof theidentiŽ ed EPNs, twoare new species; onehas been described as Steinernemapakistanense Nethi SOMASEKHAR,ParwinderS. G REWAL, Shahinaet al. 2001, while the other, Steinernema ElizabethA.B. D ENARDO andBenjamin R. S TINNER speciesis in the process of description. Pathogenicity Departmentof Entomology,Ohio State University, OARDC, ofnew indigenous EPN, S.pakistanensis was tested Wooster,OH44691,USA againstseven insect pests of common occurrence and agriculturalimportance under laboratory conditions and Biologicalcontrol is oftentouted as an ecologically safe conŽ rmed on Helicoverpaarmigera, Earias insulana, E. pestmanagement approach. This notion is based on its vittella,P apilliodemoleus, Leucinodes orbonalis, Etiella relativesafety to humans and above ground  oraand zinkenella,Hollotrichia consanguinea .Mortalitywas fauna.However, there is a paucityof data to support higher(up to 100%) on early stages of insect larvae ( H. thatlarge-scale releases of biocontrolagents do nothave armigera)ascomparedto fourthand Ž fthstage larvae. animpact on below ground food webs. W eaddressed Searchfor moreindigenous EPNs iscontinuing and thisissue using entomopathogen icnematodesand native underthe National IPM Programme,Ž eld-testingof nematodecommunity in aturfgrassecosystem as amodel. promisingspecies will be undertaken during the year Applicationof Heterorhabditisbacteriophora strain 2002.The farmers wouldbe trained for conservation GPS11, H.bacteriophora strainHP88, and H. indica ofnatural enemies including EPNs throughthe farmer signiŽcantly reduced the abundance, species richness, Želdschool (FFS) approachparticularly in cotton crop, diversity,and maturity of the nematode community by whichreceives the largest amount (60%) of pesticidesin reducingthe number of genera and abundance of plant- Pakistan. parasiticbut not free-living nematodes. In contrast to theentomopathogeni cnematodetreatments, trichlorfon, 217Heritability of the liquidculture potential of the acommonlyused insecticide in turfgrass, reduced the entomopathogenicnematodes Heterorhabditis numberof genera,abundance and diversity of nematode bacteriophora communityby adversely affecting both plant-parasitic andfree-living nematodes. The reduction in abundance O. STRAUCH, S.-A. JOHNIGK, S. HOLLMER anddiversity of plant-parasitic nematodes without any and R.-U. EHLERS¤

198 Nematology Postersessions

Institutefor Phytopathology, University of Kiel,Department of annulipes),however,made nematode penetration difŽ cult Biotechnology& BiologicalControl, Klausdorfer Str .28-36, orimpossible. For thesusceptible hosts nematodes 24223Raisdorf, Germany signiŽcantly reduced the number of adult insects ¤[email protected] emergingfrom thesoil after pupation. They could also Theinfective stage of Heterorhabditisbacteriophora developand recycle in the insect cadaver .SigniŽcant isthe mobile, but developmentally arrested dauer differenceswere foundbetween the efŽcacy of H. megidis juvenile(DJ). For commercialapplication, nematodes are and S. feltiae strainsand isolates tested against different producedin liquidculture. Prior tothe inoculation of the insectspecies. The results suggest that entomopathogen ic DJ, theirsymbiotic bacterium Photorhabdusluminescens nematodesshould be considered as potential effective iscultured. The DJ exitfrom thearrested stage (recovery) controlagents which could be safely used against many anddevelop to reproductiveadults. Recovery is aresponse treeleaf beetles and saw ies damaging urban parks and tobacterial food signals. In liquid culture the percentage forests. ofDJ recoveringfrom theDJ stageis highly variable, whichsigniŽ cantly in uences the number of reproducing 219Potential of entomopathogenicnematodes for hermaphroditesand the Ž nalDJ yields.The heritability the controlof cabbageroot  y ofthe disposition to recover from theDJ stageand of theŽ nalDJ yieldin liquid culture has been evaluated. Deena M. WILLMOTT¤ andAndrew J. H ART From ahybridstrain 30 homozygous inbred lines were establishedby inbreeding over seven generations. These inbredlines were propagatedin liquid culture several Departmentof EntomologicalSciences, Horticulture Research International,W ellesbourne,W arwick,CV35 9EF ,UK times;the percentage of recovered DJ andthe Ž nalDJ [email protected] yieldswere recorded.The calculated heritability for the ¤ DJrecoverywas low( h2 0:38).No signiŽ cant genetic D variabilitycould be detected for thistrait. In contrast, Thecabbage root  y( Deliaradicum )isthe major pest ahighheritability ( h2 0:90)was foundfor thetotal ofbrassica vegetable crops in the northern temperate numberof DJproducedDin theliquid medium. region;larvae cause extensive damage by feeding on plantroots. Chemical insecticides have traditionally been usedto control this pest. However, with the imminent 218Potential new targetsfor entomopathogenic withdrawalof organophosphorusinsecticides, alternative nematodesin biologicalcontrol of urbantree pests managementstrategies need to be developed which Marek TOMALAK mayinclude the use of entomopathogen icnematodes asbiopesticides. Under glasshouse conditions, the Departmentof BiologicalP estControl and Quarantine, susceptibilityof cabbage root  ylarvaeto ten isolates Instituteof Plant Protection, ul, Miczurina 20, 60-318 P oznan, ofentomopathogen icnematodes was examined,using Poland cauliower ( Brassicaoleracea var. botrytis) as the host [email protected] plant. SteinernemaafŽ ne (isolate179) gave the highest Manyleaf feeding pests of urban trees pupate and/ or levelof control, killing approximately 66% of larvae overwinterin the soil, the most suitable environment within28 days. The least signiŽ cant control was given for entomopathogenicnematodes. During this period by Steinernema E1and F1 (194 and GWE63) andtwo theinsects can be challenged with Heterorhabditis- or Heterorhabditis isolates(K122 and UK211) which killed Steinernema- basedbioinsecticides. Susceptibility of the approximately30% of larvae in 28 days. A second mostimportant urban tree leaf beetles and saw ies to experimentcompared the performance of S. afŽ ne and S. feltiae and H. megidis was examinedin laboratory Nemasys® (MicrobioLtd), a commercialformulation andŽ eldconditions. Last instar larvae and pupae of all of S. feltiae,overa rangeof doses (from 8000to testedleaf beetles ( Alticaquercetorum , Agelasticaalni , 64000nematodes per pot). SteinernemaafŽ ne controlled Phytodectaquinquepunctat a and Phyllodectalaticolis ) signiŽcantly more of cabbage root  y(54%)than andsome saw ies ( Acantholydaerythrocephala, Arge Nemasys® (36%)and may have potential for commercial berberidis,A. pagana )provedto be highly susceptible development. tonematode infection. A tightpupal cocoon of the remainingsaw y species( Pristiphoraabietina, Caliroa

Vol.4(2), 2002 199 Entomopathogenicnematodes (187-221)

220On the entomopathogenicnematodes belonging 221Growth andreproduction of Heterorhabditis to genera Steinernema and Heterorhabditis from the bacteriophora on Xenorhabduspoinarii RS92 Russian FarEast and comparison with entomopathogenicnematode fauna in Japan Toyoshi YOSHIGA¤ andRalf-Udo E HLERS Institutefor Phytopathology, Christian-Albrechts-University Mutsuhiro YOSHIDA Kiel,Klausdorfer Str .28-36,24223 Raisdorf, Germany NationalInstitute for Agro-Environmental Sciences, 3-1-3 ¤[email protected] KannondaiTsukuba, Ibaraki 305-8604, Japan Heterorhabditisbacteriophora isan entomopathogen ic [email protected] nematodewhich has a speciŽc symbioticrelation to Entomopathogenicnematodeswere collectedin southern thebacterium Photorabdusluminescens .Tounderstand Primoryeand southern Sakhalin, grant-aided by Gene thesymbiotic effect of Photorhabdus and Xenorhabdus BankProject of Ministry of Agriculture, Forestry and on H.bacteriophora ,thebacteria were examinedfor Fisheriesand supportedby theInstituteof MarineBiology theirpotential to support nematode growth. Axenic 1st andthe Institute of Parasitologyof theRussian Academy stagelarvae of H.bacteriophora obtainedby surface ofSciences. A totalof 197 soil samples from 34sites sterilisationof eggs that were inoculatedon lipid agar were testedfor thepresence of the nematodes using the platesalong with 24 hbacterialcultures. Heterorhabditis Galleria baitingtechnique. Forty-two nematode isolates bacteriophora grew andreproduced well not only on were detectedfrom 14sites.The nematodes were divided Photorhabdus bacteriaisolated from H.bacteriophora intosix RFLP types.Three of them were identiŽed and H. megidis but also on X. poinarii RS92 from as S. kraussei, S. feltiae and Heterorhabditismegidis Steinernemaglaseri .Nematodegrowth was notsupported andanother three were tentativelydesignated as S. spp. bythe symbiont of H. indica and by other Xenorhabdus primorye1,sakhalin1, sakhalin2 . Steinernemakraussei spp.tested. The hermaphrodites of H.bacteriphora grown was thedominant species (nine/ 14sites)in thissurvey and on X. poinarii RS92were signiŽcantly smaller than was isolatedfrom coaststo mountains. However, it has thosegrown on their own symbiont. Dauer juveniles beendetected only from twocoastal sites in Hokkaido, obtainedfrom the X. poinarii RS92culture did not retain theclosely related region with the Russian Far East. thebacteria in their intestine nor recovered from the Steinernemafeltiae was detectedonly from acoastal dauerstage in a fresh X. ponarii RS92culture. Also, grasslandin Sakhalin and also mainly from coastalsites H. megidis and H. indica as well as S.carpocapsae inHokkaido. Heterorhabditismegidis was isolatedfrom grew andreproduced on X. poinarii RS92.These results bothVladivostok and Sakhalin, but has not been isolated indicatethat X. poinariiRS92 is a Xenorhabdus bacterium from Hokkaido. Steinernema sp. sakhalin2 corresponded thatsupports the growth of both heterorhabditid and to S.sp.MY7 from Japanand S. sp. primorye1 was steinernematidnematodes. recentlyrecognised among the isolates detected from Hokkaido.

200 Nematology Postersessions

222Different developmentalresponses of virulent ¤[email protected] andavirulent isolates of Bursaphelenchusxylophilus to Monochamusalternatus Thedirect effects of root diffusates and phytohormones (auxinand kinetin) on the surface lipophilicity of the 1; 2 Takuya AIKAWA ¤, Katsumi TOGASHI and plant-parasiticnematodes Globoderarostochiensis and Hajime KOSAKA 1 Meloidogyneincognita were investigated.The uorescent lipidprobe AF18 (5-N-(octodecanoyl) amino uorescein) 1 Forestryand F orestProducts Research Institute, Tsukuba, was usedto detect surface changes. Root diffusates Ibaraki305-8687, Japan increasedAF18 uptake by G.rostochiensis while it had 2 Facultyof IntegratedArts and Sciences, Hiroshima M.incognita University,Higashi-Hiroshima, Hiroshima 739-8521, Japan no effect on .Kinetinand auxin decreased AF18uptake in G.rostochiensis ,whilethey had an ¤[email protected] oppositeeffect on M.incognita .Auxin/kinetinratio was Itis known that there is a greatvariation in virulence of alsofound to be important in triggering the surface Bursaphelenchusxylophilus, whichis thecausative agent changes,especially at highconcentrations. Whether plant ofpine wilt disease and is transmitted from wilt-killed nematodeshave auxin and/ orkinetinbinding proteins is tohealthy pine trees by the insect vector, Monochamus discussedas well as the mechanism behind the surface alternatus.Tocomparethe developmental responses to lipophilicitychanges. The implication of thesechanges in M.alternatus betweenthe two nematode isolates of theestablishment of infectionis also discussed. differentvirulence, the production of dispersalforms and thenumber of nematodes carried by newly-emerged M. 224Effect ofpopulation densities of Heterodera alternatus adult(initial nematode load) were investigated glycines race3 onleafarea, photosynthesis and yield usingpine bolts. The numbers of total nematodes, of soybean dispersaljuveniles (third-stage dispersal juvenile and 1; fourth-stagedispersal juvenile) and fourth-stage dispersal GuilhermeLafourcade A SMUS ¤ and juvenileswere muchsmaller in the avirulent isolate LuizCarlos C.B. F ERRAZ 2 thanin the virulent one around pupal chambers. Theproportions of dispersal juveniles produced in a 1EMBRAPA-CPAO, P.O.Box 661, 79804-970, Dourados, MS, populationand the proportion of third-stage dispersal Brazil 2 juvenilesdeveloping to fourth-stage dispersal juveniles ESALQ/USP –ZoologyDepartment, P .O.Box9, 13418-900, were alsosmaller in the avirulent isolate. Although the Piracicaba,SP ,Brazil [email protected] proportionof fourth-stage dispersal juveniles boarding ¤ beetleswas equalbetween the two isolates, the number of Theeffect of Heteroderaglycines race3 onphotosynthe- themwas muchsmaller in the avirulent isolate. A small sis,leaf area, and yield of susceptible soybean ‘ Embrapa initialnematode load of theavirulent isolate was ascribed 133’was studiedin two experiments carried out under toa smallerrate of reproduction and lower productive glasshouseconditions. Soybean seeds were sownin 1.5 ratesof third-and fourth-stage dispersal juveniles for the l(Experiment1) or5.0 l (Experiment2) claypots Ž lled avirulentisolate. witha sterilisedmixture of Želdsoil sand(1:1). Eight C daysafter sowing, seedlings were thinnedto oneper pot, 223Effects ofphytohormonesand root diffusates on and1 daylater inoculated with 0, 1200, 3600, 10 800, the surfacesof plant-parasiticnematodes 32400,or 97 200second stage juveniles ( Pi). Exper- D iment1 was continueduntil 45 days after inoculation 1; 1 Abdellah AKHKHA ¤, John KUSEL , whereasexperiment 2 extendeduntil harvest time. Data Malcolm KENNEDY 2 and Rosane CURTIS 3 onplantphysiological processes were takenat 10-dayin- 1DavidsonBuilding, Division of Biochemistry& Molecular tervalsduring the experiments. Data on fresh rootweight, Biology,University of Glasgow ,GlasgowG12 8QQ, UK topdry weight, grain yield, and nematode reproduction 2 GrahamKerr Building, Division of Environmental and factorwere obtainedat theend of thetrials. For bothex- EvolutionaryBiology, University of Glasgow ,Glasgow, periments,a statisticaldesign of randomisedblocks with G12 8QQ, UK sixtreatments ( Pi values)was establishedand each exper- 3 NematodeInteractions Unit, IACR-Rothamsted, Harpenden, imentalunit (one plant growing per pot) was replicated HertsAL5 2JQ,UK tentimes. There was amarkedreduction in both photo-

Vol.4(2), 2002 201 Physiology(222-243) syntheticrate and chlorophyll content, as wellas evident 1Instituteof Bioengineering and Agroecology, National yellowingof the leaves of theinfected plants. Even at the Universityof IrelandMaynooth, Maynooth Co. Kildare, Ireland lowest Pi,theeffect of H. glycines ontop dry weight or 2 Instituteof Biotechnology,University of Cambridge, T ennis grainyield were quitesevere. Also, soybean yield was CourtRoad, Cambridge CB2 1QT,UK highlycorrelated with the integrated leaf area. ¤[email protected] Whensubjected to desiccationsome nematodes can enter 225Leaf area, photosynthesis and yield of soybean astateof suspendedanimation known as anhydrobiosis. asaffected byinitial population densities of Anhydrobioticnematodes synthesise high concentrations Meloidogynejavanica ofthenon-reducing sugar trehalose during the induction ofanhydrobiosis. Trehalose is believed to protect mem- 1; GuilhermeLafourcade A SMUS ¤ and branesand proteins from desiccationdamage by replac- LuizCarlos C.B. F ERRAZ 2 ingstructural water, and to contributeto the formation of anintracellularorganic glass, which is thought to stabilise 1 EMBRAPA-CPAO, P.O.Box 661, 79804-970, Dourados, MS, thecell contents. In anhydrobiotic plants, sucrose plays a Brazil similarrole. However, several lines of evidence indicate 2 ESALQ/USP–ZoologyDepartment, P .O.Box 9, 13418-900, thatnon-reducing sugars alone are not sufŽ cient to con- Piracicaba,SP ,Brazil fer anhydrobiosis,and that additional adaptations are re- ¤[email protected] quired.T oidentifythese additional adaptations, we have Theeffect of Meloidogynejavanica parasitismon theleaf investigatedgene expression in Aphelenchusavenae un- area,photosynthesis, and yield of twoBrazilian soybean dergoinganhydrobiosis. A numberof genes were found varieties,Embrapa 133 (susceptible) and Coodetec 201 tobe upregulated in nematodes exposed to 90% relative (tolerant),were studiedunder glasshouse conditions. T wo humidityfor 24h.Of particularinterest was astronglyin- similarexperiments were setup, one continued until 48 ducedtranscript whose sequence suggests that it encodes daysafter plant inoculation, and the other until plant amemberof thegroup 3 subclassof LEA(LateEmbryo- harvest.The plantlets were individuallyinoculated with genesisAbundant) proteins. LEA proteinsaccumulate in 0,1200, 3600, 10 800,32 400or 97 200second stage responseto waterdeŽ cit in many plants but are especially juveniles ( Pi).Dataon physiological processes of abundantin anhydrobiotic plants and in maturing seeds D theplants were takenat 10-day intervals throughout the andpollen.W ewillpresent our dataon themolecular phy- experiments,whereas data on fresh rootweight, top dry logenyof the A. avenae LEA3gene and on itsinduction weight,grain yield and nematode reproduction factor in A. avenae byvariousenvironmental stresses. were obtainedat the completion of thetrials. A statistical designof randomised blocks with treatments arranged 227Dual modes of reproductionin Caenorhabditis accordingto a2 6factorialmodel (varieties Pi) was £ £ elegans establishedand each experimental unit (one plant growing perpot) was replicatedten times. V aluesof photosynthesis J. CHEN and E.P. CASWELL-CHEN rate,stomatic conductance, chlorophyll  uorescence,leaf colourand chlorophyll content were slightlyaffected by Departmentof Nematology,University of California, Davis, CA M. javanica for bothtested varieties. Conversely, soybean 95616, USA leafarea, fresh rootweight, top dry weight, and grain yield Caenorhabditiselegans isa modelsystem for studying were markedlyreduced by thenematode at thehighest Pi developmentand reproduction. The relative investment values.Also, soybean yield was highlycorrelated with the ofenergy in growth, maintenance, or reproduction is a integratedleaf area. classicquestion in the evolutionof reproductivestrategies. Here we reportthat when C. elegans isstarved, it altersits 226Characterisation of amemberof the LEAgene modeof reproduction from oviparyto vivipary wherein familyfrom the anhydrobioticnematode Aphelenchus larvaedevelop within the parent body before live birth. avenae Facultativevivipary was observedin worms placedin water,on culture plates and in soil microcosms, was 1; 1 John A. BROWNE ¤,KatharineM. D OLAN , inducedby starving late L4s, young adults, and gravid Alan TUNNACLIFFE 2 and Ann M. BURNELL 1 adults,and was reversibleby restoring food. Without

202 Nematology Postersessions food,vivipary resulted in the death of parent worms 229The kinase cascade possibly involved in becausetheir progeny appeared to consume the parent the odorantsignaling of Meloidogyneartiellia bodycontents. The larvae that resulted from vivipary 1; 2 developedto reach a normallifespan and had normal egg MariaRosaria C ORTESE ¤, Mauro DI VITO , 1 1 production,as compared to larvae arising from ovipary. Elena FANELLI and Carla DE GIORGI Larvaeproduced by viviparywere ableto enter the dauer 1 Universitàdegli Studi di Bari, via Orabona n ±4,70126 Bari, stageunder crowding and limited food but, in contrast Italy tolarvae arising from ovipary,some larvae produced by 2 Istitutodi Protezione delle Piante CNR, viaAmendola 165/ A, viviparywere ableto enter the dauerstage under complete 70126Bari, Italy starvation.W einferthat the combination of vivipary, ¤[email protected] androdioecy,and the dauer stage represent life-history adaptationsthat ensure survival of dispersal propagules Theplant parasitic nematode Meloidogyneartiellia underextreme food limitation. causessigniŽ cant economic losses worldwide by at- tackingthe roots of several cultivated crops. Although verylittle is known about the host-parasite interactions, 228Inhibition of sterolmetabolism in chemosensationprocesses might play a keyrole in the Caenorhabditiselegans by AY-9944 recognitionof the plant host thus involving specialised signaltransduction pathways. W eareinvestigating some David J. CHITWOOD aspectsof the mechanism of signal transduction using NematologyLaboratory, ARS, U.S.Department of Agriculture, molecularand biochemical approaches in order to elu- Bldg.011A, room 165B, BARC-W est,Beltsville, MD 20705, cidateroles of speciŽc proteinsin the signal cascade. In USA particular,we areinterested in RAS-Mapk pathway that [email protected] was demonstratedto be involved in olfaction in C. ele- Caenorhabditiselegans andsome other nematodes gans.Wehaveisolated and characterised in M.artiellia arecapable of attaching a methylgroup to the thecorresponding genes of let60, lin45, mek2 and mpk1 nucleusof sterols at the C-4 position. In C. elegans, thatconstitute this pathway in C. elegans.Thecompari- 4-methylcholest-8(14)-enolis the most abundant 4- sonof thesequences of thesefour genes from C. elegans methylsterolproduced, and smaller quantities of 4- and M.artiellia andkinase assays with GST -recombinant methylcholest-7-enolalsooccur .Thepurpose of this proteins mek2 and mpk1 oftheparasite conŽ rm therole studywas totreat C. elegans witha knowninhibitor oftheisolated genes in theRAS-Mapk pathways. The ap- ofsteroid C-7 double bond isomerisation in order to proachof aphosphorylationscreening of acDNA library morefully understand the pathway of 4-methylsterol of M.artiellia willallow the isolation of all the possi- formation.Thus, C. elegans was propagatedin sterile blesubstrate proteins of thelast member of thiscascade semi-deŽned medium with or without 50 mg/ lAY- mpk1. 9944.The media were supplementedwith sitosterol as a dietarysterol. Lipids were extractedfrom thenematodes 230Suitability of amathematicalmodel to describe andsaponiŽ ed; sterols were isolatedwith column the effect oftemperaturesand exposure times on chromatographyand identiŽ ed withgas chromatography. the survivalof the sugar-beetnematode Heterodera Thesterols of inhibitor-untreated nematodes contained schachtii 91.4%4-desmethylsterol plus 8.6% 4-methylsterol; the inhibitor-treatednematode sterol contained only 4.6% Trifone D’ADDABBO¤, Nicola SASANELLI, Nicola 4-methylsterol.The 4-methylcholest-8(1 4)-enol:lophenol GRECO, Vincenzo STEA andAntonio B RANDONISIO ratioin control nematodes was 9.75but only 0.71 Istitutoper la Protezione delle Piante, Sezione di Bari, C.N.R., intreated nematodes. The results are consistent with ViaG.Amendola165/ A,70126Bari, Italy apathwayinvolving the methylation of cholest-7-enol ¤[email protected] toform lophenolas an intermediate in the production ofthe Ž nalproduct of the methylation pathway, 4- Theeffect of combinationsof temperaturesand exposure methylcholest-8(14)-enol. timeson thesurvival of Heteroderaschachtii was assessed inexperiments under laboratory conditions. In the Ž rst experimentcysts in water were exposedat 25, 35, 37.5,

Vol.4(2), 2002 203 Physiology(222-243)

40,42.5, 45, 47.5, 50 or 52.5 ±Cfor amaximumof 2 h. 232Detection of changesoccurring during recovery Inthe second experiment cysts in naturally infested soil fromthe dauerstage in Heterorhabditisbacteriophora were exposedat 32.5, 35, 37.5, 40, 42.5, or 45 ±C for 1; 2 aminimumof 2 toa maximumof 2048 h. V iabilityof KatharineM. D OLAN ¤, John T. JONES and 1 eggsin cysts was assessedby hatching tests in a 3mM Ann M. BURNELL zincchloride solution. Viability in water was suppressed 1Instituteof Bioengineering and Agroecology and Department after2 hexposureat 50 ±Candinhibited after 1-2 h ofBiology,National University of IrelandMaynooth, at 52.5±C.Emergenceof juveniles from cystsin soil Maynooth,Co Kildare, Ireland increasedafter treatments at rather low temperature x 2 MBN Unit,Scottish Crop Research Institute, Invergowrie, exposuretime combinations and was suppressedat high Dundee,DD2 5DA,Scotland, UK combinations.Egg mortality started after exposures of ¤[email protected] 256 h at 40 C,32h at42.5 and 16 h at45 C. Eighty ± ± Whenconditions are unfavourable for reproductionin one,31 and 7 hofexposure were necessaryto kill 50% Heterorhabditisbacteriophora ,alonglived, non-feeding, ofthe nematode egg population at 40, 42.5 and 45 C, ± survivaland dispersal stage, the dauer juvenile (DJ), is respectively.Data Ž ttedthe models P P 10 t=.q mT / t D 0 ¡ C formed.This DJ stageis also adapted for hostŽ nding and P P 10 1T =.z pt/ for m 0:0111, q T 1 D T 0 ¡ ¡ D ¡ D andinfection. When it infects a suitablehost, the DJ 0:8238, z 2:444 and p 0:236. D D ¡ recoversand resumes growth and development. W e describea seriesof methods for improveddetection of 231Interaction of root-knotnematode infection and recovery in H.bacteriophora. Wealsodescribe some droughtstress incotton ofthe physiological changes that occur immediately afterthe onset of recovery in these nematodes as 1; 2 R.F. DAVIS ¤ and H.J. EARL revealedusing  uorescentnucleic acid binding SYTO 1USDA-ARS, P.O.Box 748, Tifton, GA 31793,USA dyes.Although recovery could be monitored using 2 Universityof Georgia, Dept of Crop and Soil Sciences, 4107 morphologicalchanges, we foundthat observation of the MillerPlant Sciences, Athens, GA 30602,USA uptakeof  uorescentlatex microspheres by recovering ¤[email protected] nematodeswas afar moresensitive and efŽ cient means Dripirrigation was usedin a Želdwith a loamysand ofdetectingrecovery. SYTO dyeswere alsofound to be naturallyinfested with Meloidogyneincognita to create usefulindicators of recovery, binding to the pharyngeal low,medium and high levels of drought stress, though glandsand genital primordia as little as 3 hafterthe rainfallminimised drought stress early in the season. onsetof recovery. The use of SYTO dyesalso indicates Fumigationwith 1,3-dichloroprope ne(0or 56 l/ ha)was thatthe pharyngeal glands produce large quantities of usedto create a rangeof nematodedensities. Three metres RNA followingthe onset of recovery,implying that these ofeach plot were handharvested and ginned to provide structuresmay produce proteins important in theinfection lintand seed yield. A subsampleof lint was analysed and/orfeedingprocess of H.bacteriophora . for Žbrequality, including Ž brelength and micronaire. Covarianceanalysis was usedto determine within each 233Isolation and characterisation of genes induced levelof droughtstress the relationship between nematode bydesiccation stress inthe insect-killingnematode densitiesat mid-seasonand seed weight, lint weight, Ž bre Steinernemafeltiae length,and micronaire at harvest.For alllevels of drought stress,seed weight ( P 0:03),lint weight ( P 0:05), Tali Z. GAL , Hinanit KOLTAI and Itamar GLAZER D D ¤ andŽ brelength ( P 0:07)decreased and micronaire Departmentof Nematology,ARO, TheV olcaniCenter ,Bet (P 0:02)increasedDas nematode populations increased. D Dagan,50250, Israel For seedweight and micronaire, the relationships with [email protected] nematodedensities were similar(alpha 0.10) among ¤ D thethree levels of drought stress (similar slopes and Theinsect-killing nematodes of the Steinernematidae intercepts),but only slope was similarfor length.The infectand kill a widerange of insect species and are decreasein lint weight with increasing nematode densities consideredamong the most promising alternatives to was greater( P 0:07)at thehigh level of droughtstress chemicalcontrol of insect pests. Steinernemafeltiae D thanat thelower levels of stress. canpartially tolerate desiccation by entering a shallow

204 Nematology Postersessions dormancyand thus may serve as a modelto study 18days the cysts were subjectedto the action of potato desiccationstress tolerance for steinernematidnematodes. rootleachates. In those cysts with an n-hexane fraction UsingcDNA subtractivehybridisation we identiŽed IS6 pre-treatmentno hatchingof J2was observed.This could genesthat are differentially expressed during exposure indicatethe presence of HIinpotatopeel. todesiccation stress. One hundred and ten genes were identiŽed, among them Late-Embryogenic-Abun dant 235Effect ofheavymetals (zinc and lead) on soil gene (Sf-LEA)andaldehyde dehydrogenase ( Sf -ALDH), nematodefauna bothknown to beinvolvedin response to water stress in otherorganisms. Furthermore, using real-time PCR, we Ludmila GRUZDEVA¤,ElizavetaM ATVEEVA and detecteda signiŽcant increment in the steady state level Tatiana KOVALENKO ofthegene transcription products upon 8 hofnematode exposureto desiccation, and further increase upon 24 h Instituteof Biology,Karelian Research Centre RAS, ofdesiccation. Future studies of desiccation tolerance, Pushkinskayast, 11, P etrozavodsk,185610, Karelia, Russia includingidentiŽ cation of additional desiccation-related ¤[email protected] genesand study of their biological roles and regulation, willshed light on thegenetic and biochemical alterations Changesof nematode fauna in soil polluted with zinc evolvedin desiccationtolerant organisms. (ZnSO4 –2500mg/ kg)and lead (PbNO 3 – 500 mg/kg ofsoil) were investigated.Faunistic diversity, nematode populationdensity, ecotrophic structure and maturity in- 234Are therehatching inhibitor factors of dexof nematode communities were analysed.Soil sam- Globoderapallida inpotatotubers? pleswere collectedmonthly in 2000 (April – Septem- 1; 1 ber) andtwice in 2001 (May and September). Simulta- Avelino GARCÍA-ALVAREZ ¤, María ARIAS , Antonio BELLO 1,JavierGutié rrez L UIS 2 and neously,an experiment on the growth of nematodes of Alejandra LÁZZARO 2 genus Cephalobus onmedium with addition of Zn and Pbin increasing concentrations under laboratory condi- 1 Dptode Agroecologí a, CCMA, CSIC,Serrano 115 dpdo, tionswas carriedout. Both salts were toxicfor nema- 28006Madrid, Spain todes.They caused a decreasein population density and 2 InstitutoUniversitario de Bio-Orgá nica, Universidad de La diversity.Trophic structure of thenematode communities Laguna,Avda Astrofí sico Sá nchez 2, 38206,La Laguna, was changed,immediately after heavy metal application. Tenerife,Spain Thenumber of representativesof Rhabditidaesharply in- ¤[email protected] creased(up to 78%, especially in the variant with Zn); Co-evolutionof potato plant Solanumtuberosum and laterthe number of polyphagous and predaceous nema- potatocyst nematode (PCN) Globoderapallida for todesdecreased. Under Zn application these groups dis- millionsof yearshas resulted in a closedinterrelationship appearedtotally from thenematode fauna. Maturity index inwhich the nematode biological cycle depends on root ofthenematode communities ( 6MI)was lowerin vari- diffusatesthat stimulate the hatching process of second antswith heavy metals (2.1-2.6) compared to thecontrol stagejuveniles (J2). Hatchingfactors (HF) arepresent not (2.7-2.8).In thelaboratory experiment, the toxic effect of onlyin roots but also in leaves and other parts of the Znand Pb was expressedin a decreasein fecundity of potatoplant, except for tubers.However, the existence nematodesand size of individualsin thepopulation. Neg- ofchemicals which inhibit the HF iswell-known, and ativeeffects were intensiŽed with increased heavy metal thesehave been found in several non-host species for concentration. PCN, suchas white mustard, asparagus or Blupeurum salicifolium .Recentlyhatching inhibitors (HI) havebeen 236Presence of Meloidogyne sp. males in identiŽed in potato root leachates, though information thickeningsof the end ofsoybeanlateral roots doesnot exist about their presence in tubers. The potato 1; 1 industryproduces thousands of tonsof potatotuber wastes Eugenia LORENZO ¤,MarceloE. D OUCET everyyear and this material could be usedas apotential and Susana SUAREZ 2 sourceof HI. Bioassayswith cysts of G. pallida have been carriedout using different fractions of peeltuber extracts 1Orientación Morfología Vegetal,F acultadde Ciencias inn-hexane, dichloromethane and Eethyl acetate. After Exactas,Fí sico-Quí micas y Naturales,Universidad Nacional

Vol.4(2), 2002 205 Physiology(222-243)

deRí o Cuarto,Agencia P ostalN ±3,5800 Rí o Cuarto,Provincia concentration;higher concentrations, if not phytotoxic, deCó rdoba, Argentina areineffective in controllingthe parasites. 2 Orientación Morfología Vegetal,F acultadde Ciencias Exactas,Fí sico-Quí micas y Naturales,Universidad Nacional 238Olfaction and odour discrimination in the insect deRí o Cuarto,Agencia P ostalN ±3,5800 Rí o Cuarto,Provincia deCó rdoba, Argentina parasiticnematode Heterorhabditisbacteriophora ¤[email protected] DamienM. O’H ALLORAN¤ and Ann M. BURNELL Soybeanplants ( Glycine max L.Merr.A 5435variety) Instituteof Bioengineeringand Agroecology and Department of from Berrotarán, province of Córdoba, Argentina, showed Biology,National University of IrelandMaynooth, Maynooth, highlygalled radical systems. Females of Meloidogyne Co.Kildare, Ireland sp.were extractedfrom insideprominent galls located on ¤[email protected] principaland lateral roots. Several lateral roots showed particularlythickened ends. Dissection of thelateral roots Thesurvival of any organism depends greatly on its revealedthe presence of several males inside them. The abilityto monitor the external environment. Organisms analysisof histologicalsections showed giant cells around havedeveloped a widerange of sensory systems to them.Thickenings at the end of soybean lateral roots detect,transduce and evaluate relevant environmental representanother element of analysisto detectnematodes signals.Olfaction is aprimaryneurosensory function and ofthe genera Meloidogyne. itremains one of the major senses by which organisms assesstheir environs. Heterorhabditisbacteriophora lives atasoil-waterinterface and so encounters many types of 237Effect ofplant defence elicitorsand cations on volatileand water-soluble molecules in its environment. reproductionof RKN ontomato Weshowthat its complex chemical vocabulary enables H.bacteriophora tobind and identify many classes S. MOLINARI ofodorant molecules. These include many host-related Istitutodi Protezione delle Piante – CNR, ViaG. Amendola chemicals,including carbon dioxide, as well as more 165/A,70126Bari, Italy complexorganic cues such as long chain alcohols. W e [email protected] show that H.bacteriophora detectsnumerous volatile and water-solublechemicals, which can be either attractant Anumberof plant defence elicitors, such as salicylic orrepellent depending on their concentration. Using acid(SA), acetylsalicylicacid (AcSA), jasmonicacid (JA) behaviouralassays, we haveidentiŽ ed several classes of andisonicotinic acid (INA), hasbeen used to test their receptorsand have investigated their afŽ nity for host- effecton root-knot nematode reproduction on tomato. relatedcues. Furthermore,tomato response to RKNwas assayedafter incubationof rootsin concentratedsolutions of K C, NaC, 239Video-assisted analysis of anhydrobioticprocess 3 2 Fe C and the Ca C chelatorEDT A.Inparticular, roots in Aphelenchusavenae onsucrose-amendedagar ofyoungseedlings of susceptibletomato were immersed substrates overnightin appropriatesolutions of SA, AcSA, JA, INA,

KH2PO4, Na2HPO4, EDTA, FeCl3 andwater as a control, Rhyosuke OHTSUBO,NobuyoshiI SHIBASHI¤ washedand immediately inoculated with over 100 and Eizo KONDO Meloidogyneincognita activeJ2 per seedlings.Inoculated tomatoplants were grownin a glasshouseat 25-27 C for SagaUniversity, Saga 849-8502, Japan ± [email protected] 6weeks.Reproduction index was determinedas number ¤ ofegg masses per root system and compared to untreated Thefungivorous nematode, Aphelenchusavenae , comes control.Previous treatment of tomatoseedlings with INA, intoanhydrobiosis under slowly drying conditions, NaC andEDT Amarkedlyreduced RKN reproduction however,the process leading to the anhydrobiotic state ontomato. The effect of different concentrations of hasnot yet been fully understood. The present study chemicalshas been also evaluated. Generally, the effect on individuallyexamined the process leading to coiling of RKN reproductionof thesingle chemical does not exceed thenematodes on agar plates with added sucrose and a50%decrease compared with untreated controls. The theirsurvival after desiccation treatment. Fourth stage maximuminhibitory effect is associated with a threshold juvenilesor young adults of A. avenae individually

206 Nematology Postersessions placedon 5% agar substrate containing 0.8 M sucrose 241V ariationin the assimilationand transpiration showedthe highest proportion of nematodesin the coiling ratein nine accessions of Passi ora inoculatedwith posture.Video-assisted image analysis indicated that the Rotylenchulusreniformis nematodesbecame quiescent within 2-3 h afterbeing 1; 1 placedon theagar plate, then gradually resumed mobility Zoraida H. SUÁREZ ¤,LigiaCarolina R OSALES , 2 2 andbecame coiled. Coiled or uncoiled nematodes were Mercedes AZKUE andMarí a AlejandraG ÓMEZ individuallytransferred, without destroying their shape, 1InstitutoNacional de InvestigacionesAgrí colas, CENIAP , toa desiccator( <20%r.h.) and exposed for 24h. Protección Vegetal Desiccationsurvival was positivelyrelated to the pre- 2 Agrometereología, Apdo 4653, Maracay 2101, Aragua, exposuretreatment of nematodeson agar .Survivalrates Venezuela were 0,3, 70, and 90% of nematodes pre-exposed ¤[email protected] for 0,2, 5, and 12 h onthe agar plate, respectively. Seekingresistance to Rotylenchulusreniformis, nine Allnematodes pre-exposed for 24h ormore revived accessions(236-2, 221, 214-1, 247-2, 241, 232-1, 219- immediatelyafter rehydration following the desiccation 1, 219-2) of Passiora edulis f.  avicarpa and 231 treatment,irrespective of whether their postures either of P.quadrangularis from thegermplasm collection of remainedcoiled or were un-coiledartiŽ cially. CENIAP were evaluated.Only accession 219-1 showed asusceptibilityreaction. An experiment to study the 240Characterisation of feeding and digestion in variationin the assimilation and transpiration rates Meloidogyneincognita betweencontrol and R.reniformis inoculatedplants was carriedout. In order to estimate the assimilation and Jane SHINGLES¤,CatherineJ. L ILLEY and transpirationrate, four measurements were madeon each Howard J. ATKINSON materialwith an infrared gas analyser (IRGA). Results demonstratedthat all accessions showed a different BiologyDepartment, University of Leeds, Leeds, LS2 9JT ,UK behaviourin the assimilation and transpiration rate ¤[email protected] betweencontrol and inoculated plants, except accessions 247-2and 231 where plants in bothtreatments exhibited Thedigestive proteinases of plant-parasitic nematodes similarresponses. Preliminary results indicate that even havebeen the target of asuccessfulproteinase inhibitor- thoughthere was aresistancereaction in eight of the basedtransgenic defence. Characterisation of the target materials,they behave physiologically differently in the enzymesof cystnematodes led to the isolation of genes presenceof thenematode. It is necessary to continue this encodingcysteine, serine and aspartic proteinases from typeof studyfor alongerperiod of timeand withdifferent femalecDNA libraries.T oextendthe work to the root- levelsof inoculum. knotnematode Meloidogyneincognita ,acDNA library was constructedfrom feedingfemale nematodes. cDNA fragmentscorresponding to proteinases were ampliŽed 242Effects ofDL-methionineon hatchingand byPCRusingdegenerate primers and were subsequently survival of Meloidogyneincognita usedto screen the library to obtain full-length clones. Miguel TALAVERA1; andT akayukiM IZUKUBO 2 cDNA clonesencoding three cysteine, two serine and ¤ oneaspartic proteinase have been identiŽ ed, some of 1Departamentode Biodiversidady Biología Evolutiva,Museo whichare predicted to be extracellular. The expression Nacionalde Ciencias Naturaes, CSIC, C/ .JoséGutié rrez Abascal,2, 28006 Madrid, Spain patternsof the genes have been characterised by 2 northernanalysis and in situ localisation,showing varying Departmentof Entomologyand Nematology, National AgriculturalResearch Center ,3-3-1Kannondai, Tsukuba, expressionpatterns at different stages of development. Ibaraki,305-8666 Japan Acysteineproteinase inhibitor expressed transgenically ¤[email protected] in Arabidopsis andpotato has been shown to have a detrimentaleffect on M.incognita .Theeffect of serine Severalconcentrations of DL-methionine in vitro and proteinaseinhibitors and inhibitor combinations is now appliedto soil were testedin the laboratory for their beingtested. effects on Meloidogyneincognita egghatching and juvenilesurvival. Solutions of 2 :5 102 mgperml reduced £ egghatching in vitro after2 daysof treatment. After 7

Vol.4(2), 2002 207 Physiology(222-243) daysin methioninesolutions, proportion of hatchedeggs Plantsecondary metabolism is responsiblefor theproduc- were reducedby 23.3%in 2 :5 10 1 mg/mland by76.4% tionof three general groups of metabolites,  avonoids, £ ¡ in 2:5 104 mg/mlmethionineconcentrations, compared terpenoidsand alkaloids, with diverse functions in plants, £ tocontrols in distilled water .Aneffect of methionine includingdefence against pathogens. The aim of thestudy solutionson juvenile activity was alsoshown after 24 h was totest members of the three groups of metabolites whenpercentage of active juveniles was reducedfrom for theireffects on plantparasitic nematodes. The nema- 99.3%in controls to 83.0% in 2 :5 104 mg per ml todespecies studied included the migratory endoparasites £ treatment.Further reductions were observedat lower Pratylenchuspenetrans and Radopholussimilis and the methionineconcentrations as timeof exposureincreased. sedentaryendoparasites Meloidogyneincognita and Glo- Whenmethionine was appliedto soil infested by M. boderarostochiensis .Thenematodes were subjectedto in incognita,reductionsin egg hatching and juvenileactivity vitro toxicitytests, chemotaxis tests and tests for theinhi- were observedat 0.1 and 1 mgof methionine per g of bitionof egg hatch. Results indicate that the  avonoids soil,either in sand or clay-loamsoils. The percentage of juglone,caffeic acid and umbelliferone are toxic to P. hatchedeggs 1 weekafter methionine application was penetrans with LD50 valuesof 90,374 and 418 ppm re- lowerin sandy than in clay-loam soils. Active juveniles spectivelyafter 72 h exposure.The monoterpenoid thy- were notfound after 1 weekin soil containing 1 mgof moland the sesquiterpenoid rishitin have LD 50 values of methionineper g ofsoil. 395and 387 ppm after 24 h exposure.Both caffeic acid andrishitin repel P.penetrans and R. similis at concen- trationsas lowas 50and20 ppmin thechemotaxis tests. 243Effects ofplantsecondary metabolites on plant Other avonoidswith repelling effects include quercetin parasiticnematodes (20ppm), resveratrol (100 ppm), kaempferol (100 ppm) andgenistein (100 ppm). Nathalie WUYTS¤, Annemie ELSEN, László SÁGI, Dirk DE WAELE and Rony SWENNEN Laboratoryof Tropical Crop Improvement, Kasteelpark Arenberg13, 3001 Leuven, Belgium ¤[email protected]

208 Nematology Postersessions

244The susceptibility of cucurbitaceousplants to largestcause of yield loss. In 2001 plant-parasitic Meloidogyneincognita (Kofoid& White,1919) nematodesaccounted for anestimated loss of 979 357 Chitwood,1949 balesvalued at 381.9 million dollars. Root-knot was responsiblefor alossof US$200.5million, reniform for 1; 1 2 Tímea ÁCS ¤, Béla PÉNZES , Mariann ELEKES , US$154.5million, and other species for US$25.5million 1 1 Antal FERENCZY and József FAIL loss.Beltwide losses due to nematodes have increased morethan 50% in the past 10 years.Most experts say that 1SzentIstvan University, F acultyof HorticulturalSciences, Ménesi ú t44,1118 Budapest, Hungary economiclosses have probably been underestimated in 2 CentralService for Plant Protection and Soil Conservation, thepast when losses due to nematodes were erroneously Laboratoryfor P estDiagnosis, Budaö rsi ú t141-145,1118 attributedto poor soil fertility, pH, or ‘ weakŽ elds’. Budapest,Hungary Improvementsin properly identifying nematode damage ¤[email protected] havebeen made, thus impacting the perceived trend of increasedlosses due to these pests. Meloidogyneincognita isan important pest of forced cucumber.As thereare no cucumber varieties resistant to Meloidogyneincognita ,theuse of cucumber plants 246 Meloidogyneincognita ,anew threat tosoybean graftedonto tolerant cucurbit species offers ahighly productionin Illinois effective,as well as an environmentallyfriendly means of control.T wocucurbit species, Syciosangulatus Harry and Jason BOND¤, Michael SCHMIDT, Alan WALTERS and CucurbitaŽ cifolia ,havebeen used by Hungariangrowers John RUSSIN asrootstock for cucumber.However, the tolerance level Plant,Soil and General Agriculture, Southern Illinois ofthese two species against M.incognita has not UniversityMailcode 4415, Carbondale, IL 62901-4415,USA beenfully elucidated. These two species, along with ¤[email protected] eightother previously untested species belonging to the Cucurbitaceae ,were evaluatedfor theirresistance to Meloidogyneincognita isan emerging threat to crop M.incognita. Testswere carriedout under glasshouse productionin southern Illinois. This pathogen has been conditions.The damage caused on theroots was assessed identiŽed in six soybean Ž elds,eight vegetable Ž elds byusing two six-graded scales (measuring size of galls andin Ž vepeach orchards. The potential impact of M. andnumber of galls). Stochastic homogeneity test was incognita tosoybean germplasm in northern latitudes usedfor statisticalanalysis. W efoundthat the galls isunknown. In 2001, four soybean varieties (Pioneer formedon theroots of S.angulatus Harry were smallbut 9481,Pioneer 9492, Gateway 493 and LS 94-3207) abundant.Galls occurring on theroots of C. Ž cifolia were were selectedand planted in infested Ž elds.Nematode notonly abundant but also large. Three cucurbit plants, populationdensities were recordedat plantingand every Cucurbitapepo Alba, Cucurbitapepo Sárga ó riás and 6weeksuntil harvest. At planting,the population density 3 Cucurbitamoschata Orange,were identiŽed aspotential of M.incognita averagedeight juveniles/ 100cm soil. rootstocksfor glasshousecucumber. Reproductionby M.incognita was higherin the plots plantedto P 9481.At harvest,the population densities (juveniles/100cm 3 soil)were 508,41, 37 and six for P 245Nematode losses in cottonin the USA 9481,P 9492,GW 493and LS 94-3207,respectively. Across thefour varieties, the increase in the population Donald J. BLASINGAME density of M.incognita was concomitantwith a linear 508Colonial Circle , Starkville,MS 39759,USA decreasein soybean yield. [email protected]

Cottonis grown on more than 15 million acres in 17 247Host suitabilityand damage thresholds of statesin the United States. Plant parasitic nematodes arablecrops to the rootlesion nematode Pratylenchus havebeen found in every state where cotton is grown. penetrans Eachyear losses to diseases and nematodes are reported E. BROMMER, T. VAN BEERS and L.P.G. MOLENDIJK tothe National Cotton Council by plant pathologist ¤ andnematologist in each state where cotton is grown. AppliedPlant Research (PPO agv),P .O.Box430, 8200 AK Comparedto other plant diseases, nematodes are the Lelystad,The Netherlands

Vol.4(2), 2002 209 Pathogenicity(244-264)

¤[email protected] William GAZAWAY Rootlesion nematodes ( Pratylenchuspenetrans ) are of 104Extension Hall, Auburn University, AL 36849,USA growingconcern in Dutch arable farming. Especially [email protected] onthe light sandy soils damage occurs in potatoes, Reniform (Rotylenchulusreniformis )androot-knot ( Me- blackroot and carrots. Dutch farmers tryto prevent the loidogyneincognita )arethe two major nematode species problemsby choosing a non-hostas preceding crop to thatattack cotton in Alabama. Nematode samples vulnerablecrops. Most data on host status and damage from Auburn’s nematodeanalytical laboratory and two thresholdswere determinedin the 1950s and 1960s. statewidenematode surveys conducted in 1990/1991and PPO startedin 1998 to reconsider the multiplication of againin 1999/ 2000revealed that reniform is the most P.penetrans onnew crops and varieties within modern widespreadand damaging nematode species in Alabama. growingpractices. The poster presents the host status of Thisspecies is responsible for over90% of the nema- importantarable- and green manure crops determined in todedamage on cotton in the state. T wenty-onepercent Želd-and in potexperiments. In addition, Ž eldresults of ofthe cotton Ž eldssampled in north Alabama and 26% initialpopulations ( Pi)andcrop yield are shown for some ofŽ eldsin central Alabama have reniform populations arablecrops. greaterthan 500 juveniles/ 100cm 3 soil.Yield losses in acottonŽ eldwith high reniform populations can be aslit- 248Effect ofthe root-knotnematode Meloidogyne tleas 10% or ashigh as 75%,depending on thepresence incognita onthe yieldof parsley( Petroselinum offavourableor adversegrowing conditions in any given crispum) year.When growing conditions for cottonin Alabama arenormal, yield losses usually average around 30-40%. 1; 1 2 R. CROZZOLI ¤, Aguirre YNDIRA and N. GRECO Statewideyield losses during the years 1999, 2000 and 1UniversidadCentral de V enezuela,F acultadde Agronomí a, 2001were estimatedto be US$14million annually. Most Institutode Zoologí a Agrícola. Apdo. 4579, Maracay, cottonproducers use nematicides to successfullymanage Venezuela reniformin cotton.A 1-yearrotation with a non-hostcrop 2 Istitutoper la Protezione delle Piante, Sezione di Bari, inAlabama has been shown to be more effective than a C.N.R., 70126Bari, Italy nematicide.However, most Alabama growers, who pre- ¤[email protected] fer togrow cotton continuously in the same Ž elds,have Ascreenhouseexperiment was conductedin 1000 cm 3 notadopted this practice. Reniform resistant or tolerant plasticpots to evaluatethe effect of increasingpopulation commercialcultivars are not currentlyavailable. Research densitiesof M.incognita onyield of parsley. Initial continuesfor moreeffective crop rotation schemes, im- nematodedensities were: 0, 0.125, 0.25, 0.5, 1, 2, 4, provednematicide application techniques and the search for reniformresistance in commercial cotton cultivars. 8,16, 32, 64, 128, 256 and 512 eggs secondstage juveniles(J2)/ cm 3 soil.The Seinhorst model, C y m D C .1 m/zP i T ,was Žttedto average plant top and total 250Investigations on Ditylenchusdipsaci damaging ¡ ¡ fresh weightand top dry weight. T olerancelimits ( T ) to carrotin Italy thenematode for topfresh weight,total fresh weightand 1 topdry weight were 0.17,0.02 and 0.05 eggs J2/cm3 Nicola GRECO , Antonio BRANDONISIO C 2 soil,respectively. The minimum relative yields ( m) were and Piero BONCORAGLIO 0.5 at Pi > 16 eggs J2/cm3 soil,0.55 at Pi > 2 eggs 1 C Istitutoper la Protezionedelle Piante, Sezione di Bari, J2/cm3 soiland 0.4 at Pi > 4 eggs J2/cm3 soil for C.N.R., ViaG. Amendola165/ A,70126Bari, Italy C C topfresh weight,total fresh weightand top dry weight, 2 SezioneOperativa 33, Assessorato Reg. Agr .eForeste, respectively.The maximum nematode reproduction rate RegioneSiciliana, V iaC.Ciano39, 97014 Ispica (RG), Italy was 37-foldat Pi 0.25 eggs J2/cm3 soil and the [email protected] equilibriumdensity Dwas 5eggs CJ2/cm3 soil. C Investigationswere undertakenin southern Italy on the epidemiologyof anApulianand a Sicilianpopulation of 249Reniform nematodes – athreat tocotton Ditylenchusdipsaci ,oncarrotunder Ž eldconditions, from productionin Alabama (USA) Septemberto the following March. The host status of someplants to the Sicilian population of the nematode

210 Nematology Postersessions was alsoassessed. In Apulia, on carrot sown in early Instituteof Zoology,NAS RA, Yerevan,, P.Sevak7, Armenia September,the nematode infection started at the end of ¤[email protected] Octoberon leaves and in early December on tap roots. Wehavestudied the rootsand the top-soilunder tomatoes, Deathof leaves and tap root rot were observedthe cucumbers,pepper, cabbage, carrot and watermelon as followingJanuary on mature plants. On carrot sown 1 wellas wheat,sweetcorn and potato. Thirty-nine species monthlater nematode infection occurred by thebeginning ofnematodesrecorded in ten crops belonged to 17 genera. ofNovember and greatly suppressed plant growth. In Themost virulent included Macropostoniaxenoplax, Sicily,on carrotsown in earlyOctober the infection was Criconemoidesmongolensis, Heterodera crucifera, Glo- obviouson aerialplant parts at the end of Novemberand boderarostochiensis, Meloidogyne hapla, M. arenaria, ontap roots in midDecember .Leafdeath and tap root rot M.incognita,T ylenchorhynchusbrassica, T .davaini,T . occurredat end of January.Early symptoms of nematode parvus,Quiniculcius acti, Helicotylenchus pseudorobus- attackswere straddledleaves, multi-bud plant crowns tus,H. eritrinae,H. conicephalus,H. californicus,H. andlight discolorations of tap root tops. The portion of digonicus,Ditylenchus destructor ,D.dipsaci,Aphelen- theplant most affected by D. dipsaci was that2-4 cm choidessubparientinus, A. ritzemabosi,Longidorus elon- belowand above ground. The Sicilian population of the gatus andothers. The nematode fauna of crops studied nematodereproduced on carrot, broad bean, oats, rye, was diverseand abundant, and was distinguishedby a Italianryegrass, celery, Amny majus and Galiumaparine . largenumber of dominant species. The species diversity ofnematodesand their population density in thetop soil 251Effect of Globoderarostochiensis onthe yieldof was relatedto the agrotechnology measures which low- potatocv. Kennebec in Venezuela eredthe number of species.

N. JIMÉNEZ-PÉREZ 1, R. CROZZOLI 2 and N. GRECO 3 1 253Pathogenicity of Pratylenchuscoffeae on UniversidadCentrooccidental Lisandro Alvarado, F acultad seedlingsof coffeecv. Mundo novo deAgronomí a, Barquisimeto,V enezuela 2 UniversidadCentral de V enezuela,F acultadde Agronomí a, 1; 2 Roberto K. KUBO ¤,RosangelaA. S ILVA , Institutode Zoologí a Agrícola, Apdo 4579, Maracay, V enezuela 2 1 3 Melissa D. TOMAZINI ,ClaudioM.G. O LIVEIRA , Istitutoper la Protezione delle Piante, Sezione di Bari, 3 2 C.N.R., 70126Bari, Italy Paulo MAZZAFERA andMá rio M. I NOMOTO Therelationship between initial densities ( Pi) of Glo- 1InstitutoBioló gico, CP 70,13001-970 Campinas, SP ,Brazil 2 boderarostochiensis pathotypeRo2 and yield of thesus- ESALQ/USP,CP9,13418-900Piracicaba, SP ,Brazil 3 ceptiblepotato cultivar Kennebec was investigatedin 30 UNICAMP,CP6109,13083-970 Campinas, SP ,Brazil dm3 microplotsat Cubiro,Lara State, during the January- ¤[email protected] Aprilgrowing season. Pi usedwere 0,0.125,0.25, 0.5, 1, Thepathogenicity of twoBrazilian Pratylenchuscoffeae 3 2,4,8,16,32,64,128,and 256eggs/cm soil.Tuber yield isolateswas evaluatedin twoglasshouse experiments on response to Pi Žttedthe model y m .1 m/zP i T and D C ¡ ¡ coffeeseedlings ( Coffeaarabica )cv.Mundo Novo. In the atolerancelimit ( T )ofpotatoto G.rostochiensis of one Žrst,the effect of initial population densities ( Pi 0, 333, 3 D egg/cm soilwas derived.Yield losses of 20and50% oc- 1000,3000 and 9000 nematodes/ plant)was studiedusing 3 curred at Pi ofeight and 32 eggs/ cm soil,respectively. P. coffeae isolatefrom Marília, SP .Thedata were Žtted Theminimum relative yield ( m) was 0.35 at Pi > 128 totheSeinhorst model. At 270days after inoculation, all 3 eggs/cm soil.The maximum nematode reproduction rate plantsinoculated with 9000 nematodes and the majority 3 was 25-foldat Pi 0:125 eggs/cm soiland the equilib- inoculatedwith 3000 nematodes were dead.Plant growth D 3 riumdensity 64 eggs/cm soil.Symptoms of thenematode andphotosynthesis were reducedwith inocula of 333and attackbecame obvious on aerial plant parts at Pi 64 1000nematodes, respectively. In the second experiment, 3 D eggs/cm soil. pathogenicityof two P. coffeae isolates(from Marília and Riode Janeiro) was comparedusing coffee seedlings. 252Parasite-host relationship and virulence of the Althoughphotosynthesis was similarlyreduced for both mostdamaging nematodes in Ž eldsin Armenia isolateswith an inoculum of 8000 nematodes/ plant,the Marília isolate caused intense darkening of theroots, leaf J. KARAPETYAN¤ and H. MKRTCHJAN chlorosisand a greaterreduction of rootand shootgrowth.

Vol.4(2), 2002 211 Pathogenicity(244-264)

Inboth experiments, reproduction ( Pf/Pi)oftheisolates confersresistance on them to the root-knot nematodes was low,suggestingthat coffee is apoorhost for both P. Meloidogynearenaria, M. incognita and M. javanica, coffeae isolates.The differential pathogenicity observed areseriously affected by these pathogens, even leaving inthe second experiment supports previously published thecrop non-viable in some cases. It has been observed datathat reported differences in morphology and host thatplants resistant to these nematodes, either because rangesbetween these isolates. theyare carriers oftheabove-mentioned gene or because theyare grafted onto a resistantstock, are susceptible principallyto attacks by M.incognita and M. javanica. 254Virulent populations of Meloidogyne in Inmost cases, this is due to a incorrectapplication vegetablecrops from Uruguay ofthese alternatives, sometimes because the plant loses 1; 2 itsresistance due to exposure to soil temperatures over J. Antonio LÓPEZ-PÉREZ ¤, Leonardo DE LEÓN and Antonio BELLO 1 27±Corby selection of virulent populations. The use ofcrop management techniques that decrease nematode 1 Dptode Agroecologí a. Centrode Ciencias Medioambientales, populations,such as biofumigation, has been regarded CSIC,Serrano 115 dpdo, 28006 Madrid, Spain asefŽ cient in the regulation of virulent populations of 2 DptoAgricultura, Rel-UITA, W .FerreiraAldunate 1229, C.P . nematodesbelonging to the genus Meloidogyne . 11.100,Montevideo, Uruguay ¤[email protected] 256Pest riskanalysis of Meloidogynechitwoodi in Sometomato varieties that are carriers ofthe Mi Norway gene,which confers resistance on them to the root- knotnematodes Meloidogynearenaria and M.incognita , Christer MAGNUSSON¤, Leon THIJSEN appearedseverely affected by these pathogens, making andRicardo H OLGADO thecrop non-viable in many cases. This happens frequentlyin the departments of northern Uruguay, TheNorwegian Crop Research Institute Plant Protection especiallySalto and Artigas, where soil temperatures Centre,Department of Entomologyand Nematology, Høgskoleveien 7, N-1432Aas, Norway oftenrise above 27 C,whichis one of themaincauses for ± [email protected] thebreak in resistance. V arious M.incognita populations ¤ from northernUruguay have been characterised, proving Meloidogynechitwoodi maybe introduced into Norway tobe virulentto resistant tomato varieties. Several pepper throughplant material and soil. The nematode may cultivarswith the N genehave also been selected easilyestablish on common crop plants. During the thatdemonstrate resistance to these nematodes. It can growingseason two generations are expected to develop beafŽ rmed that conventional crop methodologies are insouthernNorway, while conditions in thecentral parts contributingto the selection of virulent populations, wouldallow for thedevelopment of one generation. M. whichcould make the use of resistantvarieties impossible chitwoodi startsto developat 5 ±C.Plantingand sowing in inthe future. Biofumigation and other crop techniques Norwaystarts when soil temperatures reach around 4 ±C. provedeffective in the regulation of virulent nematode Theuse of plastic tunnels to increase soil temperature populationsof the Meloidogyne genus,without the use of isa commonpractice in early potatoes and vegetables, nematicides. andwould enhance nematode development. M.chitwoodi requires600-800 degree days (DD) tocomplete one 255Virulent populations of Meloidogyne in generationfrom thetime of planting, and 500-600 DD vegetablecrops in Spain for subsequentgenerations. These temperature sums are commonlyrecorded in potato growing areas of Norway.

J. Antonio LÓPEZ-PÉREZ¤, Miguel ESCUER and Qualitativedamage on potatotubers would require the re- Antonio BELLO infectionand development of thesecond generation, and isexpectedin the southern areas. Qualitative damage on Dptode Agroecología, CCMA, CSIC,Serrano 115 dpdo, 28006 carrotsdoes not require re-infection, and is expected also Madrid,Spain incentral parts. The possibility that the nematode could [email protected] ¤ causedamage to other vegetables and to cereals is an Ithas been observed that tomato varieties having the additionalconcern that supports its status as aquarantine Mi geneand pepper varieties with the N gene, which pestorganism.

212 Nematology Postersessions

257Progress of two kinds ofsymptomappearing in creasedby 0.2, 0.8 and 1.1 t/ hawhere Furadan was ap- the bodyof recovered Pinus thumbergii frompine wilt pliedto theseedbed only, to the Ž eldonly, and to both the disease seedbedand Ž eld,respectively, compared to a non-treated control.A veragedacross three farmer Želds,rice yield in- Kuniaki MATSUURA creasedby 1.1 t/ hawhere Furadan was appliedto both Forestryand F orestProducts Research Institute, Matsunosato theseedbed and Ž eldcompared to thecontrol. At harvest, 1,Kukizaki, Ibaragi 305-8687, Japan soillevels of M.graminicola inthenon-treated plots were morethan three times greater than those in the treated Pinewood nematode can cause fatal damage to Japanese plotsat both locations. Prior totransplanting, seedling black pine, Pinusthumbergii andred pine, Pinus shootheight and dry weight were signiŽcantly greater, densiora.However,not enough is known about the andsoil levels of M.graminicola signiŽcantly less, in the mechanismsleading to the death of pines killed by this treatedseedbed plots compared to the non-treated plots. organism.Hence, study of the various aspects of pine Thisis the Ž rst on-farmstudy to demonstrate a negative treeswhich are purported to represent the processes of impact of M.graminicola onricegrowth and yield during thedisease from itsearly stages through to its lethal themonsoon season in Bangladesh. stageshas helped to clarify both disease development andcausal mechanism. Thionazin or mesulfenfos was injectedinto 11-year-old P.thumbergii infectedwith 259Reaction of oliveto Pratylenchusvulnus infectionsin Italy pinewilt disease following inoculation of pine wood nematodes2, 5 and6 weeksprior to treatment with Nicola SASANELLI and Trifone D’ADDABBO nematicides.At thetimeof injectionwith nematicides, all ¤ thetrees tested were judgedas having pine wilt disease Istitutoper la Protezione delle Piante, Sezione di Bari, C.N.R., becausethey had ceased to exude pine resin. Recovery, ViaG.Amendola165/ A,70126Bari, Italy expressedas resumation of resinexudation by surviving ¤[email protected] trees,was observedin pines injected with thionazin 2 Thereaction of seven olive cultivars, Cellina di Nardò , weeksafter inoculation. By examining cross-sections of Cimadi Bitonto,Coratina, FS 17,Frantoio, Leccino and recoveredpine trees felled 2 yearsafter this experiment, Yustiand of therootstock DA 12Ito Pratylenchusvulnus twoapparently distinct symptoms were observed,one an infectionswas evaluatedin a glasshouseexperiment. earlysymptom associated with the formation of small Plantswere artiŽcially inoculated with 200 nematodes per non-water-conductingpatches, and the other associated pot.Plant growth and nematode reproduction parameters withcambium death. Both symptoms seemed to beginin were recorded.Host reaction was ratedaccording to the andprogress from thepith. reproductionfactor r Pf /Pi.Allthe tested cultivars D androotstock were goodhosts for P. vulnus,althoughthe 258Impact of Meloidogynegraminicola on yield of highestsusceptibility was shownby FS17.No statistical differencewas foundamong the P. vulnus reproduction lowlandrainfed rice in Bangladesh parameterson theother cultivars. 1; 2 1 Jon PADGHAM ¤, George ABAWI , John DUXBURY 3 and Abdul MAZID 260Impact of Pratylenchusneglectus infestationon 1Departmentof Cropand Soil Sciences wheat inWestern Australia 2 Departmentof PlantP athology,Cornell University, Ithaca, NY14853,USA Shashi SHARMA¤, Sean KELLY and Robert LOUGHMAN 3 BangladeshRice Research Institute, Rajshahi, Bangladesh Departmentof Agriculture,Baron Hay Court, South P erth, ¤[email protected] WA 6151Australia [email protected] Replicatedplots with and without the application of Fu- ¤ radan5G (40 kg/ ha)were establishedat an agricultural Pratylenchusneglectus isvery widespread in cereal- researchstation and on farmer Želdsin northwestern producingregions of W esternAustralia. A recentstudy Bangladeshto assessthe impact of Meloidogynegramini- revealedthat the average nematode population in 64% cola onlowland rainfed rice yield in the rice-wheat pro- ofthe surveyed locations exceeded the lowest level ductionsystem. At theresearch station, rice yields in- thatcauses signiŽ cant yield loss. Experiments were

Vol.4(2), 2002 213 Pathogenicity(244-264) establishedbetween 1999 and 2001 in nematode-infested for P. thornei were estimated.In addition, an exponential Želdsat Ž velocations in central and southern wheat- relationshipwas observedbetween initial density and beltsto studyimpact of thenematode on performanceof nematodemultiplication rate ( Pf/Pi). wheat.Large plots (1 :8 40 m or 1:44 20 m) were £ £ sownto susceptible wheat (cvs Machete or Cunderdin 262Infection of Arachishypogaea by Meloidogyne orBrookton). In half of the 64 plots (paired plots), arenaria mayincrease a atoxin contamination of aldicarb(T emik150G) was applied(20-30 kg/ ha)at kernels seeding.Ten weeks after sowing nematode densities inroots were assessed.Nematode density and wheat P. TIMPER¤ and D.M. WILSON yieldin the nematicide-treated and check plots were comparedusing analysis of variance. Aldicarb partially USDA ARS andDepartment of Plant P athology,University of controlledthe nematode populations. Yield enhancement Georgia,P .O.Box 748, Tifton GA 31793,USA withnematicide application was 6-14%.At onelocation, ¤[email protected] yielddifferences following use of aldicarb were not signiŽcant, partly due to inadequate nematode control. AŽeld-microplotstudy was conductedto determine Thestudy established economic damage potential of whethera atoxin levels in peanut, Arachishypogaea , P.neglectus inW esternAustralia; however, this loss were correlatedwith pod and root galling caused by assessmentcould be anunder-estimation of actualloss as Meloidogynearenaria (Ma).Theexperiment was a nematicideprovided only partial protection to plants from completelyrandomised 2 2factorialwith six replicates £ nematodeinfection. pertreatment. Factors were nematodes,plus and minus Ma,andfungus, plus and minus Aspergillus avus (Af ). 261Population dynamics of, and yield loss caused Peanutseedlings, either infected with Ma oruninfected, 2 by, Pratylenchusneglectus and P. thornei in cereal were transplantedinto 2.3 m plotspreviously fumigated cropsin Australia withmethyl bromide. Inoculum of Af was sprinkledover theplantcanopy at midbloom. Drought was inducedafter 1; 2 1 Sharyn TAYLOR ¤, Kerrie DAVIES and Danuta SZOT podset by covering plots during rain with a Žbreglass 1 shelter.Pod- and root-gall indices (1-5 scale) were SouthAustralian Research and Development Institute, GPO determinedfor allplants in the plot and averaged. Pods Box397, Adelaide, South Australia 5001, Australia from eachplot were bulked,shelled, and a subsampleof 2 AdelaideUniversity, Department of Applied and Molecular Ecology,PMB 1GlenOsmond, SA 5064,Australia kernelswas usedto determine a atoxin concentration and percentagecolonised by Af.Neitherfactor, nematodes or ¤[email protected] fungus,affected a atoxin concentration; however, there Arangeof initial soil densities of either P.neglectus was aninteraction between these factors ( P 0:003). or P. thornei was establishedin Ž eldsites in South Inplots without added fungus, there was acorrelationD Australia.This was achievedby growing replicated betweena atoxin concentration vs pod-gallindex ( P D plotsof susceptible wheat (cvs Machete and Spear), 0:001, r 0.82) and vs root-gallindex ( P 0:006, D D moderatelyresistant wheat (cv. Excalibur), moderately r 0.74).Colonisation of kernels by Af increasedwith D resistantbarley (cv. Chebec) and resistant triticale (cv. increasingpod galling ( P 0:04, r 0.42). Tahara)in the 1st year .Inthe 2nd year, these cultivars D D resultedin initial P.neglectus densitiesfor Machete, 263Yield losses of barley,oat andwheat due toroot Spear,Excalibur, Chebec and T aharaof 18,8, 4, 2and3/ g lesionnematode in South Australia soil,respectively. For P. thornei,thesecultivars resulted ininitial densities of 14, 10, 3, 1 and1/ gsoil.Plots 1; 1;2 Vivien A. VANSTONE ¤,MichelleH. R USS and were oversownin the 2nd year with an intolerant oat Sharyn P. TAYLOR 2 cultivar(Echidna). Initial ( Pi)andŽ nalnumbers ( Pf ) of nematodeswere assessedfrom soilof all plots in each 1Universityof Adelaide,Department of PlantScience, PMB 1, year.For both P.neglectus and P. thornei,asigniŽcant GlenOsmond, 5064, South Australia, Australia negativelinear relationship was observedbetween yield 2 SouthAustralian Research and Development Institute, GPO ofthe intolerant oat and initial nematode density in the Box397, Adelaide, 5001, South Australia, Australia 2ndyear .Yieldlosses of 27% for P.neglectus and 22% ¤[email protected]

214 Nematology Postersessions

Wheat (Triticumaestivum )isthe principal host of Praty- 264Response offour potato ( Solanumtuberosum ) lenchusneglectus insouthern Australia. By comparison, cultivarsto varying levels of root-knotnematode barley (Hordeumvulgare ) and oat (Avenasativa ) are mod- (Meloidogynehapla ) inoculum eratehosts. Mean yield loss for intolerantwheat in 1995- 1998was 15-20%.Trials at six sites were assessedin 2000 Luciana M. VILLANUEVA¤ andMaidina A. L INGGOY tocompare losses for barley,oat and wheat. Natural vari- Departmentof Plant P athology,College of Agriculture, Benguet ationin nematode density between and within trial sites StateUniversity, La Trinidad, Benguet 2601, Philippines allowedcomparison of plot yields over a rangeof ne- ¤[email protected] matodedensities. Final nematode density ( Pf ) was deter- Theeffect of varying levels of Meloidogynehapla minedduring grain development in mid-spring(October). inoculumon the growth and yield of four potato Negativecorrelations between yield and Pf were signiŽ- cultivarswas studiedunder glasshouse conditions. Plant cantfor barley( r 0:789-0.875),oat ( r 0:654-0.892) D D height,fresh rootand top weights, and tuber yield were and wheat (r 0:524-0.828).Y ieldloss for eachgeno- D signiŽcantly affected by the nematode. Plant height was typewas calculatedfrom themaximum genotype yield reducedby 28-45% when Pi was 1000juveniles per predictedby thelinear relationship between yield and Pf. pot,and 30-57% when the number of infectivejuveniles Meanyield loss for intolerantbarley and oat was 8.4%, presentaround young seedlings was 10000.Fresh root andfor wheat7.3%. Barley and oatare more resistant than weightwas decreasedby 35-44, 39-51, 41-50 and 37- wheat (Pf,respectively,1.0, 1.6 and 2.5 P.neglectus /g dry 46%in cvs Berolina, Igorota, Solibao and Granola, soil),but suffer comparableyield loss and are therefore as respectively,when Pi was 1000-10000. SigniŽ cant yield intolerantas wheat. Pratylenchusneglectus isaconstraint reductionwas alsonoted in the four cultivars used tocereal production in southernAustralia, and these yield when Pi was 1000-10000 juveniles per pot: 44-56% penaltiesrepresent signiŽ cant Ž nancialloss to the grower . inBerolina; 75-88% in Igorota and Solibao and 85- 100%in Granola. Based on the gall index rating used, cvsBerolina, Igorota, Solibao and Granola were rated resistant,moderately resistant, moderately susceptible andsusceptible, respectively, to M. hapla infection.

Vol.4(2), 2002 215 Parasitism(265-294)

265Novel gene discovery in plant-parasitic we successfullyampliŽ ed the chorismate mutase of nematodes Heteroderaglycines ,thesoybean cyst nematode (SCN), from cDNA extractedfrom 4-day-oldparasitic juveniles. Bryony BANKS¤, Miles ARMSTRONG, John JONES, TheSCN homologgene has been cloned and sequenced. Mark PHILLIPS and Vivian BLOK Thederived protein sequence of the SCN chorismate PlantP athogenInteractions, Scottish Crop Research Institute, mutaseshowed about 25% identity to thatof M. javanica Invergowrie,Dundee DD2 5DA,Scotland, UK andabout 55% identity to that of G.rostochiensis . TheSCN chorismatemutase is being characterised by ¤[email protected] southernblot, enzymatic analysis and in situ hybridisation Thepotato cyst nematodes ( Globoderapallida and analysis. G.rostochiensis )areeconomically important pests bothin the UK andworldwide, and cause signiŽ cant lossesto commercial potato production. As ameans 267Establishing a Lotusjaponicus -nematode ofinvestigating the complex host-pathogen interactions pathosystemto search for variations in their involved in G. pallida and G.rostochiensis infections, interactions cDNA AFLP andsuppression subtractive hybridisations Héctor CABRERA POCH1; , Andrew WARRY 1 and (SSH) usingresistant and susceptible potato cultivars ¤ Charles OPPERMAN 2 were carriedout. Novel genes expressed during invasion ofthe host plant and subsequent nematode development 1NematodeInteractions Unit, Plant-P athogenInteractions were identiŽed byharvestingplants at severaltime points Division,IACR-Rothamsted, Harpenden, Herts AL5 2JQ,UK rangingfrom 24hto14dayspost-infection. A signiŽcant 2 Departmentof Plant P athologyand Genetics, North Carolina proportionof candidate genes from theSSH matched StateUniversity, Raleigh, NC 27695,USA nematodeEST suchas F ABP-1,haemolysin, calponin, ¤[email protected] beta-1,4-endoglucanase,cytochromep450 monoxygenase andribosomal proteins. Other sequences gave no reliable Inorder to complete their life cycles, both root-knot databasematches. Localisation studies are being carried andcyst nematodes must interact with their host plants. outon these genes by in situ hybridisationto obtain Muchattention has been devoted to the nematode factors functionalinformation and levels of gene expression involvedin root penetration, migration and feeding site duringnematode invasion and development are being formation.However, very little is known about plant host investigatedusing rt-PCRand southern blot analysis. T o factorsthat are needed for thesusceptible interaction furtherquantify levels of expression during the host- tooccur because, in general, natural host plants are pathogeninteraction an analysis using real time PCR is noteasy to manipulate genetically. Lotusjaponicus has currentlyunderway. emergedin the past few yearsas a suitablemodel plant tostudyinteractions with nitrogen Ž xationsymbionts. A setof genetic resources and tools are rapidly becoming 266Cloning of the chorismatemutase homolog of availablefor L.japonicus ,includingecotypes, mutants, the soybeancyst nematode Heteroderaglycines transformationprocedures and a sequencingproject. W e haveestablished a pathosystemusing Lotusjaponicus Sadia BEKAL¤, K.N. LAMBERT and T.L. NIBLACK asa hostplant for botha plantcyst nematode and a Deptof Crop Science, University of Illinois,Urbana, IL 61801, root-knotnematode. The current search for variationin USA theinteractions between either of these nematodes and ¤[email protected] a battery of L.japonicus ecotypesand mutants will Thechorismate mutase is a nematodegland secreted eventuallylead to the isolationand cloning of plantfactors proteinthat was Žrst clonedfrom root-knotnematode responsiblefor suchinteractions. Meloidogynejavanica .Thisenzyme is extensivelystudied inroot-knot nematode and it was recentlyreported as 268Isolation of putativeparasitism genes from an EST from Globoderarostochiensis. This enzyme Globoderapallida isbelieved to be involved in nematode parasitism. 1; 1 Usingdegenerate primers to the conserved regions Héctor CABRERA POCH ¤, Andrew WARRY and ofboth known nematode chorismate mutase proteins, Charles OPPERMAN 2

216 Nematology Postersessions

1NematodeInteractions Unit, Plant-P athogenInteractions muchwork has been performed on the interaction Division,IACR-Rothamsted, Harpenden, Herts AL5 2JQ,UK betweenthe nematode and the virus, most research 2 Departmentof PlantP athologyand Genetics, North Carolina onthe nematode itself has been limited to taxonomic StateUniversity, Raleigh, NC 27695,USA studies.Antibody methodology has been successfully ¤[email protected] usedto discover parasitism genes from endoparasites. Potatocyst nematodes (PCN) aredevastating pests in the Wethereforeproduced a panelof MAbs against X. UnitedKingdom and other European countries. While index. Aseriesof secreted proteins present in different Globoderarostochiensis canbe controlled in the UK dorsalducts, which related to particular periods of byusing resistant potato cultivars, no effective resistant thenematode feeding cycle, were recognisedby one genesare available to control G. pallida populations. monoclonalantibody. Other MAbs were obtainedwhich Researchtargets for thecontrol of G. pallida include recognisethe cuticle surface, muscle Ž bres,nerve cords parasitismgenes needed to complete its life cycle. T o andoocytes. Some of these MAbs have the potential isolatethese targets, two G. pallida cDNA librarieswere tobe developed for useas nematode diagnostics. An builtfrom eitherpreparasitic juveniles (J2) andadult antibodythat recognises the virus-retention site within the (gravid)females or J2 individuals. A collectionof more nematodeodontophore may be useful for furthervirus- than1000 EST from theformer libraryhas been released nematodeinteraction studies. In future work we will intothe public domain. Data mining in this collection usewestern blots of 2D electrophoresis gels coupled to revealsEST encodingfor putativesecreted proteins with MALDI-TOF analysisto identify the secretory proteins similaritiesto T ylenchidasequences, but not tofree-living inglandcells recognised by theMAbs. C. elegans sequences,which suggests that they may have arolein plant nematode parasitism. A numberof PCN 270Peptides that bind toand inhibit cellulase of speciŽc sequenceshave no homologiesto other sequences Heteroderaglycines identiŽed bycombinatorial indatabanks. These novel sequences encode for proteins libraryscreening containingfeatures of secretedproteins, which make them 1; 3 2 candidatesfor furtherresearch into their putative role in Wanglei DU ¤, Richard HUSSEY , Thomas BAUM and PCN-speciŽc parasitismand evolution. Eric DAVIS 1

1 269Using monoclonal antibodies to discover Departmentof PlantP athology,Campus Box 7616, North parasitismproteins from the virus-vectornematode CarolinaState University, Raleigh, NC, USA 2 Departmentof Plant P athology,Iowa State University, Ames, Xiphinemaindex IA, USA 3Departmentof PlantP athology,University of Georgia, Athens, Qing CHEN1; , Rosane H. CURTIS 2, ¤ GA, USA Franco LAMBERTI 3, Maurice MOENS 4, ¤[email protected] Derek J.F. BROWN 1 and John T. JONES 1

1Plant-PathogenInteractions Programme, Scottish Crop Thefeasibility of identifying inhibitors of the products ResearchInstitute, Invergowrie, Dundee, DD2 5DA,Scotland, ofnematodeparasitism genes is beinginvestigated using UK thesecreted cellulases of Heteroderaglycines as a 2 NematodeInteractions Unit, IACR Rothamsted,Harpenden, modeltarget. The HG-eng-2 endoglucanase gene of HertsAL5 2JQ,UK H. glycines was overexpressedin a Pichiapastoris 3 Istitutodi NematologiaAgraria, C.N.R., 70126Bari, Italy host,and active recombinant HG-ENG-2 was afŽnity- 4 Laboratoryvoor Nematology, Centrum voor Landbouwkundig puriŽed. PuriŽ ed rHG-ENG-2 was usedas a targetin Onderzoek,Gent, Belgium biopanningexperiments with a commercialphage-display [email protected] ¤ combinatorial7-mer peptidelibrary. T wostringent rounds Xiphinemaindex isa plantectoparasitic nematode that ofbiopanningproduced about 1000 plaques with peptides transmitsplant viruses. Xiphenemaindex induceschanges thatbound to rHG-ENG-2. Individualplaques are now inplant cells similar to those induced by endoparasites beinginvestigated to determine the number of unique such as Meloidogyne spp. Xiphinemaindex is large peptidesthat bind to rHG-ENG-2 andto identify andcan be cultured, making it an attractive model thosethat inhibit the ability of rHG-ENG-2 todegrade systemfor someaspects of plant nematology. Although carboxymethylcellulose.One promising 7-mer peptide

Vol.4(2), 2002 217 Parasitism(265-294) thatbinds to and inhibits the activity of rHG-ENG-2 has ofmicroaspiration of cytoplasm from thepharyngeal beenidentiŽ ed to date. glandcells of tenparasitic stages of Heteroderaglycines from differenttime points in the parasitic cycle provided 271Proteins and expressed genes of the ectoparasite expressedmRNA toconstruct a glandcell-speciŽ c cDNA Xiphinemaindex libraryby long-distance-PCR. Of 2345cDNA clones sequenced,deduced protein sequences of 231cDNAs had Cleber FURLANETTO¤,DerekJ.F .B ROWN and aN-terminalsignal peptide for secretionand, thus, could John T. JONES haveroles in H. glycines parasitismof soybean. High- throughputin situ hybridisationwith probes of 112cDNA Plant-PathogenInteractions Programme, Scottish Crop clonesencoding signal peptides resulted in 40 unique ResearchInstitute, Invergowrie, Dundee, DD2 5DA,Scotland, UK clonesspeciŽ cally hybridising to transcripts within the subventral(11 clones) or dorsal (29 clones) gland cells. ¤[email protected] PSORT IIpredicted30 oftheproteins to be extracellular Xiphinemaindex ,thenatural vector of Grapevine fan leaf andten proteins as nuclear localised. In BLASTp virus,is aplantectoparasite that induces changes in host analyses,25 ofthepredicted proteins were novel.Those cellssimilar to those induced by biotrophicendoparasites proteinswith similarities to known proteins included such as Meloidogyne and Heterodera. Xiphinemaindex venomallergen like proteins, ß -1,4-endoglucanases,a islarge, readily cultured and all life stages occur outside pectatelyase, a chitinase,RanBPMs, and a cellulose- theplant making it an attractive model system for some bindingprotein. Only two of the40 gland-expressedgenes aspectsof plantnematology. W ehavebegun a programme hadhomologues in Caenorhabditiselegans . aimedat identifying genes and proteins important in thehost parasite interaction of this species. Expressed 273Activity of plantendoglucanase gene promoters sequencetags are being obtained from amixedstage innematodefeeding cells cDNA libraryof this nematode. Over 500EST have beenobtained and a varietyof genesidentiŽ ed including 1; 2 3 Melissa GOELLNER ¤, Xiaohong WANG , Ziv SHANI , proteasesand collagens. T odateno EST from genes Oded SHOSEYOV 3 and Eric DAVIS 2 acquiredfrom bacteriaby horizontal gene transfer have beenfound. T wo-dimensionalelectrophoresis has been 1Developmental,Cell and Molecular Biology Group, Levine usedto identify proteins speciŽ c tonematode portions ScienceResearch Center ,Box91000, Duke University, containingthe pharyngeal gland cells. In future work we Durham,NC 27708,USA 2 willuse MALDI-TOF analysiscoupled to the data from Departmentof PlantP athology,North Carolina State ourEST programmeto identifythe genes encoding such University,Raleigh, NC, USA 3 Kennedy-LeighCenter ,HorticulturalResearch, The Hebrew proteins. Universityof Jerusalem,Rehovot, Israel ¤[email protected] 272DeŽ ning a plant-parasiticnematode: a proŽle of Fiveendoglucanase ( Ntcel)geneswere demonstrated putativeparasitism genes expressed in the pharyngeal tobe upregulated in syncytia and giant-cells formed glandcells of Heteroderaglycines in Nicotianatabacum by Globoderatabacum and 1; 2 3 Meloidogyneincognita ,respectively.1.5 kb of the 5 0- Bingli GAO ¤, Tom MAIER , Eric DAVIS , Thomas BAUM 2 andRichard H USSEY 1 ankinggenomic DNA regionof the most strongly upregulatedtobacco endoglucanase gene, Ntcel7, was Departmentsof Plant P athology fusedto the GUS reportergene in a binaryvector 1 Universityof Georgia, Athens, GA 30602 andused to produce hairy roots of tobacco and 3 IowaState University, Ames, IA 50011 tomato, Lycopersiconesculentum , viaAgrobacterium 4 NorthCarolina State University, Raleigh, NC 27695 rhizogenes-mediatedtransformation. The 1.5 kb Ntcel7 [email protected] ¤ promoter/GUS constructwas expressedspeciŽ cally Themost evolutionary advanced adaptations for plant withinplant root tips, lateral root initials, and appeared parasitismby nematodes are the products of parasitism tobeexpressedwithin the feeding cells of root-knotand genesexpressed in thepharyngeal gland cells and secreted cystnematodes in both tobacco and tomato hosts. These throughthe stylet into plant tissue. A directmethod Žndingsagree with earlier evidence that an Arabidopsis

218 Nematology Postersessions thaliana endoglucanase( Atcel1)promoterwas activein 3 Departmentof Biology, State University, P ennsylvania,USA thefeeding cells of root-knot nematodes in transgenic ¤[email protected] tobacco.The apparent expression of the Ntcel7 promoter Thedifferentiation of syncytia of Heteroderaschachtii infeeding cells of cyst and root-knot nematodes in two (Hs) in roots of Arabidopsisthaliana isaccompanied by plantspecies suggests that these promoters may have cellwall loosening, extension and local degradation. In similaractivity in otherplant systems. orderto study whether speciŽ c expansinsare involved inthis process, RT -PCRexperiments with a syncytium- 274IdentiŽ cation and localisation of tomato speciŽc cDNA libraryand primersdifferentiating between expansingene expression in nematode-induced 17 ®- and 3 ¯-expansincDNAs of Arabidopsis were per- syncytia formed.SpeciŽ c Arabidopsis mRNAs of 8 ®- and 1 ¯- expansinswere presentin 5-7 day-old syncytia induced 1; 2 3 B. GOLECKI ¤, S. FUDALI , W. WIECZOREK and by Hs. In situ-RT-PCRexperiments indicate that ampli- F.M.W. GRUNDLER 3 Ž ed cDNA of At-Exp6is localised in the cytoplasm of syncytialcells and not in the surrounding root. These re- 1Institutfü r Phytopathologie,Universitä tKiel, 24098 Kiel, sultscould be conŽrmed with transgenic Arabidopsis pro- Germany 2 Departmentof Botany,Agricultural University, W arsaw, moter:guslines. GUS activitywas speciŽcally observed Poland insyncytiaand primordia of lateralroots. 3 Institutfü r Panzenschutz, Universitä tfü r Bodenkultur, Vienna,Austria 276IdentiŽ cation of geneexpression differences ¤[email protected] between Globoderapallida and G. mexicana by suppressivesubtractive hybridisation Inthe model system Arabidopsisthaliana/ Heterodera schachtii (Hs)itwas shownthat speciŽ c plantexpansins 1; 2 2 Eric GRENIER ¤, Vivian BLOK , John JONES , areactivated in nematode-induced syncytia. T oprove Didier FOUVILLE 1 and Didier MUGNIÉRY 1 whetherthe situation in the model system is transferable 1 tosusceptible crop plants, tomato roots were infected INRA UMR Biologiedes Organismes et desP opulations with Hs and Globoderarostochiensis (Gr),respectively. appliquée àlaProtectiondes Plantes, Domaine de la Motte, B.P.35657,35653 Le Rheucedex, France RT-PCRexperiments with primers discriminating seven 2 Mycology,Bacteriology and Nematology Unit, SCRI, tomato ®-expansinsshowed that six expansins ( Le-Exp1, Invergowrie,Dundee DD2 5DA,UK -3,-4, -5, -9 and -18) were expressedin or close to ¤[email protected] thesyncytium, 10 and 14 days after inoculation. Le- Exp5was furtheranalysed by in situ RT-PCR.AmpliŽ ed Globoderapallida and G. mexicana aretwo closely tomatoexpansin 5 cDNA couldonly be localised in the relatedspecies that are able to matebut usually develop on cytoplasmof syncytial cells but not in the surrounding differentsolanaceous plants. IdentiŽ cation of nematode roottissue. This indicates that tomato expansin 5 is genesinvolved in parasitism is one of the most critical speciŽcally expressed in syncytial cells and presumably stepsleading to the elucidation of disease resistance playsan important role in the development of syncytia mechanismsin plants. In this study, we haveused the inducedby cyst nematodes. SuppressionSubtractive Hybridisation (SSH) technique toinvestigate differences between the transcriptomes of G. pallida and G. mexicana J2.None of the cDNA 275 Arabidopsisthaliana expansingenes are fragmentsisolated in the SSH experimentsappeared expressedin syncytiainduced by tobe completely absent from theother transcriptome, Heteroderaschachtii butdifferences in expression levels of some cDNAs betweenthe two species were conŽrmed in reverse B. GOLECKI1; , K. WIECZOREK 2, D.J. COSGROVE 3 ¤ northernexperiments. Sequence analysis revealed that a and F.M.W. GRUNDLER 2 highproportion of the cloned sequences were pioneer 1Institutfü r Phytopathologie,Universitä tKiel, 24098 Kiel, genesfor whichno putative homologues were present Germany inthe databases. However, homologues of a cellulase 2 Institutfü r Panzenschutz, Universitä tfü r Bodenkultur, anda putativepathogenicity factor previously described Vienna,Austria in G.rostochiensis were isolated.cDNAs corresponding

Vol.4(2), 2002 219 Parasitism(265-294) tothese sequences were thereforecompared between G. 2 Departmentof Botanyand Plant Sciences, and Department of pallida and G. mexicana. Nematology,University of California,Riverside, CA 92521, USA ¤[email protected] 277IdentiŽ cation of putativeparasitism genes expressedin the pharyngealgland cells of Theinitiation and formation of feedingsites in plantroots, Meloidogyneincognita inducedby root-knotand cyst nematodes, involve the re- activationof cellcycle genes, resulting in multinucleated 1; 1 2 Guozhong HUANG ¤, Bingli GAO , Tom MAIER , giantcells and syncytia. The feeding site serves as the Eric DAVIS 3, Thomas BAUM 2 andRichard H USSEY 1 nutrientsource for thedevelopment and reproduction ofthe nematode. In order to extend our understanding 1Departmentof Plant P athology,University of Georgia,Athens, ofthe interaction between nematodes and plants during GA30602,USA theinfection process, we haveexamined expression of 2 Departmentof PlantP athology,Iowa State University, Ames, the Arabidopsis PROLIFERA (PRL) geneduring the IA50011,USA developmentof nematode feeding sites (NFS). PRLis 3 Departmentof Plant P athology,North Carolina State expressedin dividing cells throughout plant development University,Raleigh, NC 27695,USA andencodes an MCM proteinthought to function in the [email protected] ¤ initiationof DNA replicationduring S phase.W ewill presentthe expression pattern of PRL during formation Cloningparasitism genes coding proteins secreted from ofbothgiant cells and syncytia in the root of A. thaliana thepharyngeal gland cells and injected through the stylet seedlings. intoplant tissue is the key to understandingthe molecular basisof nematode parasitism of plants. Microaspiration ofthe cytoplasm from thepharyngeal gland cells of 43 279Direct identiŽ cation of styletsecreted proteins parasiticstages of M.incognita representingdifferent fromroot-knot nematodes by a proteomicapproach timepoints in the parasitic cycle provided expressed 1; 2 mRNA toconstruct a glandcell-speciŽ c cDNA library Stéphanie JAUBERT ¤,TerenceNeil L EDGER , 1 1 bylong-distance-PCR. Of 1200cDNA clonessequenced, JeanBaptiste L AFFAIRE , Pierre ABAD 1 deducedprotein sequences of 141 cDNAs hadan N- andMarie-Noë lle R OSSO terminalsignal peptide for secretionand, therefore, could 1INRA, UnitéInteractions Plantes-Microorganismes et Santé haveroles in root-knot nematode parasitism of plants. Végé tale, 123 Bd F .MeillandBP2078, 60606 Antibes Cedex, Of 101cDNA clonesencoding signal peptides tested France byhigh-throughpu t in situ hybridisation,probes of 21 2 Laboratoirede Pharmacologie T oxicologieINRA 180ch. de uniqueclones speciŽ cally hybridised to transcriptswithin Tournefeuille31300 T oulouse,France thesubventral (seven clones) or dorsal(14 clones) gland ¤[email protected] cells.PSORT IIpredicted18 of the deduced proteins Thestylet secretions of sedentary plant-parasitic nema- tobe extracellular and three as nuclear localised. In todesare thought to be major pathogenicity factors in- BLASTpanalyses, 19 of the predicted proteins were volvedin host invasion and feeding site induction and novelproteins. Those with similarities to knownproteins maintenance.A procedurewas developedthat allowed includeda transcriptionfactor and an avirulenceprotein. thedirect qualitative analysis of proteinssecreted by Me- Noneof the21 gland-expressedgenes had homologues in loidogyneincognita infectivejuveniles. After stimulation Caenorhabditiselegans . insemi-sterileconditions, the secreted proteins whose pI rangedfrom 5.0to 7.5were separatedby 2Delectrophore- 278Expression of Arabidopsisthaliana gene sisand the seven most abundant proteins were identiŽed PROLIFERA innematode-inducedfeeding sites bymicro-sequencing. A proteinhighly homologous to calreticulinswas identiŽed. The peptide sequences were 1; 2 Xiang HUANG ¤, Patricia SPRINGER and usedto design degenerate oligonucleotide sthatallowed Isgouhi KALOSHIAN 2 thecloningof thecorresponding Mi-crt cDNA. Transcrip- tion of Mi-crt ininfective juveniles and adults was demon- 1BASF PlantScience L.L.C., 26DavisDrive, Research strated.Expression of the calreticulin in the subventral TriangleP ark,NC 27709,USA pharyngealglands of infective juveniles was evidenced,

220 Nematology Postersessions suggestingthat calreticulin is a componentof naturally Laboratoryof Nematology,Department of Plant Science, producedstylet secretions and supporting the validity of WageningenUniversity, Binnenhaven 5, 6709 PD Wageningen, thepuriŽ cation procedure. Calreticulin is acalciumbind- TheNetherlands ingprotein involved in multiple cellular functions. It is ¤[email protected] secretedby severalanimal-parasitic nematodes and trema- Earlynodulin ( ENOD)geneshave been deŽ ned as genes todesduring parasitism. expressedin legumes during nodule formation. Recent studies,however, revealed a numberof homologues of 280IdentiŽ cation and expression analysis of pectin ENOD40sinnon-legume plants. Both in legumes and degradingenzymes from the root-knotnematode non-legumes ENOD40sarethought to play a rolein the Meloidogyneincognita developmentof vascular tissues. In situ localisationof ENOD40 transcriptsin root-knotnematode-infected roots Stéphanie JAUBERT¤,Jean-BaptisteL AFFAIRE, ofthelegume Medicagotruncatula revealedthat this plant Pierre ABAD andMarie-Noë lle R OSSO geneis expressed in and around giant cells at 6 days postinfection (dpi). T otestthe expression of ENOD40 INRA, UnitéInteractions Plantes-Microorganismes et Santé ina non-legumeupon infection with root-knot and Végé tale, 123 Bd F .MeillandBP2078, 60606 Antibes Cedex, cystnematodes, the expression of endogenous ENOD40 France promoter-gusA fusionwas studiedin nematode-infected ¤[email protected] tomatoroots at 1, 3,7and14 dpi.In the main root, GUS- stainedareas were irregularlyalternated with unstained Theestablishment of sedentary parasitic root-knot areas.The lateral roots showed staining in the root-tip. nematodesin ahostroot is dependenton theirefŽ ciency Occasionally,GUS stainingwas observedin syncytiaand ininvading the root tissue and in inducing the formation giantcells. Sections from bothGUS stainedand non- ofa permanentfeeding site. Several works have brought stainedfeeding sites were investigatedmicroscopically, evidencethat stylet secretions injected in the root tissue andthe results conŽ rmed our macroscopic observations. duringparasitism are involved in those two phases of Hence, ENOD40 expressionin tomato is basically parasitism.Secreted enzymes involved in the invasion unaffectedby the induction of feedingsites by root-knot phasecan be identiŽ ed by a candidategene approach. andcyst nematodes. Thisstrategy has previously allowed the identiŽ cation ofcellulolytic endoglucanases secreted by Meloidogyne 282Molecular cloning of chitinasegenes from incognita.Inthis study, pectin degrading enzymes were the pinewood nematode, Bursaphelenchusxylophilus identiŽed by analysing EST obtainedfrom M.incognita infectivejuveniles. T wopectate lyase ( Mi-pel1 and Mi- Taisei KIKUCHI¤ and Takuya AIKAWA pel2)andone polygalacturona se( Mi-pg)fulllength cDNAs were isolated.The deduced proteins contain a Forestryand F orestProducts Research Institute, Tsukuba predictedsecretion signal sequence. Transcription of the Ibaraki305-8687, Japan [email protected] threegenes was evidencedin pre-parasitic juveniles, ¤ sedentaryfemales and males by RT -PCR.In infective Chitinasefunction in nematodes, other than during juvenilesthe transcripts are localised in the subventral egghatch, has been the subject of interest in recent pharyngealglands. Enzyme activities were analysedby years,particularly in animal-parasitic nematodes. In this in vitro tests.This study demonstrates that root-knot studywe describethe cloning of chitinase genes from nematodesuse a batteryof cellwall degrading enzymes thepine wood nematode, Bursaphelenchusxylophilus , duringparasitism. whichis mycetophagous and phytophagous, the causal agentof pine wilt and vectored between trees by 281Expression pattern of ENOD40 intomatoroots insects.Degenerate primers derived from theconserved upon infectionwith root-knot( Meloidogyneincognita ) regionsof known chitinase sequences were usedto and cyst (Globoderarostochiensis ) nematodes amplifyfragments of putative B.xylophilus chitinase genes.Thus, two distinct fragments that encode chitinase Aneta KARCZMAREK¤, Hein OVERMANS, sequenceswere obtained.Using these fragments as Jaap BAKKER, Johannes HELDER and Aska GOVERSE probes,corresponding cDNA cloneswere identiŽed from

Vol.4(2), 2002 221 Parasitism(265-294) ourmixed-stage cDNA libraryand designated as Bx-cht- Togaininsight into how plant pathogenic nematodes par- 1 and Bx-cht-2.Sequence analysis revealed that Bx-cht-1 asitiseplants, we havebeen isolating and characterising and Bx-cht-2genes putatively encode proteins composed nematodegenes that are expressed in the pharyngeal re- of371 amino acids and 567 amino acids, respectively. gionof the root-knot nematode, Meloidogynejavanica . Thededuced amino acid sequences of these proteins Onepharyngeal gland gene we hadpreviouslycloned was showedsigniŽ cant homologies to chitinases of other determinedto be a chorismatemutase and was named nematodes.Our furtheranalyses aim at identifying the MjCM-1 (Meloidogynej avanica chorismate mutase). We timingand location of the chitinase genes expression in determinedthis enzyme was secretedfrom thenematode thenematode. asitwas initiatingits giantfeeding cells. W ehypothesised that MjCM-1 ismanipulatingthe shikimatepathway in the 283Novel pectate lyase gene from Globodera plantto thebeneŽ t ofthenematode. Since the shikimate rostochiensis pathwayin plants produces metabolites that regulate plant developmentand plant defence, we hypothesised MjCM- Urszula KUDLA¤, Ling QIN and Geert SMANT 1 maybe altering one of these processes. T otestthis hypothesiswe haveexpressed MjCM-1 insoybean hairy Laboratoryof Nematology, Department of PlantScience, rootsusing a dexamethasoneinducible plant expression WageningenUniversity, Binnenhaven 5, 6709PD Wageningen, vector.Soybean hairy roots treated with dexamethasone TheNetherlands express MjCM-1 andthe roots exhibit a distinctlack of [email protected] ¤ lateralroots. This result indicates that MjCM-1 can alter Anovelcell wall degrading enzyme was identiŽed plantdevelopment. Metabolic proŽ ling experiments are asa partof cDNA-AFLP projectthat compared gene beingconducted on the MjCM-1 expressingroots to de- expressionproŽ les in Ž vedifferent stages of Globodera termineexactly which compounds are altered by thisne- rostochiensis .OneTranscript Derived Fragment (TDF), matodeenzyme. predominantlyexpressed in infective second stage juveniles,encoded a partialopen reading frame showing 285Histochemical localisation of activeoxygen highsimilarity to bacterial and fungal pectate lyases species in Glycine max and Arabidopsisthaliana from thepolysaccharide lyase 3 family(EC 4.2.2.2). infected with the cystnematodes Heteroderaglycines Thecorresponding full-length cDNA designated Gr-Pel2 and H.schachtii harboursan open reading frame of759 bp, encoding polypeptidewith molecular mass 27kDa. Gr-Pel2 is Kristen A. LENNON¤ andThomas J. B AUM signiŽcantly different from previouslyidentiŽ ed Gr- Departmentof Plant P athology,351 Bessey Hall, Iowa State Pel1 (30%identities and 46% similarities). Sequence University,Ames, IA 50011,USA was predictedto have signal peptide for secretionat [email protected] itsN-terminal end. Digoxigenin-labe lledDNA probe ¤ hybridisedspeciŽ cally to the subventral pharyngeal Theproduction of Active Oxygen Species (AOS) glands,which are responsible for secretingall currently isconsidered to be an initial indicator and key knowncell wall degrading enzymes. Heterologous componentof theplant defence response. AOS havebeen expressionof Gr-Pel2 in Pichiapastoris isinprogress,in implicatedin direct pathogen inhibition through cell- orderto assess its biochemical properties. IdentiŽ cation wallmodiŽ cation, necrosis, direct pathogen mortality, ofanotherpectate lyase gene shows the complexity of the signaltransduction leading to defence-related gene pectinolyticsystem in nematodesecretions. expression,and programmed cell-death. The distribution androle(s) ofAOS inplant-nematode interactions have yetto be well characterised. W euseda histochemical 284Analysis of root-knotnematode chorismate approachto study the distribution of AOSinsusceptible mutaseexpressed in soybeanhairy roots andresistant interactions between Glycinesmax and Heteroderaglycines and in Arabidopsisthaliana infected Kris N. LAMBERT , E. DOYLE and J. PAINTER ¤ with H. glycines or H.schachtii . In roots of G. Departmentof Crop Sciences, University of Illinois,Urbana IL max,AOSwere associatedwith the head of the 61801, USA migratingnematode in the susceptible interaction but ¤[email protected] notin the resistant interaction. A OSwere alsonot

222 Nematology Postersessions detectedin either tissue once the nematode had initiated 287Selection for virulence in populationsof syncytiumformation. In Arabidopsis roots,AOS were Globoderapallida associatedwith nematode infection sites and possiblywith developingsyncytia. These observations were especially Mark S. PHILLIPS¤, Roy NEILSON andV ivianC. B LOK evidentin the interaction between A. thaliana and H. ScottishCrop Research Institute, Invergowrie, Dundee DD2 schachtii. Arabidopsis rootsinfected with H. glycines 5DA,Scotland, UK exhibitedsimilar staining patterns, but the incidence of ¤[email protected] nematodeinfection and feeding was lower.Patches of stainingnot directly associated with feeding nematodes Fourpopulations of Globoderapallida were reared were alsoobserved. continuouslyon hostswith resistance derived either from Solanumvernei or S.tuberosum ssp. andigena CPC 2802. After 12generations these populations were assessed 286T wonovel Arabidopsis promoterswith distinct biologicallyto determine if there was anyincrease in tissue-speciŽcities are responsive to cyst nematode virulence.Bulk DNA derivedfrom thesepopulations was infection examinedusing AFLPs toassess the effects of selection 1; 1 oninter-populationvariation. Additionally, from asubset Mitra MAZAREI ¤,KristenA. L ENNON , Steven R. RODERMEL 2 andThomas J. B AUM 1 ofthematerial, DNA was derivedfrom individualsecond stagejuveniles and was investigatedusing microsatellites 1Departmentof Plant P athology,351 Bessey Hall, Iowa State toexamine the effects of selection on inter- and intra- University,Ames, IA 50011,USA populationvariation. Biologically, selection resulted in 2 Departmentof Botany,Iowa State University, Bessey Hall, increasedvirulence, but the increase varied depending on Ames,IA 50011,USA theinitial population and the clone used. Increases in [email protected] ¤ virulencetended to bespeciŽc tothesource of resistance. Wepreviouslyisolated partial soybean cDNA clones Molecularassessments showed an effect on the genetic whosecorresponding transcript abundances are elevated constitutionshowing differences between the unselected 1dayafter infection with the soybean cyst nematode, andselected populations but showing no detectable Heteroderaglycines .Following,we isolatedfull-length associationbetween polymorphic markers and virulence. cDNAs for twoof these clones, designated Gm10.1 and Themicrosatellite data yielded allele frequencies that Gm17.1.RNA blothybridisations revealed that Gm10.1 providedevidence that selection pressures had occurred in and Gm17.1 mRNAs accumulatedin a root-preferential allfour populations. Furthermore, a principalcoordinate manner,and that their corresponding steady-state mRNA analysisderived from dissimilarityvalues based on levelsincreased over time following H. glycines infection. microsatellitedata separated the four populations into Thepredicted Gm10.1 and Gm17.1 geneproducts each twohost-speciŽ c groupingsthat were differentfrom the sharedsimilarities with the protein products of distinct original G. pallida parentpopulation. Arabidopsisthaliana geneswith unknown functions. Promotersof these Arabidopsis genes,designated At10.1 288Expression proŽ ling of the Arabidopsis -cyst and At17.1,were fusedto the GUS reportergene and nematodeinteraction transformedinto soybean hairy roots and Arabidopsis 1; 2 plants.Activity of the At10.1 promoterwas detectedin David P. PUTHOFF ¤, Dan S. NETTLETON , rootand shoot apical meristems of Arabidopsis and in Steven R. RODERMEL 3 andThomas J. B AUM 1 alltissues of soybean hairy roots. The At17.1 promoter 1 alsodirected GUS expressionto the root meristems Departmentof PlantP athology,351 Bessey Hall, Iowa State University,Ames, IA 50011,USA butadditionally was detectedin the vascular tissues of 2 Departmentof Statistics, 124 Snedecor Hall Arabidopsis At10.1 both andsoybean. Furthermore, the 3 Departmentof Botany,353 Bessey Hall, Iowa State promoterwas responsiveto infection of Arabidopsis University,Ames, IA 50011,USA by Heteroderaschachtii ,whereasthe At17.1 promoter ¤[email protected] directedincreased GUS expressionto the infection sites of H. glycines insoybeanhairy roots. Cystnematodes infect and devastate many agricultural crops.After rootpenetration, cyst nematodes induce the formationof feedingcells (syncytia) within the host roots.

Vol.4(2), 2002 223 Parasitism(265-294)

Thesesyncytia are the sole source of nutritionfor thene- version.ESTScreen searches each cDNA sequencefor matodes.Syncytium formation is accompaniedand likely a(user-deŽned) start codon, translates the sequence, mediatedby nematode-inducedchanges in plant gene ex- submitsautomatically to SignalPand retrieves the output. pression.With the availability of microarraytechnology, ESTScreenthen parses the SignalP prediction through mRNA expressionchanges of thousands of genes can auser-deŽned logical test and generates a FASTAŽle bemonitored simultaneously in order to determine the containingonly EST encodinga signalpeptide for mechanismsof biologicalprocesses-of-interest. W ehave secretion,which can be used in homology search and usedAffymetrix GeneChip technology to monitor Ara- othersubsequent analysis. This program can be a useful bidopsis mRNA changesfollowing infection by two dif- toolin the study of plant-pathogen interaction systems ferentcyst nematode species. The sugar beet cyst nema- aswell as other Ž eldsin which secreted molecules play tode (Heteroderaschachtii )readilyinfects Arabidopsis importantroles. The authors wish to thank Dr. S.Kamoun andinduces syncytia. In contrast, the soybean cyst nema- for communicatingunpublished results. tode (H. glycines)onlyrarely induces syncytium forma- tion in Arabidopsis rootswhile it retains the pre-feeding 290IdentiŽ cation of parasitismgenes from behavioursof probingand penetration. Our analysesun- root-knotnematode Meloidogyneincognita using covereda largepanel of Arabidopsis geneswith altered cDNA-AFLP,EST-analysisand GenEST mRNA levelsfollowing cyst nematode attack. Some of thesechanges were observedduring infection with both Erwin ROZE¤, Ling QIN, Jaap BAKKER and cystnematode species, suggesting a potentialinvolvement Geert SMANT inplant stress or defencefunctions. Other mRNA changes were speciŽc for H.schachtii infectiononly and, there- Laboratoryfor Nematology WUR, Binnenhaven5, 6709PD fore,are good candidates for beinginvolved in syncytium Wageningen,The Netherlands formationor function. ¤[email protected] Theroot-knot nematode Meloidogyneincognita is an 289ESTScreen: identify secretory proteins in silico obligatoryplant parasite, which has evolved a complex feedingrelationship with its host plants. Pharyngeal gland 1; 2 1 Ling QIN ¤, Theo J.A. BORM and Jaap BAKKER secretionsplay an important role in the parasitic life 1Laboratoryof Nematology, Department of PlantScience, cycle of Meloidogyne spp.They are involved in migration WageningenUniversity, Binnenhaven 5, 6709PD Wageningen, throughthe root system and induction and maintenance TheNetherlands ofthe giant cells. The cDNA-AFLP methodis adopted 2 Laboratoryof PlantBreeding, Department of PlantScience, toprovide more insight in parasitism genes involved in WageningenUniversity, Binnenhaven 5, 6709PD Wageningen, theplant-nematode interaction. Gene expression proŽ les TheNetherlands of various M.incognita stagesdiffering in pharyngeal ¤[email protected] glandactivity ( e.g.,eggs,pre-parasitic and parasitic juveniles,adult females) are being compared in acDNA- Theever increasing gene sequences in databases offer AFLP procedurebased on the restriction enzymes ApoI unprecedentedopportunity for biologiststo exploit this and TaqI.GenesspeciŽ cally expressed in the parasitic newinformation source to Ž ndanswers tovarious stagesmay represent candidate parasitism genes and biologicalquestions. The collection of agreatnumber of willbe isolated from gel.The DNA sequencesof EST (expressedsequence tags) is a usefulstarting place theisolated fragments will be compared with the M. for identifyingsecretory proteins involved in nematode- incognita EST databaseusing the computer program plantinteractions, either in the form ofpathogenicity GenEST.Furthermore, the spatial expression pattern of factorsor avirulent proteins which are targeted by thecandidates will be analysedusing whole mount in situ hostdefence mechanisms. SignalP is a robustprogram hybridisation. generallyused to identify signal peptides for secretion from proteinsequences, but it is not suitable for analysinga largenumber of EST directly.T othis 291Localisation of expressionof Le-Exp5gene in end,we havedeveloped and validated a softwaretool, feedingsites induced by Globoderarostochiensis in ESTScreen,which automates screening of thousands of rootsof susceptibletomato using in situ RT-PCR EST eitherthrough the SignalPweb server or astandalone method

224 Nematology Postersessions

1; 1 2 M. SOBCZAK ¤, S. FUDALI , B. GOLECKI , sophisticatedtransformation. The key to understand this S. JANAKOWSKI 1, G. GRYMASZEWSKA 1, W. KUREK 1, form ofparasitismis thecharacterisation of theseproteins. M. LICHOCKA 1, F. GRUNDLER 3 and W. GOLINOWSKI 1 However,the small size of plant-parasitic nematodes makesdirect analysis of their secretions extremely 1Departmentof Botany,W arsawAgricultural University, Rakowiecka26/ 30,W arsaw02528, P oland difŽcult. W epresenta methodto isolate nematode 2 Instituteof Phytopathology, Christian-Albrechts University, secretionsin sufŽcient quantities to allow direct analysis Kiel,Germany bycombining two-dimensional gel electrophoresis and 3 Departmentof PlantProtection, University of Agricultural mass-spectrometry.So far, we havebeen able to identify Sciences,V ienna,Austria severalproteins (endoglucanases and pectatelyase) which ¤[email protected] arebelieved to be involved in the invasion of the plant tissueby the nematode. Other spots from thetwo- Theexpression pattern of the tomato expansin gene Le- dimensionalgels are under investigation. This knowledge Exp5was examinedon resinand agar embedded sections willhelp us toelucidate one ofthemost fascinating forms ofsyncytiainduced by Ro1pathotype of G.rostochiensis ofparasitic behaviour between two completely different inroots of susceptible tomato cv. Moneymaker. In 1 organisms,namely a plantand an animal. and3 dayold syncytia, strong expression of Le-Exp5 was foundin cells incorporated into syncytium and in neighbouringcells. In 7 dayold syncytia, the older parts 293Cloning genes expressed in the pharyngeal ofsyncytia contained relatively less Le-Exp5 mRNA glandcells of Heteroderaglycines usingan ampliŽed thancells situated at distal parts of syncytia that had RNA technique beenrecently incorporated into it. Strong expression of 1; 2 3 Le-Exp5took place also in proliferating and dividing Xiaohong WANG ¤, Tom MAIER , Richard HUSSEY , 2 1 parenchymatouscells surrounding syncytia. These results Thomas BAUM and Eric DAVIS were gainedin experiments where in situ RT-PCR was 1 followed by in situ hybridisationwhich conŽ rmed the RT- Departmentof PlantP athology,Campus Box 7616, North PCR data. CarolinaState University, Raleigh, NC, 27695,USA 2 Departmentof Plant P athology,Iowa State University, Ames, IA, USA 292Micropreparative sampling and molecular 3 Departmentof PlantP athology,University of Georgia, characterisationof proteinssecreted by Athens,GA, USA the plant-parasiticnematode Heteroderaschachtii ¤[email protected]

1; 2 3 B. VANHOLME ¤, J. DE MEUTTER , T. TYTGAT , mRNA withinthe cytoplasm microaspirated from the 1 2 G.D.C. GHEYSEN , I. VANHOUTTE and pharyngealgland cells of parasitic stages of Heterodera 1 G.D.R. GHEYSEN glycines was immobilisedon paramagnetic beads for 1VakgroepMoleculaire Biotechnologie, F aculteit initialcDNA synthesis.T3 and T7 promoters were Landbouwkundigeen ToegepasteBiologische W etenschappen, adaptedto thegland cell cDNA andused to driveseveral UniversiteitGent, Coupure links 653, B-9000 Gent, Belgium rounds of in vitro transcription.The ampliŽ ed RNA pool 2 VIB, DepartementPlantengenetica, F aculteitW etenschappen, was usedfor cDNA synthesisand directional cloning UniversiteitGent, K.L. Ledeganckstraat35, B-9000 Gent, intoa plasmidvector for constructionof a H. glycines Belgium gland-cellcDNA librarywithout using PCR. Analysis 3 Instituutvoor Dierkunde, F aculteitW etenschappen, of90 cDNA cloneschosen at random from thelibrary UniversiteitGent, K.L. Ledeganckstraat35, B-9000 Gent, produced67 uniquecDNA sequences.Among the 67 H. Belgium glycines cDNA sequenceswere homologuesto a number [email protected] ¤ ofdifferenteukaryotic genes, including genes identiŽed in Sedentaryplant-parasitic nematodes such as Heterodera animal-parasiticnematodes and genes with potential roles schachtii haveevolved a highlycomplex relationship inhost-parasiteinteractions. The presence of apredicted withtheir host plant by redifferentiating root cells secretionsignal peptide and conŽ rmation of expressionin intospecialised feeding sites. Proteins, produced in the H. glycines pharyngealgland cells are being investigated pharyngealglands of the nematode and injected into for theseand additional clones being sequenced from the theplant cell, are thought to play a keyrole in this ampliŽed RNA H. glycines gland-cellcDNA library.

Vol.4(2), 2002 225 Parasitism(265-294)

294SSH enrichedcDNA librariesto compare thepathogen and the host species respectively. This virulentand avirulent isolates of the quarantined hasimportant implications for thesuccessful utilisation root-knotnematode Meloidogynechitwoodi ofresistance from S.bulbocastanum . Roots of S. bulbocastanum were infestedwith two isolates of M. Jane WISHART¤, Alison PATERSON, Mark S. PHILLIPS chitwoodi varyingin virulence. Plants were propagated andVivian C. B LOK intissue culture and then transferred to soil before ScottishCrop Research Institute, Invergowrie, Dundee DD2 inoculating.The infection process was monitoredby 5DA,Scotland, UK histologicalexamination of roots allowing time points ¤[email protected] tobe selected for RNA extraction.cDNA libraries Innorth-western Europe, root-knot nematodes are a se- were constructedfrom infectedroot tissue using the riousthreat for potatoproduction particularly with re- SuppressiveSubtractive Hybridisation (SSH) method whichenriches for rare transcripts.This enabled the duceduse of nematicides. Meloidogynechitwoodi causes constructionof libraries,enriched in transcriptsfrom the signiŽcant economic losses due to reduction in tu- berquality and yield of potato crops. Resistance to compatibleand incompatible interactions, at 3 and7 days M.chitwoodi hasbeen identiŽ ed in the wild tuber- post-infection.Both plant and nematode genes, which bearing Solanum species, S.bulbocastanum. Threepatho- maybe important during the host/ parasiteinteraction, typeswere identiŽed suggesting at least two differ- were identiŽed. entgenetic factors for virulenceand resistance in

226 Nematology Postersessions

295Resistance to Meloidogyneincognita in coffee: theirputative nematicidal effect. Three tropical nematode identiŽcation of molecularmarkers F-AFLP in species (Radopholussimilis , Pratylenchuscoffeae and resistantgenotypes Meloidogyneincognita )couldsuccessfully penetrate and develop in A. thaliana undermonoxenic conditions. Eight AnaPaula de AndradeA UKAR¤ and to10 weeks after inoculation reproduction ratios of ElianaGertrude Macedo L EMOS respectively16.3, 11.9 and 24 could be observed. This in vitro UNESP,Campusde Jaboticabal, Departamento de Tecnologia, systemenables the preselection of genesor gene Viade acessoProf. P auloDonato Castelani KM 5,Jaboticabal, combinationsshowing the greatest potential for nematode SP14870-000,Brazil control.Transgenic Arabidopsis plants,expressing lectins ¤[email protected] orlectin-related proteins in their roots, have already beenscreened. Results on nematode penetration and Thisstudy is about a diagnosticsystem based on multiplicationin thesetransgenic plants will be presented. molecularmarkers thatco-segregate within a geneof interest.The chosen technique was f-AFLP duetoits great powerof detectionof geneticvariability, being a dominant 297Marker-assisted selection in screeningpeanut markerefŽ cient for largeand simultaneous sampling of a (Arachishypogaea L.)forresistance to the peanut genome.A totalof 16primerscombination ECO-RI ( 3) root-knotnematode C and MSE-I ( 3)were usedto study polymorphism within 1 2; C Valeria CARPENTIERI-PIPOLO , Don W. DICKSON ¤, apopulationof 40 plants F5 (H 47-82-7-925),donated Maria GALLO-MEAGHER 3 and Dan W. GORBET 4 byInstituto Agronomico de Campinas– IAC, segregating Meloidogyneincognita for theresistance to nematode in 1 coffeetree. One hundred and twenty eight segregating loci UniversidadeEstadual de Londrina, Depto Agronomia, C.P.6001,86051-990, Londrina PR, Brazil were obtainedwith Mendelian inheritance by selecting 2Universityof Florida,P .O.Box110620, Gainesville, FL onlymarkers that segregated 1:1. These markers were 32611, USA submittedto a comparativeanalyses between the donor 3 Universityof Florida, P .O.Box11030, Gainesville, FL 32611, ofthe resistance factor of thisprogeny ( Coffeacanephora USA duplicated– C1330)and the plant group that is resistant 4 Universityof Florida, REC, Hwy 71,Marianna, FL 32446, andsusceptible (‘ bulk’resistant and ‘ bulk’susceptible). USA ¤[email protected].edu 296Nematode control by expression of lectinsin transgenicplants Arestrictionfragment length polymorphism (RFLP) markerlinked to a locusfor resistanceto Meloidogyne 1; 1 Katrien CARLENS ¤, Annemie ELSEN , arenaria race1, along with visual evaluation following Els VAN DAMME 2, Laszlo SAGI 1, Dirk DE WAELE 1 rootstaining were usedto screen four breeding and Rony SWENNEN 1 populationsand three lines of peanut in a root-knot nematodeinfested Ž eld.Coan and Florunner cultivars 1 Laboratoryof Tropical Crop Improvement, Kasteelpark were usedas the resistant and susceptible parental Arenberg13, B-3001 Leuven, Belgium controls,respectively. Genomic DNA was isolatedfrom 2 Laboratoryfor Phytopathology and Plant Protection, Willem youngleaves of these plants during the growing season, DeCroylaan 42, B-3001 Leuven, Belgium [email protected] andSouthernblot analysis was conductedusing the RFLP ¤ probeR2430E. Only line T301-1-8 was homozygous Withthe adventof transgenicmethodologies, an attractive for theresistance marker .At harvest,root systems methodto control plant parasitic nematodes is thetransfer were stainedwith Phloxine B, eggmasses counted, intoplant host species of genes encoding nematicidal andresistance phenotype scored. Field tests conŽ rmed proteins.Lectins or lectin-related proteins, for example, theRFLP markerresults. Except for T301-1-8,all arebelieved to have toxic and/ orrepellent effects on othergenotypes displayed high levels of nematode nematodes.As agoodhost for migratoryand sedentary reproduction.The RFLP probeR2430E loci linked nematodesand being considerably easier to transform tonematode resistance provided a usefulselection thanmost plants, Arabidopsisthaliana canbe usedas an methodfor identifyingresistance to thepeanut root-knot in vitro systemto rapidlyexpress transgenes and evaluate nematode.

Vol.4(2), 2002 227 Resistance,breeding (295-335)

298Understanding the geneticand molecular basis totalof four primer combinations were applied,yield- of(a)virulence in the root-knotnematode ing275 identiŽ able marker bands. Analysis of the iden- Meloidogyneincognita tiŽed marker bands was thencompleted using a hierar- chicalcluster analysis and a dendrogramproduced. The Philippe CASTAGNONE-SERENO dendrogramwas thenused to identify 16 genetically di- INRA UnitéIPMSV ,BP2078,06606 Antibes cé dex, France versespecies for furtheranalysis. These species have been [email protected] testedagainst a rangeof G. pallida populations.Histolog- icalanalysis of the resistance mechanisms involved has Usingtwo pairs of near-isogenic M.incognita lines, alsobeen conducted, focusing on the effect of the wild avirulentand virulent against the tomato Mi resistance specieson hatch, invasion, development in the root and gene,a cDNA-AFLP differentialanalysis was initiated,in productionof eggsper cyst. orderto identify genes differentially expressed between avirulentand virulent nematodes. AFLP markerswere developedon cDNA templatesdigested with HindIII and 300Host-plant responseof some Musa cultivars MseI,withall of the 256 combinations that could be fromBrazil to different speciesof root-knot generatedwith primers with two selective nucleotides. A nematodes (Meloidogyne spp.) totalof approximately84 000fragments were ampliŽed, E. COFCEWICZ 1, R.M. CARNEIRO 2, P. QUÉNÉHERVÉ 3 amongwhich 61 appeareddifferential ( i.e.,presentin the and J.L. FARIA 1 twoavirulent lines and absent in the two virulent lines). Inorder to conŽ rm thisresult, cDNA-AFLP was runagain 1 UniversityF ederalde Pelotas,C.P .354,CEP 96001-970, onthe same templates, but with all of the64combinations Pelotas,RS, Brazil thatcould be generated with primers with two selective 2 EMBRAPA,CP08223,CEP 70849-970,Brasilia, DF ,Brazil nucleotidesfor MseIandone selective nucleotide for 3 IRD, BP8006,97259 F ort-de-Francecedex, Martinique HindIII. Thisallowed us to eliminate a numberof false Musa positivebands, and 19 fragments differential between Five cultivarscommonly grown in Brazil, (triploid AAA-group,cvs Nanicã o andCaipira; triploid AAB- avirulentand virulent lines were thusselected for further group,cvs Prata and T erra;tetraploid AAAB-group, cv. analysis.The next experimental steps included cloning ofthe full-length genes, RT -PCRexperiments, etc. The Pioneira)were evaluatedfor theirsusceptibility to three Meloidogynejavanica M. in- resultswill be discussed in relation to the ability of the differentroot-knot species ( , cognita M. arenaria nematodeto overcome or notthe plant resistance gene. , )aloneand in combinations under glasshouseconditions. Four months after nematode in- oculation,plant growth parameters ( e.g.,fresh anddry 299Characterising sources of resistanceto weights,number of leaves and foliar area, macro- and Globoderapallida and G. rostochiensis identiŽed inthe micro-elementconcentrations), Ž nalnematode popula- Commonwealthpotato collection tions,reproductive factor and interspeciŽ c competition were analysed.All Musa cultivarswere foundto be hosts Lydia CASTELLI¤, Glenn BRYAN, Gavin RAMSAY, for thethree Meloidogyne species.The highest nema- Robbie WAUGH and Mark PHILLIPS todemultiplication was observedwith M.incognita on cv. ScottishCrop Research Institute, Invergowrie, Dundee, DD2 Prata.The cv. Caipira seemed the most susceptible to Me- 5DA,Scotland, UK loidogyne infestationin termsof plantgrowth parameters ¤[email protected] wheninoculated with a mixtureof M. javanica and M. in- cognita.Thecalcium content of Musa cultivarsis the only Previouslyuntested material from Professor JGHawkes’ macroelementwhich increased systematically when in- collectionof wild species of potatoes now incorporated festedwith root-knot nematodes ( e.g.,signiŽcant increase intothe CPC was screenedagainst the potato cyst nema- withcvs Prata, Caipira and Terra). Basedon their multi- todes.A totalof 56 and 53% of accessions were identi- plicationrates and on the Musa cultivar,some competition Žedas having resistance to G. pallida and G.rostochien- betweenroot-knot nematode species was observed. sis, respectively,from 198accessions. Molecular analy- sisof theresistant accessions identiŽ ed was conductedto observethe genetic relatedness and diversity of the ma- 301Evaluation of beansand cowpea genotypes for terial,using the AFLP DNA Žngerprintingtechnique. A resistanceto Meloidogynejavanica

228 Nematology Postersessions

Aline GODOY CRAVEIRO 1, João FLÁVIO VELOSO C.reticulatum were resistantto H. ciceri.Oneline each 1; 1 SILVA ¤,GeraldoE. SouzaC ARNEIRO , of C. bijugum, C.chorassanicum and C. judaicum, and Maria José DEL PELOSO 2 two each of C.pinnatiŽdum and C.reticulatum were andFrancisco Rodrigues F REIRE FILHO 3 resistantto M.artiellia .Sixlines of C. bijugum, four of C. cuneatum, 16 of C. judaicum and one of C.yamashitae 1 EmbrapaSoja were resistantto P. thornei. Because C.reticulatum is 2 EmbrapaArroz & Feijão crossablewith C.arietinum ,theseŽ ndingsindicate the 3 EmbrapaMeio Norte, CP 231,86001-970, Londrina, PR, Brazil possibilityof introgressionof resistanceto H. ciceri and M.artiellia intothe cultigen. ¤[email protected] Meloidogynejavanica isa majornematode pathogen in 303Characterisation and high-resolution genetic soybeanin Brazil.In many regions beans and cowpeas are mappingof root-knotnematode resistance genes in usedin rotation with soybeans, increasing the problem. pepper (Capsicumannuum L.): comparisonwith the Theuse of nematoderesistant cultivars in rotations with tomatoand potato nematode resistance gene locations soybeancan help reduce the damage. The reproduction of Meloidogynejavanica was studiedon 21beanand seven 1; 2 C. DJIAN-CAPORALINO ¤, V. LEFEBVRE , cowpeagenotypes. The plantswere grownon plastictubes A. PALLOIX 2 and P. ABAD 1 inglasshouse conditions, inoculated with 3000 nematode eggsand evaluated for eggproduction after 42 days. 1 INRA, UnitéInteractions Plantes Microorganismes et Santé Tomatoplants were usedas inoculum efŽ ciency check. Végé tale, 123 bd Francis Meilland, 06606 Antibes cedex, Thesame genotypes were grownin a M. javanica-infested France Želdand were evaluatedfor gallingindex. Despite 2 INRA, Unitéde Gé né tique et Amélioration des Fruits et hightolerance in Ž eldconditions, all cowpea genotypes Légumes, Domaine Saint Maurice, 84143 Montfavet cedex, permittedabundant nematode reproduction. The bean France genotypesAporé and Pot-51 were resistantwhereas the ¤[email protected] majoritywas susceptible. Inpepper, researches on genetic basis of root-knot nematoderesistance were conductedon threegenetically 302Resistance to Heteroderaciceri , Meloidogyne distantaccessions: PM687, PM217 and CM334. They artiellia and Pratylenchusthornei inwildspecies of revealedthree dominant and thermostableloci with broad- chickpea spectrumof resistance: Me3, Me1 and Me7,andtwo other M. DI VITO 1, N. GRECO 1 and R.S. MALHOTRA 2 loci: Mech1 (PM217) and Mech2 (CM334)controlling thequarantine nematode M.chitwoodi .Comparative 1 Istitutoper la Protezione delle Piante, Sezione di Bari, histologicalstudies showed that the three broad-spectrum C.N.R., ViaAmendola,165/ A70126Bari, Italy resistancegenes suppressed nematode reproduction, but 2 InternationalCenter for Agricultural Research in Dry Areas, inducedvery different response patterns in root cells Aleppo,Syria uponnematode infection. Bulked-segregant analysis was [email protected] performedto identifyAFLP markerslinked to thegenes. Chickpea (Cicerarietinum )isan important food legume Finemapping of Me3 gaveAFLP lessthan 0.5 cM throughoutthe world. Heteroderaciceri , Meloidogyne ankingthe gene. Indirect selection markers (SCAR artiellia and Pratylenchusthornei arenematodes known orCAPS) were identiŽed. Me3 locuswas localised toaffect the crop. Chickpea germplasm evaluation has onchromosome P9 on intraspeciŽ c peppermaps, in a revealedthe absence of resistance to these nematodes. syntenicregion of two other nematode resistance genes, Therefore,different lines belonging to eight wild Cicer the tomato Mi-3 andthe potato Gpa2 genes,which specieswere screenedfor theirreaction to a Syrian mappedon the short arm ofthe tomato and potato populationsof H. ciceri and P. thornei,andto an Italian chromosome12. W ealsoidentiŽ ed new AFLP markers populationof M.artiellia ina glasshouse.Out of 207, linkedin couplingto M.chitwoodi resistancegenes. The 154and 249 lines of Cicer spp.tested for theirreaction nearestwere locatedless than 1.7 cM from Mech1 (no against H. ciceri, M.artiellia and P. thornei,respectively, recombinantindividual for twomarkers), andat 1 cM 17 of C. bijugum, six of C.pinnatiŽdum , and Ž ve of from Mech2 (onerecombinant individual for onemarker).

Vol.4(2), 2002 229 Resistance,breeding (295-335)

304Comparison of two screeningmethods forearly ¤[email protected] evaluationof resistanceto Radopholussimilis in Musa germplasm Proteinsand small molecules within a soybeanroot atany given time depend on genes expressed up to Carine DOCHEZ 1, Danny COYNE 2, Michael PILLAY 1 thattime and are in uenced by the genotype and the and Dirk DE WAELE 3 environment.Earlier, we demonstratedreproducibility ofan extraction and chromatographic protocol used to 1 InternationalInstitute of TropicalAgriculture (IITA) – compareproŽ les from rootsexposed to either biotic or Easternand Southern Africa Regional Centre (ESARC), P.O. abioticstresses. W ehavechallenged cvs Williams 82 and Box7878, Kampala, Uganda 2 IITA –Ibadan,Nigeria, c/ oLambourn& Co.,Carolyn House, Hartwig,and our SCN resistantgermplasm PUSCN14 26Dingwall Road, Croydon, CR9 3EE, UK with Heteroderaglycines andanalysed the Ž ngerprints 3 KatholiekeUniversiteit Leuven, Laboratory of Tropical Crop atdifferent time intervals following inoculation. Gross Improvement,Kasteelpark Arenberg 13, 3001 Leuven, Belgium comparisonof the chromatographs showed considerable similaritybetween the inoculated and non-inoculated Plant-parasiticnematodes are one of themost important samplesof thesame cultivar. However, analyses of peak constraintsto sustainable Musa production.In the areas(four replications)showed statistically signiŽ cant lowlandsof CentralUganda, the most common nematode (P 0:05/ alterationsof speciŽc peakareas due tostress. D species is Radopholussimilis ,whileat higher elevations Allthree cultivars showed a transientincrease in the peak Pratylenchusgoodeyi ismore prevalent. The use of host 11area 2 daysafter inoculation but returned to non- plantresistance provides promising prospects towards stressedlevels at 4 daysafter inoculation. Interestingly, sustainablenematode management through conventional peak5 showedan increase in levels 4 daysafter breeding.T oidentifyresistance, reliable screening inoculation.Selected peaks were collectedand analysed methodsneed to be developed. In this study two bymass spectrometry. screeningmethods for resistanceto nematodes are compared.The Ž rst screeningmethod is based on theinoculation of individual roots placed in a cup. 306Chitin synthaseas moleculartarget in the Thesecond screening method is based on the classic plant-parasiticnematode Meloidogyneartiellia potexperiment, whereby the complete root system is Elena FANELLI1; , Mauro DI VITO 2, inoculated.The newly developed cup method requires ¤ MariaRosaria C ORTESE 1 and Carla DE GIORGI 1 lessnematode inoculum and less plant material per 1 cultivarcompared to thepot experiment. By usingsingle Universitàdegli Studi di Bari, via Orabona n ±4,70126 Bari, rootsfor inoculation,the host response to nematode Italy attackis not in uenced by differences in root growth 2 Istitutodi Protezione delle Piante CNR, viaAmendola 165/ A, ratesbetween Musa genotypes.Hybrids developed within 70126Bari, Italy thebanana breeding programme of the International ¤[email protected] Instituteof Tropical Agriculture are being evaluated for Plant-parasiticnematodes cause serious damage to resistanceagainst Radopholussimilis usingboth methods. agriculturalcrops. Molecular biology studies are focusing Nematodereproduction ratio is comparedagainst a known onplant-parasite interactions with the aim of Žndingthe resistantcheck (Y angambikm 5)anda knownsusceptible weakpoint that will permit nematode control. In spite check(V alery).The results of bothscreening methods will ofthis, several chemical compounds are still used as bediscussed. nematicidesthat are speciŽ c inhibitorsof thesynthesis of chitinwhich is present in theeggs of nematodes.Data in 305Proteomic analysis of soybeancyst nematode theliterature indicate successful applications of chemical (SCN) inoculatedsoybean roots compoundssuggesting that these may affect nematode-

1; 2 3 eggformation by interferingwith chitin deposition in the J. FAGHIHI ¤, R.A. VIERLING and V.R. FERRIS nematodeegg-shell. The prevention of chitin synthesis 1 Departmentof Entomology maybe triggered by an inhibition of the proteolitic 2 Departmentof Agronomy activationof chitin synthase zymogen or can be due to 3 Departmentof Entomology,Purdue University, W est adirecteffect on thechitin synthase. In this presentation, Lafayette,IN 47907,USA we showthat the eggs of M.artiellia docontainchitin and

230 Nematology Postersessions

havechitin synthetic activity. In our laboratory we have CarlosA. ArrabalA RIAS andSilvana R. M ARIN isolatedin the eggs of M.artiellia twodifferent portions EmbrapaSoja, CP231, 86001-970, Londrina, PR, Brazil ofchitinsynthase cDNA byRT -PCR.One of thecDNA ¤[email protected] containsthe motif QRRRW ,whichis demonstrated to beconserved in chitin synthase of different organisms. Meloidogynejavanica isa majornematode pathogen in Toprovethat the encoded protein shows chitin synthase soybeanin Brazil. The development of resistantvarieties activity,we willexpress both clones and their enzymatic isthemain goal of manybreeding programmes. Progress activitywill be tested. inselection for nematoderesistant lines can be speeded upbytheuse of molecularmarkers associatedwith genes for resistance.The objectives of this study were touse 307Evaluation of soybeangenotypes for host SSR markersto identify quantitative trait loci (QTL) suitabilityto Meloidogyneincognita race 2: an in vivo conditioningsoybean resistance against M. javanica and study todetermine their genomic location. Eighty-eight SSR 1; markerswere usedto amplify the DNA ofsoybean Hendrika FOURIE ¤,AlexanderHenrique M C lines(25 resistant and 26 susceptible) obtained from a DONALD 1, Dirk DE WAELE 2 andAlwyn Jacobus DE crossbetween BRS 133(susceptible) and PI 595099 LANGE (resistant),and selected in previous studies by galling 1 ARC-GrainCrops Institute, Private Bag X1251, numberassayed in glasshouse and infested Ž eld.HSP Potchefstroom,2520, South Africa 2 176,Satt 114 and Satt 423 showed signiŽ cant association Lab.Tropical Crop Improvement, K.U. Leuven,Kasteelpark withresistance. The QTL analysison the linkage group Azenburg13, 3001, Heverlee, Belgium indicatedthe presence of at least one gene on HSP 176 [email protected] ¤ locus(Lod 27.5). In all gene extensions between Satt 114 M.incognita race2 ispredominanton soybean in South andHSP 176(8.3 cM), the Lods were high(22.6-27.5). Africa. Sixty-ninesoybean hybrids were evaluatedfor hostsuitability to this species in the glasshouse. Each 309Ultrastructure ofresistantresponses of selected seedlingwas inoculatedwith 10 000eggs and larvae. Solanumbulbocastanum linesto infection with Assessmentswere made56 daysafter inoculation and RF- Meloidogynechitwoodi valueswere calculated.Five soybean genotypes S5995, Potties,P AN660,TXS89/ 10/29andD82-3298, identiŽ ed 1; 1 1 W. GOLINOWSKI ¤, J. JUPOWICZ , M. SOBCZAK , aspoor hosts in glasshouse screenings, were evaluated G. GRYMASZEWSKA 1, W. KUREK 1, F. ZOON 2, undernatural conditions in microplots and in the Ž eld and H. VAN DER BEEK 2 toverify their resistance. Prima 2000was includedin bothtrials as asusceptiblestandard. In the microplot trial 1 Departmentof Botany, W arsawAgricultural University, eachseed was inoculatedwith 10 000eggs and larvae at Rakowiecka26/ 30,W arsaw02528, P oland 2 plantingwhile a uniformand natural distribution of M. PlantResearch International, W ageningen,The Netherlands incognita race2 occurredin the Ž eldtrial. Meloidogyne ¤[email protected] incognita race2 eggsand larvae were extractedand Differentgenotypes of S.bulbocastanum varying in counted90 days after planting. S5995 maintained the resistanceto different isolates of M.chitwoodi were lowestnumbers of M.incognita race2 inthe above- examinedby electron microscopy. In moderately resistant mentionedtrials, followed by P AN660,TXS89/ 10/29, combinationsfeeding sites were composedof two D82-3298and Potties, and differed signiŽ cantly from tothree strongly enlarged giant cells derived from thesusceptible control (Prima 2000).Crosses between procambium.They contained condensed cytoplasm, small thesepoor-host cultivars and Prima 2000are underway to cytoplasmicvacuoles, enlarged nucleus with dispersed developa mappingpopulation for identiŽcation of genetic heterochromatin,electron translucent nucleoplasm and markersassociated with this resistance trait. hypertrophiednucleolus. In resistant combinations two differentplant responses were found.If giantcells were 308SSR markersassociated with resistanceto inducedamong procambial cells their cytoplasm was Meloidogynejavanica in soybean stronglycondensed. The central vacuole was substituted bysmallcytoplasmic vacuoles. The nucleus was enlarged Renata FUGANTI,João Flávio V ELOSO SILVA¤, andamoeboid and the nucleolus contained many

Vol.4(2), 2002 231 Resistance,breeding (295-335)

nucleolar‘ vacuoles’. If giantcells were inducedamong Johannes HALLMANN 1,KatherineG IERTH 2, corticalor pericycliccells they contained only paramural Josef SCHLANG 1, Joachim MÜLLER 1 layerof strongly osmiophilic cytoplasm. The central andRichard A. S IKORA 2 vacuolewas presentand Ž lledwith Ž brillarmaterial. 1 FederalBiological Research Center for Agriculture and Neighbouringcells contained large starch grains. Many Forestry,T oppheideweg88, D-48161 Mü nster ,Germany surroundingcells necrotised or divided hyperplastically. 2 Instituteof Plant Diseases, Nussallee 9, D-53115Bonn, Inabsolutely resistant combinations juveniles induced Germany slightenlargement of selected vascular cylinder cells. Theircytoplasm was locatedparamurally. Their central Thecrop potential to produce high yields even under vacuoleswere Žlledwith osmiophilic granules. Later on, severenematode pressure is an important economic giantcells degenerated and the juveniles were embedded traitin integrated control strategies. Plant tolerance ina layerof degraded cells. Some cortical cells were isespecially important for beetand tuber crops enlargedand contained many large starch grains. wherenematode damage at the early stages can delay ordestroy beet/ tuberformation. Breeding of tolerant cultivarsis currently limited by our poor understanding 310Effective and durable resistance against regardingfunctioning and inheritance of tolerance. plant-parasiticnematodes InŽ eldstudies and glasshouse experiments, a broad spectrumof sugar beet cultivars/ hybridswas studiedfor Sarah GOODCHILD , Peter URWIN ¤ toleranceof Heteroderaschachtii .Threecultivars/ hybrids and Howard ATKINSON were selectedfor furtherstudies on the mechanisms Departmentof Biology, University of Leeds, Leeds, LS2 9JT ,UK oftolerance: Nematop (tolerant/ resistant),Stru.1915 ¤[email protected] (tolerant,susceptible) and Penta (intolerant/ susceptible). Withincreasing nematode pressure, yield decrease in Thesuccess of transgenic approaches to plant-parasitic thetolerant cv. Nematop and tolerant hybrid Stru.1915 nematoderesistance will require an effectiveand durable was signiŽcantly less than in the intolerant cv. Penta. defence.This work aims to deŽ ne new targets that can be Tolerantplants suffered later from waterstress and the disruptedwith a deleteriousconsequence on the animal. rateof photosynthesiswas generallyhigher .Furthermore, Twocandidate genes have been selected as putative tolerantplants showed higher compensatory growth, and targets.The Ž rst isageneencoding an asparticproteinase increasedrooting depth. T olerancewas mostpronounced H. glycines isolatedfrom an libraryusing a PCR underdry summer conditions and less obvious under fragmentproduced by degenerate primers. The second humidconditions. The concept of using plant tolerance isan aminopeptidase, also isolated from an Heterodera for managingplant parasitic nematodes will be discussed. glycines cDNA libraryby heterologous probing with C. elegansEST.Recombinantproteins corresponding to both geneshave been expressed in order to conduct inhibitor 312Growing resistant sugar beets; anew assays.Plant derived aspartic proteinase inhibitors have opportunityin Ž eldsinfected with Heterodera beenshown to be effective in vitro andhave subsequently schachtii beentransformed into Arabidopsisthaliana under the controlof CaMV35spromoter. Infecting the transformed D. HEINICKE plants with H.schachtii willassess the potential of these Panzenschutzamt P ostfach910810, D 30428Hannover , inhibitorsas a defencestrategy. There are no inhibitorsof Germany aminopeptidasesdescribed from aplantsource, therefore [email protected] syntheticpeptide and antibody phage display libraries Insugar beet rotations, Heteroderaschachtii is now arebeing screened to isolate a functionalinhibitor . In keptbelow the economic threshold by growing resistant situ hybridisationexperiments indicate that the aspartic varieties.Resistant oil radish, or mustard, grown in proteinasegene is expressed intestinally whereas the autumnis not as effective as in spring. The density aminopeptidasegene is expressed in the reproductive dependentreduction after oil radish is 0.36 Pf/Pi system. comparedwith 0.64 Pf/Pi afterfallow. Even though the reductionis quite effective, the high multiplication rate 311T oleranceof sugarbeet to Heteroderaschachtii of H.schachtii onsugar beet overcomes the effect on the

232 Nematology Postersessions

Pi.Theseproblems are now overcome by resistant and 1Instituteof Biology, Karelian Research Centre RAS, tolerantsugar beet varieties. With a Pi between10 and Pushkinskayast, 11, P etrozavodsk,185610, Karelia, Russia 30 eggs juveniles/gsoil,the Pf/Pi liesbetween 0.9 and 2 Centerof AgriculturalResearch, Department of Crop C 0.7while the yield is 5-20% higher with a tolerantvariety Protection,Burg. V anGansberghelaan 96, 9820 Merelbeke, thancompared to a susceptibleone inan infested Ž eld.On Belgium [email protected] theother hand, resistant varieties earn 10% lesscompared ¤ withsusceptible ones on nematode-free land. As few Hatchingof eggsof Žvepopulationsof Globoderapallida individualsof H.schachtii canovercome the resistance inrootdiffusate of sixpotato cultivars was studied in vitro. inthe sugar beet, but not that of oil radish or mustard, Allcultivars induced hatching, regardless of theirdegree acombinationof methodswill be discussedto overcome ofresistanceto G. pallida.Thepercentageof hatchedeggs thesedisadvantages. after7 weeksvaried between populations and cultivars, butnone of thecultivarsconsistently induced most or least hatchfor allpopulations. Addition of freshly collected 313Durable resistance against Meloidogyne rootdiffusate (not stored) after 7 weeksof experiment chitwoodi and M. fallax: a dream? resultedin anew ushof hatchingand made differences betweeninduction of hatchingby thecultivars even more G.W. KORTHALS¤ and L.P.G. MOLENDIJK obscure.Results on the hatching of invasive juveniles areuseful for estimatingof the multiplication factor of AppliedPlant Research, P .O.Box430, 8200 AK Lelystad,The nematodepopulations. Multiplication of all populations Netherlands was higheston the susceptible cv. Bintje and lowest on ¤[email protected] thecompletely resistant cv. Innovator, but varied with populationfor partiallyresistant cvs Sante, Maritiema Finalgoal of EU-project DREAM (No. QLRT-1999- andCycloon. Multiplication factors were higherand 1462:Durable resistance management of the soil-borne lessvariable in closed containers than in pots in the quarantinenematode pests Meloidogynechitwoodi and M. glasshouse,which facilitated discrimination between the fallax)istodeliver crops (potato as wellas green manure cultivars.T estin closed containers could be usedto advise crops)which are resistant against the root-knot nematodes farmers whichcultivar to grow in G. pallida-infected M.chitwoodi and M. fallax.Thisposter presents the Ž rst Želds.However, correlation between multiplication in resultsof acomparisonof 15differentgenotypes of Italian closedcontainers and in theŽ eldshould be investigated. ryegrass,fodder radish and potato. These 15 genotypes were previouslyselected with colleagues from Plant ResearchInternational, Barenbrug Holding BV andP .H. 315Comparative study oftwo Heteroderaavenae PetersenSaatzucht. The host suitability of the selected resistancegenes from Aegilopsventricosa: differences genotypeswas determinedby extractingnematodes from indefence-enzymesinduction and chromosomal soilbefore planting (May 2001) and after the growing locationin wheat/ Ae.ventricosa introgressionlines season(November 2001). First results on M.chitwoodi 1 1 showedthat on average the genotypes of fodder radish M. Jesús MONTES , Isidoro LÓPEZ-BRAÑA , M. Dolores ROMERO 2, Esther SIN 3, M. Fe ANDRÉS 2, andpotato proved to be more resistant compared to all 3 testedgenotypes of Italianryegrass. However, none of the Juan A. MARTÍN-SÁNCHEZ , Mariano GOMEZ-COLMENAREJO 1 testedgenotypes reduced the populations to zero, which 1; was foundafter fallow. Meloidogynefallax was able to andAngeles D ELIBES ¤ reproducemuch more on mostgenotypes, although three 1 Dptode Biotecnologí a, ETS IngAgró nomos, UPM, Ciudad potatogenotypes had a verylow multiplication of M. Universitarias/ n,Madrid,E-28040, Spain; 2Centro de fallax. CienciasMedioambientales, CSIC, Serrano 115, Madrid, E-28006,Spain; 3CentreR+D deLleida,UdL-IRTA, Alcalde RoviraRoure 177, E-25198, Spain; ¤[email protected] 314In uence of potatocultivars with different degreesof resistanceon hatchingand multiplication Twoindependent introductions of genetic resistance ofthe potatocyst nematode Globoderapallida to Heteroderaavenae (Ha71Spanish pathotype), from Aegilopsventricosa tohexaploid wheat were compared 1; 2 Elizaveta MATVEEVA ¤ and Nicole VIAENE – the Cre2 (from Ae.ventricosa AP-1) and Cre5

Vol.4(2), 2002 233 Resistance,breeding (295-335)

(from Ae.ventricosa #10)genes. No susceptible plants themultinucleated giant cells (coenocyte) formed by M. were foundin the F 2 progenyfrom thecross between javanica infectionon these plants. The formation of a bothaccessions of Ae.ventricosa suggestingthat their syncytium-likefeeding site by root-knot nematodes is a respectiveresistance factors could be allelic. However, highlyunusual phenomenon. the genes Cre2 inH-93-8, and Cre5 insubstitution 6D/6Nv andaddition 6N v lineshave been transferred to 317Engineering coffee for root-knot nematode differentchromosome location. The induction of several resistanceusing cysteine and serine proteinase defenceresponses during early incompatible interaction inhibitors ofresistant lines carrying Cre2 and Cre5 genes has alsobeen studied. Isoelectrofocusing isozyme analysis 1; 1 2 RoxanaMyers ¤, Brent SIPES , Chifumi NAGAI , revealedchanges in peroxidase, esterase, superoxide Donald SCHMITT 1 and Howard ATKINSON 3 dismutase,glutathione reductase, shikimat dehydrogenase 1 andascorbate peroxidase activities in infected roots Universityof Hawaii at Manoa, Plant and Environmental ofresistant lines in comparison to their susceptible ProtectionSciences, 3190 Maile W ay,St. John 309, Honolulu, HI96822,USA parents.The highest differential activity between infected 2 HawaiiAgriculture Research Center ,99-193Aiea Heights anduninfected roots was foundfor theperoxidase Drive,Suite 300, Aiea, HI, 96701-3911,USA system,implicated in ligniŽ cation process. A DNA 3 Universityof Leeds, Centre for Plant Biochemistry and v marker,lacking in 6N line,was linkedto Cre2 gene Biotechnology,Leeds, LS2 9JT ,UK inH-93-8. Differences observed between Cre2 and ¤[email protected] Cre5 geneswith respect to the chromosomal location, InHawaii, the Kona coffee root-knot nematode, Meloido- detoxiŽcant enzymes induction, and behaviour against gynekonaensis ,causessevere damage to Coffeaarabica differentpathotypes, suggests that they are different CCN cv.T ypica.Due to the lack of resistantcultivars of C. ara- resistancesources for wheat. bica,geneticengineering for nematodecontrol was ex- plored.The modiŽ ed rice cystatin gene, OcI-Ä D86, and 316Comparative histology of feedingsites of thedual proteinase inhibitor gene, OcI-Ä D86/ GO/CpTI, Meloidogyneartiellia and M. javanica were usedto produce transgenic coffee for resistance to M.konaensis .Leafdiscs were transformedusing Mishael MOR Agrobacteriumtumefaciens containingthe cystatin con- Departmentof Nematology, Agricultural Research structs,then placed on G418selection medium. The 38% Organization,V olcaniCenter ,P.O.B 6,Bet-Dagan50-250, thatformed primary callus was presumedto be trans- Israel formed.Somatic embryos have not yet been obtained [email protected] from thiscallus because embryogenesis in coffee takes Theroot-knot nematode Meloidogyneartiellia causes 7-9months. In a secondtrial, somatic embryos were ob- considerabledamage to several agricultural crops in tainedfrom untransformedleaf discs. A.tumefaciens was theMediterranean region. Microscopic examination of usedto transform the embryos with the two gene con- longitudinalroot sections from twocruciferous plants, structs.Secondary embryos were producedin 37%of the Brassicaoleacea L.(cabbage)and B. rapa L.(turnip), initialexperiments under G418 selection. In addition, em- infectedwith M.artiellia ,revealedthat the feeding bryogeniccalli or somatic embryos were alsotransformed sitesconsist of several large nurse cells in the central usingparticle gun bombardment. Within 6 weeks,sec- vasculartissues, which includes the companion cells. The ondaryembryos were producedfrom thebombarded em- intracellularorganisation of thesecells closely resembled bryos,while embryogenic calli only produced more calli. thesyncytial forms. Thefeeding zone cells showed Theseresults indicate that somatic embryos are the most hypertrophyand hyperplasia and intense vacuolisation. efŽcient target tissue in the production of transgeniccof- Also,different degrees of cell wall dissolution between fee. contiguouscells were observedand only few cellnuclei andnucleoli were observed,all signiŽ cantly enlarged 318Differential gene expression between avirulent andamoeboid, as compared to those in the feeding andvirulent Meloidogyneincognita isogeniclines as cellsinduced by Meloidogynejavanica .Thefeeding monitoredby cDNA-AFLP sitesformed by M.artiellia differedconsiderably from

234 Nematology Postersessions

Cédriv NEVEU, Pierre ABAD and theroots and the Differential Display (DD) techniquewas Philippe CASTAGNONE-SERENO¤ usedto identify and isolate genes that differ between the genotypesduring nematode infection. Differentially ex- INRA UnitéIPMSV ,BP2078,06606 Antibes cé dex, France pressedgenes were identiŽed, isolated and cloned into ¤[email protected] pGEM-Tvectors for sequencing.Differentially expressed Usingtwo pairs of near-isogenic M.incognita lines, genesequences were usedto searchfor homologies.Pre- avirulentand virulent against the tomato Mi resistance liminaryanalysis identiŽ ed homology with a nucleotide gene,a cDNA-AFLP differentialanalysis was initiated,in bindingprotein, a pathogenesisrelated protein, a lowtem- orderto identify genes differentially expressed between peratureand salt responsive protein, an aquaporin protein, avirulentand virulent nematodes. AFLP markerswere andtranscription factors. developedon cDNA templatesdigested with HindIII and MseI,withall of the 256 combinations that could be 320Evaluation of maizegenotypes for resistance generatedwith primers with two selective nucleotides. A against Meloidogynejavanica and M.incognita race 3 totalof approximately84 000fragments were ampliŽed, amongwhich 61 appeareddifferential ( i.e.,presentin the NeucimaraRodrigues R IBEIRO 1, twoavirulent lines and absent in the two virulent lines). 1; 2 João Flávio V ELOSO SILVA ¤, André FRANCISCO , Inorder to conŽ rm thisresult, cDNA-AFLP was runagain José GOMES 2 andW alterFernandes M EIRELLES 3 onthe same templates, but with all of the64combinations thatcould be generated with primers with two selective 1 EmbrapaSoja; 2FAPEAGRO; 3EmbrapaMilho & SorgoCP nucleotidesfor MseIandone selective nucleotide for 231,86001-970, Londrina, PR, Brazil HindIII. Thisallowed elimination of a numberof false ¤[email protected] positivebands, and 19 fragments differential between avirulentand virulent lines were thusselected for further Theuse of nematoderesistant crops in rotationsprevents analysis.The next experimental steps included cloning lossesin susceptible crops. In Brazil corn is the main ofthe full-length genes, RT -PCRexperiments, etc. The cropused in rotation with soybean. The reproduction resultswill be discussedin relation with the ability of the of Meloidogynejavanica and M.incognita race 3 was nematodeto overcome or notthe plant resistance gene. studiedin 176and 57 maizegenotypes, respectively. The plantswere grownin glasshouse conditions, inoculated 319Differential gene expression in nematode with5000 nematode eggs and evaluated after 60 days. resistantsoybean genotypes Tomatoplants were usedas an inoculumefŽ ciency check. Theegg production of M.incognita was higherthan M. 1; 2 Júlio C. PEDROSO ¤,AlexandreL. N EPOMUCENO , javanica inall maize genotypes. Meloidogynejavanica João Flávio V ELOSO SILVA 2, Newton P. CARNEIRO 3, presentedlow reproduction in 78 genotypes. Only the Poliana K. MARTINS 1,MicheleC. B RETON 1, genotypesBRS 2114,P30F80and AG9090 were resistant SilvanaR.R. M ARIN 2 and Eliseu BINNECK 2 to M.incognita ,resultingin lowegg production. 1 UniversidadeEstadual de Londrina 2 EmbrapaSoja 321Host suitabilityin soybean cultivars for 3 EmbrapaMilho & Sorgo,Brazil the reniformnematode, 2001 tests ¤[email protected] R.T. ROBBINS 1, L. RAKES 1, L.E. JACKSON 1, Meloidogynejavanica causessigniŽ cant losses to soy- E.R. SHIPE 2, E.E. GBUR 3 and D.G. DOMBEK 4 beanproduction in Brazil. Understanding how resistant plantsrespond to nematode infection at molecular level 1 Cralley-WarrenCenter ,2601N. YoungAvenue, F ayetteville, mightindicate new strategies for thedevelopment of re- AR72704,USA sistantgenotypes. Roots of soybeangenotypes, BRS-133 2 ClemsonUniversity, P ooleAgric. Center ,Box340359, (susceptible)and PI595099 (resistant), were analysedfor Clemson,SC 29634-0359,USA differentialgene expression during nematode infection. 3 AgriculturalStatistics Laboratory, University of Arkansas, At 5daysafter germination, both genotypes were infected Fayetteville,AR, 72701,USA withnematode eggs and juveniles. Root samples were col- 4 ArkansasCrop Improvement Program, 313 Cassatt Road, lectedat 9 and12 days. T otalRNA was extractedfrom Fayetteville,AR, USA

Vol.4(2), 2002 235 Resistance,breeding (295-335)

In2001 glasshouse pot experiments, 139 soybean markers helpedto conŽ rm phenotypicseparation of ho- varietiesfrom theArkansas and Mississippivariety testing mozygousand heterozygous resistant plants. Heterozy- programs(119) and varieties submitted by Louisiana gousplants had similar levels of resistanceto root-galling extensionnematologists (20) and 34 breeding lines symptoms,but supported higher nematode reproduction from ClemsonUniversity were testedto determine comparedwith homozygous plants. This gene dosage ef- theirsuitability as hosts for thereniform nematode, fectwas moreapparent against M.incognita than M. ja- Rotylenchulusreniformis . The R.reniformis -resistant vanica,andat temperaturesof 28 ±Corhigher,when Mj- varietiesForrest and Hartwig, the susceptible variety 1 expressionwas weaker.Effective resistance was shown Braxton,and fallow- R.reniformis -infestedsoil served as inŽeldexperiments when used alone and in combination controls.T otalnumber of eggs and nematodes extracted withother cultural tactics, including use of rotationcrops from boththe soil and roots from eachpot, reproductive withresistance and manipulation of carrot planting date indices (RI Pf=P i), RI=RI of Forrest (RF), log ratio tominimiseinfection. D .log .RF 1//, and RF calculatedfrom log .RF 1/ 10 C 10 C were calculatedfor eachcultivar or breedingline in each 323New sourcesof resistanceto Rotylenchulus test.V arietieswith RF signiŽcantly greater than the RF reniformis in cotton, Gossypium spp. onForrest (1.00) were consideredsuitable hosts for R. reniformis.Inthe states variety test, 90 ofthe119varieties Forest ROBINSON, A. Edward PERCIVAL were suitablehosts when RF were comparedand 117 and Alan C. BRIDGES whenthe log ratio .log .RF 1// were compared.In 10 C theClemson test, 23 ofthe34 varietiesand breeding lines 2765F&B Rd,College Station, TX 77845,USA RF were suitablehosts when were compared,and 24when Rotylenchulusreniformis causesyield losses to cotton thelog ratio .log .RF 1// were compared. 10 C (Gossypiumhirsutum )inMississippi, Louisiana and Alabamain the USA. Croprotation and nematicides 322Characterisation of resistanceto root-knot aremarginally economic and resistance is needed but nematodesin carrot onlyweak resistance is reported in G. hirsutum, and allprevious studies represent a smallpercentage of 1; 2 3 Philip A. ROBERTS ¤, P.W. SIMON , L.S. BOITEUX , accessionsavailable within Gossypium species. We W.C. MATTHEWS 1 and T.R. MULLENS 1 evaluated ca 3000primitive accessions of G. arboreum, G.herbaceum , G.longicalyx , G. hirsutum and G. 1 Departmentof Nematology,University of California, Riverside,California 92521, USA barbadense from theUS NationalCotton Collection 2 USDA, ARS VegetableCrops Research Unit, Department of andconŽ rmed resistance to R.reniformis inthe diploid Horticulture,University of Wisconsin, Madison, Wisconsin, species G. arboreum, G.herbaceum and G.longicalyx . USA Noresistance was foundin more than 1500 accessions 3 CentroNacional de Pesquisade Hortaliças (CNPH)/Empresa of G. hirsutum.Severalof morethan 1000 accessions of Brasileirade PesquisaAgropecuá ria (EMBRAP A),CP 218, thecompatible tetraploid G.barbadense were resistant; 70359-970Brasí lia-DF ,Brazil otherswere comparableto susceptible controls; none ¤[email protected] were immune.Resistant G.barbadense were collectedin widelyscattered regions: Peru, Guatemala, Mexico and Hostplant resistance to the root-knot nematodes Meloi- StLucia. The G.barbadense accessionGB-713 had the dogynejavanica and M.incognita conferredby the Mj-1 highestresistance, followed by a secondgroup (GB-49, locusis beingincorporated into advanced breeding lines GB-264,TX-110), and a thirdgroup that was weaklybut of carrot (Daucuscarota ).Geneticanalysis of F2-F4and consistentlyresistant (TX-1347, TX-1348). Differences in BCfamiliesdetermined that the resistance is inherited originand resistance level suggest further enhancement asa singledominant gene or possiblytwo closely linked ofresistancemay be gainedthrough new combinations of genes. The Mj-1 locusis derived from thevariety Brasilia, resistancegenes. andit expresses very high levels of resistance to M. ja- vanica andalmost as high resistance to M.incognita . Toimprovethe selection efŽ ciency for the Mj-1 locus in 324Effects oftemperatureand soybean genotype on breedingprograms, closely linked STS codominant ank- Heteroderaglycines malesand females population ingmarkers have been developed. Application of these

236 Nematology Postersessions

1; 2 2 M.R. DA ROCHA ¤, T. ANDERSON and T. WELACKY resistantvarieties grown on MB disinfectedsoils. More than30% of the stocks assayed had average root-knot 1Univ.Fed.de Goiás, Esc.de Agronomia, Caixa P ostal131, indexsimilar to those of MBtreatments.More than 30% CEP 74001-970,Goiania, GO, Brazil 2 2 ofthestocks had also marketable yields (8.3 kg/ m C-58 Agricultureand Agri-F oodCanada, Greenhouse and 2 2 ProcessingCrops Research Centre, N0R1G0, Harrow ,Ontario, hybrid;8.0 kg/ m C-57and 8.2 kg/ m C-30)similar to 2 Canada thoseof MBtreatments(8.9 kg/ m /.Stocksgrown on the ¤[email protected] samesoil over 3 yearsled to anincreaseof aggressiveness in M.incognita populations.This was noticedby an Soybeancyst nematode resistant ‘ Bell’and susceptible increasein averageroot-knot index and the percentage of ‘S20-20’soybean cultivars, were pre-germinatedand infestedplants ( i.e.,inthe 1st year the C-30 hybrid had transplantedto Heteroderaglycines infestedsoil, and 0.1%of infestedplants and 0.1 of root-knotindex, while exposedto temperatures of 20, 25, 30 or 35 ±C for 10 inthe2nd year 73.3% of theplants were infestedand the daysin growth chambers. Males were countedfrom root-knotindex increased to 4.1). plantsgrowing hydroponically for 10additionaldays and femaleswere countedfrom plantsgrowing in autoclaved sandfor 30additionaldays. The male:female ratios were 326IdentiŽ cation of markersto facilitate cloning of alwayshigher than one and higher on the resistant cv. resistancegenes to Heteroderatrifolii in clover Bell.Infection by males and females was affectedby Garry M. ROSEWARNE , P.J. LARKIN temperatureand was highestbetween 26 and28 C. Total ¤ ± and E.S. LAGUDAH malepopulation was notaffected by soybean genotype. Moreabnormal males ( <800 ¹m) developedon roots PlantIndustry, CSIRO, GPO Box1600, Canberra ACT 2601, ofthe resistant cv. Bell and shorter males developed Australia; ¤[email protected] followingexposure at 35 ±C.Thefemale population was Whiteclover ( Trifoliumrepens )isa majorlegume signiŽcantly higher on the susceptible cv. S 20-20.At componentof dairy pastures throughout Australia. 35±Conlya few femalesdeveloped on both cultivars, Howeverthese plants are susceptible to a varietyof whichled to a highermale:female ratio. The number of rootendoparasitic nematodes, the most common and eggs/femalewas higheron susceptible S 20-20than on widespreadbeing Heteroderatrifolii .Nematodescan resistantBell and did notdiffersigniŽ cantly with different severelyaffect productivity and incur substantial losses to temperatures. thedairy industry. There is little resistance to H. trifolii inwhite clover, although we haveidentiŽ ed resistance 325Response ofpepper stocks to Meloidogyne incaucasian clover, Trifoliumambiguum . Conserved incognita inglasshouses in the southeast ofSpain sequencesof previously characterised resistance genes andtheir analogs are being used to identify candidate C. ROS 1, M.M. GUERRERO 2, P. GUIRAO 1, resistancegenes in T. ambiguum.Partialsequences of 2; 2 2 A. LACASA ¤, M.A. MARTÍNEZ , J. TORRES , resistancegene analogs (RGAs) havebeen isolated N. BARCELÓ 2 and A. GONZÁLEZ 2 from legumesusing DNA sequencedatabases and througha PCRapproachwith white clover .Over 200 1 FECOAM –Consejería deAgricultura,Agua y Medio RGAsequencesfrom legumeshave been analysed Ambiente,C/ Caballero,13, 30.002 Murcia, Spain phylogeneticallyand 39 sequences selected to represent 2 Protección Vegetal,Centro de Investigación yDesarrollo Agroalimentario,CAAMA, C/Mayors/ n,30.150La Alberca, distinctclusters. These individual sequences were usedas Murcia,Spain probesin a bulksegregant analysis of T. ambiguum plants thatare either resistant or susceptibleto H. trifolii. Probes ¤[email protected] thatreveal polymorphism and where subsequent analysis Commercialpepper varieties resistant to Meloidogyne ofindividuals shows a tightlinkage to the resistance incognita didnot match the standard production of phenotypewill be targeted.Diagnostic probes provide an glasshousesin the southeast of Spain. When methyl entrypoint to screen a resistant T. ambiguum genomic bromide(MB) is notusedfor soildisinfection, nematodes libraryto identify candidate resistance genes for usein becomeone of the main soil-borne pathogens. The transformationexperiments. responseof 75resistantpepper stocks was testedagainst Phytophthoracapsici and M.incognita inrelation to non-

Vol.4(2), 2002 237 Resistance,breeding (295-335)

327IdentiŽ cation of resistanceto the oatraceof manure,they were inoculatedwith 10 000eggs. The C stem nematode( Ditylenchusdipsaci )infaba bean experimentdesign was completelyrandomised with four (Vicia faba)inAustralia replicates.After 120days, the clones were evaluated bythe egg number and presence of nematodes in root, 1; 2 Sharyn TAYLOR ¤, Maria SCURRAH usingNanicã o 122clone as the susceptibility standard, 1 and Danuta SZOT attwolevels of watering.Soil samples were collectedto 3 1 SouthAustralian Research and Development Institute, GPO determinethe nematodenumber per 200cm perreplicate Box397, Adelaide, South Australia 5001, Australia for eachclone. The Nanicã o clones(90 and 122) allowed 2 InternationalP otatoCentre, Apartado 1158, Lima, P eru alargemultiplication of M.incognita ,comparedwith the ¤[email protected] othereight clones, showing susceptible reaction, but Maç ã Theoat race of stemnematode ( Ditylenchusdipsaci ) was 57showedthe lowest parasite multiplication. All the other Žrst recordedin South Australia in 1973. The principal clonesshowed some nematode resistance, depending on hostsin this region are oats and faba beans and the thewaterlevel, and it canbe seenin theresults of Nanicão onlyeconomic method for controlis the use of resistant clones.For theroot weight and aerial parts, the results crops/varietieswithin rotations. At thecommencement of showedlittle difference, compared with inoculated and researchin 1994, high levels of resistance and tolerance non-inoculatedplants, showing some difference between were identiŽed inoatbreeding material and screening has twolevels of watering. contributedto therelease of Žveresistantand tolerant oat varieties.In contrast, of the120 advanced faba bean lines 329Resistance of watermelon( Citrullus spp.) tested,only low to moderate levels of resistance were germplasmto the peanut root-knotnematode found.V ariationwas observedwithin the commercial (Meloidogynearenaria race 1) cultivarsFiord and Ascot, however, and crosses from plantsshowing superior resistance were made.Lines from Judy THIES¤ and Amnon LEVI progenyof crossesfrom Ascotproduced accessions with moderateto highlevels of resistanceand from Fiordwith USVegetableLaboratory, ARS, USDA, 2875Savannah moderateresistance following recurrent selection from Highway,Charleston, SC, 29414,USA 1994to 2000. These selections represent a signiŽcant ¤[email protected] improvementon existing material and will be used as Root-knotnematodes ( Meloidogynearenaria , M. inco- parentsto develop varieties with superior resistance for gnita and M. javanica)areserious pests of watermelon theSouth Australian Faba Bean Breeding Program. (Citrulluslanatus )inthe southern US andworld-wide. Currently,root-knot nematodes (RKN) arecontrolled in 328Reaction of bananaclones ( Musa spp.) to watermelonby pre-plantsoil fumigation with methyl bro- Meloidogyneincognita race 3 mideor other nematicides. The proposed removal of methylbromide from theUS marketin 2005has focused 1 2 3 R.C.V. TENENTE , O.A. CARRIJO , S. Silva NETO , interestin developing alternatives for managingnema- 2 2 R.D.C. SILVA and G.T. COSTA todesin vegetable crops. Resistant cultivars would pro- 1 Embrapa/Cenargen,C.P .2372(70770-900) Brasilia, DF , videan inexpensive,environmentally compatible alterna- Brazil tivefor managingRKN inwatermelon.All accessions of 2 Embrapa/CNPH, BR060,Km 09Rod. Brasilia/ Anapolis C.colocynthis (21) and C. lanatus var. citroides (88), and (70359-970),Brasilia, DF ,Brazil approximately10% of C. lanatus var. lanatus (157) ac- 3 CAMPO, RodoviaLMG 658km (38.600-000) P aracatu,MG, cessionsfrom theUSDA Citrullus germplasmcollection Brazil were evaluatedfor resistanceto M. arenaria race 1 in Theaim of this work was tostudy under glasshouse greenhousetests. The C. lanatus var. lanatus and C. colo- conditionsthe reaction of banana to Meloidogyne cynthis accessionswere susceptiblewith root gall sever- incognita race3. These clones were developedby a ityindices (GI) of4.0-9.0(scale, 1.0-9.0) and 8.0-9.0, re- BreedingProgram of Embrapa Cassava and Tropical spectively.However, 21 of88 C. lanatus var. citroides ac- FruitsCenter and Agriculture Promotion Company. cessionswere moderatelyresistant (GI, 3.0-9.0).SigniŽ - Micropropagatedoffspring were usedand 5 daysafter cantgenetic variability exists within the USDA Citrullus transplantto containers Ž lledwith sterilised soil sand germplasmcollection for reactionto M. arenaria race 1 C 238 Nematology Postersessions

and the C. lanatus var. citroides accessionsare potential ¤[email protected] sourcesof resistanceto M. arenaria race 1. Plantlipoxygenase activity is involved in plant growth and developmentby formation of fatty-acid hydroperoxides 330Multiple selection and durability of potatocyst and/orjasmonicacid. Moreover, lipoxygenases are known nematode Globoderapallida virulenceon arangeof tocontribute in regulating cell death and resistance to potatospecies pathogenicagents. In order to understand the timing of lipoxygenaseinvolvement during nematode infection, we Susan J. TURNER andColin C. F LEMING ¤ characterisedthe expression pattern of Žvegenescoding AppliedPlant Science Division, Department of Agricultureand for distinctlipoxygenase isoforms ( LOXs) in Pisum RuralDevelopment for Northern Ireland, Newforge Lane, sativum resistant(line MG103738) and susceptible (cv. BelfastBT9 5PX, UK Progress9)to thecyst nematode Heteroderagoettingiana . ¤[email protected] Theexpression pattern of the same LOX geneshas also beenevaluated in pea roots following wounding in the Aseriesof selectionexperiments on potatocyst nematode absenceof theparasite. An increased expression of each Globoderapallida ,pathotypePa1 tested the virulence lipoxygenaseisoform, in both resistant and susceptible responseof the nematode to a rangeof resistant roots,2 hafterwounding with a peakof transcript potato Solanum genotypes.Alleles conferring virulence abundanceat 6 hwas observed.Furthermore, in infected againstall four Solanum sourcesused in the study, i.e., roots LOX expressionwas followedat 24, 48, and 72 Solanumvernei, S. multidissectum,S. sanctae-rosae and hafternematode infection. The LOXsofsusceptible S.tuberosum spp. andigena were detected.Selection andresistant pea roots exhibited distinct expression for multiplevirulence against a combinationof resistant patternsindicating the occurrence of genedown- and up- sourcesresulted in the originally selected virulence regulation.Intriguingly, some LOX memberswere clearly genesbeing retained or lost. The mechanism of potato repressedduring nematode infection in the resistant cystnematode resistance differs betweenthe Solanum genotype. species.This, potentially, offers amanagementtool for controllingpotato cyst nematode levels in infestedpotato landthrough the appropriate use of resistant cultivars 332The in uences of different soybeancultivars on producedfrom arangeof Solanum-resistantspecies. the dynamicsof Heteroderaglycines race 3 Extendedselection trials on a rangeof worldwide G. pallida populationsclariŽ ed that some Solanum species Haiyan WU¤, Yuxi DUAN, Lijie CHEN and Weizhi LIU were moredurable at maintaining their PCN resistance Collegeof PlantProtection, Shenyang Agricultural University, thanothers. After 5yearsof continual selection of Shengyang,110161, P .R.China PCN populationson S.tuberosum spp. andigena no ¤[email protected] appreciableincrease in virulence had occurred, whereas on S.stenotonum and S.kurtzianum ,increasedPCN Ninecultivars were usedto study the dynamics of reproductionrates were apparentfrom thesecondselected soybeancyst nematode (SCN) Heteroderaglycines race generation.These results have clear implications when 3for differentsoybean cultivars in theŽ eldof Shenyang selectingthe most appropriate material for PCNbreeding AgriculturalUniversity, Liaoning province in China. The programmes. resultsindicated that the numbers of SCN secondstage juveniles(J2) inthe rhizosphere of Liaodou 10 reached itshighest peak, 60.4 J2, in 100 cm 3 soil,on 24 May. 331Members of the rootlipoxygenase gene family Thecyst population of soil around the root of Liaodou ofsusceptibleand resistant pea genotypes exhibit 10and Kaiyu 10 had a peakon 6 June(23.3 and 30.0 distinct expressionpatterns when infected bycyst cystsin 100 cm 3 soil,respectively), which was 1.5to nematodes 2.0times the number on Peking. There was apeakof J2in the root of soybean on 24 May for theresistant Pasqua VERONICO , Donato GIANNINO, ¤ cultivarsPI90763, Harbin small black soybean, Yingxian MariaTeresa M ELILLO and Teresa BLEVE-ZACHEO smallblack soybean and Franklin, but the peak value was Istitutodi Protezione delle Piante, C.N.R., viaAmendola 165/ A, muchlower than that of Liaodou10 andKaiyu 10. From 70126Bari, Italy thedynamics of J2,J3 and J4, soybean LiaoK89102 was

Vol.4(2), 2002 239 Resistance,breeding (295-335) resistantto invasion by SCN, PI90763was resistantto 3 UniversidadAutó noma Chapingo, Chapingo, Mé xico developmentof SCN, Liaodou10 and Kaiyu 10 were 4 UMR INRA/ENSAR, Biologiedes Organismes et des susceptiblecultivars. Populationsappliqué e àlaProtectiondes Plantes (BiO3P), BP35327,35653 Le Rheu,France ¤[email protected] 333Selection of root-knotnematode resistant sugar beet fromŽ eldplantings Wildwheat relative Aegilopsgeniculata representsa 1; 2 M.H. YU ¤ and P.A. ROBERTS valuablesource of genetic variation for improvementof diseaseand pest resistance in cultivated wheat ( Triticum 1USDA, AgriculturalResearch Service, Salinas, California aestivum).Abetterunderstanding of the resistance 93905, USA 2 Departmentof Nematology,University of California, responsesof this species against cereal cyst nematode Riverside,California 92521, USA (CCN) androot lesion nematode (RLN) isneeded. Ae. geniculata ¤[email protected] accessionsoriginating from differenteco- geographicalregions possessing useful biotic and abiotic Sugarbeet is a favouredhost for numerousspecies stressresistance traits were evaluatedfor theirreaction Meloidogyne ofnematodes, including spp.Resistance totwo populations of CCN Heteroderaavenae : Ha41 toroot-knot nematodes was identiŽed 7 yearsago, originatedfrom Franceand E125 from Syria,and to one andsince then it has been introgressed into cultivated populationof RLN Pratylenchusthornei from Mexico. sugarbeet. Preliminary observations on several breeding Resultsobtained conŽ rmed the potential value of Ae. populationswere conductedin Ž eldplots infested with geniculata asa sourceof CCN resistancefor wheat M.incognita M. javanica either or atUC Research improvement.They also demonstrated the interest of andExtension Centers, Irvine and Parlier, California. In thisspecies as a sourceof moderate resistance to P. resistantprogeny families, more than 50% of the plants thornei. Promising Ae.geniculata accessionspossessing producedhealthy taproots that exhibited no root-knot resistancetraits to cereal cyst and/ orroot lesion symptoms.In comparison, none of thesusceptible control nematodesin additionto other abiotic/ bioticstresses were plantswere free from galling.SigniŽ cant reductions of identiŽed and are presently used in wide hybridisation approximately45% or more in root weights occurred programme.They were crossedwith susceptible high- whenthese susceptible control plants were grownin yieldingbread and durum wheat CIMMYT cultivars as infestedsoil. Susceptible sugar beet suffered a higher wellas with Chinese Spring in order to transfer this sensitivityreaction to prolonged temperature ( >38 C) ± resistanceinto cultivated wheat. stressesand secondary pathogenic invasions than the resistantcounterpart. Glasshouse inoculation screenings providedreliable classiŽ cation of resistantgenotypes, but 335Durable resistance against Meloidogyne noindexof fullgrowth potential of theplants. Our results chitwoodi and M. fallax (EU-DREAM) indicatethat a productiveroot-knot nematode-resistant F.C. ZOON1; , J. BAKKER 2, W. GOLINOWSK 3, sugarbeet line with elite root yield, taproot conformation, ¤ R. JANSSEN 1, D. MUGNIÉRY 4, M.S. PHILLIPS 5, andsucrose content would be developed more readily M. SCHLATHÖLTER 6, J.G. VAN DER BEEK 1 whenresistant parents were grownand selected from and L. VAN KRUIJSSEN 7 nematodeinfested Ž elds.

1 334Resistance of Aegilopsgeniculata Roth tocereal PlantResearch International, P .O.Box 16, NL-6700 AA Wageningen,The Netherlands cystnematode ( Heteroderaavenae )androot lesion 2 Laboratoryof Nematology, W ageningenUniversity, nematode (Pratylenchusthornei ) Wageningen,The Netherlands 3 1; 2 3 Departmentof Botany (SGGW), W arsawAgricultural Maria ZAHARIEVA ¤, Julie NICOL , Higinio ROMERO University,W arsaw,Poland 4 and Roger RIVOAL 4 Laboratoirede Zoologie, INRA, Le Rheu,France 1CIMMYT,WheatProgram, A.P .6-641,06600 Mexico D.F ., 5 DeptNematology, SCRI, Dundee, Scotland, UK Mexico 6 P.H.P etersenSaatzucht GmbH, Grundhof,Germany 2 CIMMYT International,Wheat Program, PO Box39, Emek, 7 BarenbrugResearch BV ,Wolfheze,The Netherlands 06511,Ankara, Turkey ¤[email protected]

240 Nematology Postersessions

TheEU-funded project QLRT -1999-1462DREAM (Du- incorporationof resistance in important arable crops: rableResistance Against Meloidogyne )aimsto contribute potato,pepper, ryegrass and fodder radish, ii) study of tosustainable production systems by developing a strat- variationin nematode virulence and of durability of the egyfor durableresistance management for thesesoil- resistance,and iii)optimisingof production systems by bornepests. This study focuses on the two polyphagous rotationschemes. The main results expected are: resistant quarantine-organisms Meloidogynechitwoodi and M. fal- germplasm,well characterised pathogen collections, lax,whichare an important economic threat and for reliableselection and breeding methods, knowledge on whichno adequate durable alternative exists. The objec- thestability of resistance, molecular markers linked tivewill be achieved by integrating expertise in breed- toresistance and (a)virulence, knowledge on resistance ing,nematology, botany and molecular biology into one mechanismsand genes, genetic maps, and advice about project,novel by its European dimension. The project improvedrotation schemes. More information and Ž rst combinesthree areas of research: i)identiŽcation and resultscan be foundat www.eu-dream.nl .

Vol.4(2), 2002 241 Morphology,development (336-358)

336Molecular characterisation and morphometrics theglasshouse. Fifteen days after inoculation, roots were ofcereal cyst nematodes in Syriaand T urkey washedand prepared for observation.Tissues were Žxed inKarnovsky and post-Ž xed in 1% osmium tetroxide. Hussam ABIDOU 1, Sylvie VALETTE 2, Theywere pre-stainedin 0.5% uranyl acetate, dehydrated, 2 2 Jean-PierreG AUTHIER , Roger RIVOAL andembedded in Spurr .Thinsections were prepared 3 and Amor YAHYAOUI andstained for observationunder transmission electron 1 Universityof Aleppo, F acultyof Agriculture,PO Box12009, microscope.Additionally, thick sections were prepared Aleppo,Syria andstained for observationunder light microscope. 2 UMR INRA/ENSAR, Biologiedes Organismes et des Bothsusceptible and resistant plants, formed syncytia Populationsappliqué e àlaProtection des Plantes (BiO3P), withcell wall perforations, dense cytoplasm, increased BP35327,35653 Le Rheu,France endoplasmicreticulum, and increased size of nucleus. 3 ICARDA, CerealP athology,PO Box5466, Aleppo, Syria. Changesthat appear to be induced in plants with a Cerealcyst nematodes (CCN) arebecoming serious higherlevel of resistance include a layerof necrotic cerealdisease in the rainfed areas of the Mediterranean cellssurrounding the syncytia and prominent cell wall basin.A surveyof cereal Ž eldsin Syria and Turkey has appositionsin syncytium component cells near the shownthat the main cereal cyst nematodes Heterodera necroticlayer . avenae, H. Ž lipjevi and H. latipons occurfrequently. Characterisationof these nematodes was conductedon 338Morphometric variation of Rotylenchulus 14populations using PCR-RFLP analysis of the ITS reniformis geographicpopulations from regionsof rDNA andon 12 morphometric characters cotton-growingregions in the United States ofcysts and juveniles tested through a principle componentanalysis (PCA). Restrictionpatterns obtained Paula AGUDELO andRobert T .R OBBINS withfour endonucleases ( HaeIII, HinfI, ItaI, PstI) enabled differentiationof thethreecited species plus thechick-pea 217Plant Science, University of Arkansas,F ayetteville, cystnematode Heteroderaciceri .Principalcomponent AR72701,USA analysisapplied to the morphometrical parameters [email protected] followedby adirecthierarchical classiŽ cation conŽ rmed themolecular diagnostics but demonstrated a substantial Thevariability of morphological characters among variabilityin the H. latipons group. populationsof Rotylenchulusreniformis associatedwith cottonplantations in the United States is not known. A studywas conductedto determinethe range of variability 337Ultrastructural and histological changes ofdiagnostic characters in males and immature females induced by Rotylenchulusreniformis inresistant and among12 populations from differentcotton-growing susceptibleupland cotton regionsin the United States (two from Alabama,two from

1; 1 Arkansas,one from Florida,one from Georgia,two from Paula AGUDELO ¤, Robert T. ROBBINS , 1 2 Hawaii,one from Louisiana,one from Mississippi,one Kyung S. KIM andJames McD. S TEWART from SouthCarolina, and one from Texas).Specimens 1217Plant Science, University of Arkansas,F ayetteville, were extractedfrom soilby the centrifugation- otation AR72701,USA technique,and were heatnarcotised. Nematodes were 2 CropSoil & EnvironmentalScience, University of Arkansas, mountedin water and measured with the aid of a Fayetteville,AR 72701,USA compoundmicroscope. Analysis of variance was used ¤[email protected] tocompare the characters measured among populations. Severalevents that may determine the degree of Morphometricsof males and immature females from susceptibilityof cottonplants to reniform nematode occur thedifferent geographic regions of the southeastern atthe feedingsite. The objectiveof thiswork is todescribe UnitedStates were similar,but differed from those histologicaland ultrastructural modiŽ cations induced by from Hawaii.The effect of the host on the range of Rotylenchulusreniformis inresistant and susceptible variabilityof diagnostic characters was alsodetermined uplandcotton ( Gossypiumhirsutum )roots.Susceptible bycomparing measurements of the populations grown (Deltapine50)and resistant cotton materials under ontomato (Rutgers), cotton (Deltapine 50), and soybean evaluationwere inoculatedwith reniform nematode in (Braxton).

242 Nematology Postersessions

339The cellular structure ofthe female pastliterature is actually a vitellinelayer. Histochemical reproductivesystem of Meloidogyne spp. compared analysishas demonstrated a lackof chitinous material with othernematode species onthe eggshell exterior. Microscopy observations of the eggshellshows a waxyappearance and osmotic staining 1; 1 Wim BERT ¤, Ruben VAN GANSBEKE , thatis consistent with that of thevitelline layer found in 2 1 Gerrit KARSSEN , Gaetan BORGONIE othernematodes. Lectin localisation has also shown that andEtienne G ERAERT 1 theeggshell continues to develop past fertilisation with 1 Departmentof Biology,Ghent University, Ledeganckstraat 35, deliveryand integration of eggshell precursors. Further, B-9000Gent, Belgium a‘zipperlike’ structure associated with the vitelline layer 2 PlantProtection Service, Nematology Section, P .O.Box9102, oftheeggshellfor juvenileemersion was observed.Based 6700HC Wageningen,The Netherlands onour Ž ndingswe proposethat H. glycines follows the ¤[email protected] commonthree-layer structure of other nematodes and is notan exceptionas originally thought. Gonadsfrom livingyoung females, belonging to 15 Meloidogyne speciesand 80 otherspecies, were extruded tostudy the cellular structure of the female genital 341AFM proŽling of putativehydrogen bond structure.Within the genus Meloidogyne ,thespermatheca patterns onnematodecuticle surface isalways spherical and formed by a variablenumber of thick,lobe-like cells, which makes it different from any Aurelio CIANCIO otherknown nematode genus. Members of this genus Istitutoper la Protezionedelle Piante, Consiglio Nazionale arestrongly uniŽ ed by this characteristic anatomical delleRicerche, V iaAmendola 165/ A,70126Bari, Italy feature.Nevertheless a remarkableintrageneric variability [email protected] isdemonstrated; most species have 16 to18spermatheca cellswith interlaced cell boundaries while M.microtyla AtomicForce Microscopy (AFM) isanon-invasivehigh- and M.ichinohei havemore spermatheca cells with resolutionprobe microscopy useful in thestudyof biolog- atypicalcell boundaries and the spermatheca cells of the icalsurfaces in theirquasi-native state. Applications to ne- M. fallax specimensare clustered together forming lobes. matodecuticle from driedspecimens showed that atomic Whilemost species were studiedwith light microscopy, resolutionstudies are feasible, allowing the identiŽ cation thegonads of M.incognita were studiedthoroughly using ofstructuralarrangements of singlecollagen chains. AFM scanningand transmission electron microscopy. This scanningswere appliedto air or ethanoldried Xiphinema allowedus to determine exactly the structural separate diversicaudatum specimens.After scanningstabilisation gonoductcomponents and have a betterinsight in their duringlong-run sessions at room temperature and pres- actionand function. sure,atomic resolution images allowed the reconstruction ofmolecular level details. These included ordered pat- 340Eggshell ultrastructure of Heteroderaglycines : ternsof drop-likeclouds linking parallel atom chains in- histochemicallocalisation of chitinouscomponents terpretedas putativeH-bonds. Their length, shape and di- mensions,as well as theratio with the other particle mea-

B. BURGWYN¤, B. NAGEL, J. RYERSE and R.I. BOLLA surements,Ž tquantummodels of charge distribution as wellas Bohr’ s ‘hangingdrop’ model of H-bonds.These BiologyDepartment, Saint Louis University, St Louis,MO, USA arethe Ž rst atomicresolution data obtained from nema- ¤[email protected] todecuticle and the Ž rst imagingof a putativechemical Thefunction of the nematode eggshell to protect the bondproduced by AFM.The potential of thistechnology developingembryo is well conserved; however, the inhighresolution studies of biomoleculesis discussed. structureof the eggshell is varied. The most common structurethat is found amongst nematodes is a tri-layer 342Morphobiometrical and biochemical ultrastructurecomposed of alipidlayer, a chitinlayer, and characterisationof Heteroderazeae Portuguese avitellinelayer. Previous literature on theultrastructure of populations the Heteroderaglycines eggshellhas claimed that there isno vitelline layer but rather two chitinous layers. It FernandoJ.S. C ORREIA¤ isour contention that the outer chitin layer described in andIsabel M. deO.A BRANTES

Vol.4(2), 2002 243 Morphology,development (336-358)

Institutodo Ambiente e Vida,Departamento de Zoologia, speciesfound on citrus in Sã o PauloState that is beingde- Universidadede Coimbra, 3004-517 Coimbra, P ortugal scribedand named P. jaehni. Aclusteranalysis of stylet ¤[email protected] length,width and height of thestylet knobs, a, b, c,c 0 and Threepopulations of Heterodera from differentlocations Vparametersfor thesepopulations was consistentwith inPortugal were isolatedfrom Žgtreerhizosphere theidentiŽ cations. andcorn roots. Morphological and morphometrical analysisrevealed that all populations presented the two 344Scanning electron microscopy of the juvenile maindiagnostic characters of the corn cyst nematode, stagesof someWilsonematinae (Plectidae) Heteroderazeae : i)theunique two level bullae with 1; 2 characteristicarrangement and orientation; and ii) stylet Irma T. DE LEY ¤,OleksandrH OLOVACHOV 1 meanlength usually smaller than 20 ¹m.The morphology and Paul DE LEY (LM,SEM) andmorphometry of cysts and second- 1 Departmentof Nematology, University of California– stagejuveniles are given and the intrapopulation aland Riverside,Riverside CA 92521,USA intraspeciŽc variabilityare discussed and compared 2 Departmentof Zoology,Biological F aculty,Ivan Franko withother populations. Furthermore, a characteristic NationalUniversity of L’viv, Grushevsky str .4,L’ viv 79005, andreproducible phenotype of nonspeciŽ c esterase Ukraine activitywas detected,by native polyacrylamide gel ¤[email protected] electrophoresis,in thethree populations. The occurrence Lipregions of Tylocephalusauriculatus and Ereptonema of H. zeae,aneconomically signiŽ cant parasite and arcticum undergoextensive morphological transforma- originallyidentiŽ ed from corn( Zea mays L.),is reported tionsduring the nematodes developmental stages, from for theŽ rst timein Europe(Portugal). a simple Plectus-likeappearance to a muchmore com- plexset of anteriorstructures. W epresentthe Ž rst scan- 343A comparativestudy ofamphimictic ningelectron microscopy observations of thesechanges. populationsof lesionnematodes with two lipannuli Relationshipswithin Wilsonematinae based on morpho- fromcitrus, coffee, yam, and banana in Brazil logicalcladistics suggest a straightforwardhypothesis of increasingcomplexity from Plectus,whichcould be con- 1 2; Anderson S. DE CAMPOS , Jaime M. DOS SANTOS ¤, sideredancestral, to the highly ornamented condition of 3 4 Larry W. DUNCAN andRenato N. I NSERRA e.g., Tylocephalus and Wilsonema.However,small sub- 1;2UNESP/Faculdadede Ciê ncias Agrá rias e Veterinárias, V ia unit(SSU) rRNA sequencedata are not in congruence deAcesso Prof. P auloDonato Castellane s/ n,14884.900 withthis morphological hypothesis, in that Tylocephalus Jaboticabal,SP ,Brazil and Wilsonema divergedbefore species of Plectus. We 2 Univ.of Florida – IFAS,CitrusResearch and Education discussthe probablehomologies of variousstructures, and Center,Universityof Florida,700 Experiment Station Rd, Lake theimplications of differentphylogenies on characterpo- Alfred,FL 33850,USA laritywithin Plectidae. 3 FloridaDept. of Agriculture,Division of PlantIndustry, P .O. Box147100, Gainesville, FL 32614-7100,USA ¤[email protected] 345An improvedSEM technique forpreparing perinealpatterns of Meloidogyne spp. and for Acollectionof 17 amphimictic populations of Praty- studyinghost pathogeninteractions of sedentary lenchus spp.with two lip annuli collected from Dioscorea nematodes cayenensis Lam., Musa sp., Coffeaarabica L. (one from each), and Citruslimonia L.Osbeck(13 populations) has Jaime M. DOS SANTOS beenmaintained on theirhosts in a glasshouseat theUN- ESP Campusat Jaboticabal, SP ,Brazil.A comparative UNESP/Faculdadede Ciências Agrá rias e Veterinárias, V iade studywas doneon all populations with the exception of AcessoProf. P auloDonato Castellane s/ n,14884.900 Jaboticabal,SP ,Brazil eightfrom citrus,using 21 morphologic, morphometric [email protected] andderived variables obtained using SEM and light mi- croscopy.The populations from yamand banana were Piecesof galled root were Žxedin 0.05 M potassium identiŽed as Pratylenchuscoffeae (Zimmermann)Filip- phosphatebuffer, pH 7.4during 5 days.Subsequently, the jev& SchuurmansStekhoven. All others pertain to a new pieceswere washedin pure buffer and partially dissected

244 Nematology Postersessions underthe dissecting microscope, inside the buffer, to 347SEM observationson the coastalmarine exposethe posterior portion of females, juveniles inside nematode Haliplectusbickneri (Chitwood, 1956) theroots, giant cells or other details of the anatomical Swart et al., 1993 changesin the root caused by the nematodes. After that, 1; 2 thepieces were washedthree times again in the buffer G. Karssen ¤ andA.C. van A ELST andpost-Ž xed overnight in 2% osmium tetroxide. Next, 1PlantProtection Service, P .O.Box9102, 6700 HC thepieces were dehydratedin ethanol, dried in critical Wageningen,The Netherlands 2 pointdryer using CO 2,mountedonto the stubs with the WageningenUniversity, Laboratory Plant Cell Biology, structureof interestfacing up usingadhesive copper tape, Arboretumlaan4, 6703 BD Wageningen,The Netherlands sputteredwith 30 nm of gold, observed and electron- ¤[email protected] micrographedwith a scanningmicroscope JEOL JSM Thefree-living marine nematode Haliplectusbickneri 5410,operated at 15 kV .Bythis technique, perineal (Haliplectidae:Leptolaimina) was detectedin high patternsare better preserved and thereis noneedto cutthe numbersat coastal foredunes in the Netherlands and femalesin lactic acid. Juveniles inside the roots,details of examinedby Ž eldemission electron microscopy. This thegiant cells, and details of the disruption of the root provedto be very useful as most external Haliplectidae tissueswere documented. featuresare strongly reduced. The head region is tapering andincludes: six labial-, six anterior cephalic- and 346Ultrastructure ofthe feedingsite of Gracilacus fourposterior cephalic papiliform sensilla; a minute latescens Raski,1976 in timber bamboo roots stomawith 12-fold vestibulum and circular amphids. The coarslyannulated cuticle has eight longitudinal rows of 1; 2 2 R.N. INSERRA ¤, D. ACHOR , L.W. DUNCAN bodypores, between head and tail end. Annules are 3 and A. TROCCOLI ornamentedwith Ž nelongitudinal striae; additionally a doubletransverse row ofpunctations is present between 1FloridaDepartment of Agriculture,DPI, POBox147100, Gainesville,FL 32614-7100,USA theannules. In males a longventral cuticular rim runs 2 Universityof Florida, CREC, 700Experiment Station Road, from mid-bodytowards four small pre-cloacal papillae, LakeAlfred, FL 33850,USA locatedclosely together near the cloaca. The short 3 IstitutoNematologia Agraria, CNR, viaAmendola 165/ A, conoidtail is provided with a nipple-liketail tip and 70126Bari, Italy includesa spinneretwith three caudal-gland openings. ¤[email protected]..us Thevariability of someof thecharacters and the present positionof theHaliplectidae within the Chromadorida are Female Gracilacuslatescens remainattached by thestylet discussed. tothe surface of timber bamboo roots. Transmission electronmicroscope observations of the nematode feedingsite showed accumulation of callose-likematerial 348Morphological variability of bullaein vulval enwrappingthe stylet.The enwrapped stylet extends from conesof cystsof Heteroderaglycines Ichinohe,1952 thepoint of insertion in an epidermal cell wall into (Nematoda:Tylenchida) evaluated with scanning thelumen of two to three cells in the sclerenchyma electronmicroscopy andcortex. The callose deposit enveloping the stylet Paola LAX1; andMarcelo E. D OUCET 2 increasesin thickness with each additional cell wall ¤ perforatedby the stylet. Possible lignin deposit around 1Laboratoriode Nematologí a, Centrode Zoología Aplicada, thecallose is observed in perforated sclerenchyma cells. UniversidadNacional de Có rdoba, C.C. 122,5000, Có rdoba, Crosssections of theenwrapped stylet revealed the stylet Argentina 2 Facultadde CienciasAgropecuarias, Universidad Nacional tightlyencased in thecallose, which protects the affected deCórdoba, C.C. 509,5000, Có rdoba, Argentina cellsand also anchors the stylet. A syncytiumoriginates [email protected] from theinnermost cell reached by the enwrapped ¤ styletand expands into the inner cortex and stele. Cell Themorphology of bullae in two populations of H. walldissolution and pit Ž eldsare characteristics of the glycines (races 1and3), collected from differentlocalities syncytium. inArgentina, was studied.These cuticle protuberances were locatedat or under the underbridge; their size andshape were variable.Shapes varied from round

Vol.4(2), 2002 245 Morphology,development (336-358) toŽ nger-like;the round bullae were ofdifferent sizes 350Comparative structural, functional and andthe Ž nger-likeones showed variable length and expressionanalyses of the  p-11gene in nematodes thickness.Few cystshad bullae of a singletype; in 1; 1 mostspecimens of both populations the two different Nikki J. MARKS ¤,ChristyL. M OFFETT , 1 2 3 shapeswere presentsimultaneously. The number of Gunnar R. MAIR , Colin F. FLEMING ; , 1 4 thesestructures varied from scarceto highly abundant David W. HALTON ,TimothyG. G EARY , dependingon the individuals, the latter conŽ guration David P. THOMPSON 4 andAaron G. M AULE 1 beingthe most commonly observed. Although this characterwas variable,no relationship with a particular 1 ParasitologyResearch Group, School of Biologyand populationwas established.Some of the different Biochemistry conŽgurations observed would correspond to the expected 2AppliedPlant Science, Queen’ s UniversityBelfast, Belfast patternfor otherspecies. The considerable variability of BT9 7BL, UK 3 thischaracter requires observation of several specimens Departmentof Agricultureand Rural Development for NorthernIreland, Newforge Lane, Belfast BT9 5PX, UK tobe able to determinethe tendency of thesestructures in 4 PharmaciaAnimal Health, Kalamazoo, MI 49001,USA thepopulation under study. ¤[email protected]

349Discrimination among second stage juveniles of Panagrellusredivivus hascharacteristics in common with Heteroderaglycines Ichinohe,1952 of different origin Caenorhabditiselegans (size,habitat, behaviour) yet it and races hasbeen assigned to a differentclade. Caenorhabdi- tis elegans possesses22 genes (designated  ps) that 1; 2 Paola LAX ¤ andMarcelo E. D OUCET encodeFMRFamide-related neuropeptides (FaRPs), and immunocytochemicalanalyseshave revealed that some 1 Laboratoriode Nematologí a, Centro de Zoologí a Aplicada, 50%of the nerves in both worms areFaRPergic. Us- Facultadde CienciasExactas, Fí sicas y Naturales,Universidad ingchromatographic methods, two FaRPs were isolated Nacionalde Córdoba, C.C. 122,5000, Có rdoba, Argentina andstructurally characterised (AMRNA VLRFamideand 2 Facultadde Ciencias Agropecuarias, Universidad Nacional deCó rdoba, C.C. 509,5000, Có rdoba, Argentina NGAPQPFVRFamide) from P.redivivus .Functionalas- says using Ascarissuum revealedthat NGAPQPFVR- ¤[email protected] Famideinhibits body wall muscle and vulval function Secondstage juveniles of fourpopulations of Heterodera whereasAMRNAL VRFamideinhibits body wall mus- glycines from Argentinaand Brazil, belonging to races cleand stimulates vulval function. Single copies of both C.elegans p 1and3, were comparedbased on the study of their peptidesare encoded by -11a,and molec- P.redivivus morphometricalcharacters. An analysis of variance ularanalysis of the orthologous gene from showedsigniŽ cant differences among populations with revealedthat AMRNA VLRFamideand NGAPQPFVR- respectto all the characters considered, except for the Famidewere alsoco-encoded on asinglegene (designated pr p lipregion height. The possible relationships among -1) alongwith the novel peptide, AAGMRNAL VR- C.elegans p populationswere evaluatedthrough a lineardiscriminant Famide. The -11gene has been reported to analysis.Five out of the 16 charactersconsidered resulted beexpressedin threepaired head neurones and a tailneu- In situ pr p asthe most signiŽ cant in separating populations: body ron. hybridisationanalysis of -1revealed that length,body diameter, pharynx length, and ratios a and expressionwas conŽned to a singleunpaired head neuron b.Although the four populations studied showed a certain (RIS-like)with no stainingoccurring in the tail. Although  p pr p relationshipin the discriminant space, it was possibleto -11 and -1are orthologous neuropeptide genes, the P. redi- differentiatethem. In the Ž rst canonicalaxis, populations encodedpeptides appear to fulŽl differentroles in vivus C. elegans. from Argentinaand Brazil were separated;the secondaxis and showeddifferences between representatives of bothraces considered.The results obtained show that individuals 351Morphological and morphometric variations of belongingto different populations and/ orraces of H. P.loosisensu lato glycines mayrepresent different biological entities. 1; 2 3 K.M. MOHOTTI ¤, M.R. SIDDIQI , J. BRIDGE and S.R. GOWEN 3

246 Nematology Postersessions

1 TeaResearch Institute of SriLanka, T alawakele,22100, Sri offour species ( D. major Thorne,1939; D.agricolus Lanka; 2CABI Bioscience(UK Centre),UK; 3Universityof Sauer& Annells,1985; D. labiatus Peña-Santiago et al., Reading,UK; ¤[email protected] 2002; D. mariae Peña-Santiago et al.,2002)collected in Andalucía Oriental(SE IberianPeninsula) have revealed Morphologicaland morphometric variations of 11 po- aninteresting interspeciŽ c variabilityaffecting several pulationsof P.loosisensu lato were examined.Exten- features:presence/ absenceof perioralelevated liplets, oral sivemorphological and morphometric variations existed. Želddivided in six primary radial sectors but sometimes Guatemala,Iran, Japan-Kagoshima, Japan-Shizuoka, Sri withanother six smaller interradial sectors, and outer Lanka-Passaraand Sri Lanka-Talawakelepopulations margindivided in six, 12 or 18 lobes; moreover, both closelyŽ tthemorphometrics of P.loosisensu stricto. sectorsand lobes may be separatedby moreor lessdeep Scanningelectron micrographs of W estAfrica cowpea transversalor longitudinal grooves respectively. These andW estAfrica sorghumpopulations did not show sim- resultsmake possible a newapproach to the taxonomy of ilaritiesto P.loosisensu stricto. TheFlorida popula- thegenus that is briey discussed. tionexhibited morphological similarities to both P. cof- feaesensu stricto and P.loosisensu stricto. TheFlorida, Guatemala,Iran, Japan-Kagoshima, Japan-Shizuoka, Sri 353TEM observationson earlymoulting of anterior Lanka-Passaraand Sri Lanka-Talawakelepopulations body in Xiphinemadentatum (Dorylaimida: along with P. coffeae populationwere separatedby prin- Longidoridae) cipalcomponent analysis of their female and male mor- 1; 2 phometrics.The populations did overlap with no obvious Milan RADIVOJEVIC ¤ and August COOMANS clustersof populations probably because of the consid- 1PlantProtection Institute, F acultyof Agriculture, Nemanjina erablevariability of themorphometrics. The intraspeciŽ c 6,11080Beograd, Y ugoslavia variabilityobserved in the P. loosi populationscould be 2 Instituteof Zoology, F acultyof Sciences,RUG, K.L. attributedto geography, host species and host nutrition, Ledeganckstraat3, 9000Gent, Belgium agronomicpractices and origin of nematodes. Possible ¤[email protected] existenceof distinctbiological pathotypes/ strains P. loosi TransmissionElectron Microscopy (TEM) observations ishighlighted. The P. loosi populationswere conspeciŽc ontransverse sections of the anterior body in moulting witheach other and demonstrated a ‘P. loosi speciescom- Xiphinemadentatum juvenilesrevealed the following: plex’. anteriortissues are withdrawn from thehead cuticle veryearly during moulting. The space in between is 352SEM observationsof the lipregion in species of simultaneouslyŽ lledwith a liquidsubstance, produced by the genus Discolaimus Cobb, 1913(Dorylaimida: theepidermis. The epidermal cells form largechords in Qudsianematidae)with commentson theirtaxonomic thebody cavity, particularly laterally, and extend sheath- interest likeepidermal outgrowths between body cuticle and somaticmuscles. The moulting liquid passes via chords Reyes PEÑA-SANTIAGO¤, Baltasar TORRES, underneaththe body cuticle. Anteriorly, the epidermal Gracia LIÉBANAS, Joaquín ABOLAFIA tissueassumes a tubularform duringits separation from and Pablo GUERRERO theold cuticle. At thattime, it is not yet covered by newepicuticle, showing that in X. dentatum separation Departamentode Biología Animal,V egetaly Ecología, ofthe old cuticle precedes new cuticle formation. The Universidadde Jaé n, Campus‘ LasLagunillas’ s/ n, twoamphidial linings and 16 linings of papillarynerves 23071-Jaén, Spain [email protected] areleft in front of the tissues together with remnants of ¤ thelatter. These linings encircle the old cheilostoma wall The genus Discolaimus Cobb,1913 is characterised by its andform whatwas previouslydescribed as the sleeve-like peculiarlip region, expanded ( i.e.,markedlywider than membranousstructure around the old cheilostoma. theadjacent body) and with the oral Ž eldmore or less sunken,showing a sucker-likeappearance. It is a rather 354Anomalies of genitalorgans in Xiphinema homogenousgenus whose species are mainly separated dentatum (Dorylaimida:Longidoridae) bymorphometric features, so that their identiŽ cation becomesa difŽcult task. SEM studiesof the lip region Milan RADIVOJEVIC

Vol.4(2), 2002 247 Morphology,development (336-358)

PlantProtection Institute, F acultyof Agriculture, Nemanjina 6, acervicalcollar bearing numerous, posteriorly directed, 11080Beograd, Y ugoslavia spines.The collar is deected posteriorly on eitherside of [email protected] themidlateral line to form apairof cuticularlappets, the free marginof each lappet bearing a fringeof cuticular Duringa surveyof Xiphinemadentatum , numerous spines.Posterior to the lappets are two pairs of lateral individualsfrom asingleŽ eldpopulation in Y ugoslavia spines,their bases arranged more or less in tandem. In were observedunder dissecting microscope. Seven were thespecies studied, the transverse striae of the cervical foundwith an abnormal genital system, and these regionbear retrorse posterior margins. A numberof stud- were permanentlymounted and studied under light likesomatic papillae are also visible. V ariationsin form microscope.Normal males are rare, sometimeswith anddetail of the female cervical armature are of major fullydeveloped diorchic tract, but usually with only a diagnosticimportance in speciŽc determinationwithin the copulatoryapparatus. Of 137males, one is abnormal, genus Heth. havinga normalcopulatory apparatus, two abnormal testesand lacking the seminal duct. Normal females have twoopposed, equally developed genital branches, opening 356Morphometry of Pratylenchus populationsfrom througha slightlyanterior vulva (V 42:0 1:3%). Of coffee,banana, ornamental plant and citrus in Brazil some42 000 females, six were foundD to be § abnormal. 1; Onehas the genital system anatomically normal, but SilviaRenata S ICILIANO-WILCKEN ¤, 2 2 abnormallyposterior (V 71.0%).T wofemales have Mário M. INOMOTO ,LuizCarlos C.B. F ERRAZ D 3 twonormally developed systems, one system in anormal and C.M.G. OLIVEIRA position (V 45.9and 43.8%, respectively), the 1 otherposterior D(V 62.1and 65.3%, respectively). FCA/UNESP,P.O.Box 237, 18603-970, Botucatu, SP ,Brazil D 2 ESALQ/USP,Piracicaba,SP ,Brazil Anothertwo females lack central parts of thesystem, the 3 InstitutoBioló gico, Campinas, SP ,Brazil remainingdistal parts being fused proximally. The sixth ¤[email protected] femalehas a normalposterior branch, while the anterior oneis abnormally branching at parsdilatata uteri into two Themorphometric variability of four Pratylenchus distaloviducts with ovaries, one ovarybeing degenerated. populationsfrom differenthosts and regions of Brazil identiŽed as Pratylenchuscoffeae was studied.These 355SEM studies ofthe pseudolabiaand cervical populationswere extractedfrom rootsof coffee, citrus armatureof afemale Heth (Rhigonematida: (São PauloState), Aglaonema (Riode JaneiroState) and Hethidae) bananaplant (Minas Gerais State)and were cultured onalfalfa callus tissues. Baermann-extracted specimens 1; 1 Janet ROWE ¤, Rosa MANZANILLA-LÓPEZ ofeach population from calluswere killedby heat and and David HUNT 2 Žxedwith 4% formalin.T emporaryslides were madeand

1 examinedin lightmicroscope. Body length (L), excretory NematologyInteractions Unit, IACR-Rothamsted, Harpenden, poreto head end, stylet length, relative distance of vulva Herts,AL5 2JQ,UK from anteriorend (V%), bodylength/ greatestbody width 2 CABI Bioscience,UK Centre,Egham, Surrey, UK (a), bodylength/ distanceto anterior and to junction of ¤[email protected] pharynxand intestine (b), body length/ taillength (c), Heth Cobb,1898 is an unusual genus of Rhigonematida taillength/ bodywidth at anus (c’ ) were obtained.There foundin the posterior gut of tropicaldiplopods from the were statisticallysigniŽ cant differences in allparameters Americanand Australasian regions. The species studied studied. hereoriginated in a rhinocriciddiplopod from Trinidad, WestIndies. Sexes are dimorphic, the female bearing a 357Cuticular diversity within the genus Ogma complexpair of pseudolabialplates arching over the oral (Criconematidae) aperture.The pseudolabia are subtrapezoid in shape and 1; 2 areattached laterally, the remaining free marginsbeing L.R. TIEDT ¤ and Esther VAN DEN BERG fringedwith pectinate cuticular processes. These ‘ combs’ probablyact as Ž ltersduring feeding, restricting the size 1 Laboratoryfor Electron Microscopy, University of ofingestedfood particles. Posterior to the pseudolabia is Potchefstroomfor C.H.E., Potchefstroom,2520, South Africa

248 Nematology Postersessions

2 NationalCollection of Nematodes, Biosystematics Division, Cralley-WarrenCenter ,2601N. YoungAvenue, F ayetteville, ARC-PlantProtection Research Institute, Private Bag x134, AR72704,USA Pretoria0001, South Africa Longidorus [email protected] currentlyhas 132 valid species based pri- ¤ marilyon morphometrics.Many Longidorus specieshave Femalesand juvenilesof thegenus Ogma displaythe most ahighdegree of variability within their morphometrics spectacularand diverse cuticular ornamentation. Some whichleads to considerableoverlap between species and ofthe differences in the structure of a few speciesare increasesthe potential for misidentiŽcation. One way illustrated.Cuticular structure plays an important role in Anova,Student’ s tandprincipal components were ap- theidentiŽ cation of nematodespecies. Until recently the pliedto compare seven species of Longidorus found in cuticleof only females were studiedand recorded, but Arkansas (L.breviannulatus , L. crassus, L.diadecturus , itis becoming more important to take note of the fact L. fragilis,threeundescribed species) in order to screen thatthe cuticular structure of thedifferent juvenile stages for themost useful morphometrics in speciesdiscrimina- withina speciesdiffer from thoseof thefemale and even tion.Canonical analysis and hierarchical cluster analysis from eachother in most species. These differences can basedon femaleaverage morphometric characters includ- causeproblems in a populationwhere more than one ingbody length, distance from vulvaopening to headend, Ogma speciesis presentbecause, if thediversity in species lipwidth, odontostyle length, pharynx length, body di- isnot known, wrong identiŽ cations can be made. It is ameter,tail length and diameter, were usedto examine thereforeimportant to become familiar with and study themorphometric relationships and create dendrograms thesestructures and to include their descriptions in all among130 published Longidorus species (L. heynsi An- descriptionsof speciesas far asispossible.The SEM is a drassy,1970 and L.tardicauda Merzheevskaya,1951 with handytool to illustrate the cuticle variation in thisgenus. insufŽcient data were notincluded) and 94 populations from Arkansasand otherlocations by usingJump 4.0 soft- 358Statistical analysis of morphometricsof wares. Sixmajor clusters were producedfor all Longi- Longidorus species(Nematoda: Dorylaimida) dorus species.Cluster analysis may lead to identiŽcation ofunidentiŽed Longidorus species. Weimin YE and R.T. ROBBINS

Vol.4(2), 2002 249 Foodwebs, ecology, biodiversity (359-382)

359List of soilnematodes from a non-tillageŽ eldin atplanting and at harvest using a modiŽed nested Tsukuba,Japan, with acommenton the Maturity samplingdesign from twoŽ eldsin Michigan. Samples Indexand species diversity were analysedfor SCN cystsand soil texture. A strongcorrelation between cyst density and soil texture Masaaki ARAKI¤ andZakaullah K HAN was found.Coarse soil with more than 60% sand, NationalInstitute for Agro-Environmental Sciences, 3-1-3 lessthan 20% silt and less than 20% clay had Kannondai,Tsukuba, Ibaraki, 305-8604 Japan consistentlymore cysts than Ž nersoil. As deŽned by the ¤[email protected] USDA-recommendedtexture triangle, this composition correspondsto sandy loam, loamy sand and sand. Alistof soilnematodes from anon-tillage(organic) Ž eld Previouswork showed SCN preferencefor sandysoil establishedin 1981 at the National Institute for Agro- types,but it is important to note that only a portionof EnvironmentalSciences, Tsukuba, is given compared thearea deŽ ned for sandyloam in the texture triangle withthose from aconventional(inorganic) Ž eldand a isfavourable for SCN. Our resultsindicate there is a grovenearby. The soil type of the land concerned is potentialfor site-speciŽc managementof SCN basedon andosol.Barley and soybean were cultivatedin a double- soiltexture (particle size) rather than soil type. croppingsystem on the Ž elds.The non-tillage Ž eldwas amendedwith manure made yearly from litterfrom thegrove. Nematodes in 20 g soilwere extractedby 361Structure ofnematodecommunities in nine Baermannfunnel method in October,1999 and countedto typesof ecosystems eachdistinguishable taxon under compound microscope Mirjana BRMEZ , Marija IVEZIC andEmilija R ASPUDIC atmagniŽ cation of 100 .NematodeidentiŽ cation was ¤ £ donewith specimens mounted on slidesin glycerin. The Universityof J. J.Strossmayer in Osijek,F acultyof Agriculture totalnumber of speciesdetected from thenon-tillage Ž eld inOsijek,Trg sv .Trojstva3, 31000 Osijek, Croatia sofar was 55,though the number of speciesidentiŽ ed to ¤[email protected] thespecies level is rather small. The number of species from theconventional Ž eldand grove were smallerand Nematodecommunities were analysedin nine ecosys- largerthan from thenon-tillage Ž eld,respectively. This tems:winter wheat in crop rotation – standardtillage; tendencywas thesame in Maturity Indices calculated winterwheat in crop rotation – reducedtillage; corn in from thedata, but Shannon’ s H’indiceswere largestin the monoculture;sugar beet; potato – chemicalplant protec- non-tillageŽ eld.Abundance of Monhystrellapostvulvae , tion;potato – integratedplant protection; alfalfa; grass- Prismatolaimus sp., Wilsonemaothophorum , Bastiania land;poplar forest. Sampling was donein 1997and 1998, sp., Alaimus sp. and Xiphinema sp.characterised thennon- inspringand autumn, to thedepthof 15-20cm. Nematode tillageŽ eld.In contrast, the conventional Ž eldhad greater communitieswere analysedto genus level. Greatest num- Pratylenchuspenetrans and Cephalobus sp.populations. berof nematodes and number of genera were 2843ne- matodes/100g ofsoilin alfalfatreatment, and 30 genera 360The relationship between soiltype and soil inpoplar treatment. Lowest was inrow crops(216 and texturewith the spatialdistribution of Heterodera 12genera in sugar beet). Analyses of nematode trophic glycines structureshowed greatest abundance of bacterialfeeders andplant parasitic nematodes in alltreatments, and lowest

Felicitas AVENDAÑO¤, F.J. PIERCE, abundanceof omnivorousnematodes and predators. The O. SCHABENBERGER and H. MELAKEBERHAN greatestabundance of predatorswas determinedin poplar forest.Positive correlation between quantity of humusand MichiganState University, Department of Entomology,243 totalnumber of nematodes was determinedbut a nega- NaturalScience Building, East Lansing, MI 48824,USA tivecorrelation between amount of K 2Oandnumber of [email protected] ¤ genera.Statistically very signiŽ cant differences occurred Inorder to test the potential of managing soybean betweentreatments in analysing MI, PPI andPPI/ MIand cystnematode (SCN), Heteroderaglycines Ichinohe showedthe most disturbed nematode communities in row site-speciŽcally, the spatial relationship between SCN crops,the less disturbed in poplar forest, grassland and al- distributionand soil texture was exploredunder Ž eld falfa;and intermediate group was wheatwith reduced and conditions.Single-core soil samples were collected standardtillage.

250 Nematology Postersessions

362Effects ofsoilpH on Heteroderaglycines ecologicalindices based on proportions of Ž vegroups reproductionand soybean chlorosis onthe coloniser-persister scale. These data are analysed inrelation to variations in root growth and microbial Senyu CHEN and Daniel MILLER communitystructure and activity over 3 years.In 13 SouthernResearch and Outreach Center ,Universityof additionalplots, the CŽxedby photosynthesis of a 13 Minnesota,35838 120 th Street,W aseca,MN 56093,USA 24 h pulse of CO2 appliedto the herbage, was detected innematodes extracted from underlyingsoil at 8, 22, Soybeancyst nematode, Heteroderaglycines , is a 48and 69 days after the pulse. There was more 13C in destructivepest on soybean in Minnesota, a statein nematodesfrom untreatedcontrols than from limedplots. theNorth Central region in the United States, where Thesedifferences are consistent with a greatercarbon soybeanis a majorcrop. The objective of this study uxin soils treated with lime, but could not be related was todetermine the effect of soil pH on H. glycines todifferencesbetween the nematode communities. reproductionand iron-deŽ ciency chlorosis of soybeanin Želds.A surveywas conductedin a 20acre area in acommercialsoybean Ž eldin W aseca,Minnesota, to 364A new nematodegenus (Rhabditida: monitorsoil pH, density of H. glycines,andseverity of Panagrolaimidae)from pitcher plants iron-deŽciency chlorosis of soybean in 2001. Positive Paul DE LEY1; , Wim BERT 2, Andy VIERSTRAETE 2, correlationbetween either two of the three variables ¤ Irma T. DE LEY 1, Rita VAN DRIESSCHE 2 was signiŽcant. Glasshouse studies conŽ rmed that H. andHendrik S EGERS 2 glycines reproductionon susceptible soybean increased withincreasing soil pH from 5to8. The results suggest 1 Departmentof Nematology, University of California– that H. glycines mayincrease severity of iron-deŽciency Riverside,CA 92521,USA 2 ofsoybeanat higher soil pH inMinnesotaŽ elds. VakgroepBiologie, Universiteit Gent, Ledeganckstraat 35, B-9000Gent, Belgium ¤[email protected] 363Relationships between nematodeand microbial communitiesand carbon transfers in grassland soils Measurements,line drawings and scanning electromicro- graphsare provided of anewnematode species and genus, 1; 2 3 Roger COOK ¤, Lorna DAWSON , Alan GANGE , isolatedfrom pitcher uidof Nepenthes sp.from Thai- Sue GRAYSTON 2, Tony MIZEN 1, Phil MURRAY 4, land.The new genus differs from allknown nematodes in Amy TREONIS 2 and Nick OSTLE 5 havingtwo opposing and offset spermathecal pouches at thejunction of oviductand uterus. It also differs from all 1 Instituteof Grasslandand Environmental Research, knownRhabditida in having four cephalic setae instead AberystwythResearch Station, Aberystwyth SY23 3EB, Wales, ofpapillae.Phylogenetic analysis of smallsubunit rDNA UK 2 MacaulayInstitute, Craigiebuckler ,Aberdeen,Scotland, UK sequencedata robustly places the new genus within Pana- 3 RoyalHolloway University of London, Egham, Surrey, UK grolaimidae,as sister taxon to Panagrellus .Theseunusual 4 Instituteof Grasslandand Environmental Research, North nematodesresemble Panagrellus in e.g.,bodysize (1.8- WykeResearch Station, Devon, UK 2.5mm infemales, 1.3-1.8 in males)and in themonodel- 5 Centrefor Ecology and Hydrology, Merlewood Research phic,prodelphic female reproductive system with thick- Station,Grange-Over-Sands, Cumbria, UK enedvaginal walls and prominent postvulval sac. How- ¤[email protected] ever,they differ from Panagrellus inthecharacters men- At anupland Ž eldsite, plots of an established Festuca tionedabove, in thepresence of lipletsaround the mouth – Agrostis pastureand of a newlysown monoculture opening,in their comparatively longer stegostom, and in of Loliumperenne havebeen treated for 3yearswith theshape of the male spicules. Because of its aberrant nitrogenand lime or withpesticide (chlorpyrifos). These characters,inclusion of this new genus in Panagrolaimi- treatmentswere designedto produce contrasting degrees daewill entail changes to thefamily diagnosis. ofdiversityin communities of soilanimals and microbes aspart of our research on the relationships between 365Ecophysiological studies onpinewilt disease grasslandmanagement practice and the diversity of occurringin coastaldunes biologicalcommunities in soil. Nematode communities 1; 2 havebeen described by trophic composition and by Kazuyoshi FUTAI ¤,ShigenobuT AMAI ,

Vol.4(2), 2002 251 Foodwebs, ecology, biodiversity (359-382)

Norikazu YAMANAKA 2 and Fukuju YAMAMOTO 3 were conductedin three regions from 1995to 2000, 1 Laboratoryof EnvironmentalMycoscience, Graduate School comparingtwo pH levels,under twocrop rotation systems ofAgriculture, Kyoto University, Oiwake-cho, Kitashirakawa, usingcorn; one (pH5 to 5.5 in CaCl 2/ representing Sakyo-ku,Kyoto 606-8502, Japan therecommended pH range, and another (6 to 6.5) 2 Laboratoryof Revegetationand Grassland Development, representinga highpH level.In all experiments there was TottoriUniversity Arid Land Research Center ,1390, atrendof higher SCN populationdensity in plots with Hamasaka,T ottori680-0001, Japan highersoil pH. Under these conditions, 1 yearrotation 3 Facultyof Agriculture, T ottoriUniversity, 4-101 Koyama-cho withcorn reduced the SCN population,but its incidence Minami,T ottori,680-8553, Japan was stillhigher in plots with higher soil pH. ¤[email protected] Thespreading manner of pine wilt disease has been 367Edaphic microfauna in woodycrops from comparedbetween two 20 20m experimentalplots Mediterraneanenvironments establishedin Japanese black £pine forest distributed over 1; 2 coastaldunes in T ottoriUniversity Arid Land Research Avelino GARCIA-ÁLVAREZ ¤, Carlos SIMÓN , Center.Oneof the plots was fertilisedevery year since Susana C. ARCOS 1, María ARIAS 1 1996,and the other one remained with no treatmentand and Victor ESPANTALEÓN 2 servedas a control.The number of pine trees killed by pinewilt disease was far morein the fertilised plot than 1 DptoAgroecologí a, CCMA, CSIC,Serrano 115 dpdo, 28006 inthe control one, suggesting that the fertilisation might Madrid,Spain havepromoted the death of pinetrees by pinewilt disease. 2 DptoZoologí a, Univ.Autó noma, Cantoblanco, Madrid, Spain Torevealthe reason why the fertilisation promoted ¤[email protected] thepine wilt disease, mycorrhizal relationship has been examinedas a possiblecue to explain the difference. Fertilisationmight suppress mycorrhizal synthesis which Microfaunaassociated with several vineyard and olive hasbeen regarded to work as absorbing organ for pine cropsgrown in a continentalMediterranean climate treesespecially when under water stress. Diversity and undervarious soil management systems on the Spanish constitutionsof mycorrhizae formed on young pine Centralplateau were studied.Nematode trophic groups seedlingswere comparedbetween the two plots, and andmicroarthropod populations in plots under different amongthree groups of pine seedlings with different croppingsystems (non-tillage, traditional tillage, covers wateringregimes. W econcludethat mycorrhizal relations withclover, weeds or vetch)were studiedin comparison playan important role to make pine trees survive the withtwo controls (soil from anabandoned cereal crop epidemicwilt disease. anda hedgeof wild vegetation). Plant-parasitic and bacteria-feedernematodes had the highest populations in thehedge control and in plots with clover cover, while 366Effect ofsoilpH on Heteroderaglycines theirpopulations were lowerunder non-tillage. Predators populationdensity in Brazil were abundantin grape: under traditional management, undera coveringof runners and in the control group. Antonio GARCIA1; ,WaldirPereira D IAS 1, ¤ However,fewer predatorswere foundin non-tillage plots, João Flávio V elosoS ILVA 1,RobertoKazuhiko Z ITO 2 whereomnivorous predators abound, in olivewith clover andAngela Francoise C ARVALHO SANTOS 3 coverand in the control group with natural vegetation. 1 EmbrapaSoja, CP 231,86001-970, Londrina-PR, Brazil Collembolahad a higherpopulation in allalternatives in 2 EPAMIG, AfonsoRato, 1301, 38001-970, Uberaba-MG, plotswith clover cover and control groups. Acari were Brazil frequentin vineyards, except in weed-covered and non- 3 UNIFIL, JuscelinoKubtscheck, 1626, 86020-918, tilledplots and along the edge,appearing in weed-covered Londrina-PR,Brazil [email protected] oliveand the control group. Coleoptera appeared in all ¤ butnon-tilled olive plots while Diptera were invineyards, Previousobservations in growers’ Ž eldsinfested with exceptthose under tillage, and the control group, where thesoybean cyst nematode (SCN), Heteroderaglycines , onlyThysanoptera were present. suggestedthat SCN populationsremained higher for long periodsin soils with high pH. Therefore, experiments

252 Nematology Postersessions

368Diversity of soilnematodes in analpine Mediterraneanclimate, the study of nematofaunatook the dwarf-shrub heath inAustria: Žrst resultsof a2year followingissues into account: i) land use, ii)varioussoil study horizons, iii)seasons,and iv)extractingmethods. Results showthat an estimation of the taxonomic biodiversity Michael HOSCHITZ¤ and Pamela ZOLDA andindividual abundance patterns is notpossible,without Instituteof Ecologyand Conservation Biology, Althanstraß e takingall the above-mentioned items into account. 14,A-1090 V ienna,Austria Differentextracting methods show distinct scenarios, as ¤[email protected] wellas respect for seasonsand soil depth. Sampling the upper15 cmoftheA horizoninvolved a greatabundance Thesoil nematode community was investigatedin an ofsoil individuals, biomass and biodiversity. It is also alpinedwarf shrubheath in the Central Alps (1930 demonstratedthat in some instances the B horizon(not m) nearObergurgl, Austria. The vegetation on the usuallysampled for soilbiodiversity and soil monitoring acidicsoil consisted mainly of Vacciniumgaultherioides , studies)has more individuals and taxonomic diversity V.myrtillus and Rhododendronferrugineum . The aim thanthe Ahorizon.Thus, from thedata obtained, it seems ofthe study was Žrstlyto investigate the nematode thatboth the theoretical assumptions and the standard biodiversityof alpine environments. Secondly, it was techniquescurrently used for soilbiodiversity and soil- designedto serve as a basisfor investigationson the monitoringstudies do not show the complexity of soil useof nematode communities as an indicator for the nematofauna.Therefore, we needto improve present globalclimate change. A totalof 216 soil samples was methodsto develop more relevant research on these takenin July, August and September 2001. Allnematodes topics. extractedfrom bulksamples were identiŽed andcounted. Nematodesof all Ž vefeeding groups were encountered. Alistof species, species abundance, diversity indices 370Nematode abundance, diversity and trophic andthe maturity index are presented. The nematode structure inrelation to spatial heterogeneity of communityis apparently typical for alpineregions and the anaturalfen mostdominant genus was Rotylenchus sp.(T ylenchidae). Furtherplant-feeding families were Pratylenchidaeand Krassimira I LIEVA-MAKULEC Paratylenchidae.Within the predators, Mononchidae and Instituteof Ecology,P olishAcademy of Sciences,Dziekanow Anatonchidaewere encounteredand the bacterial feeders Lesny,05-092 Lomianki, P oland were representedby the Prismatolaimidae, Plectidae, [email protected] Cephalobidaeand T eratocephalidae.Genera of the Theabundance, diversity, and trophic structure of omnivorousfamilies Dorylaimidae and Aporcelaimidae nematodecommunities of a naturaltall-sedge fen in were identiŽed, along with genera of the fungivorous Polandwere studied.The sedge association was ofatufty Aphelenchidae,Aphelenchoididae and Nordiidae. structuredue to the dominance of Carexparadoxa . Soil sampleswere takenin the spring(May) and in thesummer 369Current uncertaintiesin the estimationof soil (July)within and outside of the sedge tussocks. It was nematodediversity and its use insoilquality foundthat nematode communities were moreabundant monitoring andmore diverse within than outside of thetussocks and thedifferences were morepronounced in thespring than Juan José IBÁÑEZ¤, Avelino GARCÍA-ÁLVAREZ inthe summer. Both habitats differed also in nematode and María ARIAS trophicstructure and in the relative importance of the trophicgroups. No omnivorous and predatory nematodes Dptode Agroecología, CCMA, CSIC,Serrano 115 dpdo, 28006 were foundin the soil between tussocks on the spring Madrid,Spain sampling.Bacterial-feeding nematodes predominated in jjibañ[email protected] ¤ thetwo habitats but, while within the tussocks, on both Estimationof soil biodiversity and monitoring of soil samplingdates they contributed to about 60% of thetotal qualityare two of themajor priorities of theinternational nematodenumbers in the soil between tussocks, bacterial- environmentalagenda, although their quantiŽ cation is feedingnematodes constituted 94 and 73% in spring notfree from uncertainties.This contribution analyses andsummer, respectively. The variation of nematode theseuncertainties. In an Acrisol under a continental communityparameters in such a shortdistance was

Vol.4(2), 2002 253 Foodwebs, ecology, biodiversity (359-382) explainedby the differences in some soil characteristics were greatestin forestry, successional and organic ofthetwo studied habitats. systemsin July and September compared to conventional croppingsystems ( P 0:01/.Thepopulation densities D 371Phylogenetic analysis of the phoreticassociation offungivorous and bactivorous nematodes were greater between Bursaphelenchusconicaudatus (Nematoda: inorganic systems compared to other systems at theJuly sampling, but not in September ( P 0:01/. Aphelenchoididae)and Psacotheahilaris (Coleoptera: D Cerambycidae) Temporalvariation in populationmaximum may in uence conclusionsabout the relationships of trophic groups Natsumi KANZAKI¤ andKazuyoshi F UTAI toone another. The use of organic amendments tosupply nutrients in the organic system may be Laboratoryof Environmental Mycoscience, Graduate School of responsiblefor highnumbers of free-living nematodes Agriculture,Kyoto University, Oiwake-cho, Kitashirakawa, inthese treatments. Cotton in no-till production had Sakyo-ku,Kyoto 606-8502, Japan greaternumbers of bacterial feeding nematodes than ¤[email protected] conventionaltillage ( P 0:10/.Thepresence of various D Thephoretic relationship between the nematode Bursa- speciesof plant-parasitic nematodes was relatedto phelenchusconicaudatus andthe yellow-spotted longi- thecrop grown, and was littlein uenced by tillage cornbeetle, Psacotheahilaris ,collectedat various dis- system.The entomopathogeni cnematodes Steinernema trictsin Japan were investigated.All ten subspecies of glaseri, Steinernemacarcocapsae and Heterorhabditis thebeetle examined were infectedwith the nematodes bacteriophora were detectedin all treatments. Highest ata highproportion suggesting the universality of this numbersof entomopathogen icnematodes were detected phoreticassociation. Molecular analysis based on thepar- inthesuccession and forestry treatments. tialbase sequence of cytochromeoxidase subunit I inthe mitochondrialDNA revealedthat the phylogenetic rela- 373An essayof detectionof zoogeographicunits of tionshipamong ten subspecies of the beetle was similar the territoryin baseto nematode fauna distribution: tothat among associated nematode isolates obtained from the caseof aprotectedarea in SEIberianPeninsula eachbeetle subspecies. IntraspeciŽ c vectorreplacements were supposedto occur in the isolates of B.conicaudatus 1; 1 Gracia LIÉBANAS ¤, Reyes PEÑA-SANTIAGO , ontwo islands. The phoretic association between the ne- Raimundo REAL 2, Ana Luz MÁRQUEZ 2, matodesand the beetles was assumedto be on the way JuanManuel M ARTÍN-GARCÍA 3, Joaquín ABOLAFIA 1 from non-speciŽc facultativecommensalism to speciŽ c and Pablo GUERRERO 1 obligativecommensalism. 1 Departamentode Biologí a Animal,V egetaly Ecología, Universidadde Jaén, Campus ‘ LasLagunillas’ s/ n, 372In uence of diverseagricultural systems on the 23071-Jaén, Spain populationdynamics of free-living,plant-parasitic 2 Departamentode Biologí a Animal,F acultadde Ciencias, andentomopathogenic nematodes Universiadde Má laga. 29071-Má laga, Spain 3 Departamentode Geologí a, Universidadde Jaén, Campus Stephen R. KOENNING¤ and Mary E. BARBERCHECK ‘LasLagunillas’ s/ n,23071-Jaén, Spain [email protected] CampusBox 7616, Department of Plant P athology, ¤ NC27695-7616,USA; Campus Box 7634, Department of TheidentiŽ cation of biotic regions, i.e., series of Entomology,North Carolina State University, Raleigh, operationalgeographic units (OGU) thatshare a similar NC27695-7634,USA fauna,leading to the division of the territory into [email protected] ¤ operationalzoogeographic units (OZU), was oneof the Thepopulation dynamics and densities of soil-inhabiting objectivesof a nematologicalproject addressing the nematodeswere measuredin different agricultural taxonomyand distribution of dorylaimidand mononchid systemsat theCenter for EnvironmentalFarming systems nematodefauna of the Sierra Mágina Natural Park (SE nearGoldsboro, NC, USA. Systemsstudied included IberianPeninsula). Sixty OGU ofthe UTM (Universal old-Želd succession, forestry, integrated crop/ animal, TranversalMercator) 2 2kmsquare grid were £ organic,and cotton in conventional vs no-tillproduction. classiŽed in baseto the presence/absenceof 138nematode Numbersof omnivorous and predacious nematodes speciesusing a quantitativeanalysis protocol. The results

254 Nematology Postersessions obtainedsuggest that the studied area may be divided Dipartimentodi Biologia Animale, via Androne 81, 95124 intoseven biotic (zoogeographic) regions with their Catania,Italy respectivebiotic boundaries. The biotic regions have ¤[email protected] beenenvironmentally characterised, with altitude, slope, Asurveyof nematodesof somecoastal dune ecosystems andsand and clay contents the main factors in uencing inSicily was carriedout both from afaunisticand an thedivision. The analysis also produced a faunistic ecologicalpoint of view. The species composition was characterisationof thebiotic regions. These results, which verypeculiar, with many new species, many taxa typical representa newapproach to the study of nematode ofdunesorrare, andsome others typical of coastalmarine distribution,are discussed. andbrackish environments. The ecological analysis was conductedmore deeply in two nature reserve sites, 374Nematode community and trophic structure namelyV endicariand Irminio river-mouth, with different alonga sanddune succession environmentalcharacteristics. Shannon diversity index, speciesrichness, evenness, maturity index, frequencies Roy NEILSON¤, John W. WALL andKeith R. S KENE ofc-p categories and of trophic groups were calculated ateach station. The results were comparedwith those Plantand P athogenCo-Evolution Programme, Scottish Crop from someother coastal sites with a differentdegree ResearchInstitute, Dundee, DD2 5DA,Scotland, UK; Schoolof ofenvironmental disturbance. Both biodiversity and LifeSciences, University of Dundee, Dundee, DD2 1HN, maturitywere higherin the stations where dunes were Scotland,UK betterpreserved, but the differences among the sites were [email protected] ¤ notas high as onewould expect, probably because dune Changesin below-ground nematode communities, in nematodesare adapted to survive in suchhighly changing termsof abundance,diversity, trophic structure, sex bias environments. andadult : juvenileratio were relatedto edaphic fac- torsfrom samplesites representing differing successional 376Restoration of soilnematode fauna (dorylaims stages.Nematode abundance increased along a 1kmtran- andmononchs) in the GuadiamarRiver basin (SW sectof increasingsuccession, from sandybeach through IberianPeninsula) 3 yearsafter a miningaccident activedune systems to mature dunes. As thesuccession 1; progressed,the adult nematode sex ratio changed from Reyes PEÑA-SANTIAGO ¤, 2 1 malebiased to highly female biased. Consequently, the Domingo JIMÉNEZ-GUIRADO , Gracia LIÉBANAS , 2 1 adult: juvenileratio decreased as the nematode com- Raquel MURILLO , Joaquín ABOLAFIA 2 munitybecame more female dominated. An assessment and Pablo GUERRERO ofnematode diversity using both univariate and multi- 1 Departamentode Biologí a Animal,V egetaly Ecología, variateanalysis produced contradictory results. Univari- Universidadde Jaén, Campus ‘ LasLagunillas’ s/ n, ateindices yielded inconsistent results with different in- 23071-Jaén, Spain dicessuggesting lower diversity at differentsuccessional 2 Departamentode Biologí a Animal,Universidad de Córdoba, stages.Multivariate analysis clearly separated the beach Campusde Rabanales,EdiŽ cio C-1, 14071 Có rdoba, Spain anddune heath samples from theother samples and each ¤[email protected] otherand, to alesserextent, separated the fore dunesam- As aconsequenceof themining accident which occurred plesfrom thegrey and yellow dune. Omnivorous nema- atthe Guadiamar River basin (SW IberianPeninsula) in todesrepresented the largest trophic component of thene- April1998, part of the river banks were coveredwith matodecommunity in allsuccessional stages of theterres- toxicmud containingheavy metals, and aresearchproject trialsites. A trendwas apparentof increasingnumbers of focusedon the faunistic characterisation of the area and omnivorousnematodes along the succession in relation to controlof the recolonisation process was undertaken. increasingorganic matter and/ orlitteraccumulation. Nematodefauna, particularly dorylaims and mononchs, was includedin the study. A few well-conservedand 375Analysis of nematofaunain some coastal dune non-pollutednatural areas (biocentres), and a seriesof ecosystemsof Sicily experimentalplots located in the river banks, have been surveyedover two seasons (spring and autumn) during 2 Lara ORSELLI¤ andMaria Teresa V INCIGUERRA years(2000 and 2001). One hundred and twenty seven

Vol.4(2), 2002 255 Foodwebs, ecology, biodiversity (359-382)

1; 1 soilsamples were collected,but only 99 havebeen studied Carmen SANTOS LOBATÓN ¤, Eva TRUJILLO MENA atpresent. More than 100 species constitute the fauna and Miriam CASAUS ALVAREZ 1 oftheregion, indicating a remarkablefaunistic potential 1Laboratoriode Nematología. Dpto de Fisiología yZoología, ofthis geographical area. Concerning the recolonisation Avda,Reina Mercedes 6, 41012Sevilla, Spain process,two distribution patterns, one spatial and another ¤[email protected] temporal,have been detected and are also discussed. Inthis paper we presentthe results of a studycarried outon 28 waste waters ofAndalucía (Spain)in order to 377Soil nematode community as avaluable assessthe presence of nematodes in the environs of the bioindicatorof soilecosystem recovery after mining treatmentprocess with special attention to sludge sam- spillagein southern Spain plesthat can constitute a suitableenvironment for devel- opmentof thisorganisms. Nematodes were presentin all S. SÁNCHEZ 1;2, J.A. CAMARGO 1 and A. NAVAS 2 thewaste water and 21 species belonging to 21 genera were identiŽed. These species are Alloionemaappendicu- 1 DepartamentoInteruniversitario de Ecologí a, Universidad de lata, Eudorylaimusintermedius , Brevibuccasaprophaga , Alcalá,Alcalá de Henares,Madrid, Spain Cervidelluscervus , Peloderaserrata , Cruznematriparti- 2 MuseoNacional de Ciencias Naturales, C.S.I.C., Madrid, tum, Diploscaptercoronatus , Fictorsimilis , Mesorhab- Spain ditisspinula , Neodiplogasterpissodis , Panagrobelusin- cisus, Paroigolaimellacoprophaga , Pelloditisfriderici , InApril 1998, an amountof about5hm 3 oftoxicsludge, Peloderateres , ProtorhabditisŽ liformis , Rhabditismau- withhigh concentrations of heavy metals, was released pasi, Rabditisproducta , Rhabditophanesschnederi , Ty- alongthe Guadiamar river system as a consequenceof locephalusauriculatus , Anguillulaaceti , Tylopharinx thetragic Aznalcollar accident in southern Spain. In phoetida and Xiphinemapachtaicum .Specieswere orderto evaluate the environmental impact produced by groupedaccording to theirfeeding habits and their com- theheavy metal pollution on soil health conditions, we munitycomposition were studied.The results obtained carriedout Ž eldand laboratory studies. Ten control and providenew information on theoccurrence of populations 27pollutedsoil samples were takenalong the Guadiamar androle of theseorganisms as indicators. riversidein threesampling periods (June 2000, December 2000and June 2001). For eachsoil sample, values of 379Dispersal ability of Bursaphelenchusxylophilus diversityand trophic indices were calculated,and the (Nematoda:Aphelenchoididae) and inhibition by concentrationof heavy metals (Pb, Zn, Ni andCu) was Pinus densiora branches analysed.Soil nematodes were identiŽed, at least to the genustaxonomic level. In addition, short-term toxicity Katsumi TOGASHI¤, Koji MATSUNAGA, Yoh ARAKAWA bioassayswere conductedin triplicate using adults of and Naoto MIYAMOTO Aphelenchusavenae and Cephalobuspersignis collected atthe control area. Results from Želdstudies showed that Facultyof IntegratedArts and Sciences, Hiroshima University, concentrationsof heavymetals were signiŽcantly higher Kagamiyama1-7-1, Higashi-Hiroshima 739-8521, Japan atpolluted sites than at control sites. Conversely, values ¤[email protected] ofbiological indices were signiŽcantly much lower at Todeterminethe degree of inhibition against the pollutedsites. Results from short-termtoxicity bioassays nematodedispersal by live Pinusdensi ora trees, four revealedthat Pb and Cu are mainly responsible for the Bursaphelenchusxylophilus isolatesof differentvirulence observedenvironmental impact on the soil nematode and one B. mucronus isolatewere inoculatedseparately community.In spite of the fact that concentrations of onthe upper cut ends of 2.5 and 5.0 cm long living heavymetals remained similar among sampling periods, branchsections. An increase in livesection length induced valuesof biologicalindices exhibited a cleartendency to an80% decrease in the number of passing nematodes increasesince the mining spillage. irrespectiveof isolates.That value was higherthan 45.2% for moresusceptible pine species and lower than 100% 378Nematodes as indicatorsof waste water for aresistant,interspeciŽ c hybrid.Further, to determine treatment theamong-isolate difference in vulnerability to inhibitory effectof the live trees on nematode dispersal, the Ž ve

256 Nematology Postersessions isolateswere inoculatedon the upper cut ends of boiled 381Ecology of plant-parasiticnematodes, their host andlive branch sections 5 cmlong. The vulnerability to plantsand antagonists in Europeancoastal sand inhibitoryeffect of the live trees differed among isolates. dunes: trainingopportunity for ecologists and Twoexperiments showed that virulence had no relation agriculturalbiocontrol researchers ( EcoTrain) tointrinsic and realised dispersal rates in B.xylophilus . 1; 2 Analysisof spatialdistribution of B.xylophilus 24 h after Wim H. VAN DER PUTTEN ¤, B.R. KERRY , inoculationon 25 cm long branch sections showed the K.G. DAVIES 2, R. COOK 3, M. MOENS 4, randomdispersal with diffusion coefŽ cient of 40.73 and M. FARGETTE 5, T. MATEILLE 5, J. BAKKER 6 109.4 cm2/dayfor liveand boiled branches, respectively. and H. FREITAS 7 1 NetherlandsInstitute of Ecology(NIOO-CTO), P.O.Box 40, 380Rhizodeposition and soil microbial communities 6666ZG Heteren,The Netherlands 2 inclover plants infected with the clovercyst RothamstedExperimental Station (IACR), UK 3 nematode, Heteroderatrifolii Instituteof Grassland and Environmental Research (IGER), UK 4 Centrumvoor Landbouwkundig Onderzoek (CLO), Belgium Amy TREONIS1; , Roger COOK 2, Lorna DAWSON 3, ¤ 5 Institutde Recherche pour le Developpement (IRD-CBGP), Sue GRAYSTON 3, Karen LOWES 2, Tony MIZEN 2, 3 3 3 France Brian ORD , Ruth PRIMROSE , Eileen REID 6 WageningenUniversity (WU), The Netherlands 3 andJasmine R OSS 7 Institutodo Mar(IMAR), P ortugal 1 Departmentof Biology,Creighton University, 2500 California ¤[email protected] Plaza,Omaha, NE 68178,USA 2 EcoTrainaims to examine the regulationof plant-parasitic Instituteof Grasslandand Environmental Research, Plas nematodesin natural ecosystems, which processes Gogerddan,Aberystwyth SY23 3EB, UK contributemost to nematode control, and how predictable 3 TheMacaulay Institute, Craigiebuckler ,AberdeenAB15 8QH, UK theyare. The concepts and technology resulting from thisstudy will encourage research into multitrophic ¤[email protected] interactionsin natural soils, which seriously lags behind Belowground herbivory can alter the distribution of researchon abovegroundmultitrophic interactions. Basic plant-derivedsoil organic matter and may impact on insightsresulting from EcoTrainwill also beneŽ t research soilmicrobial diversity and functioning. Invertebrate root onthe biological control of nematodes in production feedingorganisms include the larval stages of many systems,which is currently based mainly on trial insectsas well as ecto-and endoparasitic nematodes. The anderror. SpeciŽc objectivesof EcoTrain are: i/ to processof root herbivory is likely to have signiŽ cant examineand compare potential mechanisms that control impactson soilmicrobes that rely on root-derivedcarbon populationdensities of plant parasitic nematodes in asan energy source. Heteroderatrifolii ,theclover cyst nature, ii)tobridge and stimulate scientiŽ c co-operation nematode,is a sedentaryendoparasite of clover roots. betweenphytopatholog istsworking in production systems Our objectivewas todetermine how plant growth and andecologists working in natural environments, iii) to rhizospherecarbon availability change when roots are setup a trans-Europeannetwork of agronomists and infectedby parasitic nematodes and how changes in the ecologistswith a mutualinterest in soil phytopatholog y quantityand quality of rhizosphere carbon in uence the and iv)tostimulate biological control of plant-parasitic structureand function of the soil microbial community. nematodesin agriculture by learning how this works in Cloverwas grownin microcosmsin sand with or without nature.EcoTrain will use coastal sand dunes of Europe H. trifolii.Rhizodepositswere washedfrom thesand asa naturalmodel system. These dunes provide one of periodicallyfor chemicalanalysis. These solutions also thebest case studies on interactions of natural plants were appliedto plant-free grassland soil to evaluate andtheir parasitic nematodes. The natural occurrence of theresponse of the microbial communities. Changes dominantdune grasses in monospeciŽ c standsand the inrhizosphere chemistry were linkedto microbial heavysoil disturbance makes sand dunes a goodmodel responsesand will be discussed. The results contribute for agriculturalproduction systems. W ebsiteaddress: tounderstanding the indirect effects of plant-parasitic http://www.nioo.knaw.nl/projects/ecotrain/index.htm. nematodeson therhizosphere environment and microbial diversityand function in grasslands.

Vol.4(2), 2002 257 Foodwebs, ecology, biodiversity (359-382)

382Seasonal change of Caenorhabditisjaponica on of C. japonica were exclusivelyfound as clusters under the shieldbug, Parastrachiajaponensis thescutellum of almost all female bugs investigated whilethose were rare onmale bugs. During the mating, Toyoshi YOSHIGA¤, Yuji ISHIKAWA, ovipositionand egg guarding periods of the bug, from Mantaro HIRONAKA and Eizo KONDO earlyMay to June, nematode population on the bug Laboratoryof Nematology, Department of AppliedBiological remainednearly unchanged. When the Ž rst stagenymphs Sciences,Saga University, Saga 840-8502, Japan hatchedin late June, the nematode became undetectable ¤[email protected] from femalebugs. Dauer juveniles appeared on thebug at thesecond stage and were alwaysdetectable on nymphs Caenorhabditisjaponica ,aspeciesclosely related to the ofboth sexes until the bugs became adults in late July. C. elegans group,is associatedwith the shield bug, Paras- Thereafterthe number of dauerjuveniles on femalebugs trachiajaponensis showingprovisioning behaviour. T o graduallyincreased while those on male bugs decreased elucidatethe ecology of C. japonica,thenematode po- andbecame almost undetectable in February of thenext pulationon the bug was investigatedat a deciduousfor- year.Ecological relationships of C. japonica to the life estof Hinokuma Mountain, Saga, Japan from March cycle of P.japonensis willbe discussed. 2001to February 2002. During the period of reproduc- tivediapause of thebug, until early May, dauer juveniles

258 Nematology Postersessions

383Survey of the pinewilt nematodeand its vector potatovarieties cultivated in Sa Pobla-Muro(Balearic insectsin the autonomouscommunity of Galicia, Islands,Spain) in order to arrive at locally relevant northwestern Spain solutionsfor theirappropriate management. The Maris Peer shortcycle potato variety is usually planted in 1; 2 Adela ABELLEIRA ¤, Miguel ESCUER , Novemberand harvesting starts in February, while cv. 1 2 J. Pedro MANSILLA , María ARIAS Marfona,a largecycle potato variety, is usually planted in andAntonio B ELLO 2 Januaryand harvesting starts in May. In both, the highest 1 Estación Fitopatológica ’ DoAreeiro’ , Subidaa laRobleda levelof second-stage juvenile hatching takes place 2 s/n,36153P ontevedra,Spain monthsafter planting and the highestlevel of rootinvasion 2 DptoAgroecologí a, CCMA, CSIC.Serrano 115 dpdo, 28006 occursa monthlater, with the plant showing damage due Madrid,Spain tothenematode infection from thistime on. In cv. Maris ¤[email protected] Peer,nematodes cannot complete the whole cycle due to theearly harvest, while the nematode cycle is shorter in Surveyshave been carried out to detect Bursaphelenchus cv.Marfona because of the warmer temperatures,thus spp.in the Galician Community because the pine wilt givinga greaterrate of reproduction.In the same way, the nematode B.xylophilus was reportedin Portugal. Forest effectof PCN lifecycle on host-plant development and areasin this region represent almost 12% of the forest croploss has been studied to know the PCN population areasin Spain, with conifers at 7% and Pinuspinaster densitiesin which most plants can tolerate invasion and atover 25%. Besides, Galicia is one of the main routes damage,without it adversely affecting potato yield. inSpain for importedtimber .Severalsamplings were takenbetween 1999-2001. The Ž rst time,1249 samples were collectedfrom pinetrees with or withoutsymptoms, 385Survey of plant-parasiticnematode genera in takenevery 5 km 2.Theother samplings were gathered Al-Qassimarea, Saudi Arabia accordingto European Community rules, 2179 samples beinganalysed from pinetrees, sawmills and imported S. AL-REHIAYANI¤ and A. FARAHAT wood.Species were identiŽed mainly on the basis of Collegeof Agriculture& VeterinaryMedicine, King Saud morphometriccharacteristics and, when necessary, by University,AL-Qassim Branch, P .O.Box 1482, Saudi Arabia PCR-RFLPtechniques. Four species of Bursaphelenchus [email protected] were found: B. eggersi infour samples from P. pinaster Anextensive survey was carriedout in Buriedah City and P. radiata withsymptoms of decline, B.sexdentati andthe ten governorates of Al-Qassim areaSaudi Ara- and B. cf. eremus,onceeach, associated with a declining bia.The study was conductedto examine and evalu- pinetree close to sawmills, and B.mucronatus at atethe distribution of plant-parasitic nematode genera, foursawmills on P. pinaster,importedtimber from the thedegree of nematode infestation and plant nema- Ukraineand France. The vector insect for B.xylophilus , todeassociations. T wentyone nematode genera were Monochamusgalloprovincialis ,was alsocollected on foundin this survey: Meloidogyne,Helicotylenchus, importedwood from sawmills. Tylenchorhynchus,Pratylenchus, Heterodera, T ylenchu- lus,Rotylenchulus, Hemicriconemoides, Macropostho- 384Population dynamics of the potatocyst nia,T richodorus,P aratrichodorus,P aratylenchus,Tro- nematode (Globodera sp.) intwo potatovarieties phonema,Hoplolaimus, Xiphinema, Longidorus, Hemi- cultivatedin the BalearicIslands cycliophora,Aphelenchoides, Ditylenchus, Aphelenchus and Tylenchus.Themost common genera in almost all 1; 2 Raquel ALONSO ¤, Ana ALEMANY governoratesare Meloidogyne,Helicotylenchus , Tylen- and Maria FÉ ANDRÉS 1 chorhynchus and Pratylenchus .Thefrequency of ne- matodeoccurrence and the population densities varied 1 Departmentof Biology,University of the Balearic Islands, greatlybetween governorates. Meloidogyne and Helicoty- Cra.de Valldemossakm. 7,5. 07071-P almaof Mallorca,Spain lenchus dominatedon fruittrees, while Tylenchorhynchus 2 Centrode Ciencias Medioambientales, CSIC, Madrid, Spain raqxmixmail.com and Pratylenchus dominatedon Ž eldcrops. The popula- ¤ tiondensities of Meloidogyne were higheron vegetable Thebiology and population dynamics of thepotato cyst crops,especially tomato and eggplant. Unayzah, Al- nematode(PCN) havebeen studied in two commercial Mezneb,Al-Asyah governorates and the city of Buriedah

Vol.4(2), 2002 259 Distribution,population dynamics (383-443) areinfested with the highest nematode genera. The great- Inorder to ascertain the effect that agricultural practices estnumber of nematode genera is associated with date produceon themostrepresentative species of dorylaimids palm trees. Trophonema isrecordedin this study for the andmononchids in the Canary Islands, the nematode col- Žrst timein Saudi Arabia and Rotylenchulus for the Ž rst lectionat the Department of Agroecology of the Envi- timein Al-Qassim area. ronmentalSciences Center (CSIC) was reviewed,where themajority of environmentscultivated in Spain are rep- 386The cereal nematode ,Heteroderaavenae , resented.A totalof 750 samples were studied.Nineteen inAl-Qassim area, Saudi Arabia speciesof the dorylaimid order were found,the most frequentbeing: Aporcelaimellusobtusicaudatus and Be- S. AL-REHIAYANI londiratarjani (3.9%),followed by Discolaimustexanus , Eudorylaimusleuckarti , Nygolaimusbrachyuris and Oxy- P.O.Box 1482, College of Agriculture& VeterinaryMedicine, dirusoxycephalus (2.6%).Of thesamples studied, seven KingSaud University, Al-Qassim Branch, Saudi Arabia specieswere identiŽed from themononchid order, the ¤[email protected] mostfrequent being, in order: Mylonchulussigmaturus (36%), Clarkuspapillatus (13.1%) and Mononchustrun- Thecereal nematode, Heteroderaavenae , is a major catus (3.9%).The dorylaimids demonstrated greater di- limitingfactor in wheat production in the Al-Qassim area, versitywhile the mononchidspresented greater frequency, SaudiArabia. Heteroderaavenae was Žrst reportedin a sincethe latternematodes are more susceptible to changes wheatŽ eldof the Al-Bekeria governorate of Al-Qassim causedby anthropic action. Nematodes from thedory- in1987 but, over the last 5 years,it has spread in the laimidand mononchid orders are of interestbecause they majorwheat Ž eldsof mostof theAl-Qassim governorates. arebioindicators of the level of alteration that agrarian Becauseof thelack of aneconomicalternative, wheat is systemshave undergone. growncontinuously and such Ž eldpractice allowed the cerealcyst nematode population to rise above damaging 388Plant-parasitic nematodes associated with thresholds.In some heavily infested Ž eldsof Al-Qassim, asugarbeet rotationsystem in centralPortugal: thenematode population has reached 250 juveniles/ g. possibleimplications on production Nematodecontrol relies on the use of non-fumigant nematicides.Application of oxamylnematicide as a foliar Pedro BARBOSA¤ and Manuel MOTA sprayafter wheat emergence is a commonpractice and oftenresults in increases in the grain yield, but it has NemaLab/ICAM, Departamentode Biologia,Universidade de Évora,7000 É vora,P ortugal notreduced the nematode population below damaging [email protected] thresholdsfor thenext season. Recently, ploughing the top ¤ soiltwo to threetimes during summer time (solarisation), Today,at least 37% of the world’ s sugaris produced rotationwith alfalfa or avegetablecrop, and application from sugarbeet. In many growing areas, nematodes are oforganophosphate nematicides as a seeddressing have amongstthe most important pests; capable of causing givenencouraging results and have been recommended irreversibledamage and unacceptable economical crop asa managementpractice for H. avenae controlin wheat loss.Population dynamics of plant-parasitic nematodes Želdsof theAl-Qassim area,Saudi Arabia. isessential to anticipate crop damage caused by these pathogensand parasites. The Coruche region in the provinceof Ribatejo, Portugal, is the location of a 387Nematodes from the dorylaimidand mononchid majorsugar beet production region. A preliminarysurvey ordersassociated with CanaryIsland crops andstudy was conductedduring 2001 to evaluate the 1; 2 presenceof taxa and their relative abundance from Susana C. ARCOS ¤, Domingo JIMÉNEZ-GUIRADO threeŽ eldswhere a sugarbeet-corn rotation system is andAntonio B ELLO 1 inuse. The main phytoparasites collected belonged to 1 Dptode Agroecología, CCMA, CSIC,Serrano 115 dpdo the genera Meloidogyne , Helicotylenchus , Pratylenchus , 28006Madrid, Spain Tylenchorhynchus , Trichodorus and Heterodera. Two 2 Dptode Biología Animal,Universidad de Có rdoba, Campus samplingseasons were established:autumn and spring; deRabanales, EdiŽ cio C-1, 14071 Có rdoba, Spain 200g soilsamples were collectedfor nematode ¤[email protected] extraction,which was performedaccording to two

260 Nematology Postersessions methods:Baermann funnel and centrifugal  otation. stateused a standardtreatment of control. Yield losses Nematodeswere countedusing an inverted microscope were determinedby using nematicides compared to an andphotographed with a digitalcamera, from acompound untreatedcontrol. The survey has been completed on more microscope. Helicotylenchus sp.appears to be particularly than60% of the cotton producing counties and parishes numerous,and may be mainly responsible for causinga inthe USA. Themajor species found in the survey are negativeeffect on thecrop. root-knot,reniform, lance, and stingnematodes. The root- knotnematode is found in most areas where cotton is grown.The reniform nematode is prevalent in the south- 389Host rangestudy ofanundescribed eastand mid-south sections of the cotton belt. The lance Meloidogyne sp. isolatedfrom groundnut andsting nematodes are concentrated in the southeast. Thesurvey maps show the three major species present and Ivan F. BENDEZU¤ andJames L. S TARR theirrelative population levels. These maps are updated TexasA &MUniversity,Plant P athology& Microbiology annuallyand can be seenon theNational Cotton Council Dept.,College Station TX 77843-2132,USA website at http:/ /www.cotton.org/cf/nematodes. ¤[email protected] Thereproduction of a new Meloidogyne sp. (93-13) 391First records of Bursaphelenchus species in isolatedfrom groundnutin Collingsworth County (T exas) Thailand was testedagainst a rangeof different plants species andcompared to a populationof Meloidogynearenaria 1; † Helen BRAASCH ¤ andRainulf B RAASCH-BIDASAK race1. Pea cvs Progress and Early Snap were moderate hosts (RF 1:57and 1.18). The bean cvs Bountiful 1 Kantstraße 5,D-14471 P otsdam,Germany andBlue Lake D 274 had low reproductive rates (RF † died in 2001 D 0:69and 0.75) of Meloidogyne sp.93-13, whereas Pea ¤[email protected] cv.California Blackeye 5 and46, and Clemson Purple Thedamage caused by the pine wood nematode (RF 0:05,0.02 and 0.09) were resistant.Genetically D (Bursaphelenchusxylophilus )inseveral East Asian modiŽed (Roundup Ready) soyabean cvs HBKR 4660, countries(Japan, China, T aiwan)and its easy spread 4855,4920, 5588, 5920, 6020, 6600, and conventional bythe timber trade provoked scientiŽ c interestin tree- soyabeancultivars HBK 4891,5990, 5991, were also inhabitingBursaphelenchus species.Thailand is acountry tested.The best soyabean with resistance to the two withhigh temperatures which make it favourable for speciesof root-knotnematodes is HBKR 4660,followed thedevelopment of pine wilt caused by B.xylophilus . byHBKR 6020with high levels of resistance to M. Pineforest occurs naturally in small tracts in some arenaria,andHBK 5991with intermediate levels of uplandareas at altitudes between 800 and 1800 m. resistanceto M.sp.93-13. A comparisonbetween the Nativepines are Pinusmerkusii and P. kesiya. Wood conventionaland the genetically modiŽ ed soyabeans did samplescollected in North Thailand were examinedfor notshow any signiŽ cant difference ( P 0:05/ in any of D thepresence of Bursaphelenchus spp.The species found thevariables evaluated. were Bursaphelenchushylobianum, B. mucronatus as wellas twoundescribed species. The pine wood nematode 390Cotton nematodedistribution and density was notpresent in thesesamples. However, the occurrence in the USA of B.mucronatus ,aspeciesclosely related to B.xylophilus andtransmitted by thesame vectors ( Monochamus spp.), Donald J. BLASINGAME indicatesthe possible establishment of B.xylophilus in 508Colonial Circle, Starkville, MS, 39759,USA caseof its introduction to Thailand. Special quarantine [email protected] attentionshould be paidto wood imports (sawn coniferous In1986 a nematodesurvey was startedin the state timber,logs, wooden packaging material) from countries ofMississippi. A beltwideprogram was initiatedin wherethe pine wood nematode occurs. 1989under the direction of theNational Cotton Council andwith industry support. The purpose of the survey 392Nematode genera associated with mangoin was todetermine nematode species and population ZuliaState, V enezuela densities.Yield losses were alsodetermined as each

Vol.4(2), 2002 261 Distribution,population dynamics (383-443)

E. BRICEÑO 1, N. JIMÉNEZ 1, Q. QUIJADA 2 found. Meloidogyne speciesoccurred in over 30% of and G. CASTELLANO 2 farms; Nacobbus in 20%; and Globodera also in over 1 Decanatode Agronomía, Dpto.Ciencias Bioló gicas, Apdo 20%.Seed tubers are moved from thetraditional potato 400,Barquisimeto, Estado Lara, V enezuela growingareas in the higher and cool altiplano and both 2 Institutode InvestigacionesAgrí colas (INIA-Zulia). Apdo Meloidogyne and Nacobbus were foundin these seed 1316,Maracaibo, Estado Zulia, V enezuela tubers.Thirty parasitic species were foundon potato [email protected] andanother undescribed species of Globodera was also identiŽed, occurring in large numbers on the roots of WehaveidentiŽ ed nematode genera associated with Brassicarapa. mango (Mangiferaindica L.) inZulia state. Forty sam- plesof rootsand soil from productionfarms inMaraand UrdanetaMunicipality were processedwith a combina- 394Nematode genera associated with merey tionof Oostenbrink funnels and Baermann pans. Roots (Anacardiumoccidentale )inZulia state, V enezuela were groundat low speed. Results showed the presence 1 1 2 of Hemicriconemoides,Xiphinema, Meloidogyne, Hoplo- G. CASTELLANO , O. QUIJADA , N. JIMÉNEZ laimus,Rotylenchulus, T ylenchus , Trichodorus and Ty- and E. BRICEÑO 2 lenchorhynchu sinMara-Urdaneta, with the highest fre- 1 InstitutoNacional de InvestigacionesAgrí colas, INIA-Zulia, quency for Rotylenchulus,Hoplolaimus and Meloidogyne . Apdo.1316. Maracaibo, Zulia, V enezuela Abriefdescription is reported and identiŽ cation key is 2 UCLA, Decanatode Agronomí a Dptode CienciasBioló gicas, proposed. Apdo.400 Barquisimeto Estado Lara, V enezuela [email protected] 393Nematode pests ofpotatoin the mesothermic Inorder to identify the main nematodes associated with valleysof the Andean region,Santa Cruz merey (Anacardiumoccidentale )inZulia state, a survey Department, Bolivia ofnematodeswas performedin Maramunicipality farms and60 soil and root samples were taken.The samples 1; 2 John BRIDGE ¤, Ernesto MONTELLANO , were processedwith a combinationof Oostenbrink and 3 2 2 Noel ORTUÑO , Oscar DIAZ , Pablo FRANCO , Baermannfunnel equipment. Results showed the pres- 1 4 David HUNT and Janet ROWE ence of Dorylaimus,Aphelenchus, T ylenchus , Helicoty- 1 CABI Bioscience,Bakeham Lane, Egham, Surrey TW20 9TY , lenchus,T ylenchorhynchus,Criconemoides, Hoplolaimus UK and Xiphinema, with Aphelenchus asmost common fol- 2 CIAT,EjercitoNacional 131, Casilla 247, Santa Cruz de la lowed by Helicotylenchus. Finally,a briefdescription is Sierra,Bolivia reportedand an identiŽcation key is proposed. 3 FundacionPROINP A,Av.Blanco Galindo Km 12.5,Calle Prados/ n,Casilla4285, Cochabamba, Bolivia 4 IACR Rothamsted,Harpenden, Herts AL5 2JQ,UK 395Plant-parasitic nematodes associated with ¤[email protected] semerucoin ZuliaState ofVenezuela

Themesothermic valleys in the Andean region of the G. CASTELLANO 1, O. QUIJADA 1, N. JIMÉNEZ 2 SantaCruz Department occur at altitudes of 1400 to and E. BRICEÑO 2 2500m a.s.l.Potato production, both with Solanum tuberosum var. tuberosum and S.tuberosum var. andigena, 1 InstitutoNacional de InvestigacionesAgrí colas, INIA-Zulia, hasincreased in these warmer mid-altitudevalleys. Apdo.1316. Maracaibo, Zulia, V enezuela 2 IntensiŽcation of potato cropping systems has led to UCLA, Decanatode Agronomí a Dptode CienciasBioló gicas, Apdo.400 Barquisimeto Estado Lara, V enezuela increasedseverity of a complexof potato diseases [email protected] includingnematodes. Over 70farm Želdswere examined inseven valleys. All the three main nematode groups: Plant-parasiticnematodes affect production in mostcrops theroot-knot nematodes, Meloidogyne spp.(mainly M. inMara Municipality Zulia state. In order to evaluate incognita and M. javanica but also M. hapla), the thenematodes in semeruco ( Malpighiaglabra L.) crops, falseroot-knot nematode, Nacobbusaberrans , and the anematologicalsampling was performedwhere 60 soil potatocyst nematodes, Globodera spp. .mainly G. pallida androotssamples were analysed.Samples were processed but also G.rostochiensis plus G:.?/tabacum/ were witha combinationof Oostenbrinkand Baermann funnel

262 Nematology Postersessions equipment.Roots were groundat low speed. Seven Agermplasmcollecting expedition was carriedout during generawere identiŽed: Meloidogyne , Criconemoides, theautumn of 2001 to collect Andean potato cultivars Discolaimus,Dorylaimus, T ylenchus,Hoplolaimus and (Solanumtuberosum ssp. andigena)inthe province of Acrobeles. The genus Meloidogyne was foundto be Jujuy,Argentina (23 oS). Localfarming practices usually themain genus. A briefdescription is given and an involvethe purchasing or barteringof varietiesin market identiŽcation key is proposed. placesbut in a few casessome varieties are sold in majorcentres. As somenematodes are a potentialthreat for thecultivation of the potato, the detection of the 396 Heteroderamediterranea :biological speciespresent in the potato-growing areas is needed in observationswith emphasison factorsaffecting egg orderto establish quarantine methods. T wenty-Žve sites hatchingand differential responses of olivecultivars situatedat altitudes ranging from 2370to 3885 m were toinfection sampledand nematodes extracted from 100cm 3 of soil Pablo CASTILLO 1 and Nicola VOVLAS 2 withmodiŽ ed centrifugation- otation technique. Sixty- Žvegenera of nematodes were foundin the samples. 1 Institutode AgriculturaSostenible, CSIC, Apdo. 4084, 14080 Theplant-parasitic nematodes were mostlyrepresented Córdoba, Spain by Pratylenchus sp. (69%), Meloidogyne sp.(65%) and 2 Istitutodi Nematologia Agraria Applicata ai V egetali,C.N.R., Nacobbusaberrans (61%).Four new localities were ViaG. Amendola165/ A,70126Bari, Italy addedto the distribution of Globoderatabacum in the region.The bacterial-feeders were representedmainly Thein uence of temperature and olive root exudates on by Chiloplacus (77%), Rhabditis and Seleborca (42%); Heteroderamediterranea egghatch and the effects of fungivoresby Aphelenchus (96%), Ditylenchus (85%) H.mediterranea onthe growth of two olive cultivars and Aphelenchoides (57%);predators by Aporcelaimellus (Arbequinaand Picual) were investigated.Egg hatch (54%)and omnivores by Ecumenicus (35%). The occurredover a temperaturerange of 10-30 ±C and was genera Bitylenchus,Butlerius, Cervidellus, Crassolabium, optimalat 20-25 ±C.Therewere nodifferences in egg Dolichorhynchus,Enchodelus, Labronemella, Seleborca, hatchbetween sterile deionised distilled water or root Stegelleta and Zeldia arenew records for Argentina. exudatedilutions (undiluted, diluted 1 :1,and 1 :2)of Arbequinaand Picual at 20 ±C. Heteroderamediterranea reproducedon botholive cultivars in growth chambers at 398Nematodes of citrusin opennurseries and orchardsin São PauloState, Brazil 25±C.Soiland root Ž nalnematode populations, as well as totalnumber of cystsper plant and reproduction rate, were Anderson S. DE CAMPOS1, Jaime M. DOS SANTOS2; signiŽcantly higher in Arbequina than in Picual. Shoot ¤ andLarry W .D UNCAN3 dryand root fresh weights,and increases of shootheight, trunkdiameter and numbers of nodes were signiŽcantly 1;2UNESP/Faculdadede Ciências Agrá rias e Veterinárias, V ia suppressedby infectionwith 10 000eggs secondstage deAcessoProf. P auloDonato Castellane s/ n,14884.900 C juveniles/potin Arbequina but not in Picual. Heterodera Jaboticabal,SP ,Brazil mediterranea completesits life cycle (embryogenic and 3Univ.ofFlorida – IFAS,CitrusResearch and Education postembryogenicdevelopment) in about 60-70 days on Center,Universityof Florida, 700 Experiment Station Rd, Lake oliveroots. Syncytia formation and disorder of rootstellar Alfred,FL 33850,USA [email protected] structureare the main anatomical changes induced by the ¤ parasiteon olivecvs Arbequina and Picual roots. Asurveyof key-nematodes of citrus was conductedin opennurseries and orchards in Sã o PauloState Brazil. 397Survey of nematodesin Andean potatoŽ eldsof Totalsof 2518samples from 595nurseries in 99counties, northwest Argentina and1078 samples from commercialorchards in 86 countieswere examined.The nematodes were separated 3 Eliseo CHAVES¤, Eduardo MONDINO from 100 cm ofsampled soil by centrifugal  otation and Andrea CLAUSEN andfrom 10g ofsampled roots by maceration in ablenderfollowed by centrifugal  otation.The citrus INTA EEA Balcarce,C. Correo276, 7620 Balcarce, Argentina nematode (Tylenchulussemipenetrans Cobb)was found ¤[email protected] in34% of thenurseries and in 72.5%of thecommercial

Vol.4(2), 2002 263 Distribution,population dynamics (383-443)

orchards.Putative populations of Pratylenchuscoffeae PatriciaA. D ONALD¤,KennethA. S UDDUTH (Zimmermann)Filipjev & SchuurmansStekhoven were andNewell R. K ITCHEN foundin seven nurseries and ten orchards. These USDA ARS 605Airways Blvd, Jackson, TN 38301,USA; USDA populationspertain to a newspecies of lesionnematode ARS 269Ag. Engineering Bldg, University of Missouri, thatis being described and named Pratylenchusjaehni. Columbia,MO 65211,USA Mapsof the distribution of these nematodes throughout ¤[email protected] themain region of citrus production in Sã o PauloState were prepared. Heteroderaglycines (SCN) isa majorpest of soybean causingsigniŽ cant yield losses when present and when managementtechniques to reduce egg population density 399Nematode problems in Castillay Leon(Spain), arenot used. A studyinvestigated the distribution of SCN acontinentalMediterranean climate atplanting and harvest in a claypansoil Ž eldin central Missouriunder no till soybean production. Comparisons Miguel A. DÍEZ, Miguel ESCUER andAntonio B ELLO¤ were madein thenematode distribution in 1999 and 2001 DptoAgroecologí a, CCMA, CSIC,Serrano 115, 28006 Madrid, yearsof soybean production. Nematode distribution in Spain the2 yearswas comparedto yield, topsoil thickness, ¤[email protected] elevation,and soil nutrient maps. Nematode distribution was mostclosely correlated with topsoil thickness ( i.e., Plant-parasiticnematode problems were analysedin the depthto the claypan). Lower H. glycines population regionof Castillay Leon,northwestern Iberian Peninsula. densityoccurred in areaswith shallow topsoil thickness. Nematodegroups, in the order of their importance, are: i)cystnematodes, represented by Globoderarostochienis and G. pallida in potato, Heteroderaschachtii in sugar- 401T obaccorattle virus and its associatedvector beet, H. avenae incereal, and H. humili in hops; ii) the trichodoridnematodes in Portugal root-knotnematode M.incognita insugar-beetand potato; 1; 2 3 iii)virusvector nematodes, among which Xiphinema I.M. DUARTE ¤, M.T.M. ALMEIDA and D.J. BROWN index standsout, of interest in vineyards; iv) bulb and 1EscolaSuperior Agrá ria de Coimbra,Coimbra, P ortugal stemnematodes, Ditylenchusdipsaci in garlic; v) the 2 Departamentode Biologia, Universidade do Minho,Braga, endoparasiticnematodes Pratylenchus,Pratylenchoides Portugal and Zygotylenchus;vi )ectoparasiticnematodes, where 3 NematologyDepartment, Scottish Crop Research Institute Macroposthoniaxenoplax standsout for causingchlorosis (SCRI),Dundee, Scotland, UK ingrape and fruit trees ( Prunus spp.).Species from the ¤[email protected] Amplimerlinius,Anguina, Aphelenchoides, Bitylenchus, Tobaccorattle virus (TR V,genus Tobravirus),transmitted Criconema, Criconemella,Criconemoides, Crossonema, bytrichodorid nematodes, occurs as a widerange of Helicotylenchus,Hemicriconemoides, Hemicycliophora, serologicallydistinguishable strains that cause diseases Hoplolaimus,Longidorus, Macroposthonia, Merlinius, invarious economically important crops. The virus and Ogma,P aratrichodorus,P aratylenchus,Quinisulcius, itsassociated vector nematodes occur world-wide, being Rotylenchus,Scutellonema, Seriespinula, T richodorus, particularlyprevalent in Europe and North America. In Tylenchorhynchus and Xenocriconemella genera were Portugal,an investigation is being conducted to identify alsofound. V ariouscontrol alternatives are indicated, such TRVstrainsoccurring in association with their natural aspreventative practices and crop rotation, which regulate vectorspecies. The projecthas focused on potatogrowing thedevelopment of pathogenicnematode populations as areasin northern and central Portugal, with soil sampling wellas the appearance of phytopathologi calproblems. biasedin favour of areas where TR V-likesymptoms Thesealternatives must be followedby theuse of resistant havebeen reported. Nematodes were extractedfrom plantsand trap crops within an integrated production soiland phenotypically identiŽ ed. TR Vwas recovered system. from nematodesin virus transmission studies, and identiŽcation conŽ rmed using sap transmission tests, 400 Heteroderaglycines distributionwithin aŽeldin aleafsquash method with EM, ELISA, ISEM,and no-tillproduction over time RT-PCR.Trichodorids were recoveredfrequently from potatoŽ elds,and the species identiŽ ed were: Trichodorus

264 Nematology Postersessions lusitanicus , T.primitivus , Paratrichodorusanemones , P. 2 ServicioProtecció n ySanidadV egetal,Estació n Sericícola, hispanus, P. minor, P.pachydermus ,alsothree new c/Mayors/ n,30150 La Alberca, Murcia, Spain undescribedspecies and an unidentiŽed Paratrichodorus ¤[email protected] species.TR Voccurredin four of 58 (7%) soilsamples Plant-parasiticnematode problems in the region of Mur- inassociation with trichodorids: one in the north and ciain the southeastern Iberian Peninsula were analysed. theothers in the centre of the country. Molecular Thenematode groups, in the order of their importance characterisationof the TR Visolatesand their associated are: i)theroot-knot nematodes, Meloidogynearenaria , vectortrichodorids is presently in process. M.incognita and M. javanica,whichaffect horticultural cropsand fruit trees; ii)virusvector nematodes, among 402Nematodes associated with coniferouswoods in which Xiphinemaindex ,ofinterest in vineyards, stands Spain out; iii)thecitrus nematode, Tylenchulussemipenetrans , foundin citrus orchards and vineyards; iv) cyst nema-

Miguel ESCUER¤, María ARIAS andAntonio B ELLO todes,represented by Heteroderacruciferae inbroccoli, H. latipons in oats and H.schachtii in chard; v) the en- DptoAgroecologí a, CCMA, CSIC,Serrano 115 dpdo, 28006 doparasiticnematodes Pratylenchus and Pratylenchoides ; Madrid,Spain and vi)ectoparasiticnematodes, among which Macro- [email protected] ¤ posthoniaxenoplax standsout for causingchlorosis in Nematodesassociated with coniferous woods in Spain grapevines and fruit trees ( Prunus spp.).Species were were studiedin 2000-2001. Sampling was coordinated alsofound from thegenera Criconema, Criconemella, bythe Spanish Ministry of Agriculture, Fisheries and Criconemoides,Crossonema, Hemicriconemoides, Heli- Food,with the cooperation of the ofŽ cial forestry, plant cotylenchus,Merlinius, P aratylenchus,Rotylenchus and healthservices and diagnostic laboratories of severalau- Tylenchorhynchus .Variouscontrol alternatives are indi- tonomouscommunities. Seven nematode genera were cated,especially preventative practices to regulatethe de- foundon conifer trees. Laimaphelenchuspenardi , the velopmentof pathogenic nematode populations and to mostwidespread species in the Peninsula and on the preventthe appearance of phytopathologi calproblems, BalearicIslands, was foundassociated with Abies alba, followedby theuse of resistantplants and biofumigation, Pinus nigra, P. pinaster and P. pinea; Cryptaphelenchus allwithin an integrated production system. spp. and Deladenus spp.are also widespread, but their speciesare under study. Seven species of Bursaphelen- 404Distribution of Globoderatabacum chus were found, B.sexdentati beingthe most widespread inthe LaVeraregion of Spain inPeninsular Spain associated with Pinus spp. Bursaphe- lenchuseggersi and B.mucronatus appearedin northern G. ESPARRAGO 1, I. BLANCO 2 and I. PULIDO 1 Spainassociated with Abies alba and Pinuspinaster , and 1 on Pinushalepensis and Pinus spp.,respectively. Other Serviciode Investigación yDesarrolloT ecnológico, Junta de speciesare more localised: B.fungivorus on P. pinaster Extremadura,Finca La Orden, 06187 Guadajira, Badajoz, Spain inthe Cazorla mountains, Andalusia; B. leoni and B. ter- 2 CETARSA FincaLa Cañ alera, Ctra Santa Marí a delas atoespicularis Pinus restrictedto the Balearic Islands on Lomas,Km 3,510310 T alayuela,Cá ceres, Spain halepensis inMallorca and Ibiza; B.pinasteri appearing once on Pinuspinaster inExtremadura; Aphelenchoides LaV erais the principal tobacco growing area in Spain. daubidaensis , A .jonesi and Ektaphelenchusmacrobulbo- Everyyear 11 000ha ofue-curedand 2500 ha ofburley sus on P. pinaster and P. nigra inthe west, and Ž nally, tobaccoare produced in monoculture. The most important Seinura spp. on Pinus spp.from northeasternSpain. phytonematologicalproblem in thecrop is causedby root- knotnematodes. However, in August 2001 Globodera tabacum was Žrst detectedover a K326 ue-curedvariety 403Nematode problems in the regionof Murcia Želd.T oknowthe incidence of these cyst nematodes in (Spain), aMediterraneanmodel thistobacco region, a surveyof Globodera populations was carriedout during February 2002. 82 soil samples Miguel ESCUER1; , Ana CANO 2 andAntonio B ELLO 1 ¤ were gatheredin Ž eldswhere tobacco was cultivatedin 1 DptoAgroecologí a, CCMA, CSIC,Serrano 115 dpdo, 28006, 2001.Soil was analysedfor nematodeabundance (cyst Madrid,Spain andsecond juveniles). Globodera iswidely distributed

Vol.4(2), 2002 265 Distribution,population dynamics (383-443) acrossthe region. Abundance and distribution of this Investigationsabout the potential correlation between nematodein theLa V eratobacco area is shown. theoccurrence of plant-parasitic nematodes and the increasedgrape phylloxera infestation of vinesin Austrian 405Characterisation of cereal cyst nematodes, vineyardsare being carried out within a 3yearresearch Heterodera spp., inNorway project.Roots and soil adhering to the roots were investigated.In 2001, more than 100 samples in total 1; 2 Ricardo HOLGADO ¤, Stig ANDERSSON were collectedfrom eightselected vineyards in Lower and Janet ROWE 3 Austriaand Burgenland at monthly intervals from May toAugust.Subsamples of bothhighly phylloxera-infested 1 TheNorwegian Crop Research Institute, Plant Protection anduninfested grapevines were checkedfor thepresence Centre,Dept. of Entomology and Nematology, Hø gskoleveien 7, offree-living and plant-parasitic nematodes, applying N-1432Å s,Norway standardextraction methods. The predominant nematode 2 SwedishUniversity of AgriculturalSciences, Dept. of Crop genera,which were regularlydetected and identiŽ ed in Science,Box 44, S-230 53 Alnarp,Sweden 3 IACR-Rothamsted,PPI Division,Nematode Interaction Unit, infestedas well as in uninfested root samples, were Harpenden,Herts, AL5 2JQ,UK Aphelenchus sp., Aphelenchoides sp. and Helicotylenchus sp.Plant-parasitic nematodes from eightfamilies and ¤[email protected] free-livingnematodes from 14families were identiŽed Cerealcyst nematodes (CCN), Heterodera spp., are from soilsamples. Prospects for theinvestigationsin 2002 commonpests of cereal crops in Norway. Fifteen arepresented. populationswere investigatedmorphologically, through biotests,and through electrophoretic studies, using 407A surveystudy ofphytoparasiticnematodes and isoelectricfocusing and silver staining of proteins.In the the associatedhost plantsin Egypt biotestthe populations were groupedby their virulence onbarley differentials (resistance genes) V arde( Rha), I.K.A. IBRAHIM Emir (Rha“E”), Ortolan (Rha1), KVL 191 (Rha2), Siri (Rha2)andMorocco C.I. 3902 ( Rha3). Eleven Departmentof PlantP athology,F acultyof Agriculture, populationsexpressed themselves as H. avenae pathotype AlexandriaUniversity, Alexandria, El-Shatby 21545, Egypt Ha11.These were allfairly similar morphologically andelectrophoretically. The biotest further suggested Informationconcerning the occurrence and distribution twopopulations belonging to pathotype Ha12. One of ofphytoparasitic nematodes is very important to assess themwas morphologicallyand electrophoreticallysimilar theirpotential to cause economic damage to many to H. avenae;theother different in both respects, cropplants. A nematologicalsurvey was conducted indicatinga possibleundescribed species. T woother todetermine the genera and species of phytoparasitic populationsappeared to be H. Ž lipjevi,pathotype‘ West’. nematodesand the associated host plants in Egypt. The Onepopulation under study was differentin all aspects, resultsindicated the occurrence of about 56 genera and morphologically,in thebiotest and biochemically. Earlier 180species of phytoparasitic nematodes associated with studiesin Norway have also indicated the occurrence manycultivated plants, grasses and weeds. Many of ofCCN pathotypeHa51. Thus, the situation in Norway thesenematodes, e.g., Criconemella spp., Helicotylenchus Heterodera Hoplolaimus Meloidogyne regardingthe CCN isvery complicated. spp., spp., spp., arenaria, M.incognita , M. javanica, Pratylenchus spp., Rotylenchulusreniformis , Tylenchorhynchus spp., 406First year interim results of investigationson Tylenchulussemipenetrans and Xiphinema spp. are the nematodefauna in the pedobiocoenosisof grape consideredlimiting factors in crop production in Egypt. phylloxera (Dactylophoravitifolii )infested vines (Vitis vinifera)inselected Austrian vineproduction areas 408Comparison of boardingability between Michael HOSCHITZ Bursaphelenchusxylophilus and B.mucronatus on theirvectors, Monochamusalternatus and Instituteof Plant Protection, University of Agricultural M.saltuarius Sciences,P eterJordan Straß e 82,A-1190 Wien, Austria 1; 2 [email protected] Shota JIKUMARU ¤ andKatsumi T OGASHI

266 Nematology Postersessions

1HiroshimaPrefectural F orestryResearch Centre, Miyoshi, customarilykilled, a 5-10%proportion is probed,the cell Hiroshima728-0015, Japan wallsare apparently perforated; however, the nematodes 2 Facultyof IntegratedArts and Sciences, Hiroshima departprior to sap ingestion, thus leavingthese cells alive. University,Higashi-Hiroshima, Hiroshima 739-8521, Japan Additionally,transmission electron microscopy study re- ¤[email protected] vealeddifferences in the sites of retentionof serologically Bursaphelenchusxylophilus ,thecausative agent of pine distinguishabletobravirus strains in the pharynx of dif- wiltdisease, has been introduced from NorthAmerica to ferent (Para)trichodorus vectorand non-vector species, Japanand displaced the closely related, native nematode, i.e.,itrevealed three different patterns of retention.Sub- B.mucronatus ,inthe disease infestation area because sequently,immunogold labelling with antisera designed thetwo nematode species occupy the same niche in speciŽcally to identify these tobravirus strains from one thepine forest ecosystem. Both nematode species enter anotherand from otherviruses, provided unequivocal evi- thetracheal system of newly emerged adult beetles denceof thetobra-viral identity of virus-likeparticles ob- in the genus Monochamus withinpupal chambers of servedwithin the pharyngeal lumen of theexamined tri- thebeetles in xylem. T ocomparethe boarding ability chodoridspecies. oftwo nematode species on the beetles, one of the plausibleprocesses of nematode displacement, post- 410Distribution and temporal population dynamics diapauselarvae of M.alternatus or M.saltuarius of Meloidogyneincognita and Thielaviopsisbasicola in were placedindividually in artiŽ cial pupal chambers anArkansascotton Ž eld of Pinusdensi ora boltsinoculated with B.xylophilus 1; 2 or B.mucronatus . Bursaphelenchusxylophilus load on T.L. KIRKPATRICK ¤, C.S. ROTHROCK , beetleswas signiŽcantly greater than B.mucronatus load W. KINKAID 3, A. MAUROMOUSTAKOS 4, W. BAKER 5 for the two Monochamus beetles,although there was and M. DANIELS 6 nodifference in the abundance of dispersal juveniles aroundthe pupal chamber between the two nematode 1Universityof Arkansas(UoA) Southwest Research and species.These results showed an extremely low ability ExtensionCenter ,362Highway 174 North, Hope, AR 71801, of B.mucronatus toboard the beetles compared with USA 2 B.xylophilus . UoA Departmentof Plant P athology,217 Plant Sciences, Fayetteville,AR 72701,USA 3 UoA atPine Bluff, 1200 North University Drive, Pine Bluff, 409Acquisition, retention and transmission of AR71611,USA tobravirusparticles by trichodorid nematodes 4 UoA AgriculturalStatistics Laboratory, AGRX 101, Fayetteville,AR, USA 5 Eirini KARANASTASI 1,DerekJ.F. B ROWN 3, ArkansasState University College of Agriculture,P .O.Box Ian R. ROBERTS 3 and Urs WYSS 2 2340,Jonesboro, AR 72467,USA 6 ArkansasCooperative Extension Service, P .O.Box 391, Little 1 BenakiPhytopathological Institute, 8 St.Delta st, 14561 Rock,AR 72203,USA KiŽssia, Greece ¤[email protected] 2 Instituteof Phytopathology, University of Kiel, Hermann-Rodewald-Str.9,D-24118 Germany Thielaviopsisbasicola (Berk.& Broome)Ferraris, causal 3 ScottishCrop Research Institute, Invergowrie, Dundee, agentof black root rot of cotton, and the root- DD2 5DA,Scotland, UK knotnematode, Meloidogyneincognita ,frequentlyoccur [email protected] togetherin cotton Ž eldsin Arkansas. Where both Tobravirusesare naturally transmitted by plant-parasitic organismsoccur together, black root rot in seedlings is Trichodorus and Paratrichodorus nematodes.Trichodor- signiŽcantly more severe and seedling mortality is greatly idsare assumed to acquire virus particles upon sap in- increased.Infection of cottonseedlings by the nematode gestionfrom virusinfected root cells. Subsequently, these appearsto broaden the temperature range at which particlesare retained on the cuticle lining the nematode T. basicola caninfect the seedlings, and the nematode pharyngealwall and thereafter are released into further providesthe fungus access to the vascular tissue of the cellsupon salivation of pharyngealsecretions. Real-time plantthat is not normally invaded in the absence of the observationsof livingtrichodorids feeding on rootsof to- nematode.A 6.6ha cottonŽ eldin southeastern Arkansas, baccoseedlings revealed that although attached cells are witha historyof severe seedling disease and root-knot

Vol.4(2), 2002 267 Distribution,population dynamics (383-443) nematodedamage, was dividedinto 512 grids (4 30 m) nematodeand ranks second only to potato wart fungus, £ andsampled prior to planting a cottoncrop (April 2001) Synchytriumendobioticum (108countries) in a listthat andimmediately after cotton harvest (October 2001) to includesregulated insects pests and plant pathogens. The determinethe population densities of both T. basicola rankfor manynematodes has changed the past two and M.incognita ineach grid. In addition, nematode decades,which indicates that regulations are not static. populationswere monitoredin 80 arbitrarily selected For example,in 1982 Aphelenchoidesbesseyi ranked gridsat monthly intervals during the growing season. tenthand was regulatedby nine countries, but in 2002it Spatialand temporal relationships of theseorganisms and rankedsecond with 70 countries regulating it, primarily theirrelationship to variousedaphic factors are described. toprotectrice crops. In contrast, in 198211 countrieshad regulationsto exclude Meloidogynejavanica , but in 2002 nocountrylisted it asa prohibitednematode. 411Presence of Heteroderatrifolii Goffart,1932 (Nematoda:Tylenchida) in Argentina 413Biogeographical characterisation of 1 1; Paola LAX ,MarceloE. D OUCET ¤ Trichodoridaein the IberianPeninsula andEduardo V EGA 2 1; 2 1 J. Antonio LÓPEZ-PÉREZ ¤, M. Teresa ALMEIDA , Laboratoriode Nematología, Centro de Zoologí a Aplicada, 1 1 UniversidadNacional de Córdoba, C.C. 122,5000, Có rdoba, María ARIAS andAntonio B ELLO Argentina 1 DptoAgroecologí a, CCMA, CSIC,Serrano 115 dpdo, 28006 2 Estación ExperimentalRegional Agropecuaria Rama Caí da, Madrid,Spain C.C. 79,5600 San Rafael, Mendoza, Argentina 2 DptoBiologia, Universidade do Minho, Campus de Gualtar , ¤[email protected] 4710-057Braga, P ortugal [email protected] Theanalysis of rootsof Trifoliumrepens andof thesoil ¤ aroundthem, from thelocality of Malargü e, province Theexistence of two faunistic groups have been ofMendoza, Argentina, permitted the identiŽ cation of foundon analysis of the distribution patterns of the representativesof thenematode Heteroderatrifolii . White 18species from thefamily Trichodoridae that have femaleswith masses of eggsand cysts were observedon beenfound in representative crops and environments roots;second-stage larvae were extractedfrom thesoil. ofthe Iberian Peninsula. The Ž rst onerepresented by Morphologicaland morphometric studies performed on theautochthonous species, Paratrichodorushispanus , thesespecimens conŽ rm thespecies identity, which is Trichodorusazorensis, T .beirensis,T .giennensis and citedin Argentinafor theŽ rst time. T.lusitanicus ,ispresent in uncultivated and cultivated areas; T.azorensis,T .beirensis and T.giennensis 412Nematodes in international quarantine havebeen found very localised, while P. hispanus legislation iswidespread in Spain and northern Portugal and T. lusitanicus iscommonmainly in southernbut also found Paul S. LEHMAN incentral Portugal. The second one is deŽ ned by the plant-parasiticand virus vector species, P. minor, P. FloridaDepartment of Agricultureand Consumer Services, pachydermus,P .teres,T .primitivus,T .sparsus and T. DPI, P.O.Box147100, Gainesville, FL 32614-7100,USA viruliferus , in which P. anemones and T. similis could TheSociety of Nematologists currently has a USDA alsobe included,in spite of theirvery localised presence. fundedproject to list and rank exotic nematodes of Paratrichodorusanemones, P .pachydermus,T .similis regulatoryconcern to the USA. Alistof nematodes,which and T.viruliferus couldbe regarded as characteristic areregulated by international quarantine legislation in speciesfrom temperateenvironments, while P. minor, the 2002,was compiledusing the APHIS/ USDA database, mostwidespread species in subtropical crops, has also andwas comparedto Kahn’ s 1982list of the ten most beenfound in the Canary and Madeira Islands. On the frequentlyregulated nematodes. In general, in the past two other hand, P.teres,T .giennensis,T .similis,T .sparsus decadesthe number of countriesregulating nematodes has and T.viruliferus haveonly been found in Spain, while increased.For example,51 countriesregulated Globodera P.acutus,P .allius,P .nanus,P .porosus,T .azorensis and rostochiensisi sin1982, but 106 countries regulate it in T.orientalis appearedvery localised only in Portugal, P. 2002.W orldwide, G.rostochiensisi sisthe most regulated acutus,P .porosus and T.azorensis appearingonly in the

268 Nematology Postersessions

Azoresand Madeira Archipelagos. Climatic, vegetation asthe disease advanced more. Nematode reproduction andsoil type in uence are discussed. was rapidand a hugepopulation developed in the wood ofa diseasedtree. About 50 million nematodes were calculatedto be in the stem wood of a deadtree 10 m 414Association of Melilotusalbus Desr. with talland 15 cmdiameterat breast height. Nematodes were apopulationof Meloidogynejavanica (Treub,1885) distributedthroughout the dead tree. Spatial distribution Chitwood,1949 in the provinceof Mendoza, patternsof nematodes varied considerably among dead Argentina pinetrees. Aggregated distribution was demonstratedin 1; 2 thestem. Nematode populations in dead trees declined Eugenia LORENZO ¤,MarceloE. D OUCET , Susana SUÁREZ 1 and Paola LAX 2 graduallyafter reaching a maximumlevel, about 3000 nematodesper g ofdrywood, and were almostextinct in 1 Orientación Morfología Vegetal,F acultadde Ciencias Julyof thefollowing year. In this stage of thepopulation Exactas,Fí sico-Quí micas y Naturales,Universidad Nacional dynamics,dispersal third stage juveniles appeared and deRí o Cuarto,Agencia P ostalN 3,5800,Rí o Cuarto, ± theproportion of themin the whole population increased Córdoba, Argentina gradually. 2 Laboratoriode Nematología, Centro de Zoologí a Aplicada, UniversidadNacional de Córdoba, C.C. 122,5000, Có rdoba, Argentina 416Potato cyst nematodes, Globoderarostochiensis ¤[email protected] and G. pallida,inSwedish potatoŽ eldsand their Apopulationof M. javanica was detectedin thelocality relationshipsto other European populations ofMalargü e, province of Mendoza, infecting roots of Melilotusalbus .Theplant was atthe  oweringstage at Sanja MANDURIC¤ and Stig ANDERSSON thetime of analysis.Its aerialportion had a normalaspect. SwedishUniversity of AgriculturalSciences, Department of However,a greatnumber of galls containing females of CropScience, Box 44, S-230 53 Alnarp,Sweden thenematode were observedon roots.In some galls, the ¤[email protected] posteriorend of femalesprotruded beyond the tissues; in othergalls, females were insidethem. Internally, giant SixteenSwedish potato cyst nematode populations were cellswere observedin thecentralcylinder, which reduced comparedwith 19 populations from theNetherlands, thevascular tissues and also caused important anomalies UnitedKingdom, Germany and Norway as to genetic intheir arrangement. The nematodedevelops its life cycle andvirulence variability through AFLP analysisand completely,producing an egg mass. It is concluded that biotests.Obtained data indicate a largeheterogeneity, thenematode – hostrelationship is close, and that the andthe present occurrence seems to be the result of plantrepresents a goodhost for thisspecies of thegenus severalolder and more recent introductions. Swedish Meloidogyne. Thisweed is cited as analternative host of Ro1/Ro4populations of Globoderarostochiensis came aphytophagousnematode in Argentinafor theŽ rst time. from heavilyinfested Ž elds,had a generalEuropean appearance,and may be supposed to be very old 415Population dynamics of the pinewood introductions.Members of virulence group Ro2/ 3of nematode, Bursaphelenchusxylophilus , thisspecies were geneticallymore heterogeneous but indead pine trees similarto continental populations. They may have been introducedlater. Swedish G. pallida populationsof Yasuharu MAMIYA virulencegroup Pa2/ 3couldaccording to the AFLP analysisbe divided into two subgroups, one similar 5-6-8Kitanodai, Hachioji, T okyo,192-0913 Japan toa Britishpopulation and the other to continental [email protected] populations. G. pallida populationsbelonging to the Experimentalresults of inoculationtests and observations ‘British’group were purepopulations and probably fairly onnaturally infected pine trees demonstrated that the oldintroductions. Pa2/ 3populationsof the ‘ continental’ pinewood nematode was detectedrarely in wood at groupwere, withone exception, found in the starch thetime when disease symptoms appeared, such as potatodistrict where they had been selected during thecessation of oleoresin exudation from wounds.The intensivecultivation of cultivarswith Solanumtuberosum nematodepopulation in woodof adiseasedtree increased ssp. andigena resistance.They may be fairly recent

Vol.4(2), 2002 269 Distribution,population dynamics (383-443) introductions.Norwegian populations of both species taxonNematoda. At eachbeach, sampling was undertaken seemedto have backgrounds similar to theSwedish ones. atthreepoints along three proŽ les with a cuppercore of 28mm diameterto a depthof 20 cm. Each sample was dividedinto layers of 5cm.Samples for sedimentological 417Nematodes from an urbanforest in SoŽ a, analysiswere collectedand temperature, salinity, pH and Bulgaria slopeproŽ le measurements were made.Nematoda mean densityat the Enseada Beach ranged from 719.5ind. Alexander MLADENOV, Stela LAZAROVA¤ £ 10 cm 2 inDecember 98 to 1299 ind. 10 cm 2 in and Vlada PENEVA ¡ £ ¡ June99. At theUna Beach mean density ranged from CentralLaboratory of General Ecology, 2 GagarinStreet, 1113 2 751.8 ind. 10 cm¡ inFebruary 99 to 1570.1 ind. SoŽa, Bulgaria £ 10 cm 2 inNovember 99. On the Enseada Beach [email protected] ¡ ¤ in£ February 99, at the points where human density was Genericcomposition, trophic structure, diversity and higher,Nematoda was founddeeper in the sediment. In Maturityindices of nematode assemblages were studied March99 Nematodawas concentratedin the Ž rst layerof atKnyaz Borisova Gradina City Park. The study thesediment. Nematoda is beingidentiŽ ed atgenus level sitewas a60-90year-old mixed deciduous-conifero us toseekfor variationin vertical and temporal distribution. forestdominated by Quercuspedunculi ora C. Koch. Litterand soil samples: decomposition layer, ca 0.5- 419Nematodes of HaughtonCrater 1cm,and humus layer at three depths: 0-5, 5- 10and 10-25 cm were collected.Ninety nematode Peter MULLIN 1, Andrew SHUERGER 2 generawere registered.Generally, nematode abundance 1; and Thomas POWERS ¤ was lowand differed between samples and depths. 1 Bacterialfeeders were themost common, abundant and Universityof Nebraska, Department of PlantP athology,406 PSH,Lincoln, NE 68583-0722,USA diverse(36 genera) group, followed by plant feeders 2 (22genera). A cleardifference between litter and LifeSciences Program, Dynamac Co., Kennedy Space Center , FL32899,USA soilnematode assemblages was revealedin regard to [email protected] almostall parameters studied. Nematode communities ¤ from decompositionand humus layers were similarin HaughtonCrater has been called Mars on Earth due taxonomicand trophic group structure. In soil layers topresumed similarities with craters on Mars. Sixteen plant-feedingnematodes prevailed, omnivores were well kmin diameter, Haughton Crater is located on Devon representedwhile bacterial feeders decreased with depth. Island,Nunavut, Canada approximately 640 km south Noclear layer dependent patterns in generic distribution ofthe north magnetic pole. Plant communities in the were found.Maturity Indices of soil communities were craterare frequently found in isolated patches surrounded highand their values increased gradually with depth. bya coarse,rock and gravel substrate. The nematode Nematodesoil assemblages were characterisedby high communitieswithin these ‘ micro-oases’are surprisingly diversityand maturity, which could/ mightbe relatedto the diverse.T odate75 species have been identiŽ ed. age,heterogeneity and size of theforest studied although Comparedwith other nematode communities, the micro- itis situatedin anurban environment. oasisnematode community is dominatedby predatorsand omnivoresand includes an unexpectedlylarge number of 418T ourist oweffect onNematodaof Brazilian males.Outside the micro-oases, a lessdiverse community sandybeaches exists,with 58 species identiŽ ed to date, including 27 speciesnot observed within the micro-oases. Overall Adriana M. MOELLMANN¤ andThaï s N.C ORBISIER nematodeabundance ranged from 22to4129 nematodes per 100 cm3 ofsoil. All nematodes observed in this InstitutoOceanográ Žco –Universidadede Sã o Paulo,Praç a do studywere revivedfrom ananhydrobiotic state. The OceanográŽco, 191 Butantã ,Sã o Paulo,SP 05508-900,Brazil diversityand abundance of nematodesin HaughtonCrater ¤[email protected] sharplycontrasts with the depauperate communities of the Thisstudy is an attemptto evaluatethe impact of human Antarcticdry valleys. activityon theEnseada Beach during summer, comparing itwith the preserved Una Beach, through the study of the

270 Nematology Postersessions

420Phytoparasitic nematodes associated with the speciesrecovered from soiland occurred on banana and rhizosphereof lulo,mora, tree tomato and granadilla onlysix weed species, at population densities much lower plantsin Colombia thanthat of banana.From roots, R. similis was recovered from bananaonly while H.multicinctus was present GladisEmilia M ÚNERA URIBE¤ and Rafael NAVARRO onbanana and nine weed species and P. goodeyi was Apartadoaé reo 51764 Medellí n, Colombia presenton banana and only two weed species, namely, Digitariavelutina and Eleusineindica ,ataverylow count ¤[email protected] comparedto banana.No nematodes were recoveredfrom Ninegenera of phytoparasitic nematodes were found Tagetesminuta , Cyperusesculentus , Seneciodisfolius associatedwith the rhizosphere of lulo or naranjilla and Digitariascalarum ,indicatingthat these four weed (Solanumquitoense Lam.),tree tomato or tamarillo speciesare non-hosts of banananematodes. (Cyphomandra (Solanum) betacea (betaceum) (Cav.) Sendt),granadilla ( Passiora ligularis Juss.)and mora (Rubusglaucus Benth.)in different Colombian regions. 422The occurrence of burrowingnematodes The genus Meloidogyne was bothmore prevalent and Radopholus spp. ondurianand coffee in Western abundantin all the locations sampled. Meloidogyne Highlandof Vietnam incognita,M. javanica,M. hapla and M. arenaria were Chau N. NGUYEN1; , Sergei A. SUBBOTIN 2, identiŽed in the rhizosphere from luloand tree tomato ¤ Mehrdad MADANI 3, Phap Q. TRINH 1 whereas M.incognita and M. hapla were foundin the andMaurice M OENS 3 rhizosphereof granadilla. On the other hand some of theabove mentioned nematode species were foundin 1 Instituteof Ecology and Biological Resources, NCST ,12 soilborne on mora plants but M.incognita and M. HoangQuoc V ietRd., Hanoi, V ietnam javanica,Helicotylenchus sp. and Paratylenchus sp. do 2 Instituteof P arasitology,RAS, LeninskiiProspect 33, Moscow , notreproducein this plant. Trichodorus , Hemicycliophora 177071Russia and Pratylenchus doreproduce in mora plants. Other 3 AgriculturalResearch Center ,Burg.V anGansberghelaan 96, importantgenera associated with all the fruit plants 9820,Merelbeke, Belgium studiedwere Xiphinema,Paratylenchus, Hirschmanniella ¤[email protected] andCriconemella. A stronginteraction was found between Meloidogyneincognita and Fusariumsolani Burrowingnematodes associated with durian ( Durio zi- whichreduced development and growth parameters of betinus), coffee (Coffea spp:/, banana (Musaparadisiaca granadillaplants. var. sapientum),rambutan( Nepheliumlappacium ) and av- ocado (Persaeamericana )havebeen recently recorded 421Host status ofthe commonweeds ofbanana inWesternHighland V ietnam.The nematodes are widely establishmentsto banana nematodes in Uganda distributedon durianorchards and coffeeplantations; only afew specimenswere extractedfrom rootsamples of J.M. NAMAGANDA 1, I.N. KASHAIJA 1 banana,avocado and rambutan. In the infected regions, and R. MASLEN 2 Radopholus was notdetected on corn ( Zea mays), black pepper (Pipernigrum ), mango (Mangiferaindica ), citrus 1 KawandaAgricultural Research Institute, P .O.Box 7065, (Citrus spp.),pineapple ( Ananassativa ),sugarcane( Sac- Kampala,Uganda charumofŽ cinarum )andpapaya ( Caricapapaya ). Nema- 2 NRInternationalLtd, P .O.Box258, Chatham, Kent todescaused decline and death of trees in many durian ME4 4PU, UK nurserygardens and newly replanted orchards. Nematode Hoststatus to the banana nematodes Radopholussimilis , densitiesreached several thousands of individuals per g Helicotylenchusmulticinctus and Pratylenchusgoodeyi for rootsamples. Nematodes also induced some damage was studiedin 13commonweed species, in ascreenhouse incoffeeplantations ( C. arabicaand C. robusta). Morpho- experiment.Banana nematodes were recoveredfrom logicaland morphometrical comparisons of allnematode soil,roots or both in banana and nine weed species, stagesrevealed only slight differences between Radophu- butnematode population densities indicate that these lus populationsfrom durianand coffee and rom R. simi- weedspecies are poor hosts of the banana nematodes. lis. However,the molecular analysis based on sequencing Helicotylenchusmulticinctus was theonly nematode ofITS-rDNA showedsubstantial nucleotide differences

Vol.4(2), 2002 271 Distribution,population dynamics (383-443) betweenthese Vietnamese populations and R. similis and 424First report of Meloidogynepartityla on pecan in supportstheir separate taxonomic status. Georgia

Andrew P. NYCZEPIR1; ,CharlesC. R EILLY 1, 423The frequency and diversity of the cystand ¤ Bruce W. WOOD 1 andStephen H. T HOMAS 2 lesionnematodes on wheat inthe Turkish Central AnatolianPlateau 1 U.S.Department of Agriculture,ARS, SoutheasternFruit and TreeNut Research Laboratory, Byron, GA 31008,USA 1; 2 3 Julie NICOL ¤, Roger RIVOAL ,NecmettinB OLAT , 2 Departmentof Entomology,Plant P athology,and W eed 4 1 Hussein AKTAS ,Hans-JoachimB RAUN , Science,New MexicoState University, Las Cruces, NM 88003, Mohamed MERGOUM 1, Ali Faik YILDRIM 3, USA 5 6 Ahmet BAGCI , Halil ELEKCIOGLU ¤[email protected] and Amor YAHYAOUI 7 Treedecline was observedin a commercialpecan 1 InternationalWheat and Maize Improvement Center orchardin Georgia in 2000. Affected trees exhibited (CIMMYT),WheatProgram, P .O.Box 39, Emek, 06511, deadbranches in the upper canopy, stunted growth, Ankara,Turkey andfeeder roots with small galls and associated egg 2 UMR INRA/ENSAR, Biologiedes Organismes et des massestypical of root-knot nematode infection. All Populationsappliqué e àlaProtection des Plantes (BiO3P), decliningtrees that were examinedhad root systems BP35327,35653 Le Rheu,France infectedwith Meloidogyne sp.IdentiŽ cation of this 3 ANADOLU AgriculturalResearch Institute, PK 1726001, Eskisehir,Turkey nematodewas basedon two procedures. First, in 4 PlantProtection Institute, Ankara, Turkey Georgia,females were removedfrom rootgalls of 5 BahriDagdas MIKHAM, Konya,Turkey decliningtrees and identiŽ ed bydeterminingthe esterase 6 CukarovaUniversitesi, Adana, Turkey phenotypeas compared to standard root-knot nematode 7 InternationalCentre Arid Research Dryland Agriculture speciesincluding M.partityla .Secondly,galled roots (ICARDA), P.O.Box5466, Aleppo, Syria were sentto New MexicoState University in Las ¤[email protected] Cruces,where mitochondrial DNA from specimenswas extractedand compared to standard root-knot nematode InJune 2000 a surveywas conductedon theCentral Ana- species.Specimens were identiŽed as M.partityla at tolianPlateau, the major winter wheat growing region bothlocations. This is the Ž rst reportof M.partityla ofTurkey.The objective was tounderstand the distribu- from Georgia,and the third report of this nematode tionof twoeconomically important cereal nematodes, cyst outsideof South Africa. Meloidogynepartityla may be (Heterodera spp.)and lesion ( Pratylenchus spp.)Thirty endemicto North America and not South Africa. Itis soiland plant samples were collectedaround the milky believedthat this nematode entered South Africa onpecan doughstage from wheatsampled about every 50 km.Sev- rootstockimported from theUSA between1912 and entytwo percent of theroot samples and 83% of thesoil 1940.Determining the distribution of M.partityla within samplescontained cysts, and in approximately 40% of thepecan-growing regions of Georgiaand North America soilsamples, one or both lesion nematodes were found iswarranted. (P. thornei and/or P.neglectus ).Cerealcyst nematode was identiŽed to species level using both traditional morphol- 425The occurrence and geographic distribution of ogyand a RFLP PCRbased molecular method. None of Xiphinema and Xiphidorus species(Nematoda: thesamples contained the most common cereal cyst ne- Longidoridae)in Brazil matodedocumented, H. avenae.Instead,40% of thesam- plescontained H. latipons, 32% H. Ž lipjevi and 28% a mix 1; 2 ClaudioM.G. O LIVEIRA ¤,DerekJ.F .B ROWN , ofboth species. Regarding species identiŽ cation, in about Roy NEILSON 2, Ailton R. MONTEIRO 3 10%of the samples the molecular diagnostics did not andLuiz C.C.B. F ERRAZ 3 alignwith the morphological assessment. Furthermore, 60%of samples contained root-rotting fungi ( Fusarium 1 InstitutoBioló gico, Caixa P ostal70, Campinas, SP , spp. and/or Bipolaris spp.). 13.001-970Brazil; 2ScottishCrop Research Institute, Invergowrie,Dundee, DD2 5DA,Scotland, UK; 3Universidade deSão Paulo,Escola Superior de Agricultura‘ Luizde

272 Nematology Postersessions

Queiroz’, CaixaP ostal09, Piracicaba, SP ,13.418-900Brazil; ofmorphological characters under optical and scanning ¤[email protected] electronicmicroscopes. Theoccurrence and geographic distribution of Xiphinema and Xiphidorus specieswere investigatedduring a na- 427Nematode assemblages from a polararctic tionalsurvey of 16 Brazilian states between 1999 and desert 2001.Eighty-two soil samples were collectedfrom dif- 1; 1 ferenthabitats, including crop plants, grassland, savanna Vlada PENEVA ¤, Stela LAZAROVA 2 (cerrado)and typical Amazonian forest vegetation. Four- and Olga MAKAROVA Xiphinema X.brasiliense,X. costaricense, teen species ( 1 X.elongatum,X. ensiculiferum,X. ifacolum,X. krugi, CentralLaboratory of General Ecology, 2 GagarinStreet, 1113SoŽ a, Bulgaria; 2Laboratoryof CommunityStructure and X.longicaudatum,X. paritaliae,X. vulgare,X. surina- Dynamics,Institute for Problems of Ecologyand Fvolution, mense,X. brevicolle andthree distinct morphotypes of LeninskyProspect 33, Moscow ,Russia; X.americanumsensu lato tentativelyidentiŽ ed asrepre- ¤[email protected] senting X.diffusum,X. oxycaudatum and X.peruvianum ) were foundin allsurveyed states. Also, Xiphidorusyepe- Preliminaryresults are presented on the generic struc- sarayepesara , X.yepesaraparthenus , X. minor and X. tureand several ecological parameters of nematode as- balcarceanus were identiŽed from twostates; however, semblagesin relation to Žvetypesof microhabitatsfrom recognitionof X.yepesaraparthenus asa subspeciesof apolygonalpolar desert on Bol’ shevik Island, Sever- X. yepesara requiresconŽ rmation. The most frequently nayaZemlya Archipelago. Among 33 genera encoun- occurringspecies were X. krugi (46%), X.brasiliense tered,more than half were commonto themajority of mi- (22%) and X. vulgare (22%). Xiphinemalongicaudatum crohabitats(all or fourtypes). Nematode abundance was and Xiphidorusbalcarceanus constitutenew records for high in Deschampsiaborealis andblack crust microhabi- Brazil. tats(average 81 and69 specimens/g)andlow in bare soil (eightspecimens/ g).Bacterial feeders were themost di- 426 Bursaphelenchus speciesfound inmaritime pine verseand dominant trophic group (15 genera), followed in Portugal byomnivorousand plant-feeding nematodes (Ž ve and six genera,respectively). Predatory and fungal-feeding ne- 1 1 Ana CATARINA PENAS , Maria ANTONIA BRAVO , matodeswere representedin low percentage. This gen- 1 2 Joana PIRES and Manuel MOTA eralpattern differed only for assemblagesfrom baresoils wheretrophic groups were almostequally distributed. 1 Departamentode Protecçã odasPlantas, Estaç ã o Again,nematode communities of baresoils had the low- Agronómica Nacional, INIA, Quinta do Marquês, 2784-505 Oeiras,P ortugal; 2NemaLab/ICAM, Departamentode estdiversity and generic richness. The Ž rst gradientre- Biologia,Universidade de É vora,7000 É vora,P ortugal; vealedby an ordination procedure was from nematode [email protected] assemblagesof higher plant vegetation and black crust, tothese of moss communities. Along the second gradi- InMarch 1999, the Pine W oodNematode (PWN) entnematodecommunities of baresoils were separatedas Bursaphelenchusxylophilus (Steiner& Buhrer,1934) themost speciŽ c group.The genera Acromoldavicus and Nickle1970, was foundassociated with dead maritime Boreolaimus representnew geographical records. pine (Pinuspinaster Ait.)for theŽrst timein Portugaland inEurope. OfŽ cial authorities implemented an intensive 428Plant-parasitic nematodes associated with surveyin the region where the nematode was foundand mangovarieties on two rootstocksin Mara thenextended it to the rest of the country. As aresult, municipalityof Zuliastate, V enezuela thisquarantine organism was conŽrmed to beconŽned to a30km radius area in the Setú bal region, near Lisbon. O. QUIJADA 1, G. CASTELLANO 1, N. JIMÉNEZ 2 Duringthe survey other species of Bursaphelenchus were and E. BRICEÑO 2 foundassociated with P. pinaster, such as B.mucronatus , B. leoni, B. tusciae, B. hofmanni, B.hellenicus,B. 1 InstitutoNacional de InvestigacionesAgrí colas, INIA-Zulia, teratospicularis , B.hylobianum , B.sexdentati and B. Apdo1316, Maracaibo, V enezuela; 2UCLA, Decanatode pinophilus.TheidentiŽ cation was basedon observations Agronomía Dpto.de Ciencias Bioló gicas postgrado de

Vol.4(2), 2002 273 Distribution,population dynamics (383-443)

Fitopatología, Apdo400, Barquisimeto, Estado Lara, studiesare suggested, as well as correlation studies of Venezuela;[email protected] t.ve populationlevel with other soil diseases and damagelevel. Parasiticnematodes on mango ( Mangiferaindica L.) cvsHaden, T ommyAtkins, Irwin, Gleen, Keitt, Ford 430Specialities of zonegeographic distribution of andPalmer, grafted on Sinamaica and Bocao root- soilnematodes on the territoryof Russia stocks,were identiŽed from 64soil and root sam- plestaken at the Hortofruiticola Center in Zulia E.N. ROMANENKO-POPOVA state.Nematodes were extractedusing Oostenbrink and Instituteof Geography and Institute of Global Climate and Baermannfunnel equipment. The varieties grafted on Ecology,Russian Academy of Sciences,Glebovskaya ul, 20B, Sinamaicarootstocks had Hoplolaimus,T richodorus, Moscow,107258,Russia Pratylenchus,Hemicriconemoides, Rotylenchus, Heli- Thesoil nematode fauna of differentgeographic regions cotylenchus,Xiphinema, T ylenchus Meloidogyne. and The ofRussia (Karelia, Moscow and V oronezhregions and Hoplolaimus, varietiesgrafted on Bocao rootstock had Kalmykia)were investigatedduring 1992-1997. The Helicotylenchus,Rotylenchus, Xiphinema, Trichodorus baseof the nematode fauna of different soils of these and Meloidogyne ,withthe most frequent genera as Ho- regionswas theorders T ylenchida,Dorylaimida and plolaimus,Hemicriconemoides and Rotylenchus. A brief Rhabditida.The orders Enoplida and Chromadorida descriptionis reported and identiŽ cation key proposed. were characteristicof hydromorphic soils. The role anddiversity of nematodes from theorder Mononchida 429Plant-parasitic nematodes associated with increasedfrom northto south. The percentage of other papaya (Caricapapaya )indifferent producingareas nematodeorders varies in different soils. The abundance ofthe State ofBahia,Brazil ofnematodes rises in latitude-zone row ofsoil types insequence: podzol (Karelia); podzolic soils (Moscow Cecília Helena Silvino Prata R ITZINGER¤, region);chernozem soil (V oronezhregion), and decreases Rogério RITZINGER andÉ lioJose A LVES signiŽcantly in chestnutsoils of Kalmykia.The nematode faunaof intra- and azone soils of Karelia was less EmbrapaMandioca e Fruticultura,Caixa postal, 007, Cruz das Almas,BA, Brazil abundantthan those of zone soils of that region but increasesin comparison with the zone soils sequentially ¤[email protected] from northto south in other investigated regions. Thus, Inthe State of Bahia,papaya production ( Caricapapaya ) thenematodefauna of moist intra- and azone soils was six isdistributed in all microregions whereas Extremo Sul totentimes more than those of zonesoils of Kalmykia. andExtremo Oeste have the major production. Although plant-parasiticnematodes are a greatproblem in papaya, 431Histological alterations in Caricaceae roots thereare no dataon nematodeoccurrence and damage in causedby a mixedpopulation of Meloidogyne papayaplantings in the State of Bahia.This work aimed incognita Race 1 and Rotylenchulusreniformis toidentify the occurrence of plant-parasitic nematodes bycollecting soil samples in commercial plantings in LigiaCarolina R OSALES¤,ZoraidaH. S UÁREZ theExtremo Sul of Bahia, as well as in Iaç u andin an andMarí a AlejandraG ÓMEZ experimentalarea of the Federal University of Bahia, inCruz das Almas. The extraction of nematodes from InstitutoNacional de InvestigacionesAgrí colas, CENIAP , soilwas carriedout by the sieving and centrifugation Protección Vegetal,Apdo 4653, Maracay 2101, Aragua, method,and from rootssamples by theBaermannmethod. Venezuela [email protected] Plant-parasiticnematodes were reportedconsidering ¤ thefrequency and incidence for eachhomogeneous Histologicalalterations on Caricapapaya , Vasconcellea area.In general, Meloidogyne sp., Rotylenchulus sp.; cauliora and V.pubescens inoculatedwith a mixed Helicotylenchus sp., Criconemella sp., Tylenchus sp. and populationof Meloidogyneincognita Race 1 and Pratylenchus sp.were presentin all production areas. Rotylenchulusreniformis were studied.Segments of roots Thelargest diversity was observedin Porto Seguro and from eachspecies were Žxedin Craff III anddehydrated thelargest frequency for allareas by Meloidogyne sp., inatertiarybutyl alcohol series, embedded in parafŽn, cut Rotylenchulus sp. and Helicotylenchus sp.More detailed in 15 ¹msectionsand stained with quadruple Triarch’ s

274 Nematology Postersessions stainmodiŽ ed bySuárez et al.Allspecies of Caricaceae cv.T ypicain anaturallyinfested commercial Ž eldin the showedgiant cells, nuclei and nucleoli and egg mass Konadistrict on the island of Hawaii was assessed.The typical of Meloidogyne .Furthermore,the same species numbers of M.konaensis andthe concentration of K C ofplant showed also the vascular parenchyma cells Ca Mginthecoffee roots were inverselyrelated. Other C forminga syncytiuminduced by Rotylenchulus . These inverserelationships were betweenthe numbers of M. cellsare hypertrophied with granular cytoplasm, enlarged konaensis andconcentrations of Mg (roots), Cu (roots nucleiand prominent nucleoli. This research allowed andleaves) and Al (roots).Even though concentrations demonstrationthat both nematode species can establish of P, K, Mg, K Ca Mg,Mn, and Zn increased C C theirfeeding site in thesame plant. insoil between February (dry season) and May (wet season),concentrations of P,K,Mg,K Ca Mg, Cu, C C Bdecreasedin the coffee roots, and Zn and Al content 432 Meloidogyneincognita racesdistinguished by decreasedin the coffee leaves. Nematodes plus nutrient sweet potatodifferential cultivars and their deŽciencies and imbalances in thecoffee trees are causing distributionin the Kyushu Okinawaregion, Japan coffeedecline. Zen-ichi SANO and H. IWAHORI NationalAgricultural Research Center for Kyushu Okinawa 434Plant-parasitic nematodes associated with Region,Nishigoshi, Kumamoto 861-1192, Japan Ammophilaarenaria (L.) Linkin Portuguesecoastal sand dunes Tousecultivars of sweet potato with resistance 1; 1 to Meloidogyneincognita effectively,occurrence and Catarina SCHRECK REIS ¤, Helena FREITAS distributionof resistancebreaking races were examined. and Wim VAN DER PUTTEN 2 Fourpopulations of M.incognita were collectedfrom differentŽ eldsand inoculated to 24 major sweet potato 1 IMAR, Departamentode Botâ nica, Universidade de Coimbra, cultivars.Based on the reproduction rates of these 3000Coimbra, P ortugal nematodepopulations, Ž vedifferential cultivars, Norin 2 NIOO, Deptof MultitrophicInteractions, Boterhoeksestraat, No.1, Norin No. 2, T anegashima-murasaki7, Elegant 48,P .O.Box40, 6666 ZG, Heteren,The Netherlands Summer andJ-red were selected.A totalof 97 M. [email protected] incognita populationswere collectedfrom sweetpotato Želdsin central to southern Kyushu and Okinawa. Portuguesesand dunes extend for 450km, more than half Sevenraces were identiŽed from thesepopulations. In ofthe coastline. Ammophilaarenaria (marram grass) Kumamotoprefecture central Kyushu, race SP1 that isthe dominant, and eventually the only, natural plant reproducesonly on Norin No. 1 represented81% of speciesin mobile dunes. The unmatched contribution 26examined populations. In Miyazaki and Kagoshima of A. arenaria tosand Ž xationand dune formation is prefecturesouthern Kyushu, SP 2thatreproduces on relatedto its extensive root and rhizome system and bothNorin No. 1 andNorin No. 2 was predominant.In withits vertical growth, stimulated by sand deposition. Okinawa,SP 4thatreproduces on fourof the differential Instabilised dunes, where no sand accumulation occurs, hostsbut not on J-Red represented more than half of the thisspecies declines in vigour and growth. This behaviour, 12examinedpopulations. named ‘the Ammophila problem’, hasbeen largely studiedfrom nutrition,ageing, genetic differentiation tointerspeciŽ c competition,among others, but is still 433Association of Meloidogynekonaensis and unanswered.Recent studies relate this degeneration to the nutritionalstatus of Coffeaarabica thepresence of pathogenic soil organisms, like parasitic nematodes,while others believe that the cause is the D.P. SCHMITT1; , D. HURCHANIK 1, N.V. HUE 2 ¤ absenceof mycorrhizal fungi. In the presentation, we and B.S. SIPES 1 providean overview of plant-parasiticnematodes present 1 Universityof Hawaii, PEPS, 3190Maile W ay,Honolulu, inthe roots and rhizospheric soil of A. arenaria in two 2 HI9682,USA; Universityof Hawaii,TPPS, 1910East-W est dunesystems of thePortuguese coast. The data shows a Road,Honolulu, HI 96822,USA; ¤[email protected] highernumber and diversity of nematodes in the north Therelationship of population densities of Meloidogyne population,where plants are more vigorous, although only konaensis tonutritional status of rootsand leaves of coffee twoof thethree species known to parasitise A. arenaria

Vol.4(2), 2002 275 Distribution,population dynamics (383-443) arepresent. Some highlight plans for furtherresearch will soiltrap method. Three isolates of Steinernema and 59 alsobe presented. of Heterorhabditis were detected,of which 22 isolates were identiŽed as H. indica. Outof the identiŽ ed EPN, twoare new species; one has been described as 435Reaction of passionfruit genotypesto root-knot Steinernemapakistanense (Shahina et al.,2001),while nematode, Meloidogynejavanica the other Steinernema speciesis in the process of R.D. SHARMA, N.T.V. JUNQUEIRA and A.C. GOMES description.Pathogenicity of new indigenous EPN, S. pakistanensis, was testedagainst seven insect pests of EmbrapaCerrados, Caixa P ostal08223, CEP 73301-970, commonoccurrence and agricultural importance under Planaltina,DF ,Brazil laboratoryconditions and conŽ rmed on Helicoverpa Consideringthe widespread distribution and the losses armigera,Earias insulana, E. vittella,P apilliodemoleus, theroot-knot nematode, Meloidogynejavanica , causes Leucinodesorbonalis, Etiella zinkenella and Hollotrichia todifferent crops in the Cerrado region of Brazil, the consanguinea .Mortalitywas higher(up to 100%) on reactionof 11genotypes(EC-2-O Hybrido,V ermelhinho, earlystages of insect larvae ( H. armigera)ascompared IAC-comp.Hybrido, MSC, Roxo Australiano, Seleç ão tofourth and Ž fthstage larvae. Search for more DF,Longão PR-2,V ermelhão, Redondão PR-1,Roxo Fiji indigenousEPN iscontinuing and, under the National eItaquiraí) ofpassionfruit( Passiora edulis f.  avicarpa), IPM Programme,Ž eld-testingof promising species will and P. edulis)were evaluatedunder glasshouse conditions. beundertaken during the year 2002. The farmers will Five17-day old seedlings of each genotype were betrained for conservationof natural enemies including inoculatedwith initial inoculum of approximately 5250 EPNthroughthe farmer Želdschool (FFS) approach eggsand second stage juveniles per plant per pot particularlyfor thecotton crop, which receives the largest containingone kg ofsoil,and an equal number of plants amount(60%) of pesticidesin Pakistan. were maintainedas uninoculated checks. T omatovariety SantaCruz was includedas a susceptiblecheck. Eighty 437Association of plant-parasiticnematodes of daysafter nematode inoculation, indices of root-knot the familyLongidoridae with fruit cropsin Poland galls,indices of eggmasses, Ž nalpopulation densities of nematodesin soil and roots, and the percentage increase Adam SZCZYGIEL ordecrease in dry vine weights of the inoculated plants were comparedwith the uninoculated controls. The gall FruitExperiment Station, Brzezna, 33-386 P odegrodzie,P oland indexfor differentgenotypes varied from 4.6to 5.0 and [email protected] theegg mass indexfrom 0to2 andthe reproduction factorfrom 0.001to 0.061. The reproduction factor In453 soil samples, collected from rootvicinity for susceptibletomato varied from 16.95to 39.48. All ofapple, pear, plum, peach, sour and sweet cherry, genotypestested were highlyresistant to M. javanica. walnut,hazelnut, black and red currant, gooseberry and raspberry,six species of Xiphinema andeight species of Longidorus were found.Equally common Xiphinema 436Detection of entomopathogenicnematodes and specieswere X.diversicaudatum , X.brevicolle and X. theirpossible use under IPM inPakistan vuittenezi.Anotherthree species were foundonly once ortwice. The most common Longidorus species was Manzoor H. SOOMR 1, F. SHAHINA, M.A. MAQBOOL 2 elongatus L.attenuatus andIftikhar A HMAD 1 ,followedby andless frequently L. euonymus and L.leptocephalus .Theother four 1 NationalIPM Programme,National Agricultural Research speciesoccurred sporadically. Almost all species showed 2 Centre,Islamabad, P akistan; NationalNematological preferencefor somehosts and some of them also ResearchCentre, University of Karachi,Karachi, P akistan for soiltype and soil pH. Xiphinemadiversicaudatum InPakistan, studies on biology, taxonomy and efŽ cacy preferredblack currant, pear and sour cherry, Xiphinema ofentomopathogeni cnematodes(EPN) were initiated brevicolle –walnutand sour cherry, while Xiphinema in1996. Since then, during extensive surveys, over vuittenezi preferredraspberry and redcurrant. Longidorus 1000soil samples have been collected from diverse elongatus was commonlyassociated with pear, peach, habitatsof Pakistan with emphasis on Sindh Province. sourcherry, walnut, gooseberry and plum. Xiphinema Nematodeswere collectedusing the Galleriamellonella diversicaudatum showedpreference for heavysoils,

276 Nematology Postersessions while Xiphinemabrevicolle , Xiphinemavuittenezi and 2 UniversityStatistics Center ,Box30001 MSC 3CQ,New most Longidorus speciespreferred light sandy soils; X. MexicoState University, Las Cruces, NM, USA diversicaudatum and brevicolle preferredsoil pH 4.5-6.5 Theobjective of thisresearch was todetermine if either while vuittenezi and Longidoruselongatus did not show yellowor purplenutsedge altered Meloidogyneincognita preferencefor soilpH, andt other Longidorus species reproductioncompared to susceptible chile peppers. preferredsoils of pH5.6-6.5. Greenhouseexperiments were conductedin 2000 and 2001,with 240 pots planted with chile pepper ( Capsicum 438Elaboration of databaseof nematodesdetected annuum),yellownutsedge ( Cyperusesculentus YNS), D inimported plant germplasm in Brazilin 1981 purplenutsedge ( C. rotundus PNS), chile YNS, or D C to 2001 chile PNS. Sixpots from eachtreatment were harvested C at3-4 day intervals from 21through 45 days after R.C.V. TENENTE 1, V.R.V. RISSOLI 2, L. PALHARES 2 inoculationwith 5000 eggs of M.incognita .Rootsystems 2; and F.G. ALVARENGA ¤ (chile)or roots, rhizomes and tubers (YNS, PNS) were dividedand eggs extracted from half.Egg masses on the 1 EMBRAPA,RecursosGené ticos e Biotecnologia,P .O.Box remaininghalf were stainedwith phloxine B. Thenumber 2372(70849-970) Brasí lia, DF ,Brazil 2 UniversidadeCató lica de Brasí lia, QS 07-Lote01 ofeggmasses per root, eggs per egg mass, and total egg (72022-900)T aguatingaSul, DF ,Brazil productionper root system was measuredfor eachplant ¤[email protected] andsample date. Chile supported much greater nematode reproductionthan YNS orPNS, producingtwice as many Plantintroduction has been one of the most effective eggmasses per gram of rootand twice as manyeggs per actionsfor agriculturaldevelopment in the tropics. eggmass. Competition between chile and either nutsedge Coffeeand soybean in Brazil, banana in South and hadno effecton eggmass densityor numbersof eggsper CentralAmerica, sugarcane in South America and eggmass inanyhost. Reproduction occurred on rootsbut Caribbeanand grass pastures in Latin America, are notrhizomes or tubersof eithernutsedge. goodexamples of successful introductions. But the germplasmmovement involves the risks of introduction 440Weeds as hosts toroot lesion nematodes ofexotic pests that are sometimes carried by seeds (Pratylenchusneglectus and P. thornei) andpropagated vegetative materials. EMBRAP A’spost- entryquarantine is inspecting against pests, including Vivien A. VANSTONE1; ,MichelleH. R USS 1;2 nematodes,all the plant material that has been introduced ¤ andSharyn P .T AYLOR intoBrazil for researchpurposes. In this context, the 1 nematologicallaboratory did the inspection and registered Universityof Adelaide, Department of PlantScience, pmb 1, GlenOsmond, 5064, South Australia, Australia allthe nematodes found associated with germplasm from 2 differentspecies of plant and different countries. Now, SouthAustralian Research and Development Institute, GPO Box397, Adelaide, 5001, South Australia, Australia acomputationaldatabase is under construction to make [email protected] extremelyfast, easy and more efŽ cient the recovering ¤ thesedata. Other researchers that need to access such data Pratylenchusneglectus and P. thornei occurthroughout for theirwork can contactthe EmbrapaGenetic Resources thebroadacre, dryland cereal cropping zone of south- andBiotechnology Center and we willhelp them with the easternAustralia. Hosting ability of ninemonocotyledon informationthat is available. (grass) and20 dicotyledon (broad-leaf) weed species commonto cropping rotations was testedin the glasshouse.Nematode multiplication rates ( Pf/Pi) were 439Characterisation of Meloidogyneincognita assessedrelative to wheat cv. Machete (susceptible) and reproductionon yellownutsedge, purple nutsedge triticalecv. Abacus (resistant). W eedswere classiŽed as andchile pepper good(susceptible), poor (moderate) or non (resistant) hosts.Grasses differedin susceptibility to P.neglectus Stephen H. THOMAS 1,JacquelineF UCHS 1, compared to P. thornei.For eachnematode, two grass Jill SCHROEDER 1 andLeigh W .M URRAY 2 specieswere susceptible,three moderate and four 1 Departmentof Entomology, Plant P athology,and W eed resistant.Multiplication rates on broad-leaf weeds were Science,Box 30003 MSC 3BE lowerthan on the grasses. Six broad-leaf species were

Vol.4(2), 2002 277 Distribution,population dynamics (383-443) susceptibleto P.neglectus and14 resistant; one was importancedue to its extensive area and also because it moderatefor P. thornei and19 resistant. Growth of iswherecork and holm oak grow. Integrated in aPAMAF susceptibleweeds will allow increase and/ orpersistence projecta studyhas been conducted to evaluate changes ofnematodes in cropping soils, compromising use causedby some agricultural practices and its effects on ofmanagement strategies (particularly rotation with thenematode communities associated in this type of resistantcrops) employed by growersto reduce nematode ecosystem.Nematode diversity in grasslands is usually densitiesand alleviate crop losses. Monitoring susceptible high.Nematode identiŽ cation of thespecimens found in weedsin crops, pastures and fallows will aid nematode the‘ montado’of the Evora region (southern Portugal) managementand control decisions. isin progress, and some represent new genera/ species reportsfor Portugal.Related to the taxonomic work, acompilationof all plant and soil nematode genera/ 441PWN-CD: informationdatabase for species(non animal-parasitic) and relatedreferences from the pinewoodnematode Bursaphelenchusxylophilus Portugalwill be presented.

P. VIEIRA¤ and M. MOTA NemaLab– ICAM/Departamentode Biologia,Universidade de 443Plant-parasitic nematodes of maize Évora,P ortugal (Zea mays L.)inlow input agriculturein Kenya ¤[email protected] 1; 1 1 J.W. WACEKE ¤, O.J. ARIM , S.W. WAUDO Themain objective of this project is to develop an and J.W. KIMENJU 2 informationdatabase with the inclusion of text(research 1 articles,keys, bibliography), images (jpeg Ž les),video Departmentof Botany,Kenyatta University, P .O.Box 43844, 2 clips(mpeg Ž les),in order to collect in one physical Nairobi,Kenya; Departmentof Crop Protection, University of support(CD-ROM) allrelevant information related Nairobi,P .O.Box 29053, Nairobi, Kenya; [email protected] tothe pine wood nematode (PWN), Bursaphelenchus ¤ xylophilus.TheŽ rst volumepresented here covers Differentfarming systems existing in Kenya have varied thetaxonomy of the genus Bursaphelenchus . All cultivationpractices supporting diverse nematode genera majorpapers with the original species descriptions andspecies. A surveyconducted during the short rains in havebeen scanned in ‘ pdf’format. An updated and eightlocalities within two districts in Central Highlands broadbibliography has been included. In the future, a ofKenyaidentiŽ ed 12 generaof plantparasitic nematodes taxonomicalkey, based on classicaldicotomous keys but associatedwith maize. Both soil and root samples were witha betterand more intuitive presentation, will be collectedfrom therhizospere of maize at the tasseling developed.The Ž nalproduct is PCandMac compatible. stageand assayed for nematodes.Main nematode Futureeditions/ updateswill be developed, in particular generarecovered from soilsamples in decreasing relatedto the nematode vectors and the bioecology orderof prominence were Pratylenchus , Meloidogyne , ofthe nematode-vector-plant .Ultimately,researchers, Paratylenchus,T ylenchus , Ditylenchus,Heterodera and techniciansand political decision-makers will have at Scutellonema spp.The main nematode genera found theirdisposal a practicaland readily available source inmaize roots in order of decreasing prominence ofinformation on this extremely important pest and were Pratylenchus , Hemicycliophora , Tylenchorhynchus , pathogen. Tylenchus, Ditylenchus , Paratylenchus and Meloidogyne spp.In both root and maize samples, Pratylenchus was themost predominant genus occurring in 100 and 96% of 442A checklistof nematodesfrom plant and soil soiland maize root samples, respectively. A coefŽcient of in Portugal determination( r2 0:83/ was obtainedbetween number D of Pratylenchus spp.recovered from thesoiland that from P. VIEIRA¤, M. MOTA and C. COSTA theroot. Soil samples collected in 63% of the localities ICAM, Departamentode Biologia, Universidade de É vora, hadabove critical infestation level of two Pratylenchus 7000É vora,P ortugal spp./cm3 ofsoil.Root necrosis differed signiŽ cantly with [email protected] ¤ locality7 showingthe highest root necrosis. Insouthern Portugal the grassland ecosystem known as‘ montado’has a particularecological and social

278 Nematology Postersessions

444Biofumigation tests: changesin the population problem.Poultry litter was evaluatedunder Ž eldcondi- of Globodera spp. andin the enzymaticactivity of tionsfor themanagement of Meloidogyneincognita on soilsadded with organicamendments pumpkin (Cucurbitamoschata )cv.Soler. The amend- mentwas appliedto a root-knot-infestedsoil at rates of B. de AYMERICH VADILLO¤ and J. LOPEZ ROBLES 0,7.3 and 14.5 kg/ plant;in addition, a treatmentwith Edafologiay QuimicaAgricola, F acultadde Ciencias, phenamiphosat 1.5 gai./ plant,was includedto determine Universidadde Burgos,Plaza Misael Bañ uelos s/ n,09001 theeffectiveness of theamendment. The experiment was Burgos,Spain conductedduring two cropping seasons. Final soil and ¤[email protected] rootpopulations of root-knotnematode were determined Smallbiofumigation tests were carriedout in pots duringeach cropping season. Results showed that poultry withsoil naturally infected by Globodera spp., using litterreduced population densities of M.incognita , and differentamendments as fumigants: seaweed of the improvedplant development, and crop yield. genus Gelidium,remainsfrom gardening(grass, dry leaves)and potato peel ( Solanumtuberosum ).Soilwithout 446Evaluation of the efŽcacy of mixingtops of amendmentwas usedas control. The treated soil was foragegrasses with soilto control Meloidogyne watered,sealed with black plastic, and kept under incognita and Heteroderaglycines controlledconditions of humidityand temperature for 30 days.The value of theratio Ž nal/initialpopulation of the C.R. DIAS-ARIEIRA and S. FERRAZ Globodera spp.was calculated,as was thevariation in ureasic,phosphatasic and deshydrogenasic activity at the UniversidadeF ederalde Viçosa, Depto de Fitopatologia, Viçosa, MG, Brazil,36571-000 endof the treatment. The nematode population fell by [email protected] 34%for thesoil amended with Gelidium, 56% for the sampleswith grass and potato peel, compared with 5% To control Meloidogyneincognita and Heterodera for thecontrol. This fall in the population is causedby the glycines, Brachiariabrizantha, B. decumbens,P anicum liberationof nematicide substances by the amendments maximum cv. Guiné, Andropogongayanus cv.Planaltina, andan increase in the chitinolic micro ora and total andsoybean tops were mixedone per se withsoil and edaphicbiomass, re ected in the increase in enzymatic leftthere to deteriorate. In trial 1, 8 gofthe grass activityof theamended soil in relation to thecontrol. topswere eachmixed into soil in 2.5 l pots,and 30 dayslater one indicator tomato or soybeanseedling was 445Poultry litter for the managementof root-knot establishedper pot. Meloidogyneincognita eggs and egg nematodeon pumpkin inPuertoRico masses and H. glycines femaleswere countedrespectively 45and 30 days later. In trial 2, infested pots were 1; 2 José A. CHAVARRÍA-CARVAJAL ¤ , Edwin ACEVEDO , plantedwith the grass seedlings: half were cultivated Carlos FLORES 3, Nidia VYCENTE 1, for 30days and their tops mixed with the soil; in the Luis SILVA-NEGRÓN 1 and Evelyn ROSA 1 otherhalf cultivation lasted 30 moredays and evaluation was doneas indicated. Results in trial 1 showedno 1 Departmentof CropProtection, Mayagü ez Campus, Universityof PuertoRico, P .O.Box 9030, Mayagü ez, treatmenteffects on nematodes survival. In trial 2, only 00681-9030,Puerto Rico 37females of H. glycines were foundper root system 2 Departmentof Agronomy and Soils of B.decumbens while689 and 547 were countedin 3 Departmentof Horticulture,Isabela Substation, Isabela, soybeanroots cultivated for 60and30 days,respectively. 00662,Puerto Rico Andropogongayanus cv.Planaltina was theonly grass ¤[email protected] amongall tested to allow M.incognita toreproduce. Butoverall there was noeffect of mixing any of these Accumulationof solidwastes from humanactivities and foragegrasses with soil on the efŽ ciency of therotation agro-industriesis a seriousproblem in Puerto Rico that incontrolling M.incognita and H.glycines. representsan environment hazard and leads to signiŽ - cantpollution of soils,waterways, and lakes. The proper useand disposal of these materials in agricultural soils, 447Effect ofrepeateduse oforganicamendments throughtheir application for managementof phytonema- andbiofumigation plus solarisationon glasshouse todes,could be very useful for Žndingsolutions to this pepper nematodesin the southeast ofSpain

Vol.4(2), 2002 279 Organicamendments (444-448)

M.M. GUERRERO 1, C. ROS 2, P. GUIRAO 2, 448Nematode-suppressive effects ofcomposts 1; 1 1 A. LACASA ¤, M.A. MARTÍNEZ , M.C. MARTÍNEZ , against Meloidogynejavanica on tomato N. BARCELÓ 1, M. ONCINA 1, A. CANO 1 1; 2 and A. GONZÁLEZ Yuji OKA ¤ and Uri YERMIYAHU 1 Protección Vegetal,Centro de Investigación yDesarrollo 1 Divisionof Nematology 2 Agroalimentario,CAAMA, C/Mayors/ n,30.150La Alberca, SoilChemistry and Plant Nutrition, Gilat Research Center , Murcia,Spain M.P.Negev85280, Israel 2 FECOAM –Consejería deAgricultura,Agua y Medio ¤[email protected] Ambiente.C/ Caballero,13, 30.002 Murcia, Spain Suppressiveeffects of two composts, cattle manure [email protected] ¤ compost(CMC) and grape marc compost(GMC), onthe Whenmethyl bromide (MB) isnot used for soil root-knotnematode Meloidogynejavanica were tested disinfection, Meloidogyneincognita becomesone of the in pot and in vitro experiments.No root-galls were mainsoil-borne pathogens in the southeast of Spain foundon tomato roots grown in soils containing 10 (Murciaand south of Alicante). Nematode incidence or25% (v/ v)CMC, and very few were foundon afterseveral organic amendments (7 kg/ m 2 fresh sheep thosegrown in soil containing 50% GMC. SigniŽcant manure(FSM) 3 kg/m2 chickenmanure; FSM at 7 reductionsin galling index were alsofound in tomato C kg/m2 0:5 kg/m2 soybean our;EFO at7 kg/m 2 of plantsgrown in soils containing smaller concentrations C ureaat 25 kg/ m 2/ was comparedwith MB treatments ofthis compost. Chemical analysis of the composts inC commercial and experimental glasshouses. The effect andleachates from thesoils showed that the CMC ofthe reiterated use of FSM and chicken manure was containedhigher concentrations of N– NH 4 and higher alsostudied. The three amendments had root-knot index ECvaluesthan the GMC, whilethe latter had higher N– and M.incognita incidencesimilar to MB treatments. NO3concentration.The water extractof theCMC showed However,marketable yield was signiŽcantly lower for highnematicidal activity to the nematode juveniles and urea(8.1 kg/ m 2/ andchicken manure (8.6 kg/ m 2/ than for lessactivity toward the eggs in vitro.Theextract of soybean our(9.6kg/ m 2/ orMB(9.7kg/ m 2/.Marketable theGMC showedweaker nematicidal activity to the yieldof 1st year biofumigation-solari sationtreatments juvenilesand eggs. W ashingcomposted soils with excess was signiŽcantly lower than those of the2nd (11.3kg/ m 2/ waterbefore nematode inoculation and tomato planting and3rd year (11.2 kg/ m 2/ andthatof MBtreatments(11.2 ledto better plant growth, but the nematode-suppressive kg/m2/.Similarresponse was observedfor theroot-knot effectwas lowered.These results suggest that high index(1st year, 4.5; 2nd year, 1.8; 3rd year, 1.3; MB,1.5) nitrogenconcentrations, especially N– NH 4, and high EC andthe percentageof infestedplants (1st year, 90.0%; 2nd valuescontribute to thenematode suppressiveness of the year,43.3%; 3rd year, 40.0%; and MB, 36.7%). composts.

280 Nematology Postersessions

449Assessment ofDiTerato control plant-parasitic theplant-parasitic species. The results revealed that there nematodesin vineyards in Chile arerecognisable nematicidal potentials of the tested algal extracts.Some of the tested extracts caused more than Erwin ABALLAY¤ and Andrea RIVEROS 90%mortality of thetested nematodes. The egg-hatching Facultadde CienciasAgronó micas, Universidad de Chile, test,which was conductedon the plant-parasitic species Casilla1004, Santiago, Chile only,proved that some algal extracts were ableto prevent ¤[email protected] ordisruptthe embryonic-develop mentof theeggs in the testednematode species. Duringthe 1999/ 2000and 2000/ 2001seasons, trials were carriedout to evaluate the performance of thebiological nematicideDiT erafor thecontrol of plant-parasitic 451The fungicide effect ofthe biologicalnematicide nematodesaffecting grapes, Vitisvinifera L., mainly ‘Novibiotec7996’ Xiphinema index,X.americanum s.l. and Meloidogyne Mouldi M. B’CHIR andY oussoufR AISSY spp.Trials were carriedout in the central region on ¤ vineyardsfor wineproduction, and for tablegrape ResearchCentre Al Rumais,P .O.Box150, P .C.328,Sultanate productionin anorthernregion of Chile.Grape varieties of Oman includeSauvignon Blanc, Cabernet Sauvignon and ¤[email protected] ThompsonSeedless, respectively. DiT erawas applied Novibiotec7996 was registeredin Tunisia as abiological througha dripirrigation system at the rate of 40 l/ haat nematicideovicide, efŽ cient in different nematode species thebeginning of spring root  ushingand results were control.It was tested in vitro andunder plastic shelters comparedwith a chemicalstandard and an untreated against Pythium sp.associated with cucumber crops, and control.The effect of thetreatments was assessedtaking inopen Ž eldconditions on citrus and mango orchards intoaccount variations in number of nematodes per infested,respectively, with Fusarium, Lasiodiplodia and 3 250 cm of soil, (Pf/pi),andalso pruning weight. The Ceratocystis sp.in thesultanateof Oman.The Novibiotec Žnalresults showed no statistical differences between the 7996comparative mode of actionon nematodeeggs and chemicalstandard and the DiT eratreatments; however fungiorgans will be presentedand discussed. withrespect to theuntreatedcheck, signiŽ cant differences (P 0:05/ were noted,with control percentages D between77.4 and 91.5% at 30 days after applications. 452 In vitro pathogenicityof Monacrosporium Yieldand pruning weight also were signiŽcantly different robustum to Rotylenchulusreniformis (P 0:05/ inrespect to the untreated plants. DiTera D EduardoRoberto DE A. BERNARDO offers anewbiorational option for controlof nematode and Jaime M. DOS SANTOS populationson grapesin Chile. ¤ UNESP/Faculdadede Ciências Agrá rias e Veterinárias, V iade AcessoProf. P auloDonato Castellane s/ n,14884.900 450The toxic and the antagonisticeffects ofmarine Jaboticabal,SP ,Brazil algalnatural products onthe embryonicdevelopment, ¤[email protected] juveniledevelopment, and survival of nematodes Thereniform nematode ( Rotylenchusreniformis ) is an

Fawzia ABDEL-RAHMAN¤ andRohaiah S ALAH importantpathogen of many of the most important Braziliancrops such as soybean, cotton, beans, cowpea, BiologyDepartment, T exasSouthern University, Houston, passiora, papaya, tomato, lettuce, among others. The Texas77004, USA successfulbiological control of pests in the upper [email protected] ¤ partsof the plants and the environmental concern Thenematicidal activity of the methanol extracts of aboutthe continuous use of nematicides have the tenspecies of marine brown algae collected from the effectof promoting the interest in biological control RedSea coast were testedagainst different nematode ofnematodes, increasing considerably the economic species,including some free-living and plant-parasitic potentialof this activity. Among the nematophagous species.The test was conductedon the adult stages of fungiwith potential for commercialutilisation as thefree-living nematodes, the adults and/ orthejuveniles agentsof biological control of nematodes are the ofthe parasitic species, as well as the egg-masses of species of Monacrosporium. Inthis study, the predatory

Vol.4(2), 2002 281 Biologicalcontrol (449-488)

1; 2 capacity of Monacrosporiumrobustum tothe reniform Myoung Rae CHO ¤, Chun Keun LIM , nematode (Rotylenchulusreniformis )was evaluated in Heung Yong JEON 1, Yong Moon CHOI 1 vitro.Concentratedsuspension of J2, J3, J4, males and andMyoung Soon Y IEM 1 youngfemales were transferredto the cultures of the 1 Divisionof HorticulturalEnvironment, National Horticultural fungusin Petridishes with 2% water-agarand incubated ResearchInstitute, Suwon 441-440, Korea o at 26 Cinthe dark. In periods of 24, 48 and 72 h 2 Departmentof AgriculturalBiology, Kangwon National afterthat it was observedthat the percentage of captured University,Chunchon 200-701, Korea nematodeswere, respectively,81.3, 99.87 and 100%. ¤[email protected] Theadhesive branches as the capture structure of the Asurveywas conductedon the distribution of Pas- fungusand captured nematodes were alsodocumented by teuria spp.associated with plant-parasitic nematodes in scanningelectron microscopy. glasshousesin Koreafrom 1996to 2000. Among the 372 soilsamples assayed, 20 samples had plant-parasitic ne- 453Selection and diversity of Pasteuriapenetrans matodeswith Pasteuria spp.endospores attached. Ne- isolatesin relationto Meloidogyne spp. fromcoffee matodesfound with Pasteuria endosporesattached, host plants,and their localities were: Meloidogyne spp. on ori- ReginaM.D.G. C ARNEIRO¤, Myrian TIGANO, Camila entalmelon at Sungju,Cheju, and Y eoju; Helicotylenchus LOPES JORGE andAna Cristina O LIVEIRA TEIXEIRA sp.on whitepotato and carrot at Cheju; Meloidogyne sp. onsoybean at Cheju; Heterodera sp.on unknown host EMBRAPARecursosGené ticos e Biotecnologia,CP .02372, plantat Chuju; and Aphelenchus sp.on garlicand radish at 70849-970,Brasí lia, DF ,Brazil [email protected] Cheju.Morphological characteristics of endosporesfrom ¤ Meloidogyne spp.observed under electron microscope Pasteuriapenetrans isolatesfrom differentgeographical were thesame as the previous descriptions of Pasteuria areaswere testedfor theability of endospores to attach penetrans. tosecond-stage juveniles and to colonise females of differentpopulations of Meloidogyne spp.from coffeeand 455Biology and ultrastructure of anovelbacterial othercrops. Our resultsconŽ rm thatspore attachment parasiteof nematodes testcannot be used as theonly criteria for screeningfor 1; 2 theefŽ ciency of the bacterial isolate against nematodes. Aurelio CIANCIO ¤, Michelle CERMOLA , Thepercentage of femalesinfected with endospores and Renée FAVRE 2 andGaetano G RASSO 3 theendospore concentration/ mlin 100 females were 1 Istitutoper la Protezione delle Piante, Consiglio Nazionale thebest approaches for screeningthe highly aggressive delleRicerche, V iaAmendola 165/ A,70126Bari, Italy isolates.Using these parameters, it was possibleto 2 Istitutodi Genetica Buzzati Traverso, Consiglio Nazionale selectone isolate Pp12 for thethree populations of M. delleRicerche, V iaG. Marconi10, 80125, Naples, Italy paranaensis andthe isolate Pp10 for thefour races 3 CSATA, Valenzano,Bari, Italy of M.incognita from coffee.For theother species M. ¤[email protected] hapla, M. exigua, M.graminicola and M.mayaguensis , Anovelgram-negative bacterial parasite of free-living theisolates were notcompatible. In this study, there andplant parasitic nematodes was discoveredin southern was clearevidence that the greatest parasitism occurred Italy.The bacterium is a septaterod with a uniqueshape whenthe isolates of P.penetrans were exposedto resultingfrom the atteningof bothcell ends. It attacks species of Meloidogyne geneticallyclose to those from vermiformstages producing a lethaldisease. Parasitised whichthe bacteria populations were originallyisolated. nematodesmay be observed easily at low magniŽ cation RAPD analysiswas appliedto Ž ngerprintthe genomes becauseof thecreamy-yellow colour they assume. After of P.penetrans isolates.T wenty10-mer oligonucleotide adheringto thehost, the bacterium eventually penetrates primersof arbitrary sequence were usedon this study. itsbody, giving rise to a multiplicativephase spreading Ahighlevel of homogeneity was observedamong the theinfection and Ž llingthe whole nematode body with isolates. cells.The bacterial cells are released at nematode death andcadaver decomposition, infecting new hosts in the 454Distribution of Pasteuria spp. associatedwith soilthrough direct contact and cuticle adhesion. Atomic plant-parasiticnematodes in Korea forcemicroscopy (AFM) ultrastructuraldata of livecells

282 Nematology Postersessions showedthat the bacterium is covered by an adhesive Twosingle egg mass linesof Meloidogyneincognita matrixresponsible for attachment.SEM andTEM were were developedand from eachof these lines freshly usedto investigate the bacterium ultrastructure, and hatchedsecond-stage juveniles (J2) were collectedfrom showedelongated cells measuring 4-5 0.5 ¹m. threeindividual egg masses. Three separate replicates £ Attemptsto culture the bacterium on common nutritive of20 individual J2 from eachegg mass were exposed mediafailed. The pathogen was discoveredin association toendospores of a populationof Pasteuriapenetrans withfour distinct Meloidogyne spp.populations in originallyisolated from apopulationof Meloidogyne southernItaly, but it was alsoobserved in free-living javanica andthe endospores adhering to each J2 Cephalobidae.The bacterium appears to be a novel were counted.One population of M.incognita became invertebratepathogen and will probably represent a encumberedwith between nine to more than 100 furthertool in nematode biological management. endosporesper J2 while in the other population this rangedfrom zeroto less than 70 endosporesper J2. The distributionof the numbers of endospores attaching to 456Attachment tests of Pasteuriapenetrans to individualJ2 was thereforevery different and in one clonallines of Meloidogyneincognita populationof M.incognita thenumber of endospores were highlyskewed with the majority of J2having fewer Keith G. DAVIES¤ and Rosa H. MANZANILLA-LÓPEZ thanten endopores and one or two J2 having more than NematodeInteractions Unit, IACR Rothamsted,Harpenden, 30.The experiment was repeatedand similar results were Hertfordshire,AL5 2JQ,UK obtained.The results will be discussed in relation to clonal ¤[email protected] variationand their implications.

Twosingle egg mass linesof Meloidogyneincognita were developedand from eachof these lines freshly 458The in uence of Tagetespatula on Pratylenchus hatchedsecond-stage juveniles (J2) were collectedfrom penetrans andthe growthof roserootstocks threeindividual egg masses. Three separate replicates Lutgart DE WAEL ,Geraerard H ENDRICKX of20 individual J2 from eachegg mass were exposed ¤ andMaurice M OENS toendospores of a populationof Pasteuriapenetrans originallyisolated from apopulationof Meloidogyne javanica andthe endospores adhering to each J2 CropProtection Department, Agricultural Research Centre, Burg.V anGansberghelaan 96, 9820 Merelbeke, Belgium were counted.One population of M.incognita became [email protected] encumberedwith between nine to over 100 endospores ¤ perJ2 while in the other population this ranged from zeroto less than 70 endospores per J2. The distribution Thein uence of growing Tagetespatula cv.Single Gold ofthe numbers of endospores attaching to individual J2 orItalianryegrass, and of applicationsof aldicarb (0.9 g 2 2 was thereforevery different, and in one population of a.i./m /,matalaxyl(0.075g a.i./ m /,or12-monthsfallow M.incognita thenumbers of endospores were highly was investigatedon Pratylenchuspenetrans in a sandy skewedwith the majority of J2 having fewer thanten soil.Nematode numbers decreased signiŽ cantly after endosporesand one or two J2 having more than 30. Tagetes butincreased after Italian ryegrass. The fungicide Theexperiment was repeatedand similar results were hadno effecton the nematode while the effect of aldicarb obtained.The results will be discussedin relationto clonal was onlytemporary. Fallow resulted in a decreaseof P. variationand their implications. penetrans.Toestimatethe effect of the treatments, the stemdiameter and plant weight of subsequently grown roserootstocks Rosadumetorum cv.Laxa were measured. 457Attachment tests of Pasteuriapenetrans to SigniŽcant increase in both plant parameters was obtained clonallines of Meloidogyneincognita after both Tagetes andaldicarb. The other treatments had no effect. Keith G. DAVIES¤ and Rosa H. MANZANILLA-LÓPEZ

NematodeInteractions Unit, IACR Rothamsted,Harpenden, 459Field evaluation of the effect of Verticillium Hertfordshire,AL5 2JQ,UK chlamydosporium on Meloidogynearenaria race 3 ¤[email protected]

Vol.4(2), 2002 283 Biologicalcontrol (449-488)

M. Clara V. DOS SANTOS, Isabel M. DE O. ABRANTES, tothe cuticle within the sheath. T wenty-sixdays after SoŽ a DOS S. DA R. COSTA egression,numerous unattached spores were observedbe- andM. SusanaN. DE A. SANTOS tweenthe 2nd and 3rd stage cuticles of someensheathed nematodes;however, no sporeswere observedwithin the Institutodo Ambiente e Vida,Departamento de Zoologia, Universidadede Coimbra, 3004-517 Coimbra, P ortugal bodiesof those IJ3. When spore-encumbered IJ3 were [email protected] usedto infect D.abbreviatus larvae,no spores were de- tectedon juvenile or adultmale or femalenematodes up Theconcern and restriction on nematicideuse require the to96 h afterinsect death. However, moulted cuticles of developmentof alternative nematode control strategies. IJ3with attached spores were recoveredfrom dissected Thenematophagous fungus Verticilliumchlamydospo- insectcadavers. Studies are ongoing to determine the life rium hasshown potential as a biologicalcontrol agent for cycleof theorganism relative to the nematode and insect. root-knotnematodes in pot and microplot experiments. A 2-yearŽ eldexperiment was conductedat Coimbra, Por- tugal,to study the effect of V.chlamydosporium on Me- 461EfŽ cacy of Pochoniachlamydosporia var. loidogynearenaria race3. T omatocv. Joker, susceptible chlamydosporia and Paecilomyceslilacinus on tothe nematode, and cabbage, as apoorhost, were used biocontrolof Heteroderaschachtii onsugarbeet inthe summer and winter seasons, respectively. The ex- Seddigheh FATEMY perimentcompared the use of V.chlamydosporium with thenematicides oxamyl and methyl bromide. Nematode NematologyDepartment, Plant P estsand Diseases Research 1 populationdensities, gall index and number of eggsg ¡ Institute,P .O.Box1454-19395, T ehran,Iran rootwere higherafter the tomato crop. The fungus was ableto colonise only the tomato roots and was detectedin Differentisolates of Pochoniachlamydosporia var. soiland in nematode eggs only after 2 years.Under the chlamydosporia and Paecilomyceslilacinus were highly conditionsof this experiment, the fungus was noteffec- effectivein controlling nematodes on water agar . tivein decreasing nematode levels. A longerperiod would Aglasshouseexperiment was conductedusing P. beneeded for thefungus to be established and effective lilacinus andfour isolates of P.chlamydosporia var. againstroot-knot nematodes. Further research, exploring chlamydosporia (Po1,Po3, Po4, Po5). Seedlings of sugar environmentalfactors that also affect survival and prolif- beetwere plantedin steam-sterilisedsoil and, at thesame erationof thefungus in soil,is neededto increasecoloni- time,100 g cerealsinfected with different fungi, and 5 g sationof soiland roots by thefungus. oxamylwere incorporatedinto the soil of appropriate treatments.After 27days, plants were inoculatedwith 4000juveniles of H.schachtii andpots were arranged 460 A Pasteuria-likeorganism on an randomlyin a glasshousefor 2months.There were entomopathogenicnematode parasite of Diaprepes signiŽcantly fewer cystson the roots treated with fungi abbreviatus andoxamyl than untreated nematode-infected plants; the smallestnumbers observed were onPo1 treated roots. F.E. EL-BORAI, L.W. DUNCAN and D. DUNN Freshweight of theseplants signiŽ cantly increased with Universityof Florida,Citrus Research and Education Center , additionof fungi and oxamyl. Nematodes multiplied 700Experiment Station Rd, Lake Alfred, FL 33850,USA nearly22-fold on untreatedplants, 11-fold on P.lilacinus Duringa Želdstudy of endemic entomopathogeni cne- andPo3 treatments and between two to Ž ve-foldon matodesin Florida, Pasteuria-likespores were frequently plantstreated with Po1, Po4 and Po5,respectively. Percent foundadhering to 3rd stage juveniles (IJ3) of Steinernema control of H.schachtii numbers(eggs/ gsoil)were 46% sp.emerging from cadaversof theweevil Diaprepesab- for Po3,52% for P.lilacinus ,78%for Po4,87% for Po1 breviatus.Cadaverswith emerged IJ3 were maintained and90% for Po5,respectively. in2 mlwater in sealed Petri dishes and observed peri- odically.Numbers of unattached spores in the water in- 462Production of Pasteuriapenetrans ‘in vivo’ in creasedover time. Spores were neverfound attached to the soilswith different texturesand in the sand IJ3sheath (2nd stage cuticle), whereas most exsheathed 1; 2 2 IJ3were encumberedwith several hundred spores. A few C.B. GOMES ¤, L.G. FREITAS , S. FERRAZ , (<1%)ensheathed IJ3 had one to severalspores attached R.D.L OLIVEIRA 2 and W.B. SCIVITTARO 1

284 Nematology Postersessions

1 EmbrapaClima T emperado,C.P .403,96001-970, P elotas-RS, were analysedby probit analysis. Chrysanthemum roots Brazil were themost toxic with an LC50 of 1.0 mg/ cm 3 fol- 2 UniversidadeF ederalde Viçosa, Depto Fitopatologia, lowedby black-eyedSusan with an LC50of 3.5mg/ cm 3. 36571-000,V içosa-MG, Brazil Sesameroots showed some activity but no mortalitywas ¤[email protected] observedin the marigold, castor bean or crotalaria root Themass productionand attachment of Pasteuria bioassays.Leaves of marigoldand black-eyed Susan had penetransto Meloidogynejavanica secondstage juveniles LC50values of 8.0 and 9.0 mg/ cm 3,respectively,while (J2) were evaluatedin soils of different textures and leaftissue of theremaining plants showed little toxic ac- inriver sand, in the glasshouse. T omatoseedlings were tivity. transplantedto pots containing those substrates. The 5 upper5 cmlayer of the substract was infestedwith 10 464Occurrence and biocontrol potential of endospores/gsoilof P.penetrans ,andeach plant was potato-associatedbacteria inoculatedwith 2000 J2. Seventy days later, the plant rootswere harvested,dried, weighed and ground. The Johannes HALLMANN 1,AnnekathrinF AUPEL 1, numberof endospores/ gofroot, and per plant were Annette KRECHEL 2 andGabriele B ERG 2 estimated.Soil samples were collectedfrom upperlayer 1 andfrom thebottom of each pot, dried, and infested Instituteof PlantDiseases, Bonn University, Nussallee 9, with600 J2 of M. javanica toevaluate the endospore D-53115Bonn, Germany 2 Institutefor Molecular Physiology and Biotechnology, percolationof P.penetrans .Moreendospores/ plantwere RostockUniversity, Albert-Einstein-Str .3,D-18051 Rostock, producedin sandy soils compared to clay soils or river Germany sand.Negative correlation was observedbetween P. penetrans productionper plant and the soil clay content. Plant-associatedbacteria are able to improveplant growth Soilswith lighter texture allowed higher attachment of P. andhealth, but little is known about their diversity penetrans tojuveniles of M. javanica thanheavier ones. andantagonistic potential against disease complexes Mostof theendospores were leachedto thebottom of the consistingof plant parasitic nematodes and soil-borne potby thepercolating water in the sand. fungi.In a cooperativeproject, bacteria were isolated from therhizosphere, endorhiza, phyllosphere and endosphereof potatocv. Cilena at Bonn and Rostock, and 463Nematicidal activity of selectedplant tissues to studiedfor theirantagonistic potential against Verticillium Xiphinemaamericanum dahliae/Rhizoctoniasolani (Rostock)and Meloidogyne incognita (Bonn).The bacteria were characterisedbased John M. HALBRENDT¤ andJames A. L AMONDIA ontheir fatty acids using standard procedures for GC- PennState University, 290 University Drive, Biglerville, FAMEandthe Sherlock Microbial IdentiŽ cation System PA17307,USA; The Connecticut Ag. Expt Station, (MIDI). Bacterialrichness and diversity were determined 153Cook Hill Road, Windsor ,CT06095,USA tocompare population dynamics depending on location, [email protected] ¤ habitatand growth stage. Up to 10% of the culturable Freeze-driedleaf and root tissue of six plants includ- bacteriashowed antibiosis against V. dahliae and R. ingedible mum ( Chrysanthemumcoronarium ), black- solani.Outof these isolates, 20% gave signiŽ cant eyed Susan (Rubeckiahirta ),marigold( Tagetesminuta control of M.incognita ingreenhouseexperiments. Main ‘polynema’), sunn-hemp( Crotalariajuncea ), castorbean bacterialgenera containing isolates with antagonistic (Ricinuscommunis )andsesame( Sesamumindicum ) were activitywere Curtobacterium and Streptomyces for the assayedfor nematicidalactivity against the dagger ne- rhizosphereand phyllosphere and Pseudomonas for matode (Xiphinemaamericanum ).Thetissue was pul- theendosphere and endorhiza. In 2001, samples were verisedand mixed with dry sterile sand in concentrations takenat three different growth stages (establishment, thatranged from 0.5-5.0mg/ cm 3 sandfor rootsand 5.0- owering,maturity). Differences in bacterial community 20.0 mg/cm3 sandfor leaves.Nematodes were transferred structureand antagonistic activity will be shown. The to2 mlsample cups in 250 ¹lsteriledistilled water fol- potentialof indigenous antagonistic bacteria to suppress lowedby 1.5 cm 3 oftheplant/ sandmix. The cups were nematode/fungaldiseases complexes is discussed. sealedand incubated for 24hat24 ±C.Allnematodes were recoveredand the numbers of alive and dead nematodes

Vol.4(2), 2002 285 Biologicalcontrol (449-488)

465Study ofcombineduse ofantagonistplants and with Agrobacteriumradiobacter .Theroleand importance arbuscularmycorrhizal fungi on Meloidogyne ofthe bacteria-nematode relationship on the host are incognita on tomato discussed.

M.C. JAIZME-VEGA , A.S. RODRÍGUEZ-ROMERO, ¤ 467Isolates of the generalistparasite Pochonia P. GARCÍA DE LA ROSA, P. AFONSO MORALES chlamydosporia havespeciŽ c interactionswith their and L.A. BARROSO NÚÑEZ nematodehosts InstitutoCanario de Investigaciones Agrarias (ICIA), Dpto Protección Vegetal,Apdo 60 38200La Laguna, T enerife,Spain B.R. KERRY¤, C.O. MORTON, T.H. MAUCHLINE ¤[email protected] and P.R. HIRSCH Organicdisease management programmes are being in- NematodeInteractions Unit, IACR-Rothamsted, Harpenden, creasinglyresearched in order to limitthe environmental Hert,AL5 2JQ,UK pollutioncaused by standard agricultural methods. Cur- ¤[email protected] rently,much research is focusing on nematode control Pochoniachlamydosporia isan opportunistic,facultative usingantagonist plants with nematicidal properties such parasiteof cyst and root-knot nematode eggs, fungi asaromatic species which produce essential oils with ac- andmolluscs, but individual isolates demonstrate host knowledgednematicidal and fungicidal properties, and speciŽc interactions.Genetic variation was examinedin the Tagetes spp.with a knownsuppressive effect on par- acollectionof 54 isolates from diversegeographical asiticnematodes. Glasshouse trials were setup to eval- regionsand several nematode hosts using ERIC PCR. uatethe combined effects of inoculationwith the mycor- Incluster analysis, isolates formed groups on the basis rhizalfungi Glomusmosseae or Glomusmanihotis and the oftheir original host; those from cystnematodes varied presenceof eitherthe aromaticherb Lavanduladentata or morethan those from Meloidogyne spp.The fungus Tagetesminuta ontomato infected with the root-knot ne- producesan alkaline serine protease, VCP1, during matode Meloidogyneincognita .Theresults show signif- infectionof nematode eggs. The gene encoding VCP1 icantdifferences in nematode reproduction between my- was sequencedand cDNAs from sixisolates from corrhizaland non-mycorrhizal plants with an important differentnematode hosts examined. Translated cDNA reductionin nematode levels in the plants inoculated with sequencesin isolates from cystand root-knot nematodes arbuscularmycorrhizal fungi and cultured in the presence consistentlyhad two amino acid polymorphisms at ofantagonistplants. positions65 and 99 in the binding region of the enzyme.Such differences may contribute to host 466Occurrence of Rhizobiumradiobacter associated preference.Isolates from cystor root-knot nematodes with Heteroderaavenae eggsin T unisia were addedalone and together to soil in which tomato orpotato plants infected with Globoderapallida or 1; Najoua KACHOURI-NAMOUCHI ¤ Meloidogyneincognita were grown.Isolate abundance andMouldi M. B’C HIR 2 was increasedonly in compatible combinations and the speciesof nematode in uenced the proportions of the 1 INRAT,2049Ariana, Tunisia mixedisolates. Pochoniachlamydosporia has complex 2 INAT 1082Cité mahrajè ne, Tunis, Tunisia interactionswith its hosts that may in uence its biological [email protected] ¤ controlefŽ cacy. Populationsof cerealcyst nematode, Heteroderaavenae , collectedfrom variouscereal regions in Tunisia were 468Comparative study onthe nematicidalactivities associatedwith several bacterial and fungal strains. ofgarlicand mustard oils Scanningelectron microscopy study has demonstrated the occurrenceof a bacteriumwithin the cyst and eggs of P.S. KING and R. RODRÍGUEZ-KÁBANA thepopulation nematode collected from Zaghouanregion. Thisbacterium has been puriŽ ed through micro-culture Deptof Entomologyand Plant P athology,Auburn University, ofdisinfected eggs on agar media. The bacteria was Alabama36849, USA thenidentiŽ ed using ampliŽ cation of ‘ rrs’gene coding Thenematicidal properties of mustard and garlic oils theARNr 16Sas Rhizobiumradiobacter synonymous were comparedto those of allyl isothiocyanate in a

286 Nematology Postersessions glasshousestudy. Five percent (v/ v)acetone solutions ofaction against other parasites (trematodes, fungi and ofallyl isothiocyanate, garlic oil and mustard oil were bacteria).It was alsodemonstrated in severalexperiments preparedand each delivered at rates of 0.1, 0.2, 0.4 thatconcurrent applications of chitinolytic agent or and0.8 ml a.i./ kgsoil that was naturallyinfested with achitinolyticactivity-inducing agent, and sulphide Rotylenchulusreniformis .Treatedsoils and untreated orsulphide-producin gagentsfrom microorganismsor controlwere placedin pots in the glasshouse. Pre-plant chemicalcompounds, required ranges or doses of soilsamples for nematodeanalysis were collected1 week applicationsigniŽ cantly less than any of thecompounds afterapplication of thematerials. The pots were planted appliedindividually, in order to achieve an effective withsoybean ‘ Young’and grownfor 8weeks.Plants were controlof the nematodes. This indicates that there is a thenremoved and nematodes were extractedfrom soil synergisticeffect in the combination of chitinases and androots. Fresh shoot weights were determined.Pre-plant sulphidefor thecontrol of nematodesand other parasites. samplingresults indicated that all three materials were Theresults are subject of a patentapplication on any activeagainst the nematode when applied at rates >0.2 antiparasiticcomposition obtained from themetabolites ml/kgsoil. Nematicidal activities of allyl isothiocyanate of Corynebacteriumpaurometabolum strainC-924, as andmustard oil were muchhigher than that of garlic wellas some other possible antiparasitic compositions oil.In the end-of-experiment sampling, populations of andbiocontrol microorganisms producing chitinases and reniformnematodes in soil were lowestin soils treated sulphideconcurrently. withthe two highest rates of allylisothiocyanate and the 0.8ml rate of mustard oil; numbers in soil with garlic 470Population of Corynebacteriumpaurometabolum oildid not differ from thecontrol. Nematode populations strainC-924 in soilstreated with HeberNem inthe roots followed the same pattern as for thesoil samples,except that the 0.8 ml rateof garlicoil reduced 1; 1 2 Jesús MENA ¤, Liuven VELOZ , Ramón P. VÁZQUEZ , nematodenumbers compared to the control. Treatments Manuel EXPÓSITO 1, Marieta MARÍN 1, withallyl isothiocyanate and garlic oil had no effect on Eladio SALAZAR 1, Licette LEÓN 1, shootweight; those with mustard oil at >0.1ml/ kgsoil Yaisa COCA 1, Yamilka RAMÍREZ 1, resultedin improved shoot weights. Results indicate that Gustavo RODRÍGUEZ 1,ArmandoT. H ERNÁNDEZ 1 garlicoil does not have the nematicidal properties shown andEulogio P IMENTEL 1 byallylisothiocyanate and mustard oil. 1 Centrode Ingeniería Genética y Biotecnología, P.O.Box387, Camagüey 70100, Cuba 469Mechanism of actionof Corynebacterium 2 LaboratorioProvincial de Sanidad V egetalde Camagü ey, paurometabolum strainC-924 on nematodes Cuba ¤[email protected] Jesús MENA1; , Eulogio PIMENTEL 1, ¤ Aprocedureto re-isolate bacterium Corynebacterium Armando T. HERNÁNDEZ 1, Liuven VELOZ 1, paurometabolum strainC-924 from ferriticsoils treated Ramón P. VÁZQUEZ 2, Licette LEÓN 1, withHeberNem, was developed.Strain C-924 is theactive Juan D. MENCHO 3, Yamilka RAMÍREZ 1, agentof HeberNem, a newbionematicide formulated Merardo PUJOL 1 and Carlos BORROTO 1 asa concentratedand humectant powder. It was tested 1 Centrode Ingenierí a Genética y Biotecnología, P .O.Box 387, inglasshouses on several crops, such as pepper and Camagüey 70100, Cuba tomato.HeberNem applicationswere carriedout by 2 LaboratorioProvincial de SanidadV egetalde Camagüey, meansof the irrigation system in doses from 1to5 Cuba kg/ha.A selectiveculture medium was developed,taking 3 Universidadde Camagü ey, Cuba intoaccount antibiotic combinations and also based on [email protected] ¤ bacteriacolony chromogenesis for potassiumtelurite Simultaneousproduction of hydrogen sulphur and reduction,so the differentiated re-isolation from soil chitinasesby Corynebacteriumpaurometabolum strain C- samplesof thespeciŽ c strainC-924 was performed.The 924was demonstrated.It happens in sufŽcient amounts to resultsconŽ rmed the effectiveness of the bacterium in impedeegg hatching and produce death in juveniles of thecontrol of Meloidogyneincognita ,whenit is in a severalspecies of plant and animal parasitic nematodes. concentrationhigher than 10 5 ufc/gofsoil.It was proven Thisresult permitted further study of the mechanism thatsoil populations of C-924 diminished progressively

Vol.4(2), 2002 287 Biologicalcontrol (449-488)

1; 2 3 andit was notpossible to detectit intreatedferritic soils K.M. MOHOTTI ¤, S.R. GOWEN and J. BRIDGE between60 and 120 days. It was concludedthat for the 1 TeaResearchInstitute of SriLanka, T alawakele,22100, besteffectiveness of theproduct, a Žrst applicationshould Sri Lanka bedone7 daysbefore planting and a secondone 30 days 2 Universityof Reading, UK afterit, using doses between 2 and5 kg/ha. 3 CABI Bioscience(UK Centre),UK ¤[email protected] 471Examination of rhizosphere-associated Themorphology and morphometrics of mature en- microbesfor production of compoundsactive against dospores of Pasteuria populationsrecovered from Sri plant-parasiticnematodes Lankannematodes belonging to several taxonomic groups were evaluated. Pasteuria populationsoriginating from Susan L.F. MEYER1; , Daniel P. ROBERTS 2, ¤ Meloidogyne and Pratylenchus from Australia,Japan, James K. NITAO 1 andDavid J. C HITWOOD 1 Kenyaand UK were alsoincluded in thestudy for com- 1 USDA, ARS, NematologyLaboratory, Bldg 011A, Rm 165B, parison.A widevariation in spore morphometrics was BARC-West,10300 Baltimore Avenue, Beltsville, MD evident. Meloidogyne yieldedcomparatively large spores 20705-2350,USA while Pratylenchus and Radopholus generatedsmall 2 USDA, ARS, SustainableAgriculture Systems Laboratory, spores;intermediate sizes were seenwith Xiphinema. Sri Bldg001, Rm 140,BARC-W est,10300 Baltimore Avenue, Lankanendospores possessed morphological changes fol- Beltsville,MD 20705-2350,USA lowingattachment onto nematodes. The endospores that [email protected] ¤ originatedfrom Sri Lankansoils had the three distinct In vitro studiesidentiŽ ed fungiand bacteria that produce forms (I-III). PPKB exhibitedthe very rare Form I;PPKB compoundsactive against plant-parasitic nematodes. attached to M.brevicauda retainedthe sporangium wall Assaysof fungus culture Ž ltrateswere conductedwith coveringthe surface. PPPT exhibitedrough intact pe- Heteroderaglycines (soybeancyst nematode: SCN) and ripheralŽ bresand the central body of the endospores Meloidogyneincognita (root-knotnematode: RKN). The was attachedto Pratylenchusloosi afterthe sporangium testedŽ ltratesexhibited a lowcorrelation in activity walland the exosporium were sloughingoff. PPNW en- againstSCN comparedwith activity against RKN. Only dosporespossessed peripheral Ž bresas Ž ne,delicate, re- 5.1%of thetested isolates produced culture Ž ltratesthat markablylong and prominent hairy structures. Spores of stronglyinhibited egg hatch of bothSCN andRKN ( i.e., PPKenyaisolated from P. goodeyi andPPBotesdale iso- egghatch was 40%or less than egg hatch in potato lated from M.ardenensis were foundto be smallest and dextrosebroth controls), 3.9% strongly inhibited SCN largest,respectively, described so far amongthe mem- egghatch only, and 13.4% inhibited RKN egghatch bers of the Pasteuriapenetrans group.PP-a alsoshowed (butnot SCN). Percentagesof Ž ltratesthat inhibited ahighermean endospore diameter . secondstage juvenile mobility were 0.8%(affected both SCN andRKN), 0.4%(SCN only),and 1.2% (RKN 473Endophytic behaviour of eggparasitic fungi in only).Compounds with broad-spectrum activity can be roots identiŽed without testing speciŽ c nematodetaxa, but identiŽcation of compounds with more speciŽ c activity E. MONFORT, J.J. BORDALLO, J. SALINAS, shouldbe tested against the target nematode. Similar L. ASENSIO and L.V. LOPEZ-LLORCA experimentswith rhizosphere-inhabit ingbacteria and RKN identiŽed isolates producing compounds active Departamentode Ciencias Ambientales y RecursosNaturales, againstRKN. OneŽ ltratethat suppressed egg hatch was Universidadde Alicante, Aptdo correos 99, 03080, Alicante, selectedfor furtherstudy; size-fractionation of theculture Spain Žltrateshowed that the active component is largerthan >3 Therhizosphere is an important zone for theactivity kDa in size. ofnematophagous fungi since plant-parasitic nematodes commonlyattack plant roots. Micro-organisms are found 472Morphological and morphometric innumbersten to 20 times(occasionally 100 times)higher characteristicsof SriLankan populations of mature thanin the root-free soil. The presence of nematode- endospores Pasteuria trappingfungi in the rhizosphere of agricultural plants hasbeen studied. The capability to colonise the roots was

288 Nematology Postersessions notadvantageous for thebiocontrol potential of isolates nematodedamage and stimulate plant growth. In 2001, onroot-knot nematode ( Meloidogyne spp.),in contrast TerraPywas successfullymarketed in the mediterranean tothe egg-parasiticfungus Verticilliumchlamydosporium , regionas aplantgrowth and yield-enhancingagent in veg- whereroot colonisation was suggestedto be aprerequisite etables. for successfulbiological control. W ehaverecently showncellular details of barley root colonisation by 475Ultrastructural study byTEM ofeggsand V.chlamydosporium .Itseems that the fungus is not juvenilesof nematodestreated with Corynebacterium recognisedby the plant, similar to mycorrhizae and paurometabolum strainC-924 non-pathogenicendophytes. W ehavealso found signs 1; 1 oflate induction of plant defence responses (3 weeks Eulogio PIMENTEL ¤, Jesús MENA , afterinoculation) such as phenolic droplets and papillae Abel HERNÄNDEZ 1, Ramón P. VÁZQUEZ 2, similarto howother fungal antagonists of plantpathogens Marina FERNÁNDEZ 2, María C. DE LA ROSA 1, modulateplant defences. W ethereforehypothesise a Ivon MENÉNDEZ 1, Liuven VELOZ 1, Licette LEÓN 1 doublemode of action for V.chlamydosporium and andV ivianaF ALCÓN 1 perhapsother nematophagous fungi: i)directparasitism onnematodeeggs; and ii)immunisationagent. W epresent 1 Centrode Ingeniería Genética y Biotecnología, P.O.Box387, ourinitial results on plant defence modulation by V. Camagüey 70100, Cuba chlamydosporium andshow examples of its potential to 2 LaboratorioProvincial de Sanidad V egetalde Camagü ey, controlroot pathogens. Cuba ¤[email protected]

474The potential of TerraPy ® toreduce nematode Changesin the ultrastructure of Haemonchus spp. eggs, damage treatedwith Corynebacteriumpaurometabolum strain C- 924,were observedby TransmissionElectron Microscopy 1 2 MULAWARMAN , Johannes HALLMANN , (TEM).Alterations in the eggs of Haemonchus spp. 3 1 Doris BELL andRichard A. S IKORA exposedto the action of thebacterium, were detectedafter 1 Instituteof Plant Diseases, Bonn University, Nussallee 9, 12htreatment.Regions normally containing lipids were D-53115Bonn, Germany occupiedby determinedvacuolar spaces, which separated 2 FederalBiological Research Center ,Toppheideweg88, thelipidic and chitinous layers. Closely bound to these D-48161Mü nster ,Germany spaces,numerous foldings produced in theexternal layer 3 CognisDeutschland, Henkelstrasse 67, D-40551 Dü sseldorf, ofthe eggs were observedas the main damage suffered Germany ineggshell. These structures were notobserved in the TerraPy stimulatesmicroorganism activity in thesoil and eggswithout C-924 treatment. It was alsoobserved promotesgrowth of plant-beneŽcial microorganisms such that C.paurometabolum strainC-924, caused the death asmycorrhiza and plant growth promoting rhizobacteria of Meloidogyneincognita secondstage juveniles (J2). (PGPR). As aresult,damage caused by plantparasitic ne- After 24h exposureto C-924, vacuoles observed for matodesis reduced.In greenhouse experiments, Meloido- Haemonchus spp.eggs were smallerthan for the M. gyneincognita infestationon tomatowas signiŽcantly re- incognita J2.This situation could be related to greater ducedfollowing a soildrench with TerraPy at 2 kg/ha.In- susceptibilityof M.incognita J2to gases emitted by the creasingapplication rates up to 200kg/ hafurtherreduced bacterium.Studies on toxic gas production, proteolytic nematodedamage. At thesehigher rates, plant growth was activity,as well as the capacity of C.paurometabolum alsosigniŽ cantly enhanced. Y ieldof cherrytomatoes was strainC-924 to degradechitin and collagen are proposed. 50%higher when treated with 200 kg/ haTerraPy thanin thecontrol. The increase in yield was causedby more 476The relationship between glucosinolatesin fruitsper plant. However, fruit weight was notaffected. canola (Brassicanapus )andtheir ability to control Similarly,for commercialtomato production in Turkey the plant-parasiticnematodes, Pratylenchusneglectus anincrease in thenumber of owers isreported as major and Meloidogynejavanica factorof yieldenhancement. Due to its positive effect on 1; 2 3 beneŽcial microorganisms, coapplication of TerraPywith Mark POTTER ¤,EkaterinaR IGA , Graham STIRLING mycorrhizaor Pseudomonas inoculumwill further reduce and Sharyn TAYLOR 4

Vol.4(2), 2002 289 Biologicalcontrol (449-488)

1AdelaideUniversity, Department of Applied and Molecular creasedplant health, shoot and root weights and greening Ecology,PMB 1GlenOsmond, SA 5064,Australia were alsonoted in multiple Ž eldtrials, prompting inves- 2 WashingtonState University, P .O.Box646430 W ashington tigationsinto microbial changes in DiT era ® treatedsoils StateUniversity, Pullman, W ashington99164, USA ingreenhouse and under Ž eldconditions. V ariousstud- 3 BiologicalCrop Protection Pty Ltd, 3601 Moggill Rd Moggill, iesincluding FDA hydrolysis,enzymatic activity, pH and Queensland4070, Australia conductivitymeasurements, and determination of soil, 4 SouthAustralian Research and Development Institute, GPO roottissue and rhizosphere microbial populations demon- Box397, Adelaide, 5001, Australia stratedDiTera ® increasedgram-positive microbial popu- ¤[email protected] lationsespecially in therhizosphere and soil. These direct Thetoxicity and susceptibility of canola( Brassicanapus ) andindirect effects on nematodesand the soil rhizosphere plantscontaining different root levels of glucosinolates contributesto theoverall beneŽ cial response of plantsto were testedagainst the root lesion ( Pratylenchusneglec- DiTera®. tus)androot-knot ( Meloidogynejavanica )nematodesin glasshousetrials. Plants containing higher root levels of 478Effect ofmycorrhizalarbuscular fungus 2-phenylethylglucosinolate were signiŽcantly more re- Glomusmanihotis onmicropropagatedbanana sistant to P.neglectus andmore toxic as they degraded cultured onsoilnaturally infested bylesion nematode inthe soil. This conŽ rms previousŽ ndings,and suggests under microplotconditions thathigh root 2-PE glucosinolatecanola crops should pro- videa diseasebreak effect against this nematode. High A.S. RODRÍGUEZ-ROMERO¤, M.C. JAIZME-VEGA 2-PE glucosinolatetissues were alsomore toxic to popu- and L.A. BARROSO NÚÑEZ lations of M. javanica,butplants showed no signiŽ cant advantagein termsof resistanceto the nematode. In fact, asigniŽcant and positive relationship between glucosino- InstitutoCanario de InvestigacionesAgrarias (ICIA), Dpto latelevels and nematodenumbers generated was observed Protección Vegetal,Apdo 60, 38200 La Laguna, T enerife,Spain inone variety, suggesting that the elevated presence of the ¤[email protected] glucosinolateactually gave the nematode an advantage. Thedisease break effect of thecrop against the root-knot Theroot lesion nematode Pratylenchusgoodeyi Sher nematodemay be underminedby itssusceptibility, and so &Allenis considered an important nematode pest of cautionshould be used in crop selection in areas where commercialbananas in the Canary Islands. Arbuscular theroot-knot nematode is a problem. mycorrhizalfungi (AMF) areobligate symbionts that colonisethe roots of most cultivated plant species. It 477Examination and disclosure of the multifaceted hasbeen demonstrated that early mycorrhizal inoculation modesof actionof DiTera ®,abiologicalnematicide promotesplant growth and increases the tolerance ofbanana to attack by nematodes. However, most Linda REHBERGER¤, Bassam SHAMMO interactionstudies between both microorganisms have and Prem WARRIOR beendeveloped during the nursery phase where most conditionsare under control. In this trial we study ValentBioSciences Corporation, 6131 RFD OakwoodRoad, theeffect of the interaction between the AMF Glomus LongGrove, IL 60047,USA manihotis and Pratylenchusgoodeyi onmicropropagated ¤[email protected] bananaGrande Naine, under microplot conditions. This Researchlaboratories such as IACR-Rothamsted, Impe- experimentalmethod is the nearest to Ž eldconditions. rialCollege, UK andAuburn University, AL, were in- Severalsamples were takenat differenttimes to evaluate volvedin the investigation into the complex activities mycorrhizaleffect and nematode infection. Six months foundin DiT era ® (a biologicalnematicide) on plant- aftertransplanting to microplotsthere was stilla positive parasiticnematodes, during 1997-2000. Egg hatch inhi- effectof AMF .Thiseffect decreased at the end of trial, bition,muscle paralysis, feeding inhibition, depletion of after18 monthsin themicroplots. At Žnalharvest, plants lipids,and changes in sensory perception affecting activ- showedimportant levels of nematode infection and the itiessuch as host/mate-Žnding were affectedby exposure consequentdamage. to DiTera®.Inmost instances these activities were found tobe irreversible. In addition to nematode activities, in-

290 Nematology Postersessions

479The elaboration of biologicalcontrol methods of second-stagejuveniles (J2) andeggs; ii)metabolitespro- nematode-,virus-, fungi-, bacteria-complexinfections ducedby thefungus affect the J2 viability and egg hatch- onleguminousplants ing. In vitro parasitismability was presented,to different extents,by all the strains tested: IMI206040 (designated 1; 2 N.D. ROMANENKO ¤, B.F. TARAVALLI WT)andits transgenic strain P-2 whichcontains multi- 2 and V.G. ZAEC plecopies of theproteinase gene prb1,strainsT -203(Y), 1 Instituteof Parasitologyof RussianAcademy of Sciences, T-44,T -35and T -315.A transgenicstrain of IMI206040 Moscow117071, Leninskii prosp. 33, Russia carryinga fusionof the prb1 promoterwith the green 2 RussianUniversity of Friendshipof Peoples,Moscow ,Russia uorescentprotein (GFP) genewas usedto demonstrate ¤[email protected] thatthis gene is turned on duringthe interaction between thefungus and the nematode. Culture Ž ltratesof WT and Nematodes,virus, fungi and bacteria and their associ- P-2 paralysedand killed the M. javanica J2,as well as ationswere studiedin different type phytocenoses of otherplant-parasitic nematodes. The activity on M. java- Moscowon leguminousplants. The complex seed infec- nica J2was heatsensitive and restricted to the low mole- tionsincluded two to Ž vespecies, mainly of fungi and cularweight fraction (less than 3 kDa).The higher mole- bacteria.The root and abovegroundplant organ-infections cularweight fraction, which contained the proteases, did included,as arule,Ž vemore pathogens, consisting mainly notdirectly affect the J2. However, a ladderof proteases ofnematodes, fungi, bacteria and viruses. The meth- between20-31 KDa, whichpresented optimal proteolytic ods,dates and efŽ ciency of Russianbiological pesticides activityat lowpH range,was positivelycorrelated to the (planrhiz,trichodermin) and nematicidal strains of bac- nematicideactivity of thevarious strains Ž ltrates. teriaantagonists ( Bacillus, Pseudomonas )andbacteria whichsecrete antibiotic substances ( B.polymixa,Enter- obacter spp.)were studiedas ecologically safe methods 481Evaluation of Polishisolates of nematophagous torestrict the quantity and harmful in uence of complex fungifor the controlof the root-knotnematode infections(nematodes, viruses, fungi, bacteria) on legu- (Meloidogynearenaria (Neal) Chitwood) in vitro and minousplants grown under the conditions of theMoscow inglasshouseconditions region.The in uence of differentspecies of plantprede- cessorson numberand species of nematodesof different Danuta SOSNOWSKA ecologicaland trophical groups in theprocess of growing leguminousplants in seven-Ž eld crop-rotation were stud- Instituteof PlantProtection, Department of Biocontrol& ied. Quarantine,ul. Miczurina 20, 60-318 P oznan,P oland [email protected] 480Biocontrol of plant-parasiticnematodes by Tricodermaharzianum –possiblerole of proteases Theeffects of different species of nematophagous Ponchoniachlamydosporia chlamydosporia 1; 2 fungi: var. , Edna SHARON ¤, Alfredo HERERA-ESTRELLA , Ilan 3 1 Paecilomyceslilacinus and Cylindrocarpondestructans CHET andY itzhakS PIEGEL on Meloidogynearenaria eggsand juveniles were tested 1Departmentof Nematology,Agricultural Research in vitro.Allspecies of fungi were isolatedfrom eggs Organization(ARO), TheV olcaniCenter ,P.O.B.6, Bet-Dagan ofsugar beet nematode ( Heteroderaschachtii ) and 50250,Israel controlled M. arenaria population in vitro.IsolatesVc- 2 Centrode Investigació n yEstudiosAvanzados, Plant 1 of P.chlamydosporia and P.lilacinus parasitised90% Biotechnologyand Genetic Engineering Unit, Apartado, ofeggs, while C.destructans parasitisedonly 50%. C. Mexico destructans infectedmore than 19% of juveniles at a 3 OttoW arburgCenter for Agricultural Biotechnology, F aculty o temperatureof 25 C, while P.chlamydosporia infected 3- ofAgricultural, F oodand Environmental Quality Sciences, The P.lilacinus HebrewUniversity of Jerusalem, Israel 10%and none was infectedby .Thepotential of C.destructans asabiologicalcontrol agent against M. ¤[email protected] arenaria inglasshouse conditions is under investigation Strainsof the fungal biocontrol agent Tricoderma andresults will be presented. harzianum canaffect the root-knot nematode Meloido- gynejavanica byvarious modes: i)directparasitism on

Vol.4(2), 2002 291 Biologicalcontrol (449-488)

482Endotoxin activity of lipopolysaccharide out.The data have shown that the treatment of seeds produced by Moraxellaosloensis ,abacterium oftomatoes with water solutions (0.5 mg/ ml) of associatedwith the nematode Phasmarhabditis thesubstances stimulated development of plants and hermaphrodita ,againstthe slug Derocerasreticulatum suppressednematodes. The size of galls and females of nematodesin testedplants were muchless than in control Li TAN¤ andParwinder S. G REWAL plants.The treatment by eñ disone stimulated formation Departmentof Entomology, The Ohio State University, W ooster, ofmales in a population.The most effective were the Ohio44691, USA furostanolglycosides, solamargine and tomatosid. These ¤[email protected] reducedinfestation of rootsby nematodes and increased Moraxellaosloensis isa gram-negativebacterium as- durationof developmentof theparasites 1.5-2 times. The sociatedwith Phasmarhabditishermaphrodita , a slug- combinationsin the concentration used did not affect parasiticnematode that has prospects for thebiological nematodes in vitro.Itis possible that the action of controlof molluscpests, especially the grey garden slug somesteroids (in particular eñ disone) is caused by their Derocerasreticulatum .Thisbacteria-feeding nematode participationin biosynthesisand metabolismof moultand actsas a vectorto transport M.osloensis intothe shellcav- sexualhormones of nematodes,as itresultedin change of ityof theslugwhere the bacteria multiply and kill the slug theirage qualiŽ cation-sexual structure. within1 to2 weeks.W ediscoveredthat M.osloensis pro- ducesendotoxin(s) highly toxic to the slug when injected 484Effect ofculturalpractices on the management intothe shell cavity. Injection of puriŽed lipopolysaccha- of Meloidogyne spp. onglasshousecrops by the ride(LPS) from M.osloensis culturesinto the shellcavity nematophagousfungus Verticilliumchlamydosporium causedslug death with an estimated LD 50 of 48 ¹g per slug.No contact or oral toxicity of the LPS tothe slug Veerle VAN DAMME¤, Annemie HOEDEKIE, Wouter was detected.Isolated lipid A portionfrom theLPS was VANHOVE and Nicole VIAENE toxicto the slug after injection into the shellcavity, but the polysaccharideportion of the LPS was not.Further, we AgriculturalResearch Centre, Crop Protection Department, semi-quantitatedthe LPS as6 107 endotoxinunits per Burg.V anGansberghelaan 96, 9820 Merelbeke, Belgium £ mgand detected endotoxin activity in thelipid A portion ¤[email protected] butnot in thepolysaccharide portion by limulus amoebo- Severalisolates of Verticilliumchlamydosporium , an egg- cytelysate assays. This appears to be theŽ rst reportof an parasitisingfungus of Meloidogyne spp.,were selected activebiological toxin against molluscs. for applicationin commercial vegetable production in glasshouses.For thispurpose, the fungus must be 483Research on the inuence of Žveplantsteroid compatiblewith cultural practices such as fertiliser substances onplant-nematoderelationships (tomato – use,soil type and pesticide application. Glasshouse Meloidogyneincognita ) experimentsin pots were performedto study the effects ofthose practices on the efŽ cacy of the biocontrol 1; 1 2 Z.V. UDALOVA ¤, S.V. ZINOVIEVA , I.S. VASILIEVA agent.Second-stage juveniles (J2) of Meloidogyne spp. 2 and V.A. PASESHNICHENCO were usedas inoculum and tomato was thehost plant. 1 Instituteof Parasitology,Russian Academy of Sciences, Fertilisersdid not have negative effects on the efŽ cacy Leninskiipr .,33,Moscow ,119071,Russia of V.chlamydosporium .Additionof the fungus together 2 BakhInstitute of Biochemistry,RAS, Leninskiipr .,33, withinorganic, organic-mineral and pure organic fertiliser Moscow,119071,Russia causedup to 75, 76 and 86% reduction in the number ¤[email protected] ofJ2 per 100 cm 3 soil,respectively. Without fertiliser, Itis known that plant steroids, with various chemical theeffect of V.chlamydosporium onthe nematode structures,can in uence the growthand moult of parasitic populationwas notsigniŽ cant. Of thefour soil types nematodesand their hatch from eggs.The estimation tested,organic heat-sterilised soil had a negativein uence ofŽ veplant steroid substances (solasonine, eñ disone, onthe performance of V.chlamydosporium .Incontrast, solamargine,tomatosid and furostanol glycosides) on addition of V.chlamydosporium toorganic untreated soil, nematodeand plant in thehost-parasitic system (tomato – mineraluntreated soil and mineral heat-sterilised soil root-knotnematode Meloidogyneincognita )was carried reducedbetween 71 and 82% of the numbers of J2

292 Nematology Postersessions per 100 cm3 soil.The in uence of pesticide use on the TerraPy Ag® stimulatesmicrobial activity in thesoil and performanceof V.chlamydosporium willbe discussed. promotesgrowth of beneŽcial plant organisms. The effect ofTerraPy Ag ® onnematode densities, gall rating, and yieldwas evaluatedin a plastichouse. T omatocv. Durinta 485Compatibility of Corynebacterium was plantedin Marchand harvested in July 2001. TerraPy paurometabolum strainC-924 with Nemacurand Ag® was appliedat a totaldose of 20 ml/ m 2 per plot, otherchemical pesticides splitand applied in two ways: i)monthlyapplications at 1; 1 1 arateof 5 ml/m 2 for 4monthsstarting at planting, and Ramón P. VÁZQUEZ ¤, Jesús MENA , Liuven VELOZ , 2 Yamilka RAMÍREZ 1, Gilda JIMÉNEZ 2, ii)applicationat a rateof 10 ml/ m inthe 2nd and 3rd Victor M. PICO 2, Licette LEÓN 1 monthafter planting. Untreated plots served as control. andEulogio P IMENTEL 1 Finaldensities, egg production, and gall rating were lower (P < 0:05/ inplots treated monthly with 5 ml/m 2 of 1 Centrode Ingenierí a Genética y Biotecnología deCamagü, TerraPy Ag® thanin those left untreated. Densities in soil P.O.Box 387, CP 70100,Camagü ey, Cuba androot, and gall rating were unaffectedby TerraPy Ag ® 2 LaboratorioProvincial de SanidadV egetalde Camagüey, inthe 2nd and 3rd month after planting. The number of Cuba fruitsand their average fruit weight were higher( P < ¤[email protected] 0:05/ inplots treated monthly with 5 ml/m 2 of TerraPy Ag® thanin those untreated. Percent yield increase in Interactionswith chemical pesticides are required dur- thoseplots was 30%with respect to untreatedones which ingthe registration process of biological products. Bac- provideda proŽt of18030euros/ ha. terium Corynebacteriumpaurometabolum strainC-924 isa representativecase, which has been reported pre- viouslyas a newnematicidal agent (patent application 487Observations on the lifecycle of in vitro cultures PCT/NL95/00271).A selectionof the eleven chemical of Pasteuriapenetrans pesticides,more widely used in agriculture, were tested 1; 2 in vitro.Potexperiments to conŽ rm in vitro assays are James H. WHITE ¤, John F. GERBER , 1 1 shown,and positive results were obtainedspeciŽ cally for Thomas E. HEWLETT , Kelly S. SMITH , 1 1 Nemacur(phenamiphos). Interaction tests with chemical Judith S. PHILLIPS , Susan T. GRISWOLD and 1 pesticideswere developedunder controlled conditions, Amanda K. MORRIS growthinhibition zones of C-924 appear when interac- 1 EntomosLLC, 445SW 35 th Terr.,Suite310, Gainesville, tionswith different chemical products were performed. FL32608,USA Threetest concentrations of chemical pesticides were 2 CDG Laboratories,Inc., Gainesville, FL, USA used for in vitro assays:work-Ž eld, higher and lower con- ¤[email protected] centrations,so irregularities in Ž eldcondition when chem- icalsare used were takeninto account. Different results Studiesof the life cycle of Pasteuriapenetrans in were observedwhen using different chemical agents, so Meloidogyne spp.have employed light microscopy, broadsensitivity range and resistance degrees are shown. scanningelectron microscopy and transmission electron Potresults and in vitro testsshowed the possibility of us- microscopy.These studies used P.penetrans infected ing both C.paurometabolum strainC-924 as abionemati- juvenilesthat were inoculatedon plants, sequentially cidalformulation and Nemacur in combination,for nema- andtemporally retrieved from rootsand destructively todecontrol in IntegratedPest Management. sampled.The close association between P.penetrans cells andnematode tissue allows for observationsof onlylarge 486Evaluating the effect ofTerraPyAg ® on structures, i.e.,mycelialballs, thalli, and endospores. densitiesof Meloidogynejavanica andyield of tomato Developmentof in vitro culturetechniques allows detailed ina plastichouse observationsof early, small growth structures of this bacterium.Early growth stages of this bacterium were

Soledad VERDEJO-LUCAS¤ and Magda GALEANO recordedusing light and scanning electron microscopy. Theseearly growth stages progressed in in vitro cultures IRTA, Ctra.de Cabrilss/ n.08348 Cabrils, Barcelona, Spain to classic P.penetrans structures,similar in sizeand shape ¤[email protected] tothosereported from in vivo studies.

Vol.4(2), 2002 293 Biologicalcontrol (449-488)

488Sensitivity of Meloidogynejavanica and againstsecond stage juveniles of Meloidogynejavanica Tylenchulussemipenetrans toisothiocyanates in and Tylenchulussemipenetrans .Therelative toxicity of laboratorybioassays ITC(inascending order) was: phenyl < ethyl< butyl

294 Nematology Postersessions

489Effect ofOxycomon growthof tomatoand bromidenematode control. In nine of the assays carried reproductionof Meloidogyneincognita outsince 1998 no signiŽ cant differences were found TelopicEC andmethyl bromide in M.incognita average Safdar A. ANWAR¤ and M.V. MCKENRY¤¤ root-knotindex (Telopic: root-knot index 0.5. MB: D root-knotindex 0.5),marketable yield (T elopic:9.4 Universityof California,Riverside CA 92521,USA D kg/m2. MB 9.3 kg/m2/ orplant growth (T elopic ¤[email protected] 137.3 cm. MBD 143.5cm). However,the proportion D ¤¤[email protected] D of M.incognita infestedplants (21.1%) on the mixture Singleor multipleapplications of Oxycom,peroxyacetic treatmentswas signiŽcantly higher than that of MB acidplus various biocontrol agents, were comparedon 15- (5.1%).When application was repeated,marketable yield day-oldsusceptible tomato inoculated with 1500 second andplantgrowth at the3rd year were similarbut root-knot stagejuveniles (J2) M.incognita. Fortypots were dipped indexwas signiŽcantly higher on Telopic(21.1%) than on into2500 ppm (v/ v)Oxycom,and 20 inwater.T wentyof MBtreatments(0.6). the40 potsreceived four additional Oxycom applications at10-day intervals. Plants were harvestedat 60 days afterinoculation to assess the effect of treatments on 491Chemical options for the managementof plantgrowth, nematode development and reproduction. Heteroderaschachtii insugar beet under Želd Asingletreatment of Oxycom just prior to nematode conditions inoculationsigniŽ cantly increased tomato top weight Saad L. HAFEZ andSundararaj P ALANISAMY butnot other growth parameters. Associated with plant ¤ growthbeneŽ t was asigniŽcant increase in the number Universityof Idaho,P armaResearch and Extension Center , ofroot-knot females. Multiple treatments of Oxycom 29603U ofILane,P arma,Idaho 83660, USA signiŽcantly reduced leaf area, top weightand root weight ¤[email protected] whilesigniŽ cantly increasing the numberof galls,females AŽeldexperiment was conductedat the Parma Research andJ2 per plant or perg ofroot.This study demonstrates andExtension Center, Parma, Idaho,to evaluate the thatOxycom stimulates plant growth and earlier fruiting efŽcacy of Temik at different rates and timing on the whilehastening nematode development. Proper timing sugarbeet cyst nematode management in sugarbeet. The andfrequency of applicationsis important.No treatment experimentwas laidout in a randomisedblock design reducednematode population levels in this60-day trial. withseven treatments each of Ž vereplications. T emik was appliedat planting on 31 May and Counter was 490Dichloropropene with chloropicrinsoil appliedat planting to individual plots (6 rows 50 ft). £ disinfectionfor nematode control in sweet pepper Sugarbeet seeds were plantedon 3 Marchin rows 22” crops apart.Application of Temikat planting of sugarbeet or 3 monthsafter planting signiŽ cantly increased the beet yield 1 2 2; P. GUIRAO , M.M. GUERRERO , A. LACASA ¤, ascompared to the untreated control. In all treatments, C. ROS 1, M. ONCINA 2, M.C. MARTÍNEZ 2, applicationof Temik @ 33lb/ acreat planting increased M.A. MARTÍNEZ 2, C. BELTRAN 2 and J. TORRES 2 thebeet yield and beet vigour .Percentsugar yield was maximumat the rateof Temik14 lb/acreat plantingor the 1 FECOAM-Consejería deAgricultura,Agua y Medio Ambiente,C/ Caballero13, 30003 Murcia, Spain splitapplication (at planting 20 lb/acre+ postapplication 2 Protección Vegetal,CIDA, Consejería deAgricultura,Agua y 13lb/acre).Temik performed better than Counter in terms MedioAmbiente, c/ Mayors/ n,30.150 La Alberca, Spain ofbeetyield, percent sugar and beet vigour . ¤[email protected] Inpepper glasshouses of the Southeast of Spain, 492Evaluation of nematicidesfor the management Meloidogyneincognita and Phytophthoracapsici are the of Meloidogynechitwoodi onpotatoin Idaho mainsoil pathogens. Methyl bromide (MB 98:2)hasbeen Saad L. HAFEZ andSundararaj P ALANISAMY usedfor soil-bornepathogens in the last 17 years. The ¤ mixture1,3-dichloroprope ne(60.5%) and chloropicrin Universityof Idaho,P armaResearch and Extension Center , (33.3%)(Telopic EC) appliedwith PE plasticat 50g/m 2 29603U ofILane,P arma,Idaho 83660, USA bydrip irrigation was assayedas an alternative to methyl ¤[email protected]

Vol.4(2), 2002 295 Chemicalcontrol (489-499)

1; 1 2 AŽeldexperiment was conductedat the Parma Research A. LACASA ¤, M.M. GUERRERO , C. ROS , andExtension Center, Parma, Idaho,to study the efŽ cacy P. GUIRAO 3, P. BIELZA 3, J. TORRES 1, T. DE PACO 3, ofMocap and T emikin combination with V apamand J. CONTRERAS 3, R. MOLINA 4 and M. TORNÉ 4 fosthiazateor Vydate for controlof Columbia root-knot nematodein potato. Fosthiazate and Mocap treatments 1 Protección Vegetal,Centro de Investigació n yDesarrollo were surfacebroadcast on 4 May2001 and Telone II Agroalimentario,CAAMA, c/Mayors/ n,30.150La Alberca andV apamwere appliedon 27 March and 2 April, (Murcia),Spain 2 respectively.Potato cv. Russet Burbank seed pieces were FECOAM-Consejería deAgricultura, Agua y Medio plantedon 18Aprilin rows 3ftapart. Five months after Ambiente,C/ Caballero,13, 30003 Murcia, Spain 3 Producción Agraria,ETSIA, UniversidadP olitécnica de planting,the tubers were hand-harvestedfrom 15ft of Cartagena.P aseoAlfonso XIII, s/ n,30.203Cartagena-Murcia, themiddle two rows ofeachplot and weighed. Y ieldof Spain tubersunder different treatments indicated that there is 4 Dow AgroSciencesIberica, S.A., c/ Cerrode Castañ ar ,72B, anincrease in market yield and total yield in different 28034Madrid, Spain combinationsof Temikand Mocapas comparedto control ¤Alfredo.Lacasa@carm,es plot.Percent of nematodeinfected tubers in treated plots rangedfrom 3.9to 13.4. Lowest level of nematode TheefŽ cacy of 1, 3 dichloropropene(60.5%) plus infectionwas observedin the treatmentof 37.5gal V apam chloropicrin(33.3%) (T elopicEC) appliedby drip (3tier shanks) + 2galMocap. irrigationhas been evaluated in commercial pepper glasshousesin the southeast of Spain as an alternative 2 493Comparative efŽ cacy of Fosthiazate tomethyl bromide (BM 98:2 at 60 g/ m / Meloidogyne formulationsfor the managementof Meloidogyne incognitacontrol.The followingtreatments were assayed: chitwoodi 500,400 and 300 l/ hawith polyethylene plastic (PE); 300and 200 l/ hawithVIF plastic(Virtually Impermeable

Saad L. HAFEZ¤ andSundararaj P ALANISAMY Film).Plastic set transversally and perpendicularly to the driplines were assayedat 500 and 400 l/ harates. All Universityof Idaho,P armaResearch and Extension Center , treatmentswere comparedto MB98:2at 60 g/m 2 applied 29603U ofILane,P arma,Idaho 83660, USA withPE anda non-treatedcontrol. The best results were ¤[email protected] obtainedwith 400 and 500 l/ haapplied with PE. No TwoŽ eldexperiments were conductedto study the signiŽcant differences were foundin the percentage of efŽcacy of Fosthiazateformulations along with V apamon infestedplant, M.incognita root-knotaverage index, plant thecontrol of Columbiaroot knot nematode( Meloidogyne heightand commercial yield. The use of VIF enhanced chitwoodi)inapotatoŽ eld.Fosthiazate was appliedat the disinfectionefŽ cacy and allowed rate reductions. No rateof 8.6or 11.5pt/ acrein the autumn of 1999or spring signiŽcant differences were foundbetween transversal of2000alone or alongwith V apam(37.5 gal). Potato cv. andlongitudinal settings. Longitudinal plastic setting RussetBurbank was plantedon 21March 2000. Inthe Žrst makeseasier commercial Telopic EC applicationand experiment,maximum marketable yield was recordedin reducescost. theplots where V apamwas appliedin autumn 1999 and fosthiazatein thespring 2000. In the second experiment, 495Effect ofnematicideson Pratylenchuszeae maximummarketable and total yield was observedin Graham1951 in sugarcane in Goias, Brazil theplot treated with V apamand Fosthiazate in spring 1; 1 2000before planting. Among all treatments, Fosthiazate V.O.F. MACHADO ¤, P.M. FERNANDES , 2 2 withV apamincreased the marketableyield and total yield R.A.B. SOARES , P.F.M. OLIVEIRA 1 withthe lowest nematode infection. Percent of nematode and M.R. ROCHA infectionranged from 0-1.9and 0-17.4 in the Ž rst and 1 secondexperiments, respectively. Escolade Agronomia/UFG, CampusSamambaia, Caixa Postal131, CEP 74001-970,Goiâ nia-GO, Brazil 2 JallesMachado S/ A,FazendaSã o Pedros/ n,Rodovia 494EfŽ cacy of dichloropropenewith chloropicrinto GO-080,km 71,5,Zona Rural, Caixa P ostal04, CEP control Meloidogyneincognita onpepper crops:rate 76380-000,Goiané sia-GO, Brazil ofapplication and effect ofplasticsetting ¤[email protected]

296 Nematology Postersessions

Nematicideswere appliedin theŽ eldspertaining to Jalles were increasedan average of 227kg/ hainallMessenger MachadoS/ A,municipalityof Goiané sia-GO, infested andaldicarb combinations compared to aldicarb alone. with Pratylenchuszeae Graham,1951. The experiment However,in Alabama, Messenger did not increase cotton was conductedduring the period 03/ 1999to 09/ 2000, yieldsover aldicarb. In Test 2 inMississippiand Alabama, utilisingthe randomised complete block design with four theapplications of Messenger aldicarb(0.59 kg a.i./ha) C repetitions.The treatments utilised were asfollows: i) averagedan increased seed cotton yield of 69.9 kg/ ha Check; ii)Terbufos50G 60 kg/ ha; iii)Terbufos150G overaldicarb (0.59 kg/ ha)alone. Messenger did not 20 kg/ha; iv)Aldicarb150G 10 kg/ ha; v)Carbofuran increasecotton yields in either location when applied in 50G60 kg/ ha; vi)Thiamethoxam250WG 20 kg/ ha.The combinationwith aldicarb at 1.2 kg a.i./ ha.Messenger parametersof evaluation were thepopulation density of incombinationwith thiamethoxam treated seed increased nematodes(specimen/ 10g ofroots and specimen/ 100g yieldin Alabama but not in Mississippi. ofsoil) determined at 3, 6 and9 monthsafter planting andyield data of the variety RB72454, determined 497Abamectin effect onroot-knotnematode after18 months. The nematicides showed efŽ ciency in populationin melonhybrids reducingpopulation level of plant nematode up to 6 monthsafter their application and differed from thecheck W.A. MOREIRA¤ and F.R. BARBOSA andthiamethoxam treatment. The grain yield increase was signiŽcant as a resultof plant nematode population EmbrapaSemi-Arido, C.P .23,CEP 56.300-970,P etrolina,PE, Brazil reduction.The highest yield gains of 8.72 and 8.58 t/ ha [email protected] were obtainedfor Terbufos150G and Carbofuran 50G ¤ treatments,respectively. TheefŽ ciency of abamectin on root-knot nematode populationswas evaluatedin three melon hybrids, Rochedo,Gold Mine and AF 646.The experiment was 496Evaluation of Messengerin combinationwith carriedout in glasshouseconditions with two completely aldicarbor thiamethoxamfor management of randomisedfactors and two independent treatments, with Rotylenchulusreniformis on cotton fourreplicates, each replicate comprising a plasticpot 1; 2 withtwo plants. Three levels of abamectin,equivalent to K.S. MCLEAN ¤ and G.W. LAWRENCE 27,36 and45 lofa.i./ha,were used.Naturally nematode 1Departmentof Entomology and Plant P athology,209 Life infestedsoil, with initial populations determined as 30 Science,Auburn University, Auburn, AL 36849,USA secondstage juveniles (J2)/ cm 3,was used.V ariables 2 Departmentof Entomology and Plant P athology,103 Clay root-knotindex (0-4) andnematode reproductive factor Lyle.Mississippi State University, Mississippi State, MS 39762, (Rf Pf=P i/,inthe soil were evaluated.Root-knot USA D [email protected] indexwas signiŽcantly lower in chemicaltreatments, with ¤ highestreductions in population development of J2being Messenger,a hairpinprotein, was examinedin Alabama obtainedwith 45 l/ ha.The highest nematode population andMississippi for itseffect on R.reniformis population incidencewas observedin Gold Mine hybrid. developmentand cotton yields. Tests were established inlocations which were incotton production and had 498Controlling parasitic nematodes in ahistoryof reniform nematode infestation. Messenger anestablishedvineyard in Cyprus was appliedwith a CO 2 chargedbackpack Ž eldplot spraysystem as a broadcastspray using a 6ftboom John PHILIS with8002E  atfan tip nozzles calibrated to deliver 95 l/ha.In T est1, Messenger was appliedat the two leaf SpyrosStavrinides Chemicals Ltd, P .O.Box21278, Nicosia stage(2LF), pinhead square (PHS), fullbloom (FB), 1505,Cyprus [email protected] and/or3weeksafter full bloom (FB 3)incombination C withaldicarb (0.85 kg a.i./ ha).In T est2, Messengerwas Xiphinemaindex, Mesocriconema xenoplax and Paraty- appliedat 2LF ,PHS, FB,and/ orFB 3incombination lenchushamatus areall pathogenic to grapevine ( Vitis withthiamethoxam (300 g a.i./100kg Cseed)treated seed vinifera L.).A trialin anestablishedvineyard (cv. Sultan- oraldicarb (0.59 or 1.2 kg a.i./ ha)applied in-furrow at ina)during 1999-2001 has shownthat theorganophospho- planting.In T est1, in Mississippi, seed cotton yields ruscontact nematicide cadusafos (as Rugby ®) can effec-

Vol.4(2), 2002 297 Chemicalcontrol (489-499) tivelycontrol these nematodes whereas yields were also 1ESAB increased.It was appliedas granules and in liquid for- 2 UPC, Comted’ Urgell,187. 08036 Barcelona, Spain mulation(microcapsules) in spring 1999 and 2000, at the 3 IRTA, crta.Cabrils s/ n,08348Cabrils, Barcelona, Spain rateof 3.5g a.i.per sq. m. ofactuallytreated area. Gran- ¤[email protected] uleswere appliedin 8 cmwide bands, near and at both Theeconomic impact of resistant tomato was evaluated sidesof the plants (incorporated) while microcapsules ina plastichouse infested with the nematode for three were appliedthrough irrigation in small basins around consecutivegrowing seasons starting in 1999. Treatments ® eachplant. Carbofuran (as Furadan )atthe rate of 5.5g includedsusceptible (cv. Durinta) and resistant (cv. a.i.per sq. m. was alsoapplied using the same method. Monika)tomato cultivated in methyl bromide fumigated Therewere fourtreatments, including the control, repli- soilor insoilinfested with Meloidogynejavanica . Methyl catedthree times. The soiltype was clayloam. T otalnum- bromidewas appliedat a rateof 75 g/ m 2 in October berof plants was 144.Nematode control performance 1998at acostof 2.44euros/ m 2.Tomatoeswere cultivated ofcadusafos, on an overall mean of 3 years,was high, from Marchto July, and there were 2.9plants/ m 2. whereasyields were signiŽcantly increased between 20.7- Eachtreatment was replicatedfour times. Nematode 21.9%.The proŽ t:treatment cost ratio for thisnematicide densitieswere determinedat the beginning and end reachedfour while no chemical residues were foundin ofeach crop. Yield was assessedin eight plants/ plot thefruit. Carbofuran was lesseffective than cadusafos in weeklyfor 6weeks.Initial densities were 480and 660 controllingthe nematodes or increasingyields, under the secondstage juveniles (J2)/ 250cm 3 soil,and changed to conditionsof thistrial. 10356and 190 J2/ 250cm 3 soilafter growing susceptible andresistant tomato, respectively, for threeconsecutive 499Economic impact of resistant tomato cultivars seasons.The average net proŽ t ofcropping resistant asanalternativeto methyl bromide to control tomatoin infested plots was 30000euros/ haper season Meloidogynejavanica withrespect to the susceptible cultivar in nematode infestedplots, and 10600euros/ haperseason with respect 1; FrancescoXavier S ORRIBAS ¤, tothe resistant cultivar in fumigated plots. The average César ORNAT 2, netloss by cropping susceptible tomato in infested plots Soledad VERDEJO-LUCAS 3 and Magda GALEANO 3 was 21,200euros/ hawith respect to those cropped in fumigatedplots.

298 Nematology Postersessions

500The effect ofsomelegumes on stimulating resultedin a signiŽcant improvement of garlic yields. egg-hatchto reduce densities of soybeancyst Soilsolarisation for 8weeksresulted in the reduction to nematode undetectablelevels of the nematode in the upper 20 cm layerof soil, even in heavily infested soils. Rates of Satoshi AIBA diseaseprogress were greatlyreduced in solarised plots, NationalAgricultural Research Center Nematology Laboratory, withyield increments of 50-140% over the unsolarised 3-1-1Kannondai, Tsukuba, Ibaraki, 305-8666, Japan controlplots. Soil solarisation was alsohighly effective [email protected] ina secondconsecutive crop of garlic, with signiŽ cant improvementsin yield and garlic quality. Hatchingof soybeancyst nematode, Heteroderaglycines , isstimulated by legumeroot-exudates. Non-host legumes alsosecrete these substances and can be used to 502Hot waterdrench treatments forthe controlof Radopholussimilis in Rhapis andFishtail palms decreasesoil egg-densities. Root exudates of soybean, azukibean, kidney bean, peas, two red clovers, white Albert ARCINAS1; , Brent SIPES 2, Arnold HARA 3 clover,alfalfa, sun hemp, Chinese milk vetch, sweet ¤ and Marcel TSANG 4 pea,lupine and mimosa were testedfor stimulating egg-hatch.Kidney bean was themost effective legume 1Universityof Hawaiiat Manoa, Dept of Plant and andstimulated hatching of 66% of eggs assayed. The EnvironmentalProtection Sciences, 3190 Maile W ay,St John nematodereproduced on soybean, azuki bean, kidney Lab313, Honolulu, HI 96826,USA 2 beanand Chinese milk vetch. The non-host sun hemp CTAHR, Deptof Plant and Environmental Protection stimulated58.8% egg-hatch and was themost effective in Sciences,3190 Maile W ay,Honolulu, HI 96826,USA 3 BeaumontAgricultural Research Center ,461W .Lanikaula reducingsoil egg-densities. Alfalfa, mimosa and pea did Street,Hilo, HI 96720,USA notstimulate hatching. In Ž eldplots the cultivation of sun 4 CANFRM, Universityof Hawaii,Hilo, 200 W .KawiliStreet, hemp (Crotalariajuncea and C.spectabilis )orredclover Hilo,HI 96720,USA decreasedsoil egg-densities by 57-64% compared to ¤[email protected] fallowcontrol plots. Parasitic fungi attacked 14.9-18.1% Exportersof potted nursery stock face strict quarantine ofeggs present in susceptible or resistant soybean Ž eld regulationsagainst Radopholussimilis ,theburrowing plotsand 30.6-43.6% in egg-hatch stimulating legume nematode(BN). Currentlyno quarantine treatments to Ž eld plots. disinfestplants of BN areapproved. Therefore, hot waterdrench treatments were investigatedfor possible 501Comparison of physicaland chemical methods quarantineutility. Rhapisexcelsa and Caryotamitis palms ofcontrolling Ditylenchusdipsaci ingarlic crops were inoculatedwith 5000 mixed life stages of BN 14 weeksprior to treatment. Hot water drench treatments 1; 2 M. Fe ANDRES ¤ and Javier CABRERA were appliedat 50 ±Cfor 0-16min. In Rhapis palms, a 1DptoProtecció n Vegetal,Centro de Ciencias moderatelygood host,a 16minhot waterdrench achieved Medioambientales,CSIC, Serrano 115, Madrid 28006, Spain 99.6%mortality of BN. InFishtail palms, a poorhost, 2 DptoProducció n Vegetal,E.U.I.T .A.Universidadde alltreatments longer than 10 minat 50 ±Cachieved100% Castilla-laMancha Ronda de Calatrava s/ n,13004Ciudad nematodemortality. Probit regression estimates were used Real, Spain toestimateL T 99,resultingin 16.9 and10.3min for Rhapis ¤[email protected] andFishtail, respectively. However  2 goodness-of-Žt testsfor deviationfrom observeddata was signiŽcant Theeffectiveness of soil solarisation and oxamyl ( 2 21:136, df 3, P < :0001/ for Rhapis. The high applicationsagainst Ditylenchusdipsaci ongarlic crops D D efŽcacy of hot water drenches for thecontrol of BN is were evaluatedunder Ž eldconditions in Central Spain. approachingthe Probit 9 standardof 99.9968%mortality Oxamyltreatments were: dipping garlic cloves in water requiredfor UnitedStates Department of Agriculture solutionof Vydate at 3 ml/kgand at 1.5 ml/ kgdoses, approvalas aquarantinetreatment. andVydate added in granule formulation (0.8 g/ m) to thefurrows atplanting time. All treatments achieved a signiŽcant reduction in the rate of disease progress and 503Forage pearl millet for the controlof theŽ nalincidence of plant death by D. dipsaci, which Pratylenchuspenetrans andpotato early dying disease

Vol.4(2), 2002 299 Integratedmanagement (500-534)

Guy BÉLAIR¤, Yvon FOURNIER, Nathalie DAUPHINAIS (‘plátanos’ ) andhorticulturalcrops under cover. The main and O.P. DANGI nematologicalproblems in order of theireconomical im- portanceare: root-knot ( Meloidogyneincognita and M. ja- Agricultureand Agri-F oodCanada, 430 Gouin Blvd., vanica), cyst (Globoderapallida and G.rostochiensis ), St-Jean-sur-Richelieu(Quebec), J3B 3E6, Canada virusvector ( Longidorus,P aratrichodorus,T richodorus ¤[email protected] and Xiphinema),endoparasitic( Pratylenchusgoodeyi ), Root-lesionnematode Pratylenchuspenetrans causes citrus (Tylenchulussemipenetrans )andectoparasitic ne- yieldlosses in potato Ž eldsin Quebec. In some areas, matodes(Criconematidae, Hoplolaimidae and T ylenchi- ithas been found associated with Verticilliumdahliae dae).It is also indicated that the environmental charac- andthe potato early dying disease (PED). UnderŽ eld teristicsof theCanary Islands climate and soil conditions conditions,we evaluatedforage pearl millet (FPM) as permitthe design of integratedand ecological production arotationcrop for controllingnematode populations, systemsbased fundamentally on theuse of non-chemical reducingPED symptomsand potato yield losses. In 1999, alternativessuch as crop rotation and fallow ,soilamend- a1-yearcrop rotation experiment was establishedon three ments,biofumigation, plantation date and other cultural commercialŽ eldsand where potato cv. Superior was practices,sanitation, plants grown on natural substrates, grownthe following year .FPMsigniŽ cantly reduced by theuse of resistant varieties, grafting, biological control 89%the number of P.penetrans inthe soil and increased agentsand solarisation. These alternatives prevent pest by11% the subsequent potato yield when compared to anddiseases from becomingserious problems and obtain oats.In 2000, again on a commercialpotato Ž eld,we highquality agricultural products. assessedthe usefulness of combiningFPM rotation with preplantsoil fumigation (fall in-furrow application of 505Non-chemical alternatives for the controlof methamsodium 148 l/ ha).The three following treatments Meloidogyneincognita in Uruguay were made:1/ rye fumigation,2/ FPMalone, 3/ FPM C fumigation.FPM alone performed as good as the rye 1; 2 C Leonardo DE LEÓN ¤,J.AntonioL ÓPEZ-PÉREZ fumigationtreatment. FPM fumigationtreatment 2 C C andAntonio B ELLO reducedby 98% P.penetrans soilpopulations, reduced by69% PED symptomsand increased by 31% the 1 DptoAgricultura, Rel-UITA, W .FerreiraAldunate, 1229, C.P . subsequentpotato yields when compared to rye C 11.100,Montevideo, Uruguay fumigation. 2 DptoAgroecologí a, CCMA, CSIC,Serrano 115 dpdo, Madrid, Spain [email protected] 504Integrated and ecological nematode ¤ managementin Canary Island crops Amongother problems caused by soil-borne pathogens, 1; 1 lossesare greatest from theroot-knot nematode Meloi- Antonio BELLO ¤, María ARIAS , Miguel ESCUER 1,J.AntonioL ÓPEZ-PÉREZ 1, dogyneincognita inUruguay intensive horticulture. Sev- Avelino GARCÍA-ÁLVAREZ 1, Javier LÓPEZ-CEPERO 2, eralnon-chemical alternative control methods have been Ildefonso ACOSTA 1 and Rafael SANZ 1 tested:biofumigation, using various organic amendments andgreen manures, resistant varieties, plant covers, trap 1 DptoAgroecologí a, CCMA, CSI,Serrano 115 dpdo, 28006 cropsand other crop techniques, all within a programme 2 Madrid,Spain; CooperativaNuestra Señ ora de Abona,38589 ofintegrated production. Resistant varieties are used in Arico,Santa Cruz de Tenerife,Spain tomatocrops; however, we musttake into account that re- [email protected] ¤ sistancecan be inefŽ cient when soil temperature is higher TheCanary Islands are characterised by Mediterranean than 27±Candparticularly when virulent nematode po- andsubtropical climatic conditions, its seasonal nature pulationsexist. These alternatives are being applied to withhot summers andmild winters represents an eco- severalcrops and production zones in Uruguay, gener- tonebetween temperate and tropical regions. Integrated allywith similar, and sometimes even better, effective- andecological nematode management are analysed. For nessthan conventional phytosanitary products. Moreover, themost part, Mediterranean agrosystems are represented farmers Žndno difŽ culty in applying these alternatives bypotato, vegetables, citrus and fruit trees and vineyards. sincethey have been developed through a participativere- Coastalenvironments are largely represented by bananas searchprocess.

300 Nematology Postersessions

506Reaction of bacterial-feedingnematodes to pig the2 yearswas comparedto yield, topsoil thickness, manurewith antibioticand manure without antibiotic elevation,and soil nutrient maps. Nematode distribution was mostclosely correlated with topsoil thickness ( i.e., Ewa DMOWSKA¤ andKrassimira I LIEVA-MAKULEC depthto the claypan). Lower H. glycines population Instituteof Ecology,P olishAcademy of Sciences,Dziekanow densityoccurred in areaswith shallow topsoil thickness. Lesny,05-092 Lomianki, P oland ¤[email protected] 508Computer modelingof integratedcontrol of potatocyst nematodes Thestudy was carriedout in three experimental plots ingrassland (10 m 2 each)treated in three different Martin J. ELLIOTT1; , James W. MCNICOL 2, ways: i)100l ofwater, ii)100l ofmanure from ¤ David L. TRUDGILL 1 and Mark S. PHILLIPS 1 pigsfed on forage with antibiotic ZnBacitracin, iii) 100l ofmanure from pigsfed on forage without 1 ScottishCrop Research Institute, Dundee, DD2 5DA, antibiotic.Nematode communities were studied6 and12 Scotland,UK weeks,and 10 months after the treatment. In our study, 2 Bioinformaticsand Statistics Scotland, Scottish Crop bacterial-feedingnematodes belonged to Ž veorders: ResearchInstitute, Dundee, DD2 5DA,Scotland, UK Areolaimida,Dorylaimida, Monhysterida, Rhabditida and ¤[email protected] Teratocephalida.In every treatment, on all sampling occasionsnematodes belonging to the order Rhabditida Acomputerprogram modelling the integrated manage- visiblydominated. The following parameters were used mentof PCN hasbeen developed based on equationscali- for evaluationof the effect of two kinds of manure bratedand validatedwith Ž eldtrial data. Population trends onbacteriovores: i)totalabundance of bacterial-feeding anddamage are presented for uptosixrotations, and the nematodes, ii)Diversityindex (H 0gen), iii) Maturity controlmeasures can be varied for eachpotato crop. Cur- index (MI), iv)numberof genera in order Rhabditida. rently,it isan effectiveeducational tool for demonstrating Abundanceand dominanceof particulargenera belonging theeffects of soiltype and of varyingrotation length, ne- toRhabditida in three plots were alsoanalysed. Obtained maticideeffectiveness and the resistance and/ ortolerance resultsshowed that the values of analysedparameters as ofthecultivar being grown. T odevelopits predictive po- wellas the response of particular genera of Rhabditida tential,the relationship between PCN populationdensity, differedin three plots: untreated, and treated with two damageand multiplication rates are being calibrated for kindsof manure,from pigsfed on foragewith and without speciŽc Želds.Nematicide effectiveness and rates of PCN antibiotic. populationdecline between potato crops are also being determined.Slider controls are used to vary the parameter estimatesto producea leastsquares ‘ bestŽ t’line and the 507 Heteroderaglycines distributionwithin aŽeldin newparameters are inserted into the main management no-tillproduction over time program.The advantage of thisapproach is thatrelatively crude/variabledata, based on asmallnumber of samples, PatriciaA. D ONALD¤,KennethA. S UDDUTH andNewell R. K ITCHEN canbe used to calibrate the models and avoid ’ conver- gence’problems. USDA ARS 605Airways Blvd, Jackson, TN 38301and USDA ARS 269Ag Engineering Bldg, University of Missouri, Columbia,MO 65211,USA 509The interaction between anarbuscular ¤[email protected] mycorrhizalfungus andmigratory endoparasitic nematodesin Musa genotypesdiffering in host plant Heteroderaglycines (SCN) isa majorpest of soybean response causingsigniŽ cant yield losses when present and when managementtechniques to reduceegg population density Annemie ELSEN¤, Rony SWENNEN and Dirk DE arenot used. A studyinvestigated the distribution of SCN WAELE atplanting and harvest in a claypansoil Ž eldin central Missouriunder no-till soybean production. Comparisons Laboratoryof TropicalCrop Improvement, K.U.Leuven, were madeof thenematodedistribution in 1999and 2001, KasteelparkArenberg 13, 3001, Leuven, Belgium yearsof soybean production. Nematode distribution in ¤[email protected]

Vol.4(2), 2002 301 Integratedmanagement (500-534)

Arbuscularmycorrhizal fungi (AMF) areobligate sym- beanand cowpea genotypes supplied additional, valuable biontsof plantsthat biotrophically colonise the root cor- informationon theapplication of increasedresistance for texand develop an extramatrical mycelium which helps theprevention of root-knotnematode population build-up theplant acquire water and mineral nutrients from the incropping systems. Finally, use of a covercrop such soil.AMF alsomay protect plants against soil-borne asV etivergrass can contribute signiŽ cantly to root-knot pathogens,including nematodes. T wo Musa genotypes, nematodecontrol, particularly where livestock forms an i.e.,GrandeNaine as susceptible genotype and Pisang integralpart of afarmingsystem. Integration of someor JariBuaya as resistant genotype, were selectedfor their allof these control strategies that are currently available knownhost plant response to twonematode species. The canalready contribute signiŽ cantly to household food migratoryendoparasites, Radopholussimilis and Praty- securityof developing farmers withminimal additional lenchuscoffeae ,areimportant pests in Musa. The AMF inputcosts and effort. canprotect both Grande Naine and Pisang Jari Buaya againstthese nematodes, since nematode reproduction 511Grapevine management, virus vector nematode was signiŽcantly suppressed by the AMF .Inthe case of populationsand GFL V:anapproachto disease P. coffeae theAMF was ableto reduce the damage in the epidemiology roots,caused by thenematodes. For R. similis, no reduc- tionof damage was observed.It appears that the nema- 1; 2 2 Jesús FRESNO ¤, María ARIAS andAntonio B ELLO todesreduced the frequency of mycorrhizationwithout re- ducingthe intensity of themycorrhizal association. Myc- 1INIA-SGIT,DptoBiotecnologí a, Ctrade la Coruña Km 7.5, orrhizationresulted in a signiŽcant better plant growth, 28040Madrid, Spain evenin thepresence of R. similis and P. coffeae. 2 DptoAgroecologí a, CCMA, CSIC,Serrano 115 dpdo 28006 Madrid,Spain [email protected] 510The challenge towards integrated control of ¤ root-knotnematodes for developing agriculture in Factorsaffecting GFL Vtransmissionin vineyards from South Africa theCanary Islands have been studied for morethan 10 years.The special agricultural characteristic of grape Hendrika FOURIE¤,MichaelJacobus S CHOEMAN, MbaliyethuBenedicta M TSHALI, cropsin Lanzarote, La Palma, El Hierro andsouthern CarolineMakgabo L ESWIFI, Tenerife,which use traditional methods of grapevine HendrikFrederik R IEKERT selection,have prevented the dispersal of the virus by andAlexander Henrique M CDONALD nematodes.Moreover, no problemsoccur with nematodes inthosevineyards growing on volcanicsubstrates except ARC-GrainCrops Institute, Private Bag X1251, P otchefstroom, inGran Canaria,because there vineyards are localised 2520,South Africa inthe Laurisilva where high humidity exits. However, [email protected] ¤ newcrop techniques could contribute to an increase in Root-knotnematodes are omnipresent in most resource- infection,especially irrigation and non-controlled plants poorareas of South Africa. Asurveyconducted in introducedin new plantations. Xiphinemaindex and Žveprovinces conŽ rmed the extent of this problem, X. italiae populationsare increasing. Moisture in the whiledemonstrations of signiŽ cant crop yield losses soiland the steady year-round temperature on the island alsoaccentuated an urgent need for developmentand increasethe viral infection. Besides, Ž lmwater isa applicationof integrated,effective nematode management goodvehicle for nematodeand viral dispersal, especially strategiesto enable sustainable food production. On- onslope farms withirrigation. Both vectors and GFL V farm trials,where alternative, low-cost nematode control arewidespread in the whole archipelago, disseminated strategieswere evaluated,revealed treatments with a fair mainlyin new plantations through infected rootstocks, as potentialfor implementationby resource-poor farmers aconsequenceof theintroduction of non-certiŽed plants. toreduce root-knot nematode numbers signiŽ cantly. A Thegreatest focal points for bothpathogens appeared in signiŽcant reduction of root-knotnematodes by meansof irrigatedvineyards or areaswith the highest soil moisture, croprotation, using crops popular to small-scalefarmers wherethe viral infection is maintained over the years. ina particularcropping sequence was establishedin a multi-seasonalmicroplot trial. Resistance screening of dry

302 Nematology Postersessions

512 Meloidogyneincognita controlby biofumigation replicatedtrials at several sites in eastern Australia. In the plus solarisationon glasshousepepper cropsin short-term(2-3 years), tillage and residue management the southeast ofSpain affectedplant-parasitic nematodes ( Pratylenchus , Tylen- chorhynchus , Paratrichodorus , Paratylenchus ).Microbial 1 1; 2 M.M. GUERRERO , A. LACASA ¤, C. ROS , feeding,substrate ingesting and certain predatory ne- 2 3 P. GUIRAO , A. BELLO, J.A. LÓPEZ-PEREZ , matodeswere moreabundant and diverse in less cul- M.A. MARTÍNEZ 1, M.C. MARTÍNEZ 1, N. BARCELÓ 1 tivatedsoils, while fungivorous nematodes were more and J. TORRES 1 abundantand diverse in cultivated soils. Omnivorous 1 Protección Vegetal,Centro de Investigación yDesarrollo nematodesshowed few differenceswith cultivation but Agroalimentario,CAAMA, c/Mayors/ n,30.150 La Alberca – were lessabundant where more organic residues were re- Murcia,Spain tained.There were moremicrobivores and they were con- 2 FECOAM –Consejería deAgricultura,Agua y Medio centratedcloser to the surface where residues were re- Ambiente,C/ Caballero,13, 30.002 Murcia, Spain tained;fungivores, by contrast, were deeperin the soil. 3 Dptode Agroecología, Centrode Ciencias Medioambientales, Therewere nodifferences in the Maturity Index. Over CSIC,C/ Serrano115 apdo 28.006 Madrid, Spain longer-terms(5-10 years), rotation from wheatto pas- ¤[email protected] turedecreased the abundance of mostplant-parasitic ne- Meloidogyneincognita isone of the main soil borne matodes (Pratylenchus , Tylenchorhynchus ,Criconemati- pathogensin pepper glasshouses of thesoutheast of Spain. dae,T ylenchidae),but not Ditylenchus or Dorylaimoides , Inthis area pepper may be considered a monoculture whichincreased. Helicotylenchus populationswere un- grownover more than 90% of the 1800 ha total changed.Fungivores, predators, omnivores and substrate cropsurface. Timing of the biofumigation-sol arisation ingesterswere moreabundant in continuous cropping; mi- applicationas anematodecontrol method was assayedin crobivoresand algal feeders more abundant after rotation. commercialand experimental greenhouses. EfŽ cacy was Short-termmanagement practices such as cultivationand evaluatedby measuringthe percentage of infectedplants, residueretention affect soil nematodes, and by implication averageroot-knot index and marketable yield in relation theenergy pathways in the soil, in different ways from tomethyl bromide (MB). When a mixtureof fresh sheep longerterm practices such as rotation. manure(FSM) at7kg/m 2 andchicken manure at 3 kg/m 2 was usedas an organicamendment applied from theend 514Effect ofhot-water treatment onnematode ofAugustto the beginningof September,marketable yield speciescomposition and densities in plantain roots in 2 2 (9.4 kg/m inAugust and 12.0 kg/ m inSeptember) was shortand long fallow (Central Province,Cameroun) similarto that of MB(9.9kg/ m 2 inAugust and 12.2 kg/ m 2 inSeptember). However, the percentage of M.incognita Kim JACOBSEN1,§,* Stefan HAUSER 1 and infectedplant was lowerin biofumigation treatments. Dirk De WAELE 2 Whenapplications were initiatedat the beginning of 1 IITA HFC, BP.2008,Y aoundé(Messa), Cameroun Octoberand November,biofumigation treatments differed §presentaddress: INIBAP/ CARBAP,BP.12438,Douala, signiŽcantly from MBtreatmentsin all the parameters Cameroun measured. 2Laboratoryof Tropical Crop Improvement, Catholic Universityof Leuven(K.U.Leuven), Kasteelpark Arenberg 13, 513Changes in nematodefaunas under different 3001Leuven, Belgium short- andlong-term management regimes ¤[email protected] Rootnematodes were identiŽed as a majorcause of Mike HODDA1; and Emma BROOS 2 ¤ yieldloss in plantains. The objective of this study was 1CSIRO Entomology,GPO Box1700, Canberra ACT 2601, todetermine the effects of plantain variety, fertiliser Australia application,hot-water treatment (20 min at 52 ±C) of 2 FarmingSystems Research Centre, University of Western suckersbefore planting and fallow length, on the root Sydney-Hawkesbury,Richmond NSW 2753,Australia damageparameters and nematode numbers in plantain ¤[email protected] roots.The experimentwas setup on-farm as arandomised Differenttrophic groups of soilnematodes were affected completeblock design with 64 plots, each of 25 plants. bybothshort-term and long-term agronomic practices in Rootsof Žveplants per plot were sampledat 15 months

Vol.4(2), 2002 303 Integratedmanagement (500-534) afterplanting. Preliminary results indicate that variety relyon an interaction between sludge toxicity, and the andfertiliser application had no signiŽ cant effect on changesin temperatureand pressure. nematodenumbers or rootdamage parameters. Hot-water treatmentsigniŽ cantly reduced total nematode numbers 516Nematode management in subsistence farming inboth fallow types, although fallow length had no in Mexico signiŽcant effect on nematode numbers. SigniŽ cantly higherroot necrosis indices (RNI) were observedin plants Rosa H. MANZANILLA-LÓPEZ inlong fallow plots compared to plants in the short fallowplots. However, the total number of functionalroots 16ColeswoodRoad, Harpenden, Herts, AL5 1EQ,UK [email protected] was signiŽcantly more in the long fallow plots, giving anoverall signiŽ cantly better health of the root system, Factorssuch as geography, climate, vegetation, biogeog- asindicated by the non-damaged root index (NDRI raphyand human settlement have contributed in deŽ n- D numberof functional roots (100-RNI)). Hot-water ingMexican rural and socio-economic scenarios. Mexi- £ treatmentdid not signiŽ cantly reduce the RNI intheshort canagriculture has two main approaches: the campesino fallow,althoughit did signiŽ cantly increase the NDRI in andthe agro-industrial, both with contrasting character- bothfallow types. isticsrelated to production and management strategies againstpests and diseases. In campesino and subsistence 515Sanitation efŽ cacy of acombinedthermal Ž lter systems,low external-input strategies that have proved ef- pressand vacuum dryer on potatocyst nematode and fectivein the management of themost common and im- wildoat in the productionof VEAS-Biosolids portantplant-pathogenic species of nematodes in Mexico (i.e., Meloidogyne spp., Nacobbusaberrans , Pratylenchus Christer MAGNUSSON 1, Paul SAGBERG 2 spp. and Punctoderachalcoensis ),includeproduction of and Bonsak HAMMERAAS 1 plantand tree seedlings free ofnematodes,physical con- trolmethods, rotations, organic amendments, antagonis- 1 TheNorwegian Crop Research Institute, Plant Protection ticplants, suppressive soils and maintenance of biodiver- Centre,Department of Entomology and Nematology, sity.Most of these practices are considered to be envi- Høgskoleveien 7, N-1432Aas, Norway 2 VestfjordensAvlø psselskap (VEAS), Bjerkå sholmen 125, ronmentallyfriendly but, although subsistence agriculture N-3470Slemmestad, Norway canproduce enough food to feed families or smallcom- munities,it maybe adverselyaffected by socio-economic Thesoil conditioner, VEAS-Biosolids, is produced factorssuch as availabilityof goodland, adequate water, throughcompression, heating and vacuum drying of migration,pressures on landuse, shortened rotations, crop sewagesludge at theVEAS wastewatertreatment plant in valueand changes in cultural patterns, thus compromising Slemmestad,Norway. The sanitation efŽ cacy for potato sustainability. cystnematode (PCN), Globoderarostochiensis and wild oat, Avena fatua,was evaluatedfor theŽ nalproduction step,the combined thermal Ž lterpress and vacuum dryer . 517Interactions between vinylhouse tomato IPM Germ carriers,each carrying three nylon bags with 20 measuresfor control of Meloidogyneincognita and nematodecysts and one bag with 100 seeds of wildoat, Fusariumoxysporum f. sp. lycopersici were placedin three Ž lterpresses. Germ carriers were Takayuki MIZUKUBO , T. TAKEHARA, K. ITO, S. AIBA eitherexposed to sludge for 4days,or kept dry before ¤ and T. NAKAYAMA thetest. The test pathogens were exposedto pressures of uptoseven bars at 35 oC,followedby heatingto 80 oC and NematologyLaboratory, Department of Entomologyand vacuum at 0:92barduringcontinued heating. Nematode Nematology,National Agricultural Research Center ,National ¡ vitalitywas studiedby recording juvenile hatch, by AgriculturalResearch Organization, Tsukuba 305-8666, vitalitystaining with New BlueR, andby recording Ibaraki,Japan [email protected] nematodedevelopment on susceptiblepotato. V iabilityof ¤ wildoat was examinedby tetrazolium test. There was a Hotwater (HW), Pasteuriapenetrans (PP), Glomus 100%efŽ cacy in the sanitation of PCNandwild oat. For sp.R10 (Arbuscular mycorrhiza, AM) andan avirulent bothorganisms pre-exposure to sludgeseemed to reduce Fusariumoxysporum (AF) were appliedto 2 m 2 vinyl vitalityand viability. The mechanism of sanitation may houseplots where Meloidogyneincognita and Fusarium

304 Nematology Postersessions oxysporumf. sp. lycopersici were present.The experiment soilbiological and chemical properties were alsoseen. used a 23 factorialdesign with three replicates to identify Thenewly planted tea in mechanically uprooted Ž elds interactionsbetween IPM measures.Untreated plots and exhibitedsigniŽ cantly greater root biomass. fumigated(D-D 52% chloropicrin40%) plots were C thecontrols. T omatoseedlings were plantedand grown 519The effect ofcroprotation and tillage systems from Marchto July 2001 (15 weeks). HW completely onsoilnematode trophic structure suppressed Fusarium wiltof tomato. HW andPP-AM suppressedroot galling and juvenile population ( P < Eduardo MONDINO, Eliseo CHAVES 0:01/,withsynergistic interactions between HW andPP- andGuillermo S TUDDER AM (P < 0:05/.Whenapplied together HW PP-AM £ rootgalling was similarto that of thefumigated plots. AF Facultadde Ciencias Agrarias, Universidad Nacional de Mar appliedwith HW was associatedwith increased galling; delPlata, C. Correo276, 7620 Balcarce, Argentina however,this effect diminished when AF ,HW,andPP- [email protected] AMwere appliedtogether .HWandPP-AM yieldedthe Theeffect of the use of crop – pasturerotation and most fruit (P < 0:01/ withoutan interaction between tillagesystems (conventional tillage – CTandno-tillage thetwo measures. When applied together, HW PP-AM –NT) onsoilnematode communities was studiedduring £ and HW AF PP-AM hadyields 28 and39% greater 1yearin along-termcrop rotation and tillagesystem Ž eld £ £ thanthe untreated controls, respectively. The interaction experimenton a TypicArgiudoll at Balcarce, Argentina. betweenHW andPP-AM, aswell as increased galling Fourseasonal samplings were doneand nematodes were associatedwith AF andHW ,willbe discussed. extractedfrom soilusing the centrifugation- otation technique.The trophic structure was dominatedby 518Mechanisation has added beneŽ ts innematode bacteriafeeder and plant feeder nematodes and the managementin tea highestnematode abundance was foundin winter. The highestplant feeder nematode abundance was shownin 1; 2 Keerthi MOHOTTI ¤, Maithri LIYANAGE pasturein autumn ( P < 0:05/,followedby crop – andNissanka N AVARATNE 1 pasturerotation. Tillage system did not signiŽcantly affect theabundance of such trophic group. The fungivores 1TeaResearchInstitute of SriLanka, T alawakele,22100, Sri Lanka abundancewas lowestin crop rotation under NT in 2 GreatW esternEstate, T alawakele,22100, Sri Lanka autumn.The bacteriovores were lowestin the pasture in springand were notaffected by the tillage system. The ¤[email protected] maturityindex (MI) was affectedby treatmentsin spring Replantinghas paramount importance in sustaining (P < 0:05/.Thepastureand crop – pasturerotation under theproductivity of tea. This involves uprooting old CTshowedthe highest MI value.The diversity index teafollowed by rehabilitation of soils using grasses. (H0)was affectedby the treatments in winter and spring Mechanicaluprooting of old tea bushes has recently (P < 0:05/. The H0 was lowestfor crop– pasturerotation beenintroduced to the industry owing to shortage of underCT inwinter. The H 0 was lowestunder pasture in workers,cost factor and timing of replanting operation spring.A highernematode diversity richness was detected etc.,overconventional manual uprooting. The present incomparisonto previousreports from thisregion. exercisecompared the effect of mechanical and manual uprootingon management of populations of the root lesionnematode, Pratylenchusloosi inold tea Ž elds. 520The harmful in uence of associationsof Mechanicaluprooting left signiŽ cantly low amounts of nematodesand mono- and complex virus infections rootfragments and the soils possessed low residual onpotatoplants and the effectivenessof ecological populationsof P. loosi comparedto that in manually safecontrol methods of these pathogens uprootedsoils. The growth of rehabilitation grasses S. NASROLLANEJAD 1, O.O. BELOSHAPKINA 1 and was superiorin mechanically uprooted Ž eld.It resulted N.D. ROMANENKO2,* ina greaterproportion of organic matter addition andsigniŽ cant elevation of densities, activation and 1 K.A. TimiriyazievAgricultural Academy, Moscow establishmentof nematode biocontrol agents, especially 2Instituteof P arasitologyof Russian Academy of Sciences, free-livingnematodes, in the soil. Improvements in Leninskiiprosp. 33, Moscow ,Russia

Vol.4(2), 2002 305 Integratedmanagement (500-534)

¤[email protected] reproduceswell on plant-pathogenicfungi,although both nematodespecies did well on R. solani and A.bisporus. Theharmful in uence of associations of longidorid andtrichodorid nematodes and four viruses (TR V, 522Soil solarisation, nematicide and chicken X,S,M)andassociations from twoto six virus compostfor root-knot nematode control in carrot in andnematode components exercised considerable and Brazil reliablein uence on the dynamics of growth (height andnumber of shoots) and yield of tubers of potato 1; 1 ClaudioM.G. O LIVEIRA ¤,RobertoK. K UBO , plants.The largest synergetic in uence of complex Mario M. INOMOTO 2,Flávia R .A.P ATRÍCIO 1, trichodoridand longidoridnematodes and virusinfections 3 1 Osvaldo CABRAL andBenedito C. B ARROS was foundin cases of four, Ž veand six virus 1 andnematode components. The antivirus action of InstitutoBioló gico, Caixa P ostal70, Campinas, SP , twobiopesticides (planrhiz and trichodermin) and six 13001-970Brazil 2 Universidadede São Paulo,Escola Superior de Agricultura physiologicalactive agents (narciss, immunocytophyt, F- ‘Luizde Queiroz’ , CaixaP ostal09, Piracicaba, SP ,13418-900 760,F-1153, immunal, and nikfan) by spraying above- Brazil groundorgans of potato plants in Ž eldconditions in 3 EMBRAPAMeioAmbiente, Caixa P ostal69, Jaguariú na, SP , comparisonwith untreated control were studied.Different 13820-000Brazil nematicidaland antivirus actions were foundin most ¤[email protected] bioagentsthat exercised positive in uence on yield of AŽeldexperiment was performedin sandy soil, naturally potatotubers and dynamic growth of potatoplants. infestedwith Meloidogyneincognita and M. javanica, inPiracicaba, SP ,Brazil,to assess the effectiveness of 521Population growth rates of two fungal-feeding soilsolarisation, chicken compost (20 t/ ha),carbofuran nematodes, Filenchusmisellus (Tylenchdiae)and 100G(40 kg/ ha),and combinations of these treatments Aphelenchusavenae (Aphelenchidae),on ten fungal for thecontrol of root-knotnematodes in carrotcv. Aline. isolates Soilsolarisation was performedover 69 days during thewarmest monthsof summer of 2000/ 2001,using a Hiroaki OKADA¤ and Ikuo KADOTA 100 ¹mpolyethyleneplastic Ž lmas soil covering. The soiltemperature in solarised plots was 8-12 ±C higher NationalAgricultural Research Center for T ohokuRegion, thanin the nonsolarised. Maximum soil temperatures Arai,Fukushima City, 960-2156, Japan were recordedin solarised soil combined with chicken ¤[email protected] compostand reached 77, 56 and 46 ±C,atdepths of 5, 10and 20 cm. Population densities of M.incognita and Toclarifydifferences in reproduction characteristics be- M. javanica were controlledby both soil solarisation tween Filenchusmisellus (familyT ylenchidae)and Aphe- aloneand combined with chicken compost or carbofuran. lenchusavenae (Aphelenchidae),their population growth Carrotcv. Aline yield was signiŽcantly increased by rateswere measuredafter 40 days incubation at 25 oC combinationof solarisation and chicken compost plus onten isolates of fungi representing nine species. Fi- nematicideapplication before planting. lenchusmisellus growthrates were largeon Agaricusbis- porus,Chaetomium cochlioides, Chaetomium globosum , Chaetomiumfunicola , Coprinuscinereus, Pleurotus os- 523Current managementstrategies employed treatus and Rhizoctoniasolani ,andsmall on Pythiumulti- againstthe reniformnematode ( Rotylenchulus mum, Fusariumoxysporum f. sp.conglutinans and Fusar- reniformis)incottonproduction in Louisiana, USA iumoxysporum f. sp.cucumerinum .Incontrast, A. ave- 1 2 nae growthrates were largeon P. ultimum, R. solani, F. Charles OVERSTREET , Edward C. MCGAWLEY and 3 oxysporum f. sp. conglutinans,F .oxysporum f. sp. cuc- Boyd PADGETT umerinum and A. bisporus,andsmall on C.cochlioides, 1 LSU AgCenterResearch and Extension, Department of Plant C.globosum,C. funicola , C. cinereus and P.ostreatus . Pathologyand Crop Physiology, 302 Life Sciences Building, Theseresults suggest that F. misellus,whichreproduces BatonRouge, LA 70803,USA wellon saprophyticfungi and mushrooms, may occupy an 2 Departmentof PlantP athologyand Crop Physiology, 302 Life ecologicalniche different from thatof A. avenae, which ScienceBuilding, Baton Rouge, LA 70803,USA

306 Nematology Postersessions

3 NortheastResearch Station, 212 Macon Ridge Road, seedlingswere transplantedto 2.5 dm 3 bagsof sterilised Winnsboro,LA 71295,USA sandyloam inoculated with two eggs secondstage C ¤[email protected] juveniles/cm 3 soil.Samples were collectedat 30 and Reniformnematode has become a majorpest of cotton 60days. W efoundstatistically signiŽ cant differences inLouisiana during the past decade. Since 1990, 40% of betweentreatments in all variables studied except for the18 477samples processed by theNematode Advisory fungiin root samples and bacteria in rhizosphere and Servicerevealed detectable levels of reniformnematode. roots.The lowest Ž nalpopulation level and multiplication Nematicidesand crop rotation are the primary methods rateswere recordedfrom thecarbofuran treatment, as well ofmanagement. In-furrow applicationof aldicarbat 0.59 asthehighest values for FRW,FAWandDA W.However, kg/hahasbeena producerstandard since the 1980s.Based thistreatment also caused the least development of fungal on35 Želdtrials, plots receiving an in-furrow application andbacterial colonies in the rhizosphere, rhizoplane and ofaldicarb (0.59 kg/ ha)produced an average 262 kg/ ha roots.The treatments studied showed a lowcapacity for moreseed cotton than non-treated plots. The addition of controlling M.incognita onguavaseedlings. 1,3dichloropropene at 28 l/ hato the producer standard rateof aldicarbincreased yields over the grower standard 525Distribution of Globoderarostochiensis in by326 kg/ hain six Ž eldtrials. Approximately 25-30% the centralregion of Russiaand new controlmethods ofthe cotton crop is rotated to a non-hostor resistant cropsuch as corn, grain sorghum, or soybean.Only Ž ve I.O. POPOV and N.D. ROMANENKO¤ ofthe 69 soybean varieties recommended for Louisiana havebeen reported as resistant against this nematode. Instituteof Parasitology,Russian Academy of Sciences, Populationsof reniformnematode averaged 2682 per 500 Leninskiipr .33,Moscow ,117071,Russia [email protected] cm3 ofsoil in 217 Ž eldsin Rapides Parish after cotton ¤ followedby 1 yearof corn. Populations of reniform Considerableinfection of potatoplantation by Globodera 3 nematodeaveraged 9558 per 500 cm in158 Ž eldsafter rostochiensis was foundin industrialand scientiŽ c Želds cornfollowed by 1yearof cotton. indifferentregions of Russia(Karelia, Moscow, Vladimir andV oronezhregions). Cysts of G.rostochiensis were 524Biological and chemical control of Meloidogyne foundat seven of 20 investigated potato crops. The incognita andits effect onsoilmicro ora and growth inuence of biological agents and bioactive substances of Psidiumguajava seedlingsin ZuliaState, V enezuela (i.e.,biologicalpesticides, planrhiz and trichodermin;new biologicallyactive strains of Bacillus, Pseudomonas and 1; Evelyn PÉREZ-PÉREZ ¤, others;chitin containing substances narciss and agrochyt; Ana Maria CASASSA-PADRÓN 2;3, Merylin MARIN 3, andnew Russian physiologically active substances, Casilda GONZÁLEZ 1, Dubia CHIRINOS 1 and Luis oligophurostanozid,immunocytophyt ,compactinand SANDOVAL 2 lovastatin)on numberand life activity of G.rostochiensis 1 CentroFrutí cola del Zulia-CORPOZULIA, Maracaibo, cystsin soil and on potato plants growth and yield, Venezuela aftertreating tubers and plants with these biologically 2 Universidaddel Zulia, F acultadde Agronomí a, Institutode activeagents, was studied.Nematicidal activity of InvestigacionesAgronó micas threephysiologically active substances (two inhibitors, 3 Departamentode Botánica, Maracaibo, Apto 15205, compactinand lovastatin, and one resistance stimulator, Venezuela immunocytophyt)was studiedin experiments in vitro. ¤[email protected] Inhibitorsof plant growth (compactin and lovastatin) Theeffects of Trichodermaharzianum , Paecilomyces demonstratedstimulating in uence on the release of the lilacinus andcarbofuran on Meloidogyneincognita and infectivejuveniles from G.rostochiensis cysts. fungaland bacterial populations in the rhizosphere, rhizoplane,roots and soil were studiedon Psidium 526Development and transfer of technologyfor guajava seedlings.The effects of the nematode on fresh bio-intensivenematode management in Southern rootweight (FRW), fresh aerialweight (F AW)and India driedaerial weight (DA W)were alsoevaluated at the CentroFrutí cola del Zulia-CORPOZULIA. Sixty-day-old M.S. RAO andP .ParvathaR EDDY

Vol.4(2), 2002 307 Integratedmanagement (500-534)

Divisionof Entomology and Nematology, Indian Institute of SDS were evaluated.Treatments were fallsubsoiling HorticulturalResearch, Bangalore, India (depthof 46 cm), conventiontillage, and no tillage. In eachtreatment, 16 varietieswere plantedin 2000 and 32 Increaseddemand in the domestic and international mar- varietieswere plantedin 2001. The varieties varied with ketfor thehorticultural produce has led to intensive culti- regardsto SCN andSDS resistance.In both years, SCN vationof horticulturalcrops in variousstates of India. This reproductionwas higheron SCN-susceptible varieties. hasresulted in theincrease of root-knotnematodes ( Me- Eggdensities of SCN were higherin plots receiving loidogyne spp.)to analarming extent affecting adversely subsoilingtreatments at one site in 2000 and at both thecrop growth and productivity. No single method of sitesin 2001. Across varieties,subsoiling reduced the controlof nematodes and no single component of man- severityof SDS atbothlocations in 2000and 2001. Root agementof nematodeswas effectivein the management of colonisationby FSG differedamong varieties but was nematodeson anyhorticultural crop on asustainableba- notaffected by the tillage practices. In 2000 and 2001, sis.Biocontrol agents such as Paecilomyceslilacinus , Tri- soybeanyield was higherin plots receiving the subsoiling chodermaharzianum , Verticilliumchlamydosporium and treatment. Pasteuriapenetrans arenot effective individually in the managementof nematodes under Ž eldconditions. Simi- larly,endomycorrhiza ( Glomusmosseae or G.fascicula- 528Crop rotation options for management of tum),botanicals(neem or castor or pongamia cakes) or Pratylenchusneglectus incereal-basedproduction anytrap crop, antagonistic plant or anycultural practice systemsin Western Australia isnot effective individually in the Ž eld.Hence, we in- Shashi SHARMA , Sean KELLY and Robert LOUGHMAN vestigatedthe possibilities of integrationof thesecompo- ¤ nentsfor themanagement of thenematodes on horticul- Departmentof Agriculture,Baron Hay Court, South P erth turalcrops on asustainablebasis. The investigations have WA 6151,Australia resultedin the development of integratednematode man- [email protected] agementstrategies of economicallyimportant nematodes ¤ ofthisregion by integratingbiocontrol agents, botanicals Pratylenchusneglectus isa keynematode parasite andendomycorrhizae under Želdconditions by exploiting ofcereals in W esternAustralia. T oidentifycrop thesynergistic interaction between P.lilacinus neem C rotationoptions for nematodemanagement, cultivars cake G. mosseae; P.lilacinus castor cake G. fas- ofbarley (Stirling), oat (Dalyup), wheat (Nyabing), ciculatumC ; T.harzianum neemCcake G. mosseaeC ; V. C C canola(Dunkeld and Karoo), chickpea (Heera), fababean chlamydosporium neem cake G. mosseae combina- C C (Fiord),Ž eldpea (Dundale) and lupin (Merrit) were sown tions.The data on the adoption of thetechnology, percent in 1999 in 40 1.8m plotsin a randomisedblock increasein the yield and the reduction in the use of chem- designwith six £replications in anematode-infestedŽ eld. icalsis presented. Nematodenumbers in soil before planting and in roots after10 weekswere assessed.Wheat cv. Machete was the 527In uence of tillageon soybeanpathogens in highlysusceptible check. All the plots in 2000 were sown Illinois withwheat cv. Cunderdin, and in 2001 with the same cropas in 1999. Nematode data were log-transformedfor John RUSSIN¤ and Jason BOND analysis.Field pea and faba bean supported signiŽ cantly Plant,Soil and General Agriculture, Southern Illinois fewer nematodesthan other crops. Previous year crops University,Mailcode 4415, Carbondale, IL 62901-4415,USA signiŽcantly in uenced nematode infestation and the fewest were extractedfrom plantsin plots previously ¤[email protected] plantedto faba bean and most in plotspreviously planted Fusariumsolani f. sp. glycines (FSG), thecausal agent of towheat. In 2001, infestations were leastin faba bean soybeansudden death syndrome (SDS), and Heterodera andŽ eldpea and greatest in chickpea, oats and canola cv. glycines arethe two most important soybean pathogens in Karoo.Faba bean and Ž eldpea were identiŽed as suitable Illinois.Sudden death syndrome is generally more severe cropoptions to aid in the rotational management of P. inareas with greater soil compaction and/ orwaterlogged neglectus. conditions.At twolocations, the effects of three tillage practiceson pathogen populations and foliar severity of

308 Nematology Postersessions

529T rapcropping of potatocyst nematodes using Initialnematode density ( Pi)immediatelyafter sowing in resistantSolanaceae potato clones 1999was 5-11 P.neglectus /gdrysoil at Miltaburra and 1-3/gatY andra.In 2000, Pi atMiltaburrawas 8-25/gand Susan J. TURNER¤ andTrevor J.G. M ARTIN atY andra1-3/ g.Final nematode numbers ( Pf ) were de- AppliedPlant Science Division, Department of Agricultureand terminedduring grain development in spring (September- RuralDevelopment for Northern Ireland, Newforge Lane, October).In both years at both sites, Pf decreasedas BelfastBT9 5PX, UK rateof appliedphosphorus increased. In 1999, phospho- ¤[email protected] rusrates of 20-30kg P/hareduced P.neglectus density by Inrecent years the levels of Globoderapallida in potato 30%compared to plots with no addedphosphorus. Sim- landhas been steadily increasing, such that some land is ilarly,in 2000, nematode levels at Miltaburra were 37% nowconsidered unproductive for continuedcommercial lowerand at Y andra32% lower with 20 kgP/hacompared tocontrolplots. The cropping soils of theEyre Peninsula potatoproduction. The concept of using Solanaceae potatoclones as biological control agents for PCNwas areinherently nutrient deŽ cient, and the rainfall low (av- investigated.The criteria for suchclones were thatthey erageannual rainfall 275 mm atMiltaburra and 400 mm havefull resistance to all known PCN pathotypes(both G. atY andra,with 200 mm and320 mm, respectively,falling Pratylenchusneglectus rostochiensis and G. pallida),havethe ability to stimulate duringthe growing season). in this highlevels of PCN hatch,and be tolerant to high levels regionis a signiŽcant impediment to cereal production, ofPCN invasion.A 3-yearŽ eldtrial was undertaken andnutrition can play a rolein the management of these thatevaluated clones that fulŽ lled these criteria. These nematodes. clonesincluded wild potato species from SouthAmerica, advancedbreeders lines and potato cultivars. Overall, 531Nematodes on strawberriesin Southern Spain: thetest clones signiŽ cantly enhanced the annual natural effects onyieldand control of Meloidogynehapla declinerate of ca 19%under Northern Ireland conditions, 1; 1 2 witha rangefrom 26to 54%. Comparison of yield, J.M. VEGA ¤, J.I. PÁEZ , J.M. LÓPEZ-ARANDA , 3 3 1 whennematicides were applied,conŽ rmed that several J.J. MEDINA , L. MIRANDA and F. MONTES clonescould tolerate extremely high levels of PCN (up 1 Laboratoriode Sanidad V egetal,CAP-Junta de Andalucí a, 1 to1000 eggs g ¡ soil).These trials have conŽ rmed that 41089Montequinto (Sevilla), Spain thebasic principle of using such material is a sound 2 CIFAMálaga, CAP-Junta de Andalucía, 29140 Churriana wayof enhancingPCN decline,and may be anattractive (Málaga), Spain alternativefor cleaningup land under the ‘ setaside’ 3 CIFALasT orres-Tomejil,CAP-Junta de Andalucí a, 21800 scheme,or for organicpotato production. Moguer(Huelva), Spain ¤[email protected] 530Effect ofappliedphosphorus on population Theprovince of Huelva,with more than 7500 Has, isthe densityof rootlesion nematode ( Pratylenchus strawberrylargest producer area of Spainand one of the neglectus) largestof Europe. The Laboratorio de Sanidad V egetal ofSeville has been receiving disease strawberry plants 1; 1;2 Vivien A. VANSTONE ¤,MichelleH. R USS , from thisregion during the last Ž fteenyears, and the 3 2 Alison FRISCHKE andSharyn P .T AYLOR parasiticnematode species detected were Meloidogyne 1 Universityof Adelaide, Department of Plant Science, PMB 1, incognita, M. arenaria, M. hapla, Pratylenchuspenetrans , GlenOsmond, 5064, South Australia, Australia Ditylenchusdipsaci and Aphelenchoidesfragariae . Until 2 SouthAustralian Research and Development Institute, GPO nowa systematicsurvey on incidence and severity of Box397, Adelaide, 5001, South Australia, Australia nematodeproblem was notmade in this area, but during 3 MinnipaAgricultural Centre, Box 31, Minnipa, 5654, South afouryears (1998-2001) assays made on two localities Australia,Australia (Cartayaand Moguer) on chemicals and non chemicals ¤[email protected] alternativesto Methyl Bromide, M. hapla was the most Trialsat two sites on the Eyre Peninsula of South Aus- importantphytopatholog icalproblem. Its presencewas traliawere sampledin 1999 and in 2000 to determine ef- explainedby the use of plantlets from highelevation fectof phosphorusapplication on densityof Pratylenchus nurserieslocated on Castile-Leon (Central Spain). The neglectus intrial plots of thesusceptible wheat cv. Frame. incidenceand severity of root knotted plants and the

Vol.4(2), 2002 309 Integratedmanagement (500-534) numberof nematodespresent were negativelycorrelated Departmentof Entomology and Plant P athology,Auburn withsome agronomic traits and effect of M. hapla on University,Auburn, Alabama 36849-5409, USA cropproduction and fruit quality was studied.Although ¤[email protected] Solarization,helped with some chemicals or associated A3-yearŽ eldstudy was initiatedin central Alabama in a toBiofumigation, exerted some nematode population cottonŽ eldinfested with Meloidogyneincognita to assess control,only Chloropicrin alone or with DD proved thelong-term effects of selected cropping sequences on similarefŽ ciency to MethylBromide. nematodepopulations and yields of cotton. In the Ž rst year,six blocks were setup, each containing 24 rows 532Effect ofinorganicphosphate fertiliserson on 400 centers.The six blocks were plantedwith cotton the efŽcacy of an arbuscularmycorrhiza fungus (CT), soybean(SB), pearlmillet (PM), corn+ velvetbean againsta root-knotnematode on pyrethrum (CR+VB)and pearl millet + velvetbean(PM+VB). In the2nd year the Ž rst twoblocks were maintainedwith 1; 1 2 J.W. WACEKE ¤, S.W. WAUDO and R. SIKORA cottonand soybean monoculture. The remaining four blockswere splitwith 12 rows ofcotton and 12 rows 1 BotanyDepartment, Kenyatta University, P .O.Box43844, ofsoybean. In the 3rd year, each block was planted Nairobi,Kenya withthree cotton cultivars (SG125, SG747, DP458) both 2 Instituteof Plant Diseases, University of Bonn,Nussallee 9, non-treatedand treated with 2.0 lb a.i./ Aofaldicarb D-53153,Germany intwo row plots.This setup allowed for thefollowing [email protected] ¤ croppingsequences: (CT -CT-CT), (SB-SB-CT), (PM-SB- Effectsof triplesuper phosphate (TSP) andsingle super CT),(PM-CT -CT), (CR+VB-SB-CT),(CR+VB-CT -CT), phosphate(SSP) fertiliserson efŽ cacy of a Glomus (SG+VB-SB-CT), (SG+VB-CT-CT), (PM+VB-SB-CT), sp.(isolate KS 14)against Meloidogynehapla were and(PM+VB-CT -CT). Treatmentwith alcicarb across all determined.The fertilisers were appliedat the rate of 1 croppingsequences resulted in a 34%reduction in M. and 2 ¹g/gsoilat the time of fungus inoculation. T wo incognita anda 64%increase in yield compared to non- monthslater, plants were inoculatedwith the nematodes. treatedcotton monoculture. Most non-treated rotation Plantgrowth and nematode disease parameters were sequencesdid not show a reductionin M.incognita determined2 monthsafter nematode inoculation. The comparedto non-treated cotton monoculture except for fertilisersat both levels improved plant growth in all (SB-SB-CT) and(SG+VB-SB-CT) whichshowed 30 and treatments.The fungus,in general, improved plant growth 41%reductions, respectively. All non-treated rotation onits own or in the presence of nematodes but not sequencesaveraged a 20%increase in yield compared inthe presence of fertilisers. Both fertilisers at both tonon-treated cotton monoculture. The best performing levelswere moreeffective in improving plant growth treatedrotation sequences were (PM-SB-CT), (PM-CT- thanthe fungus. The fungus suppressed nematode disease CT),(PM+VB-SB-CT) and(PM+VB-CT -CT) resulting severityunlike the fertilisers. The suppressive effects of in26, 21, 26 and 21% increases in yield, respectively, thefungus on thenematodes were, inmost cases, reduced overand above what was obtainedwith treated cotton bythe fertilisers. In addition, the fertilisers signiŽ cantly monoculture. reducedroot colonisation of pyrethrumby thefungus. The nematodes,on theother hand, did not haveany signiŽ cant 534Induced resistanceof plantsto root-knot effectson rootcolonisation by thefungus or on itsability nematode toimprove pyrethrum growth. The presence of nematodes infertiliser or fertiliser-fungus-treated plants, however, S.V. ZINOVIEVA¤, N.I. VASYUKOVA, signiŽcantly reduced pyrethrum growth. N.G. GERASIMOVA, Zh.V. UDALOVA, G.I. CHALENKO and O.L. OZERETSKOVSKAYA

533T oassessthe long-termeffects ofselected Instituteof P arasitologyRAS andBach Institute of Biochemistry croppingsequences to control Meloidogyneincognita RAS, Moscow,Russia,119071 Moscow ,Leninskyprospect, 33 [email protected] andincrease the yieldof cotton ¤ Incurrent investigations the mechanisms of induced C.F. WEAVER¤, R. RODRÍGUEZ-KÁBANA plantresistance to parasitic nematode were studied. and D.G. ROBERTSON Theactivation of protective mechanisms in plants, in-

310 Nematology Postersessions hibitingthe growth of parasitic nematodes, was stud- andstimulated the generation of reactiveoxygen species. iedin the system tomato – Meloidogyneincognita . Themixture of elicitors with signal molecules (salicylic Thebiogenic elicitors (arachidonic acid and water- acidand methyl ether of jasmonic acid) caused more solublechitosan) stimulated the development of in- signiŽcant immunostimulation effect in tomato. The vadedtomato plants. Arachidonic acid and chitosan in- treatmentof tomato seeds by elicitors and mixture of ducedthe accumulation of phytoalexinsin the tissues of elicitorsand signal molecules signiŽ cantly suppressed the hostplants; decreased the total content of free sterols numberof gallsand eggs produced and increasedduration andchanged their composition, producing adverse ef- ofnematodedevelopment. The data obtained suggest that fectson infesters; activated the chitinase, -glucanase, thenatural mechanisms and those induced by biogenic lipoxigenase,peroxidase, phenylalanine ammonia lyase; elicitorsin tomatoto the nematode have the same origin.

Vol.4(2), 2002 311 Animalparasitic nematodes (535-543)

535Nematodes parasites of somefreshwater Žshes characterof vulval apinfemale,length of spicules,and in Zimbabwe distancefrom theirtip to the tip of the barbs in males. Datawere subjectedto statistical analysis. Our Žndings Ben N. DUBE¤ and N.A.G. MOYO revealedthe presence of three species: Haemonchus Universityof Zimbabwe,Dept of BiologicalSciences, P .O.Box longistipes in camel, H.contortus in sheep and H. placei MP167,Mt. Pleasant, Harare, Zimbabwe ingoatsand cattle. Statistical Ž ndingsbased on oneway ¤[email protected] ANOVAandTukey post hoc showedthat the length of Theobjective of this study was tosurvey and docu- spiculesof worms collectedfrom sheepis signiŽ cantly mentthe various nematode parasites that infect some differentwith those of goats and cattle ( P < 0:05/. freshwater Žshspecies of economic importance in Zim- Meanwhile,as far asthe left and right hook distance babwe.V ariousspecies of fresh waterŽ shTilapia ( Ore- from thetip of the spicule is concerned, there was also ochromismacrochir , O.mossambicus , O.niloticus ), Trout asigniŽcant difference between H. placei of goats and (Salvellinusfontanalis) ,GreenHappy ( Sargochromisco- cattlewith that of sheep ( P < 0:05/.Thediscriminant dringtonii)andAfrican catŽ sh ( Clariusgariapinus ) were functionanalysis demonstrated further that the three amongsome Ž shspecimens collected from thelakes groupsof worm couldbe considered as three different (Chivero,Manyame and Kariba). Fish collections were strains. madeduring the 12 monthsof Januaryto December2001. Freshlycollected Ž shspecimens were takento thelabora- 537Nematodes of wildcavicorns in the faunaof toryand all gastrointestinal tracts, liver, vascular system Uzbekistan andŽ shgill contents were removedand examined for the presenceof nematodeparasites. Excised nematode speci- A.E. KUCHBAEV, F.D. AKRAMOVA, D.A. AZIMOV, menswere Žrst preservedand sent to SouthAfrica marine A. PAZILOV and B. RUZIEV laboratoriesfor experttaxonomic examination and iden- tiŽcation. Preliminary results indicate that the most com- Instituteof ZoologyUzbek Academy of Sciences, T ashkent, monnematode parasites associated with the gastrointesti- Republicof Uzbekistan naltracts and liver from almostall Ž shspecies collected are Capillaria spp., Camallanus spp., Contracaecum spp. Helminthsof the class Nematoda are widespread in the and Ascaris spp.The Acanthocephalus group of parasites biogeocenosesof Uzbekistan, parasitising all vertebrate were alsoencountered and identiŽ ed. It was alsonoted speciesinhabiting aquatic and terrestrial ecosystems. thatin some Ž shspecies, nematode parasite counts cor- Wildcavicorns in the fauna of Uzbekistan comprise relatedwith the level of pollutionin the respective water sixspecies, namely, Gazellasubgutturosa Guldenstaedt, bodies. 1780, Saigatatarica Linnaeus1766, Caprafalconeri Wagner,1839, C. ibex Linnaeus,1758, Ovis ammon karelini Severtzov,1873 and O.orientalis Gmelin, 536Comparative study of Haemonchus species of 1774.The above species inhabit deserts, semi-deserts different ruminantsof Iran andare important components of the corresponding biogeocenoses.As aresultof a studyon parasitic A. ESLAMI1; , S. BOKAIE 2 and S. MESHGI 1 ¤ worms ofwild cavicorns, 54 nematodes were recorded, 1 Departmentof P arasitology ofwhich six are Ž rst recordsin Uzbekistan. Diversity 2 Divisionof Epidemiology,T ehranUniversity, V eterinary ofhelminth fauna in wild cavicorns is as follows: O. Faculty,P .O.Box14155-6453, T ehran,Iran ammon, 43; O.orientalis , 41; G.subgutturoza , 35; [email protected] ¤ C. sibirica, 33; C.falconeri , 29; and S. tatarica, 22. Haemonchus speciesis reported from sheep,goats, Undernatural conditions, cavicorn species show, between cattle,camel and wild sheep, as well as man in themselvesand domestic animals, close biocenotic links. Iran.IdentiŽ cation of Haemonchus speciesin domestic Thispromotes circulation of helminths in wild animals. ruminantsmight be very useful in the study of natural Wildanimals play a certainrole in epidemiology of a population,especially when two (or three)species are numberof helminthoses and take part in distribution of sympatric.In the recent investigation, ten males and ten diseasesamong domestic animals and humans. females of Haemonchus spp.collected from sheep,goat, cattleand camel were examinedfor totalworm length,

312 Nematology Postersessions

538Populatiom ecology of the family viableup to next grazing period. According to our Protostrongylidae,Leiper 1926, parasites of animals results,strongyle larvae can be transferred to deep soil layersby earthworms and can remain viable during the A.E. KUCHBAEV, F.D. AKRAMOVA wholewinter period. T oavoidsheep infections, the main and R.R. KARIMOVA prophylacticprinciple is separate grazing of lambsfrom Instituteof Zoology, Uzbek Academy of Sciences, T ashkent, ewes becausethey are the main infection sources in Republicof Uzbekistan spring. Nematodesof thefamily Protostrongylids are adapted to therespiratory system of animals inhabiting mountain 540Histology of the anteriorintestine of Chabertia ecosystems.They are widely represented in ruminants ovina ofthe Palaearctic. Fifteen species of Protostrongylids G. HOV NANYAN and S. MAKARYAN inhabitmountain ecosystems of Uzbekistan, which are ¤ thehabitats of cavicorns. Dominant species of proto- Instituteof Zoology,4 SevakStreet, Y erevan,375014, Armenia strongylidsof cavicornsare representatives of the genera ¤[email protected] Protostrongylus , Cyctocaulus and Muellerius. The eco- Histologicalstudies of Chabetiaovina widespreadin logicalniche for therecorded species of nematodes is sheephave never been conducted in Armenia. Chabertia therespiratory system, which witnesses high adaptive po- ovina were obtainedfrom largeintestines of freshly tentialof the helminths. T errestrialmolluscs of genera slaughteredsheep and Ž xedin Barbagallosolution before Xeropicta,Pseudonapaeus, Bradybaena , Macrochlamys , histologicalpreparation. After azanstaining the cuticle Pupilla, Derocerasetc. ,participatein life cycles of pro- ofthe buccal capsule (BC) andpharyngeal funnel (PF) tostrongylids,in which the 2-3 stage larvae populations turnbright red while other tissues in the anterior end develop.Helminths parasitise animals within the system turnblue. The pharyngeal gland duct is clearly visible parasite-host.A vailabilityof certainhelminth complexes ontransverse sections. Haematoxylin-eosin stains all ismainly predetermined by theavailability of correspond- tissuesinto various shades of red: the lightest are BC inghost groups and microclimatic conditions in thegiven andPF cuticles.Clearly outlined are the structure and biogeocenosiswhich provide the development of all the tissueborders of the pharyngeal musculature and dorsal phasesof theparasitic worm ontogeny. pharyngealgland. After ferroushematoxylin staining thecuticle of the BC andPF turnblack, while other 539Population dynamics of sheep digestivetract tissuesin the apical part take various shades of brown. strongylosesin Estonianclimate conditions Longitudinalsections clearly show the lamellar structure oftheBC cuticle,which is notmentioned in thedrawings E. MÄGI and A. KAARMA ortextualdescription of Chabertiaovina .Thusmethods usedprovided new histological results for Strongylidae EstonianAgricultural University, Kreutzwaldi 62, T artu5104, Estonia andcomplemented each other . [email protected] Theobjective of thisstudy was toinvestigatepopulation 541Changes in prepatent periodsand species dynamicsand viability of strongylelarvae on pasturesin compositionof Oesophagostomum spp. isolatesover naturalconditions. According to our data, strongyloses of time sheepdigestive tract are the most widespread parasitic H. TALVIK1; , A. JOACHIM 2 and A. KARIS 3 diseasesin Estonia. Our observationshave shown that ¤ pasturescontaminated with parasites the previous year 1 EstonianAgricultural University, Kreutzwaldi 62, T artu arethe main sources of invasion.In order to estimatethe 51014,Estonia 2 UniversitätLeipzig, An den Tierkliniken 33, Leipzig, Germany viabilityof strongylelarvae on sheeppastures, Ž vetrials 3 withlambs were carriedout in different pasture regimes. Universityof T artu,V anemuise46, T artu,Estonia [email protected] Alllambs were submittedto faecal examinations for ¤ numbersof parasiteeggs once a week.Our observations FourŽ eldisolates of Oesophagostomum spp. were haveshown that although most strongyle larvae did not collectedfrom Estonianpig farms in1997 and used for surviveduring the winter period, some of themremained inoculationof eightpiglets (2000 L 3 larvaeper pig). Four

Vol.4(2), 2002 313 Animalparasitic nematodes (535-543) newpigs were inoculatedwith the second generation directionof thermokinetic behaviour depended on prior larvaeof one isolate kept in the fridge for 3years. experienceof environmentaltemperature. L3 cultured at Oneextra pig was inoculatedwith 5000 L 3 larvae from 25±Ctendedto move towards higher temperatures from theprimary Ž eldisolate to follow the egg excretion releasingpoints between 22 ±C and 29±C.At releasing ofthe two species by PCR analysis.All four Estonian points of 30±Corabove,most L3 moved little and showed Želdisolates contained Oesophagostomumdentatum and nodirectional response. At 20 ±Corbelow, few orno O.quadrispinulatum .Prepatentperiods of both nodular L3moved towards the zone of higher temperature. L3 worm speciesvaried from 18to 43 days. Fairly constant culturedat 20 ±Ctendedto migrateto a hightemperature eggexcretion of bothspecies was demonstratedfrom day arearegardless of temperature,and those cultured at 30 ±C 19to day 82 post inoculation by PCR analysis.Second didnot respond to the temperature gradient. generationworms showedshortened prepatent periods andshift towards predomination of O. dentatum (80-85% vs 38%in the Ž rst experiment).It is unclear whether this 543Mono- and mixed infection of ratswith larvae phenomenonis the consequence of the long storage of of Trichinellaspiralis Owen, 1835and T. larvaein the fridge or somekind of hostresponse. pseudospiralis Garkavi,1972

H. ZANGINYAN¤, A. ASATRIAN and M. HARUTUNYAN 542The behaviour of the infectivethird-stage larvae Instituteof Zoology,NAS RA, Yerevan,P .Sevak7, Armenia ofthe animal-parasiticnematode, Strongyloidesratti ¤[email protected] 1; 2 1 H. TOBATA-KUDO ¤, H. KUDO and I. TADA Theexperiment was undertakento testmale rats of Vistar line.Animals were splitup into three groups of Ž ve 1 Departmentof P arasitology,Graduate School of Medical animalseach. Groups I andII were treatedas control Sciences,Kyushu University, Fukuoka, Japan 2 Departmentof Anatomy, University of Occupationaland andinfected with larvae of Trichinellaspiralis and T. EnvironmentalHealth, School of Medicine,Kitakyushu, Japan pseudospiralis ina doseof 20larvae/gbodyweight of the ¤[email protected] rat.Group III was treatedas trial group and infected with acombinationof two Trichinella speciesin a doseof ten Weareanalysing the behaviour of Strongyloidesratti larvaefrom eachspecies per g bodyweight. The autopsy infectivelarvae (L3), a parasiteof a rodent,for studying was doneon day 35 postinfection. It was estimatedthat themechanism of the amphid functions using in vitro theformation of larvaein control animals was complete. agarosetracking assay method. In the present study, Trichinellaspiralis larvaewere encapsulatedand those we showthe behaviours of S. ratti L3, both on a of T.pseudospiralis hadtheir ends twisted or curved. sodiumchloride (NaCl) gradientand on a temperature Thestage of infection in control animals was 15-20 gradient.On a NaClgradient, two types of chemokinetic larvae/gbodyweight and eight larvae/ gbodyweight in behaviorwere seen:a unidirectionalavoidance movement trialanimals. These Ž ndingssuggest that mixed infection inunfavourable conditions ( ca 80mM NaCl) and a ofrats holds back the development of Trichinella and randomdispersal movement in favourable conditions. lowerstheir survival rates. The reason is obviously an Trackpatterns were straightin the avoidance movement interspeciŽc competitionof Trichinella species. butincluded multiple changes of direction and loops in thedispersal movement. On a temperaturegradient, the

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