Nematology,2000,V ol.2(7), 739-803 Symposiumabstracts

Desiccation stress of insect-killingnematodes inducesthe accumulation of a novel heat-stable protein

Solomon AHARON 1, Salomon RAFFI 2, Paperna ILAN 1 and Glazer ITAMAR 3 1 Departmentof AnimalSciences, F acultyof Agricultural, F oodand Environmental Quality Sciences of theHebrew University of Jerusalem,Rehovot 76100, Israel 2 Departmentof Virology,ARO, TheV olcaniCenter ,BetDagan, 50250, Israel 3 Departmentof Nematology,ARO, TheV olcaniCenter ,BetDagan, 50250, Israel Presentedat the 25th International Nematology Symposium, Herzliya, Israel, 2-7 April 2000

Infectivejuveniles (IJ)ofentomopathogenicnematodesof cumulatedto high concentrations by organisms that nat- thefamilies Steinernematidae and Heterorhabditidae can urallysurvive dehydration. A secondcycle of IJ dehy- activelylocate, infect and kill, with the cooperation of a drationdid not alter the RCL ofDesc47 (79.3% for the symbioticbacterium, a widerange of insect species. Both Žrst cycleand 73.3% for thesecond cycle). Desc47 re- Steinernema and Heterorhabditis passthrough four juve- tainedits high RCL (69.7%)in rehydrated active IJ for nilestages in theinsect host before maturing. Only the IJ 3days,reaching 51.2% of its initial RCL onlyafter a (third-stagelarva) can survive outside the insect host and week.No homology to otherknown proteins was foundby movefrom oneinsect to another.These parasitoid nema- mass-spectrometryelectrospray-ion-trap analysis. How- todesare considered among the most promising alterna- ever,from Žveobtained sequences of theprotein (ranging tivesto chemical control of insect pests. However, their from 10to21 aminoacids), the 21-amino-acid peptide N sensitivityto desiccation stress and other environmental VASDAVETVGNAAGQAG(D/T)AVshowed constraintsreduces their Ž eldefŽ cacy. Therefore, in or- 82%similarity and 59% identity to the Caenorhabditis derto understand better the molecular mechanisms which elegans lateembryogenesis abundant (LEA) homologue elicitresistance to desiccation stress in entomopathogeni c protein,and 79% similarity and 53% identity to theLEA nematodes,we identifya novelwater-stress-related pro- group3-like protein from thefern Oncleasensibilis . LEA teinwhich is induced in Steinernemafeltiae (IS-6) dur- proteinsare a diversegroup of water-stress-related pro- ingthe process of inducingthe IJ intoa quiescentanhy- teins,heat-stable, that are expressed in maturing seeds, drobioticstate. The protein was foundto be heat-stable undercold stress and duringwater deŽ cit in the vegetative witha molecularmass of47 kDa (designated Desc47). Desc47accumulated about 10-fold (from 7.84 1.85 to tissuesof higherplants. In plants, accumulation of LEAis 74.09 4.35%relative content level [RCL]) § indehy- alsoaccompanied by theaccumulation of solublesugars drated§clumps of IJ, whichlost 34.4% of theirinitial wa- suchas sucrose. These water-stress-related proteins and tercontent (WC) (from 65.1 1.7% to 42.7 0.72%) solublesugars have been suggested to play a majorrole ina desiccation-tolerance-inducing§ treatment (97% § rela- indesiccationtolerance. Our Žndingsin thepresent study tivehumidity [RH] for 3days).The appearance of Desc47 mayindicate a commonmolecular mechanism in nema- was accompaniedby trehaloseaccumulation (from 300to todesand plants which elicits resistance to desiccation 600mg trehalose/ gprotein).Trehalose is knownto beac- stress.

c KoninklijkeBrill NV ,Leiden,2000 739 ® Symposiumabstract

Root-knot nematode project expressed sequencetag (EST)

Pierre ABAD, Philippe CASTAGNONE-SERENO andMarie-Noë lle R OSSO INRA, UnitéSanté Vé gé tale et Environnement,Nematology Department, B.P .2078,06606 Antibes Cedex, France Presentedat the 25th International Nematology Symposium, Herzliya, Israel, 2-7 April 2000

Withthe aim to identify root-knot nematode genes in- begrouped in three main categories: i)sequenceswith volvedin plantparasitism, e.g.,migrationin the root tis- nohomologyin databases; ii)genesencoding housekeep- sue,feeding cell induction, virulence phenotype, we are ingproteins such as those involved in translation or key developinga largescale EST project(5000 clones). A enzymesin metabolic pathways, and, more interestingly; cDNA library(Zap vector) was constructedfrom invasive iii)genesputatively involved in nematodeparasitism e.g., second-stagejuveniles of Meloidogyneincognita . Before genesencoding proteins involved in protection against ox- sequencing,the quality of thelibrary was checkedby an- idativestress. The proportion of genes encoding poten- alyzingthe number of primaryrecombinants ( > 106) and tiallysecreted proteins was evaluated.These preliminary thelength of theinserts ( > 1kb).Here we reportthe data resultswill be discussedin relationto current knowledge obtainedfrom 300sequenced clones. The mRNAs could ofplant-pathogeninteractions.

Effect of temperature and pH on development of nematode trapping fungi

Aniko ANTAL 1, Erzsebet DORMANNS-SIMON 2 andC. JohannesK OK 3 1 PannonUniversity of Agricultural Sciences Georgikon F aculty,Department of Entomology,Deak F .str.57,8360 Keszthely, Hungary 2 CsongradCounty Plant Health and Soil Conservation Station, Biological Control and Quarantine Development Laboratory, P.O.Box 99, 6800 Hodmezovasarhely, Hungary 3 IPO ResearchInstitute for Plant Protection, P .O.Box9060, 6700 GW Wageningen,The Netherlands Presentedat the 25th International Nematology Symposium, Herzliya, Israel, 2-7 April 2000

Thepurpose of thesestudies was todeterminethe effectof ofpracticallyall strains was fast.At 35 °C,development environmentalfactors on potentialbiological agents. The stoppedand some strains were damagedby this tem- experimentswere carriedout by nematodetrapping fungi perature.Altogether it was establishedthat Monacrospo- belongingto Arthrobotrys and Monacrosporium genera, rium isolatespreferred lower temperatures compared with suitablefor biologicalcontrol of Meloidogyne nematodes. Arthrobotrys isolates.Concerning the effect of pH, the Thetrials were carriedout in vitro onPDA plates.The ex- bestgrowth was observedat pH 7. The isolates did not aminedtemperatures ranged from 5to35 °C, in 5°C inter- growat pH 3,4,and 5. At pH6,fastdevelopment was ob- vals.The pH intervals ranged from 3to11.At 5 °C the iso- latespractically did not grow .At 10 °C,growthwas poor, servedinitially but then growth slowed down. At pH8,9, exceptfor the Monacrosporium strainsthat showed bet- 10,and11 developmentwas slowand differences between teractivity. In the interval from 15to 30 °C,thegrowth isolateswere notsigniŽ cant.

740 Nematology Symposiumabstract

Inuence of climaticand crop management conditions on Xiphinema index populations and GFLVspread inCastilla-La Mancha (Spain) vineyards

María ARIAS 1, Jesús FRESNO 2 andAntonio L ÓPEZ-PÉREZ 1 1 DptoAgroecologí a, Centrode Ciencias Medioambientales, CSIC. Serrano 115 dpdo, 28006 Madrid, Spain 2 Dptode MejoraGené tica y Biotecnología, SGIT-INIA,Ctra de la Coruña Km 7.5,28040 Madrid, Spain Presentedat the 25th International Nematology Symposium, Herzliya, Israel, 2-7 April 2000

A study on Xiphinemaindex andGFL Vincidencehas banding,symptoms appear, GFL Vdispersaland X. in- beencarried out in Castilla-La Mancha (Spain) vineyards, dex populationsincreases occur, while in those under tra- wherethe climatic conditions and traditional crop man- ditionalmanagement disease and pathogens remain sta- agementprevent the pathogen development. Many plants blefor years.On the other hand, in vineyards with virus diedbecause of thedry climatic conditions (rainfall rates andnematode presence, only isolated wood and fanleaf about300 mm) in1988-1996. Therefore, different irri- symptomsin smallpatches were observedin dry periods, gationsystems were introducedto grow plants and in- theyellow mosaic and veinbanding symptoms becoming creasetheir production. The in uence of climaticand crop patentsince 1996 to 1999 when the climatic conditions managementconditions are analyzed by comparing the situationin vineyards with drop irrigation or under tra- changed(rainfall rates over 600 mm). Itis concludedthat ditionalmanagement and between different rainfall peri- onintroducing irrigation, the risk of thesepathogens in- ods.Results show that in vineyards where irrigation had creasingmust be considered before the introduction of beenintroduced, yellow mosaic, sometimes plus vein- newmanagement techniques.

Durable resistance management of soil-borne quarantine nematode pests Meloidogyne chitwoodi and M. fallax

Jaap BAKKER 1,WladyslawG OLINOWSKI 2, Richard JANSSEN 3, Joka KLAP 4, Didier MUGNIÉRY 5, Mark S. PHILLIPS 6,MichaellaS CHLATHÖLTE R 7 andJ.G. Hans V AN DER BEEK 3 1 Laboratoryof Nematology,W ageningenUniversity and Research Centre, P .O.Box 8123, NL-6700 ES Wageningen,The Netherlands 2 Departmentof Botany(SGGW), F acultyof Agriculture, W arsawAgricultural University, Rakowiecka 26/ 30,P-02-528 W arsaw, Poland 3 PlantResearch International, W ageningenUniversity and Research Centre, P .O.Box16, NL-6700 AA Wageningen,The Netherlands 4 BarenburgResearch, Duitsekampweg 60, NL-6874 BX Wolfheze,The Netherlands 5 Laboratoirede Zoologie,Institut National de la Recherche Agronomique, Domaine de laMotte,P .O.Box29, F-35160 Le Rheu, France 6 Departmentof Nematology,Scottish Crop Research Institute, Invergowrie, DD2 5DA Dundee,United Kingdom 7 P.H.PetersenSaatsucht Lundsgaard, Streichmü hler Strasse 8, D-24977Grundhof, Germany Presentedat the 25th International Nematology Symposium, Herzliya, Israel, 2-7 April 2000

TheEU-funded project QLRT -1999-1462DREAM world-wideproblem of theuse of very toxic chemicals to (DurableResistance Against Meloidogyne )aimsto con- controlthe soil-borne pests. This study will use the two tributeto sustainable production systems by developing quarantineorganisms Meoidogynechitwoodi and M. fal- astrategyfor durableresistance management for these lax whichare an important economic threat and for which soil-bornepests. This is necessary in order to solve the noadequatedurable alternative exists. The objective will

Vol.2( 7),2000 741 Symposiumabstract beachieved by integrating expertise in breeding, nema- resistantgermplasm; well characterised pathogen collec- tology,botany and molecular biology into one project, tions;reliable selection methods; knowledge on thestabil- novelby its European dimension. The project combines ityof resistance, molecular markers linked to resistance threeareas of research: i)identiŽcation and incorporation and(a)virulence;reliable tools for breedingmethods, re- ofresistancein important arable crops; ii)studyof dura- bilityof theresistance and iii)optimisingproduction sys- sistantgermplasm, genetic maps, and advice about im- temsby rotationschemes. The main results expected are: provedrotation schemes.

Development and evaluation of new sampling methods for Želdswith infestation foci of potato cyst nematodes ( Globodera rostochiensis and G. pallida)

Thomas H. BEEN andH. CorrieS CHOMAKER PlantResearch International, P ostbox16, 6700 AA, Wageningen,The Netherlands Presentedat the 25th International Nematology Symposium, Herzliya, Israel, 2-7 April 2000

Anextended version of thecomputer program ‘ SAMPLE’ acontrolmeasure without noticeable yield reductions in was usedto develop new sampling methods for thede- a1:3year cropping frequency of potatoes. In that way tectionof infestationfoci of potatocyst nematodes ( Glo- the‘ worstcase scenario’ used in earlier approaches, re- boderarostochiensis and G. pallida). Bycombining a quiringlarge soil samples, could be avoided.Several new modelfor themedium scale distribution of cysts,which versionsof highperformance detection methods were de- providesthe expected population densities at each posi- veloped.For ware potatogrowers sampling methods were tionwithin the focus, and a modelfor thesmall-scale dis- developedwhose intensity (amount of soil collected) de- tributionwithin square meters, sampling procedures can pendon thecropping frequency of thefarmer. SAMPLE besimulated. SAMPLE 4alsotakes into consideration V4can be considereda toolto developsampling methods thebivariate normal distribution of theparameters (length ondemand for everypossible combination of character- andwidth gradient) of the medium scale distribution to calculateaverage detection probabilities over the range isticsrequired for useby seed and ware potatogrowers ofpossible shapes of a standardinfestation focus (cen- (recommendationsfor optimumcontrol measures leading tralpopulation density 50 cysts/ kgsoil).This infestation tomaximum returns, IPM) andby governments (legisla- focusis smallenough to use resistant potato varieties as tion,quarantine and export protection).

742 Nematology Symposiumabstract

Biofumigation, solarization and nematode control

Antonio BELLO,José-Antonio L ÓPEZ-PÉREZ, Luisa DÍAZ-VIRULICHE and Rafael SANZ DptoAgroecologí a, Centrode CienciasMedioambientales, CSIC. Serrano 115 dpdo, 28006 Madrid, Spain Presentedat the 25th International Nematology Symposium, Herzliya, Israel, 2-7 April 2000

Theaction of gasesdischarged by the decomposition of caldynamics, which is a problemto all mobile organisms, organicmatter is studied for thecontrol of nematodes aproblemcaused when the soil is heated in polarisation. andother phytopathogen icsoil-borne organisms. Effec- Resultsof theapplication of biofumigationtechniques in tivenessis similarto conventional biofumigants, differing extensivecrops are shown under low temperature condi- from polarisationin that temperatures over 30 °C are not tionsand without the application of plastic covers. This required.Therefore it can used in differentseasons of the yearand in areas having low temperatures, as wellas on isdifferent from polarisation,although both techniques extensivecrops. On the other hand, biofumigation acts in canbe complementary, increasing their effectiveness in depth,in thecase of nematodessolving problems of verti- thecase of phytopathogenicnematodes.

Gonad morphology and evolutionary relationships within plant parasitic nematodes

Wim BERT andEtienne G ERAERT Universityof Gent,Zoological Institute, Ledeganckstraat 35 B-9000Gent, Belgium Presentedat the 25th International Nematology Symposium, Herzliya, Israel, 2-7 April 2000

Themorphology of thefemale reproductive system is im- nadmorphology for eachgenus; this sheds a newlight portantin nematode systematics. In this study the cellu- onthe evolutionary relationships of this group. Within larmorphology of the gonads of 60 different species of thegenera, all species share typically similar gonad mor- tylenchidsis describedand compared. Previous research phologyexcept for Meloidogyneichinohei thatis differ- suggestedthat uterine cells arranged in three rows repre- entfrom other Meloidogyne species.DNA studiesalso senta derivedcondition within T ylenchoidea;this is sup- demonstratedthis species as being quite divergent from portedin thisstudy. According to ourresults the Cricone- the other Meloidogyne species.The results, obtained by matoideaappear to have a closecommon ancestor with ourmethods to study the cellular morphology of there- Tylenchidae.The economically very important sedentary productivesystem, lead to a betterunderstanding of the endoparasiticnematodes display markedly different go- relationshipswithin the plant parasitic nematodes.

Vol.2( 7),2000 743 Symposiumabstract

Functional analysis of gene expression ingiant cells

David BIRD 1, Ann GREENE1,†, Hinanit KOLTAI 1, Jennifer SCHAFF 1, Jessica WATKINS 1 and Joseph KIEBER 2 1 PlantNematode Genetics Group, Department of Plant P athology,North Carolina State University, Raleigh, NC 27695,USA 2 Departmentof Biology,University of NorthCarolina, Chapel Hill, NC 27599,USA †Deceased Presentedat the 25th International Nematology Symposium, Herzliya, Israel, 2-7 April 2000

WehaveidentiŽ ed a largesuite of genes expressed in tionas well as uorescencemicroscopy. T oassemblereg- Meloidogyneincognita- inducedtomato giant cells, and ulatorycascades it will be necessary to establish func- areinterested in the regulatory cascades they deŽ ne. tionalrelationships ( e.g.,bygenetic epistasis), and we Thesesequences are available in GenBank. Because root- havebegun to examine expression of selected giant cell knotnematodes produce biologically-act ivecytokinin, genesin various backgrounds created by transgenic ab- oneapproach we arepursuing is to map the spatial lation.Null mutations established by anti-sense expres- andtemporal induction of cytokinin-responsive promot- sionor co-suppressionare conŽ rmed by sensitive, in situ ers duringgiant cell initiation, and during gall formation. transcriptlocalization. The use of constitutive(35S), giant Our initialexperiments have employed the IBC6 promoter (Plant Cell 10,1009-19, 1998) drivingGFP expressionin cellspeciŽ c (TobRB7)and glucocorticoid-in duciblepro- tomatohairy roots. T oaccountfor thedifferential accu- moters (Plant J. 11,605-12, 1997) will permit the role of mulationof detectable GFP mRNA vs GFP protein(10 thesegenes in the inductionand maintenanceof thepatho- min vs 10hr) we areusing in situ transcriptlocaliza- logicalstate to beestablished.

How nematode secretions work inplant cells

Teresa BLEVE-ZACHEO 1, A. LEONE 2, L. ROBERTSON 3, M.T. MELILLO 1 and C. FENOLL 1 1 Istitutodi NematologiaAgraria, Amendola 165/ A,70126Bari, Italy 2 Departamentode Biologia, UAM, 28049Madrid, Spain 3 DepartamentoMedio Ambiente, UCLM, AvdaCarlos 111, S/ N,45071T oledo,Spain Presentedat the 25th International Nematology Symposium, Herzliya, Israel, 2-7 April 2000

Theavailability of secretionsobtained by stimulation of solutionof adjoining cell walls. Ultrastructural analy- J2juveniles of Globoderarostochiensis allowedus toin- sesof syncytiademonstrated cytological changes such as vestigate,using the microinjection technique, whether se- granularcytoplasm, abundant mitochondria and endoplas- cretionshad the capacity to interact with injected cells. micreticulum, plastids with starch grains, small vacuoles, Corticalcells in the differentiating zone of root apices andenlarged amoeboid nuclei. These cytological changes ofpotato and tomato explants were injectedwith a so- were similarto those induced by feeding nematodes on lutioncontaining 0.02-0.4 m g/m lofsecretions added to hostroots. Microinjection of eitherbuffer alone or heat- Luciferyellow as a tracer.Injected roots collected and inactivatedsecretions did notgiveany syncytialinduction. Žxedat different times after injection were analysedat lightand electron microscope level. Sectioned roots ob- Thesepreliminary studies provided the Ž rst directexper- servedat light microscope revealed that the least amount imentalevidence that artiŽ cial injection of nematodese- ofinjected secretions were sufŽcient to induce a syn- cretionscan interact with host cells and can induce a struc- cytium,as a conglomerateof fusedprotoplasts with dis- turesimilar to feeding sites.

744 Nematology Symposiumabstract

Virulenceand avirulencein root-knot nematodes inCrete on tomato

Vivian C. BLOK 1, Miles R. ARMSTRONG 1,EmmanuelA. T ZORTZAKAKIS 2 and Mark S. PHILLIPS 1 1 Departmentof Mycology,Bacteriology and Nematology, Scottish Crop Research Institute, Invergowrie, Scotland DD2 5DA,UK 2 PlantProtection Institute, National Agricultural Research F oundation,P .O.Box 1802, 71110, Heraklion, Crete, Greece Presentedat the 25th International Nematology Symposium, Herzliya, Israel, 2-7 April 2000

Ina surveyof vegetablegrowing areas of Crete,the ma- derivedfrom thesame founder populations. W ehavebeen jorityof the infestations were foundto be Meloidogyne comparingexpressed gene sequences of bothvirulent and javanica.Mostof these populations were avirulenton avirulentpopulations using cDNA AFLPs andsuppressive Mi tomatocarrying the gene;however a few virulentpop- subtractivehybridisation to identify pathogenicity factors. ulationswere alsoidentiŽ ed. In an AFLP comparisonof Wehavealso been using these techniques to identifyhost bothvirulent and avirulent populations, a veryhigh mean similaritybetween these populations was found,suggest- geneswith altered expression during the compatible and ingthat both virulent and avirulent populations probably incompatibleresponses.

Neuropeptide type sequences and activities in Globodera spp.

Vivian C. BLOK 1,KonstantinaB OUTSIKA 1, Lydia CASTELLI 1, Mark S. PHILLIPS 1, Geert SMANT 2 and John T. JONES 1 1 Departmentof Mycology,Bacteriology and Nematology, Scottish Crop Research Institute, Invergowrie, Scotland DD2 5DA,UK 2 NematologyDepartment, W ageningenAgricultural University, W ageningen,The Netherlands Presentedat the 25th International Nematology Symposium, Herzliya, Israel, 2-7 April 2000

Theanatomy of the nervous system of nematodes is was shownto be expressedin thenerve ring using in situ conserved,generally conforming to that described for hybridisation. Caenorhabditiselegans .However,there is an appar- Thefeasibility of altering nematode behaviour by ex- entlyrich diversity in putative signalling molecules. ogenousexposure to neuropeptides was alsoassessed in SeveralESTs were identiŽed with high homology to aplateassay. An increase in movement of PCN J2was neuropeptide-typesequences during a smallscale project observedwhen exposed to oneof thepeptides suggesting with G.rostochiensis .FMRF-likemotifs occur in these sequencesand protease cleavage sites for processingneu- thatplant parasitic nematodes may be vulnerableto small ropeptidesare present. This sequence was alsofound to peptidemolecules and that such peptides may be useful beexpressedin G. pallida.Oneof theneuropeptide genes for novelcontrol applications.

Vol.2( 7),2000 745 Symposiumabstract

Bursaphelenchus speciesin conifers inEurope and theirdetermination

Helen BRAASCH 1, Kai METGE 2 andW olfgangB URGERMEISTER 2 1 Departmentfor National and International Plant Health, Kleinmachnow Branch, F ederalBiological Research Centre for Agriculture andF orestry,Stahnsdorfer Damm 81,D-14532 Kleinmachnow ,Germany 2 Institutefor Plant V irology,Microbiology and Biosafety, F ederalBiological Reseach Centre for Agriculture and F orestry,Messeweg 11,D-38104 Braunschweig, Germany Presentedat the 25th International Nematology Symposium, Herzliya, Israel, 2-7 April 2000

Informationon the distribution and reliable identiŽ ca- torsfor thedistribution of Bursaphelenchus species are tionof thevarious Bursaphelenchus speciesin European thepresence of hostplants and the availability of suitable conifertrees is of increasingimportance due to the recent insectvectors. Different Bursaphelenchus speciesdistrib- introductionof the pinewood nematode ( B.xylophilus ) utionhas been found in Central and Southern Europe. The intoPortugal and the survey in Europenecessary to con- speciesare grouped by theirmorphological and molecu- trolits possible spread. Results from anEU-supportedre- larbiological features. Appropriate identiŽ cation methods searchproject on European Bursaphelenchus spp. (Fair basedon selectedmorphological characters and rDNA se- CT95-83)and a listof European species with details quencedifferences revealed by ITS-RFLP techniqueare ontheir distribution and hosts are presented. Key fac- emphasised.

Resistance of vegetable crops to Meloidogyne spp.: suggestion for acrop rotation system

ReginaM.D.G. C ARNEIRO 1, Onivaldo RANDIG 2 andAngela Diniz C AMPOS 2 1 EMBRAPA/RecursosGené ticos e Biotecnologia,C.P .02372,70849-970 Brasí lia, DF ,Brazil 2 EMBRAPA/ClimaT emperado,C.P .403,96001-970 P elotas,RS, Brazil Presentedat the 25th International Nematology Symposium, Herzliya, Israel, 2-7 April 2000

Agreenhousetest was carriedout to determine the repro- Híbrido T ongo,spicy dry cress andkale cv. Manteiga are ductivepotential, resistance or susceptibilityof Meloido- resistantto M. arenaria race2. Squashes( Cucurbitapepo ) gynejavanica, M. incognita race 3, M. arenaria race 2 and cvsCaserta and Branca de Virgí nea, melons ( Cucumis M. hapla,on24vegetablecrops belonging to 14 botanical melo)cvsHí brido Piel de Sapoand Y ellowStart, spinach species.Sweet peppers ( Capsicumannuum ) cv. All Big (Spinaceaoleracea )cvsViroFlay and Nova Zelâ ndia, cu- andHí brido T ongo,spicy dry cress ( Rorippanasturtium ) cumbers (Cucumissativus )cvsSMR58and Marketer and and kale (Brassicaoleracea var. acephala)cv.Manteiga lettuce (Lactucasativa )cv.Crespa Rá pida are resistant to areresistant to M.javanica. Cauliowers ( Brassicaoler- acea var. botrytis)cvsT eresópolis Gigante and Bola de M. hapla.Acroprotation system alternating these veg- Neve,spicy dry cress andkale cv. Manteiga are resistant etablesis suggestedfor thegrowers in Rio Grande do Sul to M.incognita race3. Sweet peppers cvs All Big and State,Brazil.

746 Nematology Symposiumabstract

Screeningthe Commonwealth Potato Collection for novel sources of resistance to the Potato Cyst Nematode Globodera pallida

Lydia CASTELLI, Gavin RAMSAY, Sharon J. NEILSON and Mark S. PHILLIPS ScottishCrop Research Institute, Invergowrie, Dundee, DD2 5DA,UK Presentedat the 25th International Nematology Symposium, Herzliya, Israel, 2-7 April 2000

Professor JackHawkes’ collection (the Birmingham tantof thetwo species in theUK. Atotalof 10seedlings PotatoCollection) of wild species of potatoes has been for eachoutbreeding accession and 3 seedlingsfor each recentlyincorporated into the Commonwealth Potato Col- inbreedingaccession were tested.Forty of the outbreed- lection,supplementing the range of speciesheld. The ma- ingand 28 ofinbreedingaccessions were foundto beuni- terialoriginates from severalexpeditions to the Americas formlyresistant, with a further17 segregating for resis- andrepresents a potentiallyvaluable source of resistance tance.These accessions were from 34differentspecies of tothe potato cyst nematodes Globoderarostochiensis and wildpotato, of which11 arebelieved to benovelsources, G. pallida,bothof whichare serious pathogens of pota- followingan extensive search through all available data- toes.The germplasm screened in thisstudy included taxa baseson knownsources of resistantto G. pallida.To iden- mainlyfrom Mexico,Bolivia, Argentina and Peru. Seedsfrom 199accessions, comprising 66 different tifygenotypes with broad spectrum resistance the acces- speciesand focusing predominantly on materialfrom the sionswill also be screenedfor resistanceto G.rostochien- BirminghamPotato Collection, have been screened for re- sis aswellas thetemperate root knot nematode Meloidog- sistance to G. pallida (Pa 2/3)whichis the more impor- ynechitwoodi.

Soilpopulations dynamics of Heterodera schachtii ,using resistant cultivars of sugar-beet

Georges CAUBEL 1,CatherineP ORTE 1 and Hervé MARZIN 2 1 InstitutNational de laRecherche Agronomique, Zoologie, 35650 Le Rheu,France 2 LaboratoireNational Protection Vé gé taux, 35650 Le Rheu,France Presentedat the 25th International Nematology Symposium, Herzliya, Israel, 2-7 April 2000

In France, Heteroderaschachtii affectthe level and reg- uenceon soil H.schachtii populationdensities of these ularityof sugar-beetyields, especially in short rotations. cultivars,resistant or not, is always correlated with Pi. Since1996, sugar-beet cultivars with resistance conferred At lowinitial infestations, Ž nalpopulations ( Pf ) are in- by Betaprocumbens areregistered. In infested soils, root creasedand at very low infestations, below 5 juveniles/g, yieldincrease is 10-20%. The objective of thisstudy was multiplicationmay be high, even with a resistantculti- toevaluate the population effect over multiple years at var.Reproductionrate was negativelycorrelated with Pi differentrepresentative locations. In uence of resistant acrossenvironments of bothresistant and susceptible cul- sugar-beeton nematode population dynamics was inves- tigatedin 14 naturally infested Ž elds,with a rangeof tivars.The relative performance of resistant and suscep- initialpopulation densities ( Pi), usingsusceptible (Carat, tiblecultivars can be reliablypredicted based on preplant Roberta)in comparisonwith resistant cultivars (Evasion, eggdensities. By useof aresistantcultivar, soil infestation Nemakill,Mercure, Anema). Results indicated that the in- decreasedonly at Pi higherthat the economic threshold.

Vol.2( 7),2000 747 Symposiumabstract

Magnetic capture of nematodes using antiserum and lectincoated Dynabeads

Qing CHEN, Lee ROBERTSON, John T. JONES, Vivian C. BLOK, Mark S. PHILLIPS andDerek J.F .B ROWN Mycology,Bacteriology and Nematology Unit, Scottish Crop Research Institute, Invergowrie, Dundee, DD2 5DA,Scotland Presentedat the 25th International Nematology Symposium, Herzliya, Israel, 2-7 April 2000

Theeffective use of integratedmanagement strategies re- ingto the entire surface of thenematode was aprerequi- quiressimple, rapid and reliable identiŽ cation and quan- sitefor efŽcient extraction that was relatedto strengthof tiŽcation of theplant parasitic nematodes present in a soil. bindingof theantisera to thenematode (as determinedby Unfortunately,plant parasitic nematodes are small and ex- immunouorescence microscopy). Factors affecting the tremelydifŽ cult to identify. Previous studies have demon- extractionefŽ ciency when using lectin coated beads are stratedthe potential for antibody-assistedidentiŽ cation of currentlyunder further investigation. This study expands nematodes.W epresentan extension of this technology, theuse of Dynabeads from celland molecular biology usingantibody or lectin coated magnetic beads (Dyna- toparasitology and shows, in principle, that Dynabeads beads)to recover target nematodes from samples.First, coatedwith a probeof suitable speciŽ city can be usedto we identiŽed lectins and antisera that bound speciŽ cally extractspeciŽ c nematodesfrom testsamples. This tech- andreproducibly to thewhole surface of Globoderaros- nologymay ultimately be usefulin “non-expertsystems” tochiensisand Meloidogynearenaria .Wefoundthat bind- for detectionand quantiŽ cation of nematodespecies.

Stem nematode resistance inwhite clover and variation innematode virulence

Roger COOK andK. AnthonyM IZEN Instituteof Grassland and Environmental Research, Aberystwyth, SY23 3EB, Wales,UK Presentedat the 25th International Nematology Symposium, Herzliya, Israel, 2-7 April 2000

Stemnematode ( Ditylenchusdipsaci )iswidespread on opedwhite clover populations for useas resistant(R) and whiteclover ( Trifoliumrepens )ingrassland in England susceptible(S) standardsin experiments and developed andW ales.White clover populations and cultivars are het- tworesistant cultivars. Before releasing these, we arein- erogeneousfor manycharacters. Resistant germplasm has vestigatingvariation in virulence characteristics of white beenselected by assessingsymptoms after inoculation of cloverstem nematode populations. Resistant clovers se- meristemswith nematodes. T wentyseven cultivars and 4 lectedwith a UKpopulationwere alsoresistant to popu- breeders’lines had a rangefrom 0to30% resistantplants. lationsfrom Franceand New Zealandbut were fullysus- Thevery large leaved cv. Aran had64% resistant plants. ceptibleto a populationfrom Switzerland.This appears Noneof thisgermplasm is suitable for usein long-term, tobeevidencefor variationin virulence in naturalpopu- grazedswards withoutfurther selection. W ehavedevel- lationsof thenematode.

748 Nematology Symposiumabstract

Bacterialsymbionts in Xiphinema americanum s.l.

August COOMANS 1 and Tom VANDEKERCKHOVE 2 1 BiologyDepartment, Institute of Zoology, University of Gent, Belgium 2 BiologyDepartment and Department of Biochemistry, Physiology and Microbiology, University of Gent, Belgium Presentedat the 25th International Nematology Symposium, Herzliya, Israel, 2-7 April 2000

Thebest known example of this phenomenon is Wol- theyaggregate near the anterior pole and apparently be- bachiapipientis inparasitoidwasps and other arthropods. comeenclosed in the descendants of the E-founder cell. Wolbachia hasalso been found in some nematodes, but in Indeed,endosymbionts are stored in intestinal cells, but Xiphinemaamericanum s.l.other symbionts are present. notingenitalcells of atleast the Žrst threejuvenile stages. Theybelong to a newclade of V errucomicrobia.In the Thetransfer to theovaria is supposedto occur during the femalethe bacteria occur in the intestine and the ovaria; theyare most obvious in the ovarial wall. A numberof lastmoult of thenematode. Molecular analyses show that thementer the maturing oocytes where they further di- thesymbionts are species speciŽ c andthat the symbiosis videtill about 250 bacteria are present in the egg. There isaveryold and stable one.

Selectionof Belgian Verticilliumchlamydosporium isolates for the control of Meloidogyne javanica ingreenhouse crops

G. DE DEYN 1, T. DERYCKE 1, M. MOENS 1 and N. VIAENE 1 1 AgriculturalResearch Centre, Department of CropProtection, Burg. V anGansberghelaan96, 9820 Merelbeke, Belgium Presentedat the 25th International Nematology Symposium, Herzliya, Israel, 2-7 April 2000

Weobtained92 isolatesof Verticilliumchlamydosporium , tomatoroots in the glass house, 8, 10, 12 and 15 weeks afungusparasitizing nematode eggs, from 46soilsamples aftersoil infestation. In a testperformed in spring, 4 to takenat 11 locations in Belgium. Fungal isolates should 48%of eggswere empty.In summer, 32 to 61% of eggs bevirulent, establish well in the host plant rhizosphere were empty.The ability of 23 isolates to colonise plant andbe easily produced when used in an integrated con- rootswas assessed in vitro ongerminating barley seeds. trolstrategy. In vitro virulenceagainst Meloidogyneja- Colonisationranged from 0.5to 87.6% of the total root vanica often isolates was testedusing egg masses. The length.Fifty isolates were tested in vitro for theirdicty- percentageof infectedeggs per egg mass variedfrom 49 ochlamydosporeproduction in a wheat:sandmixture. At to96%. T estegg masses should be similar in composi- 25°Ctwentyisolates did not produce any dictyochlamy- tion,and contain few emptyand fully developed eggs. W e dospores.The isolates scoring well for allthree criteria comparedthe composition of 50eggmasses produced on were selectedfor pottests.

Vol.2( 7),2000 749 Symposiumabstract

Measurement of the infection potential of nematode-transmitted virus insoil

Ate DE HEIJ 1, Frans C. ZOON 1, F. Astrid DE BOER 1 and Cees J. ASJES 2 1 PlantResearch International, P .O.Box 16, 6700 AA Wageningen,The Netherlands 2 BulbReseach Centre, P .O.Box85, 2160 AB Lisse,The Netherlands Presentedat the 25th International Nematology Symposium, Herzliya, Israel, 2-7 April 2000

Tobaccorattle virus (TR V)istransmitted by trichodorid anELISA. Themost probable number (MPN) ofvirus- nematodesand causes considerable losses in owerbulbs transmittingnematodes per unit soil was calculatedby andpotato. QuantiŽ cation of theinfectionpotential of soil likelihoodapproximation of the number of positive bait (IPS) isneeded to assessthe risk of infectionin cropsand plantsin each dilution. MPN valuesin Ž eldexperiments effectsof treatmentsin Ž eldand laboratory experiments. withtulip and gladiolus explained about 45% of thevari- Countingvector nematodes is oflittleuse, because not all ancein the fraction diseased plants. Sources of variance nematodesare viruliferous. Therefore a methodwas de- inthe method itself, in the sampling and in disease ex- signedto estimatethe number of nematodesable to trans- mitTR Vtoa baitplant. Soil samples were dividedover pression,are studied in orderto decidewhether develop- potsin replicateddilution series. Petunia bait plants were mentof IPS measurementfor riskanalysis at Želdscale is grownin eachpot andsubsequently root sap was testedin worthwhile.

Biochemicalcharacterisation of pharyngeal secretions from Heterodera schachtii stage 2juveniles

Jan DE MEUTTER 1, Tom TYTGAT 2, Greetje GHEYSEN 1, August COOMANS 2 andGodelieve G HEYSEN 1,3 1 VIB, Deptof PlantGenetics, University of Gent,Belgium 2 Instituteof Zoology,University of Gent, Belgium 3 Facultyof Agriculturaland Applied Biological Sciences, University of Gent,Belgium Presentedat the 25th International Nematology Symposium, Herzliya, Israel, 2-7 April 2000

Heteroderaschachtii belongsto the group of sedentary culturingand the puriŽ cation techniques were upscaledto plantparasitic nematodes which causes a lotof agricul- assurea sufŽcient supply of material.A rapidnematode turaldamage. In the roots of ahostplant stage 2 juveniles sterilisationmethod, based on a disinfectanttreatment, inducespecialised feeding cells, most probably by inject- was developedwhich did not compromise the production ingsecretions from thepharyngeal glands. Despite many ofsecretions. An efŽ cient way for producingsecretions efforts,the triggersresponsible for thisfeeding site forma- anddifferent concentration methods for therecovery of tionare still unknown. This research is aimedto gain in- secretoryproteins in dilute solutions are presented and sightinto the composition and the function of thepharyn- evaluated.The methodology presented should allow se- gealsecretions from Heteroderaschachtii .Thereforethe quencingof proteinsseparated by PAGE.

750 Nematology Symposiumabstract

Distribution of Bursaphelenchus xylophilus inpine logs and consequences for sampling

Lutgard DE WAEL 1, Nancy DE SUTTER 1, Renaat MOERMANS 1, Marc LINIT 2 andMaurice M OENS 1 1 AgriculturalResearch Centre, Crop Protection Department, Burg. V anGansberghelaan 96, 9820 Merelbeke, Belgium 2 Universityof Missouri-Columbia, Department of Entomology,Columbia, Mo, 65211, USA Presentedat the 25th International Nematology Symposium, Herzliya, Israel, 2-7 April 2000

Thepine wood nematode (PWN), Bursaphelenchusxy- woodcurls was skewedto theright (mean: 14.76; median: lophilus isvectoredby Monochamus spp.and is reported 4.00;st. dev.: 28.96). The proportion of theinfected sam- tocause severe pine wilt in several Asian countries and pleswas 0.63.The presence/ absenceof PWN was related partsof NorthAmerica. In the EuropeanUnion, the nema- toobservable characteristics (grub holes, colour, knots todeis classiŽ ed as aquarantineorganism. T odevelopap- andresin) of the plank. The highest probability for de- propriatesampling strategies, nematode distribution was tectingPWN isrelatedto grey stained wood parts (0.80), monitoredin 10 PWN-infected Pinussylvestris logs (3- followedby thepresence of agrubhole (0.77). The low- 8mlong)cut into planks. A totalof 5387samples were estprobability was observedin parts with obvious resin collectedwith a joinerdrill and extractedwith a technique presence(0.38). The combination of characteristics did basedon centrifugation.The concentration of nematodes notincrease the probability of detection. Sampling sites inthe trees did not follow any consistent pattern over the notshowingany of theaforementioned characteristics still trees.The distribution of nematode numbers per 5 gof hada probabilityfor detectingPWN of0.65on average.

Pathogenicity of the root-lesion nematodes Pratylenchusneglectus and P. thornei on faba bean ( Vicia faba L.)

Mauro DI VITO, Giovanni ZACCHEO and Fabio CATALANO Istitutodi Nematologia Agraria, C.N.R., 70126Bari, Italy Presentedat the 25th International Nematology Symposium, Herzliya, Israel, 2-7 April 2000

Anexperiment was undertakento investigatethe effect of 256specimens/cm 3 soil.Data of fresh topweightof plants arangeof inoculumdensities of twoItalian populations Žttedthe Seinhorst’s model y = m + (1 ­ m)zP ­ T . Ac- of Pratylenchusneglectus and P. thornei onthe growth cordingto thismodel, tolerance limits ( T )offababean to 3 offaba bean in pots. Eighty-four clay pots of 1000 cm P.neglectus and P. thornei were 2and2.4 specimens/ cm 3 each,Ž lledwith steam sterilized sandy soil, were used soil,respectively. Minimum relative yields ( m) of faba for eachnematode species. Pots were sownwith a single beanwere 0.4and 0.5 for fresh topplant weight for P. pregerminatedseed of fababean cv. Aquadulce. At plant neglectus and P. thornei,respectively,and occurred at Pi emergencea suspensionof juvenilesand adults of oneof 3 thenematode species was pouredinto six holes around > 128specimens/ cm soil.Maximum reproduction rates seedlingroots to give initial population densities of 0, (Pf/Pi)were 17.9and 3.9-fold, at lowest Pi, for P. neglec- 0.03,0.06, 0.125, 0.25, 0.5, 1, 2,4,8,16,32, 64, 128 or tus and P. thornei,respectively.

Vol.2( 7),2000 751 Symposiumabstract

Mapping of the pepper Me3 gene conferring heat-stable resistance to root-knot nematodes (Meloidogyne spp.) and comparison with the tomato- and potato-nematode resistance genes location

Caroline DJIAN-CAPORALINO 1, Lucette PIJAROWSKI 1, Ariane FAZARI 1, Manuella SAMSON 1, Laurent GAVEAU 1, Véronique LEFEBVRE 2, Carole CARANTA 2, Anne BLATTES 2, Alain PALLOIX 2 and Pierre ABAD 1 1 UnitéSanté Vé gé tale et Environnement,INRA, Antibes,France 2 Unitéde Gé né tique et Amélioration des Fruits et Légumes, INRA, Montfavet,France Presentedat the 25th International Nematology Symposium, Herzliya, Israel, 2-7 April 2000

Me3,singledominant gene of the PM687 line of Cap- DHprogeny,conŽ rmed its closed position. An other re- sicumannuum, confersheat-stable resistance to root-knot sistancegene to RKN, presentin PM687 Me4, was shown nematodes(RKN). Wehavelocated it using doubled- tobelinkedto Me3,nearly10 cMfrom it.Inorder to local- haploid(DH) linesand F 2 progenyfrom acrossbetween ize Me3 and Me4 ontheintraspeciŽ c pepperlinkage map, thesusceptible cultivar Y oloW onderand the highly re- twoAFLP markerswere mapped.The Me3 nearestmarker sistantline PM687. Bulked-segregant analysis with DNA linkedto resistance was 2.7cMfrom RFLP-CT135.PCR- bulks,from susceptibleor resistant DH lines,was per- speciŽc markersobtained from twoAFLPs willbe usedin formedto identify AFLP markerslinked to Me . Four 3 pepperbreeding for marker-assistedselection. W einvesti- repulsion-phaseand two coupling-phase markers comple- Me mentaryto the HindIII and MseIsiteswere locatedbe- gatemap position homologies between 3 andtwoother tween0.5 and 26.3 centimorgan (cM) onbothsides of the nematoderesistance genes, Mi-3 and Gpa2 whichmapped inthetelomeric region of theshort arm ofthetomato and gene.Analysis of the F 2 progeny(162 plants) with the coupling-phasemarker located 0.5 cM from Me3 in the potatochromosome 12.

Effects of ediblefungi oyster mushroom ( Pleurotus ostreatus ) on Aphelenchus avenae and Steinernema feltiae

Ewa DMOWSKA Instituteof EcologyP olishAcademy of Sciences, Dziekanow Lesny, 05-092 Lomianki, P oland Presentedat the 25th International Nematology Symposium, Herzliya, Israel, 2-7 April 2000

Theoyster mushroom is able to capture and consume timated.These two species were analysedbecause they nematodes.Hyphae of olderfungi produce nematotoxin. couldplay important roles in commercialgrowing of the Whennematodes come in contact with this substance, oystermushroom. A. avenae,afungal-feedingnematode, theybecome immobilised. Then hyphae penetrate into ne- canbe apestof ediblemushroom, and the entomopatho- matodebody and digest it. Some authors proved that the genicspecies S. feltiae couldbe appliedin biologicalcon- oystermushroom is capable of destroying the bacterial trolof pestsof theoyster mushroom,  iesbelonging to the feedingnematode, Panagrellusredidivivus ,andprepar- asiticlarvae of several animal parasites. In this study family Sciaridae.Theresults of thelaboratory tests indi- survivalof two species, Aphelenchusavenae and Stei- cated that A. avenae was moresusceptible than S. feltiae nernemafeltiae ,affectedby oyster mushroom, was es- tothenematotoxin produced by theoyster mushroom.

752 Nematology Symposiumabstract

New Musa hybrids with partial resistance to Radopholus similis

Carine DOCHEZ, Paul R. SPEIJER†, John HARTMAN and Dirk VUYLSTEKE InternationalInstitute of Tropical Agriculture (IITA), Eastern and Southern Africa Regional Centre ( ESARC), P.O.Box 7878, Kampala,Uganda †Deceased Presentedat the 25th International Nematology Symposium, Herzliya, Israel, 2-7 April 2000

Several Musa hybridswere testedfor nematoderesis- ingall vermiform developmental stages and sexes. From tanceand compared to thereference cultivars Y angambi- thesecounts, the reproduction ratio was calculated.All km5 (resistant),Gros Michel(partially resistant) and genotypesshowed a lowerreproduction ratio than V alery. Valery(susceptible). Genotypes included the diploid hy- Thegenotypes Gros Michel,2521S-31 and -47, 1411S- brids2521S-31, -47 and -50, 1411S-2 and -10, 2569S- 10,2569S-2, 2094S-1 and TMHx 660K-1 showed a re- 1and-2 and the tetraploid hybrids 2094S-1 and TMHx productionratio that was notsigniŽ cantly different from 660K-1.Genotypes were plantedin wooden boxes con- Yangambi-km5. Genotypeswith low reproduction ratios tainingsterilised sawdust. From eachplant, three roots support low R. similis densitiesand are therefore interest- were selectedand a plasticcontainer was placedaround eachof them.Inoculation was doneby pouring a suspen- inggenotypes for furtherevaluation. Except for theEast sionof 50 females of Radopholussimilis oneach indi- Africanhighland banana hybrid TMHx 660K-1, all the vidualroot. Eight weeks after inoculation, the roots were otherhybrids have the highly R. similis-resistantPisang harvestedand the number of nematodescounted, includ- JariBuaya in theirpedigree.

Thescope and limitations of acomputer programme for the integrated control of potato cyst nematodes

Martin ELLIOTT 1, Mark S. PHILLIPS 1, James W. MCNICOL 2 andDavid L. T RUDGILL 1 1 ScottishCrop Research Institute, Invergowrie, Dundee, DD2 5DA,UK 2 Biomathematicsand Statistics Scotland, SCRI, Invergowrie, Dundee, DD2 5DA,UK Presentedat the 25th International Nematology Symposium, Herzliya, Israel, 2-7 April 2000

Fieldtrials at different sites prepared with plots span- astheduration and rates of PCNdeclineduring the years ninga rangeof populationdensities of potatocyst nema- betweenpotato crops. The programme has demonstrated tode (PCN, Globoderapallida )provideddata for devel- thedifŽ culties many farmers facein managing G. palli- opinga computer-basedprogramme for modellingPCN da,theneed to prevent small populations from increasing populationdynamics and yield losses. Potato yields and andthe problems of decreasinglarge populations. The ef- PCN multiplicationwere welldescribed by density de- fectivenessof integratedmanagement has been shown to pendentequations which were madepotentially predic- bevery sensitive to differences in the percentage “ kill” tiveby incorporatingfunctions for consistentsite and cul- bynematicides and rates of population decline between tivardifferences in tolerance. The current programme al- potatocrops. Even quite low levels of partial resistance lowsfor differencesin site and cultivar tolerance, yield havebeen shown to be useful when combined with ne- potentialand resistance. The effects of differentmanage- maticides.The challenge now is to devise realistic sam- mentstrategies can be examined by changing the effec- plingstrategies for generatingdata to allow the model to tivenessof nematicides and/ orresistantcultivars as well beappliedin speciŽ c situations.

Vol.2( 7),2000 753 Symposiumabstract

Development of aseptic culturesystems of Radopholus similis for in vitro host-pathogen studies

Annemie ELSEN, Ruth STOFFELEN and Dirk DE WAELE Laboratoryof Tropical Crop Improvement, K.U. Leuven,K. Mercierlaan92, 3001 Heverlee, Belgium Presentedat the 25th International Nematology Symposium, Herzliya, Israel, 2-7 April 2000

Radopholussimilis isone of the most damaging nema- ductionratio increased up to 223.2 after 12 weeks. Re- todesin bananas. Chemical control is currentlythe most sultsof agreenhousetest showed that R. similis did not usedmethod, but nematode control through genetic im- loseits pathogenicity after culturing on alfalfa callus. In provementis widely encouraged. The objective of this asecondpart the infection and reproduction of thenema- studywas toestablish an aseptic culture system for R. todescultured on the callus was studiedon both in vitro similis andto determinewhether R. similis caninfect and bananaplantlets and A. thaliana.Itwas shownthat R. sim- reproduceon in vitro bananaplantlets and Arabidopsis ilis couldinfect and reproduce on both banana and A. thaliana. Ina Žrst parta suitableaseptic culture system thaliana.Furthermore,nematode damage was observed was developedusing alfalfa callus. Radopholussimilis inthe root system of bothhosts. These successful infec- couldpenetrate and reproduce in thecallus: six weeks af- tionsopen new perspectives for rapid in vitro screening terinoculation with 25 femalesthe reproduction ratio was for resistanceof the existing banana cultivars and anti- 26.3and all vermiform stages were present.The repro- nematodeproteins expressed in A. thaliana.

Aphelenchoidoidea nematodes of phytopathological interest and their distribution inSpain

Miguel ESCUER, María ARIAS andAntonio B ELLO DptoAgroecologí a, Centrode CienciasMedioambientales, CSIC. Serrano 115 dpdo, 28006 Madrid, Spain Presentedat the 25th International Nematology Symposium, Herzliya, Israel, 2-7 April 2000

Previousreferences of plantparasitic nematodes belong- A.fragariae and A.ritzemabosi havebeen found in sev- ingto the superfamily Aphelenchoidoid eain Spain have eralSpanish areas. A.ritzemabosi hasonlyappeared in La beenreviewed and new samples have been studied, due PalmaIsland, in theCanary Island. Quarantine nematodes totheir interest as pathogens to rice crops, edible mush- A. besseyi,apathogenof ricecrops, and Bursaphelenchus rooms,ornamental plants, strawberry crops and pine tree xylophilus,causalagent of pinewiltdiseases affecting sev- woodlandsand for havingquarantine species. Aphelen- eralconifer species and recently reported from Portugal, choidesbicaudatus, A. blastophthorus,A. composticola, havenot been found.

754 Nematology Symposiumabstract

VAMfungi increasenematode resistance, growth, and nutrient uptake of Želd grown apple trees

Thomas FORGE, Andrea MUEHLCHEN, Clemens HACKENBERG, Gerry NEILSEN andThierry V RAIN Agricultureand Agri-F oodCanada, P aciŽc Agri-FoodResearch Centre, Summerland, BC, VOH 1ZO,Canada Presentedat the 25th International Nematology Symposium, Herzliya, Israel, 2-7 April 2000

Theeffects of V esicularArbuscular Mycorrhizal (V AM) mosseae and G.intraradices supportedsigniŽ cantly fewer fungion the development of Pratylenchuspenetrans and P.penetrans perg rootthan G.etunicatum and G. clarum, growthof Ottawa 3 applerootstock were studied.Six butwere notdifferentfrom controls.Colonisation of roots speciesof fungiwere evaluatedin twogreenhouse exper- byV AMwas notaffected by nematode inoculation. G. iments,and two species were selectedfor a2-yearŽ eld mosseae and G.intraradices increasedrootstock growth experiment. Glomusmosseae increasedtotal dry weights andnutrient uptake in the Ž eld.After twoyears in the ofnematode-inoculatedand non-inoculated plants in both initialgreenhouse experiments, and G.intraradices,G. Želd,numbers of P.penetrans perg rootand per 50 ml versiforme and G.etunicatum eachincreased dry weights rootzone soil were signiŽcantly lower for G. mosseae- inone of the two experiments. Plants inoculated with G. inoculatedplants than for controlplants.

Cuticlestructure among the Tylenchs

Etienne GERAERT Departmentof Biology, Gent University, Ledeganckstraat, 35, 9000 Gent, Belgium Presentedat the 25th International Nematology Symposium, Herzliya, Israel, 2-7 April 2000

Itis wellknown that cuticle ultrastructure differs among tionallayering of thecortex as well in Criconema,Hemi- thevarious tylenchs. When the cuticle is rather thin a sim- cycliophora as in Tylenchulus. Inthe family Hoplolaimi- ilarstructure is foundthroughout the several systematical daeand in the subfamily Heteroderinae the thick cuticle groupings.When the cuticle is thicker or becomesthicker istheresultof anadditional layer or layersunderneath the somespecial patterns are found that can be usedto charac- terisesome systematical subdivisions: in the superfamily striatedlayer .Inall other tylenchs the striated layer is the Criconematoideaa thickcuticle is the result of an addi- basallayer .

Vol.2( 7),2000 755 Symposiumabstract

Ultrastructure of syncytia after micro-injection of barnase

Wladyslaw GOLINOWSKI 1, Petra VOSS 2, Miroslaw SOBCZAK 1, Stephan OHL 3 andFlorian M.W .G RUNDLER 2 1 Departmentof Botany, W arsawAgricultural University, Rakowiecka 26/ 30,02528 W arsaw,Poland 2 Instituteof Phytopathology,Hermann-Rodewald Str .9,24118 Kiel, Germany 3 ZENECA-MOGEN, Einsteinweg97, 2333CB Leiden, The Netherlands Presentedat the 25th International Nematology Symposium, Herzliya, Israel, 2-7 April 2000

Syncytiainduced by Heteroderaschachtii in roots of Ara- readyone day after microinjection, degenerating syncytia bidopsisthaliana were usedfor microinjection.As acon- were found.Their protoplasts were stronglyaccumulated trol,lucifer yellow (L YCH) dissolvedin PBS was taken. sothat only remnants of cytoplasm were foundin wide After injection,L YCHspreadwithin the syncytia and was areasof the former syncytium.Four days after injection, readilytaken up by the associated juveniles. At theul- thetreated syncytia contained only membraneous mater- trastructurallevel, these control injections were foundto ial,vesicles and osmiophilic granules. Degeneration was causeconspicuous swirling and accumulation of thesyn- strictlyconŽ ned to the treated syncytia. In some cases de- cytialendomembranes and the formation of numerous generationof bothsyncytia and nematodes was retarded. vesicles.Injections of barnasetogether with L YCHcaused Thecytoplasm of thesesyncytia appeared to be stillalive amortalityof about70% of theassociatednematodes. Al- butwas almostfree ofribosomes.

Pathogenic potential of the root-knot nematode, Meloidogyne incognita , on rice inV enezuela

Nicola GRECO 1, Renato CROZZOLI 2, Franco LAMBERTI 1 andAntonio B RANDONISIO 1 1 Istitutodi NematologiaAgraria, C.N.R., ViaAmendola165/ A,70126Bari, Italy 2 Institutode Zoología Agrícola, F acultadde Agronomía, Universidad Central de V enezuela,Apdo 4579, 2101-A Maracay, V enezuela Presentedat the 25th International Nematology Symposium, Herzliya, Israel, 2-7 April 2000

Anexperiment was conductedin a glasshouseto relate shootweight and tiller height of ricewere 8,6.6,6.6 and densitiesof three V enezuelanpopulations of Meloido- 4,2.4 and 5.5 eggs and juveniles/ cm 3 soilfor theZulia, gyneincognita withrice growth. Nematode densities were Barinasand Lara populations, respectively. A minimum 0,0.25, 0.5, 1, 2, 4, 8, 16, 32, 64, and 128 eggs and relativeyield of 0couldhave been achieved at 520-420, 3 juveniles/cm soilfor theZulia population and 0, 2,8,32, 420-256,156-226 eggs and juveniles/ cm 3 soil,for plant 3 64,and 128 eggs and juveniles/ cm soilfor theBarinas weightand shoot height, for theZulia, Barinas and Lara andLara populations. Tiller height was recordedat Ž ve populations,respectively. Nematode maximum reproduc- plantstages and fresh plantweight and nematode popu- tionrates were 50,19.5, and 57-fold while equilibrium lationsin roots and soil were determinedat harvest. The 3 Larapopulation was themost aggressive; plants in pots densitieswere 119,256, and 62 eggs and juveniles/ cm inoculatedwith 128 nematodes/ cm 3 soilwere yellowing soil,respectively, for thepopulations from Zulia,Barinas atemergence and died 40 dayslater .Tolerancelimits for and Lara.

756 Nematology Symposiumabstract

Toxicity of 1,3-dichloropropene (TeloneII )to potato cyst nematodes

Ivan GROVE andPatrick H AYDOCK PlantNematology and Soil P estsGroup, Crop and Environment Research Centre, Harper Adams University College, Newport, Shropshire,TF10 8NB, UK Presentedat the 25th International Nematology Symposium, Herzliya, Israel, 2-7 April 2000

TeloneII, asoilfumigant containing 94% 1,3-dichloro- mentby 25.26mg.litre/ soil.day1,3-D (equivalent to 50.5 propenein both the cis and trans isomers,is applied in l/haŽeldapplication). In conclusion, both air-vapour and theŽ eldat 225 l/ hausing specialist equipment. Experi- soil-waterphase fumigation with Telone II, showshigh mentsdetermined air-vapour and soil-water phase toxic- toxicityto PCN. TeloneII willcontrol PCN atdosages Globodera ityof T eloneII to spp.Air-vapour phase fu- belowthe current Ž eldapplication rates when used in a migationat 4.37 mg.litre.day 1,3-D almost completely in- controlledenvironment. Field applications of T eloneII, hibitedhatching of amixed G.rostochiensis and G. pal- lida populationover 14 weeks. In a sandyloam soil the however,should remain at 225l/ hauntilfurther research mixed G.rostochiensis and G. pallida populationwas pre- canidentify whether there is scope to reduceapplication ventedfrom hatchingfor upto 7 weeksinto the experi- ratesfor commercialuse.

Ahighly speciŽc cDNA library from nematode-induced syncytia

FlorianM.W .G RUNDLER, Petra VOSS andPiotr S. P UZIO Institutfü r Phytopathologie,Universitaet Kiel, Hermann-Rodewald-Strasse 9, 24098Kiel, Germany Presentedat the 25th International Nematology Symposium, Herzliya, Israel, 2-7 April 2000

Anewmethod for theisolation of speciŽ c RNA from distincthybridisation signals. The cDNA ofpyk20,a gene nematodefeeding structures (NFS) inducedby cyst ne- from A. thaliana whichis highly active in NFS, was used matodeswas developed.With the aid of a microcapil- asaprobe.Gene expression could be monitoredby North- larythe cytoplasm was extractedfrom NFS of Heterodera ernblotting and RT -PCRwith isolated RNA equivalentto schachtii in roots of Arabidopsisthaliana .Inorder to de- 25to50and 3 to5 NFS, respectively.Using a PCRbased termineRNA quality,total RNA was isolatedand then hy- systema highlyspeciŽ c cDNA librarywas constructed bridisedwith a 18SrDNA probe.The detection limit for whichconsisted of approximately2.5 ´ 106primary re- theisolated RNA was determinedby probing total RNA combinants.This library opens anewdimension in the ef- of15,25 and 50 NFS with18S rDNA. Theresults indi- fortsto cloneplant genes which are speciŽ cally regulated catedthat an equivalentof 15NFS issufŽ cient to produce in NFS.

Vol.2( 7),2000 757 Symposiumabstract

Use of the green uorescent protein to study the localization of antagonistic bacteria innematode-infested plants

Johannes HALLMANN 1, Andrea QUADT-HALLMANN 1, W.G. MILLER 2, Ste E. LINDOW 3 andRichard A. S IKORA 1 1 Institutfü r Panzenkrankheiten, Nuß allee 9, 53115 Bonn, Germany 2 USDA-FoodHealth and Safety Unit, Albany, CA, USA 3 Departmentof Plantand Microbial Biology, 111 Koshland Hall, University of California, Berkeley, USA Presentedat the 25th International Nematology Symposium, Herzliya, Israel, 2-7 April 2000

Plant-associatedbacteria have been reported to beantag- theendophytic phase of colonization, especially in the onisticto plantparasitic nematodes making them suitable presenceof plant-parasiticnematodes. For thefollowing candidatesfor biologicalcontrol strategies. Modes of ac- studieswe utilizedthe green  uorescentprotein as abac- tioninclude: competition for nutrientsand space, produc- terialmarker to detect, identify and localize the introduced tionof metaboliteswith nematicidal activity and bacterial- antagonisticbacteria. The plasmid pGT -trpcontaining a mediatedinduction of plantdefense mechanisms. In gen- trppromoter-GFP transcriptional fusion was introduced eral,these antagonistic bacteria have been considered to intothe biocontrol strain Rhizobiumetli G12 by mobi- bethorough colonizers of the rhizosphere, however, re- lization,and studied in Meloidogyneincognita -infested centstudies have shown that some isolates colonize the potato and Arabidopsis plants.Bacterial colonization was endorhiza.W oundson theroot surface caused by thene- evaluatedusing epi uorescent and confocal laser scan- matodeseem to play an important role in bacterial col- ningmicroscopy. The plasmid pGT -trpwas stablefor onizationand plant-nematode-ba cteriainteractions. They increaseleakage of root exudates which stimulates mi- morethan 80 bacterialgenerations without selection and crobialpopulation growth in the rhizosphere, and also thegreen  uorescenceit conferred allowed detection of favoursbacterial entry into the plant tissue. Whereas rhi- singlebacterial cells. The presentationwill summarize the zospherecolonization by antagonistic bacteria has been resultson bacterialcolonization with emphasis on nema- studiedintensively in the past, very little is knownabout todefeeding sites.

Anew speciesof cyst nematode ( Heterodera sp.) on Panicum coloratum in Egypt, with observations on closelyrelated speciesdescribed on various grasses and ricein Florida and Louisiana inthe United States

Zafar A. HANDOO 1 and I.K.A. IBRAHIM 2 1 USDA, ARS, NematologyLaboratory, Beltsville, MD 20705-2350,USA 2 Departmentof PlantP athology,F acultyof Agriculture,Alexandria University, El-Shatby, Alexandria, Egypt Presentedat the 25th International Nematology Symposium, Herzliya, Israel, 2-7 April 2000

In1997, an undescribed cyst nematode ( Heterodera sp.) ofthis species. It differs from H.leuceilyma in having closelyrelated to H.graminophila Golden& BirchŽeld, second-stagejuveniles (J2) witha shorterstylet (stylet 1972 and H.leuceilyma Di Edwardo& Perry,1964 was length23 (22.5-23.5) vs 26 (23-28 m m)) andin the shape foundassociated with Qasabagrass roots ( Panicumcol- ofJ2stylet knobs, anchor-shaped vs prominentlyrounded. oratum)growingnear date palm in Alexandria, Egypt. Thenew species differs from H.graminophila in that it Soilsamples around roots also yielded several juveniles hasJ2 with longer bodies and with longer, more hya-

758 Nematology Symposiumabstract linetail termini than those described from Louisianaand lengthin the Florida population of H.graminophila is Floridapopulations: body length 516 (485-550) vs 430 shorterthan that of thenew species from Egypt18.2(17.6- (380-460 mm)inthe Louisiana population on barnyard 18.6) vs 23(22.5-23.5 m m). Themale specimens pre- grass (Echinochloacolonum ), and391 (374-412 mm) in viouslyreported in H.graminophila and H.leuceilyma theFlorida population on roots of Panicumrigidulum , were absentin thesamples from Egypt.The known distri- hyalinetail terminus 40 (35-43) vs 32 (25-38 m m) in the butionin Alexandria, Egypt is restricted to theoriginal site Louisianapopulation and 27 (23-31 mm)inthe Florida thusfar .Becausethis species is limitedin distribution,its population.The new species also has bullae in thecysts, economicimportance in rangeland grasses and cultivated whichare absent in H.graminophila .Also,the J2 stylet cropssuch as riceis not known.

Yieldloss instrawberries caused by Aphelenchoides blastophthorus

Solveig HAUKELAND SALINAS and Kari BREKKE NorwegianInstitute of CropProtection, Department of Entomologyand Nematology, Hogskoleveien 7, 1432Aas, Norway Presentedat the 25th International Nematology Symposium, Herzliya, Israel, 2-7 April 2000

Aphelenchoidesblastophthorus isecto- and endoparasitic Symptomswere notexpressed in the Ž rst year;however, onleafand bud tissues of scabiousand some other plants; samplesof runners demonstrated that nematodes were italso readily reproduces on fungi. A.blastophthorus was present.The following spring symptoms were recorded recognizedfor theŽ rst timeas a problemin Norwegian asdistorted/ darkergreen leaves, and decreased growth. strawberryproduction in 1972. In recent years this ne- Inthis experiment, damage was mostsevere on Honeoye matodeappears to be increasingly important in straw- plantswhereas the Korona plants appeared less affected. berryproduction in some parts of the country. Symp- As theseason progressed, immediate symptoms became toms,runner-production and yield was comparedfor the lessclear and by July/ Augustit was difŽcult to differen- strawberryvarieties: Honeoye, Inga, Jonsok, Korona and tiateinfested and non-infested plants. For allthe varieties Zephyr.CertiŽ ed plants were artiŽcially infested with yieldloss and the number of runnersper plant was signif- 100nematodes per plant and planted outdoors in con- icantlyreduced in infected plants compared to uninfected Žnedmicroplots. Planting took place at the end of May. controls.

Vol.2( 7),2000 759 Symposiumabstract

Biological control agents against soilborne plant diseases and nematodes

Rüdiger HAUSCHILD, Johannes HALLMANN andRichard S IKORA Institutfü r Panzenkrankheiten, Phytomedizin in Bodenö kosystemen, Universitä tBonn, Nussallee 9, D-53115Bonn, Germany Presentedat the 25th International Nematology Symposium, Herzliya, Israel, 2-7 April 2000

Adiseasecomplex comprised of root-knot nematodes developnew approaches to control the disease complex (Meloidogyne spp.)and the wilt pathogen Fusariumoxys- ontomato by enhancement of antagonistic microorgan- porum f. sp. lycopersici causessevere economic losses isms.In a Žrst phase,the impact of different rhizobac- intomato production in the middle east and world wide. teriaon reduction of Fusarium and Meloidogyne attack Controlof thisdisease syndrome has been accomplished hasbeen analysed and effective strains selected. Remark- for decadesonly on thebasisof methylbromide soil fumi- ablereduction of bothpathogens by antagonisticbacteria gationwhich will ofŽ cially be bannedfor usein theyear 2001.Finding new, effective and environmentally safe al- willbe presented.Further experiments are on theway to ternativemanagement systems therefore is amajorchal- improveformulation and application techniques of there- lengefacing plant pathology. In our project we attemptto spectiverhizobacteria.

Theanalysis of light reected from the potato crop as adiagnostic assay for infection by potato cyst nematodes

William HEATH 1, Patrick HAYDOCK 1, Andrew WILCOX 1 andKenneth E VANS 2 1 PlantNematology and Soil P estsGroup, Crop and Environment Research Centre, Harper Adams University College, Newport, Shropshire,TF10 8NB, UK 2 IACR-Rothamsted,Harpenden, Herts, AL5 2JQ,UK Presentedat the 25th International Nematology Symposium, Herzliya, Israel, 2-7 April 2000

Potatocyst nematodes invade the roots of developing witha rangeof PCNdensities.Preliminary results indicate potatoplants, damaging the root structure and reducing thatPCN infestationcan be detected by spectral analy- theuptake of nutrients by the plant. Current methods of sis.Spectral readings from aŽeldexperiment correlated determiningPCN populationdensities are both expensive NormalisedDerived V egetationIndex (NDVI) withroot andtime consuming, providing only estimates of theaver- invasion (R 2 = .825, N = 138,cv. Estima) and was agepopulation levels within Ž eldswithout measurement supportedby a concurrentpot experiment ( r2 = .866, ofvariabilityor meansof identifying developing foci of N = 285,cv. Pentland Dell). NDVI isa broadmeasure infestation.Nematode infestation may alter both the struc- ofplant condition and a furtherexperiment is underway tureand chemical content of leaves.As eachelement in topursue spectral response features speciŽ c toPCN in- aleafre ects light at different wavelengths, correlation fection.Further development of theanalysis is envisaged ofa changein canopy re ectance with nematode infes- usingairborne remote sensing, leading to a comparisonof tationmay provide a methodof identifying foci of in- groundand airborne readings with data recorded from a festation.A hand-heldspectroradiometer has been used plannedseries of highresolution, infra-red environmental torecord canopy re ectance from potatoplants infected monitoringsatellites.

760 Nematology Symposiumabstract

Transformation with Agrobacterium rhizogenes inducesnematode resistance in Raphanus sativus roots

Petra HEINEN 1, Inke NITZ 1, Piotr S. PUZIO 1,WladyslawG OLINOWSKI 2, Miroslaw SOBCZAK 2, Stephan OHL 3 andFlorian M.W .G RUNDLER 1 1 Instituteof Phytopathology,Hermann-Rodewald Str .9,24118 Kiel, Germany 2 Departmentof Botany, W arsawAgricultural University, Rakowiecka 26/ 30,02528 W arsaw,Poland 3 ZENECA-MOGEN, Einsteinweg97, 2333CB Leiden, The Netherlands Presentedat the 25th International Nematology Symposium, Herzliya, Israel, 2-7 April 2000

Raphanussativus plants(cv. Siletina) susceptible to He- tiesof hairy roots. The structure of uninfected wildtype teroderaschachtii were transformedeither with wildtype roots(WT) andtransformed roots (WTHR andBSHR) Agrobacteriumrhizogenes (WTHR), orwith A. rhizo- was identical,but syncytial elements in WTHR andBSHR genes thatcarried the barstar gene fused to the promoter hypertrophiedonly slightly. The endoplasmic reticulalum rolD(BSHR). Inthe wildtype roots a susceptibleresponse (ER)ofyoungsyncytiawas foundto be lessabundant than withthe formation of females was observed,while in inWTbutno otherstriking differences occurred. Entering hairyroots only males reached the adult stage whereas theJ3stage, the syncytial cytoplasm of WTHR andBBHR manyjuveniles stagnated in theJ2 and J3 stage. This re- sponsealso occurs in resistant plants ( e.g. cv.Pegletta). becomeheavily stained and granular.They contained only Nodifference between WTHR andBSHR could be ob- few ERmembranesand other organelles while their cell servedso the response appears to bebasedon theproper- walldid not thicken. Later the syncytia fully degenerated.

Expression of anematode responsive promoter of A. thaliana in potato

Petra HEINEN 1, Piotr PUZIO 1, Stephan OHL 2 andFlorian M.W .G RUNDLER 1 1 Institutfü r Phytopathologie,Hermann-Rodewald Str .9,24118 Kiel, Germany 2 ZENECA MOGEN, Einsteinweg97, NL-2333 CB Leiden,The Netherlands Presentedat the 25th International Nematology Symposium, Herzliya, Israel, 2-7 April 2000

Apromoterwith high activity in nematode induced syn- ablybetween different lines and between different plants cytiawas taggedin Arabidopsisthaliana .Promoter:: gus withinone line. The most striking result was astrong constructsof different length were examinedfor their gusexpression upon wounding in bothroot and shoot tis- expressionin A. thaliana duringdevelopment and af- sue.In addition to the histological determination of pro- terinfection with Heteroderaschachtii and Meloido- moteractivity,  uorimetricgus assays were performed gyreincognita ,respectively.Based on these results pro- withplant material of twotransgenic potato lines. Respon- moterconstruct ppyk20-Cwas chosenfor furtherappli- sivenessto treatment with plant hormones was shown. cations.Potato plants cv. Desiree were transformedwith appyk20-C::gusconstruct viaA. tumefaciens . Regen- Thepromoter is currently tested for applicationin differ- eratedpotato plants were testedfor gusexpression in entanti-nematodestrategies. Potato was transformedwith rootsand shoots. In contrast to the expression in Ara- ppyk20-C::barnase/rolD::barstarand genes with nematici- bidopsis,expressionpatterns in potato varied remark- daleffects. Preliminary results will be shown.

Vol.2( 7),2000 761 Symposiumabstract

Extraction of free-living nematode stages from soilwith zonal centrifugation

G. HENDRICKX AgriculturalResearch Centre, Department of Crop Protection, Burg. V anGansberghelaan 96, 9820 Merelbeke, Belgium Presentedat the 25th International Nematology Symposium, Herzliya, Israel, 2-7 April 2000

Whenusing conventional centrifugation for extractionof Wateris addedto 200ml soilin abeakeruntil a volume nematodes,the extracted volume is limited; maximum of2lisreached. Of thissoil suspension, 1 liscentrifu- 50ml of soilcan be extracted. An apparatus was devel- gated.At theend of the process, nematodes are collected opedthat is ableto process100 ml ofsoil.The extraction ina100ml beaker. The apparatus can also be usedto ex- withthe apparatus is based on theprinciple of zonalcen- tractnematodes from asuspensionof blendedroots. Ex- trifugationand is fullyautomated. The machineis built as atablemodel and is 0.6 m high,1 mwideand 0.6mdeep. tractionswith the apparatus are reproducible, reliable and Allindividual parts, as wellas the control by aPLC(Pro- havea higherefŽ ciency that the conventional centrifuga- grammableLogic Controller), are contained in thisspace. tiontechniques.

SigniŽcant interceptions and other new or unusual plant-parasitic nematodes recordedin England 1996-1999

Sue HOCKLAND InvertebrateIdentiŽ cation T eam,Plant Health Group, Sand Hutton, Central Science Laboratory (CSL), Y orkYO41 1LZ, UK Presentedat the 25th International Nematology Symposium, Herzliya, Israel, 2-7 April 2000

TheInvertebrate IdentiŽ cation T eamin the Plant Health T.leviterminalis and Xiphinema spp.,all associated with Group,CSL, identiŽ es plant-parasitic nematodes and penjingfrom China, Helicotylenchustunisiensis on orna- otherinvertebrate pests for thePlant Health and Seeds In- mental Allium from Uzbekistan ,Hoplotylus cf. montanus spectorate,pest control consultants, farmers andgrowers. from Bhutan, Rotylenchulus cf. reniformis on Schefera Aselectionof notablenematode identiŽ cations made on cana from Cuba,and Scutellonemabradys on yams from importedmaterial is summarised here. The international SouthAmerica. Meloidogynechitwoodi hasbeen inter- tradeof ornamental plants has continued to grow in the ceptedin imported ware potatoes,but the pestremains ab- lastfew years,and comprises the most important route for sentfrom theUK. Theoccurrence of other,non-statutory, entryof non-indigenousspecies. Amongst notable inter- ceptionswere Aphelenchoidesbesseyi , various Helicoty- root-knotnematodes in importedpotatoes, such as M. ja- lenchus spp.including H.dihystera,Hirschmanniella mu- vanica,hasstimulated research into improved diagnosis cronata,T ylenchorhynchus cf. annulatus,T . cf. coffeae, toolsfor thisfamily.

762 Nematology Symposiumabstract

Improving detection and identiŽcation of root-knot nematodes inpotatoes by immunochemicaltechniques

Sue HOCKLAND, Chris DANKS, Sarah DUFF, John BANKS, Mark DELANEY, Louise MACLEOD and Sue CRACKNELL PlantHealth Group, Central Science Laboratory ( CSL),Sand Hutton, Y orkYO41 1LZ, UK Presentedat the 25th International Nematology Symposium, Herzliya, Israel, 2-7 April 2000

TheEuropean Plant Health Directive and phytosanitary gated. Meloidogynechitwoodi and M. hapla were used regulationsof many countries list several plant-parasitic asmodelspecies. An ELISA hasbeen developed incorpo- nematodesas quarantinepests. IdentiŽ cation is routinely ratingmonoclonal antibodies prepared by immunisation doneusing traditional microscope techniques to distin- ofjuvenile and female nematodes. This has enabled de- guishkey morphological features. This has always pre- tectionof nematodesin potatotubers and tomato root ex- sentedparticular difŽ culties because nematodes often tractsat levels not previously detected by visual inspec- havefew distinguishingcharacters, and the variability tion,but it is not yet able to distinguishbetween species. ofsome of these makes positive identiŽ cation difŽ cult. At CSL,the use of immunological techniques as reli- Associatedwork on improving electrophoretic tools for able,robust tools to assist in the detection and iden- diagnosisof M.chitwoodi willalso be illustrated. This tiŽcation of plant-parasitic nematodes has been investi- workis funded by MAFF (PlantHealth Division).

Cyst nematodes Heterodera spp. incereal Ž eldsin Norway —preliminary results

Ricardo HOLGADO 1, Christer MAGNUSSON 1, Stig ANDERSSON 2 and Janet ROWE 3 1 TheNorwegian Crop Research Institute, Plant Protection Centre, Dep. of Entomology and Nematology, Hø gskoleveien 7, 1432Å sNorway 2 SwedishUniversity of Agricultural Sciences, Dep. of PlantProtection Sciences, P .O.Box 44, S-230 53 Alnarp, Sweden 3 ICAR-RothamstedDep. of Entomology and Nematology, Harpenden Hertfordshire AL5, 2JQ,UK Presentedat the 25th International Nematology Symposium, Herzliya, Israel, 2-7 April 2000

Severalspecies of cystnematodes in the genus Heterodera isan increase in the damage in cereals in recent years. attackcereals. The most common, and maybe the most Earlierstudies have demonstrated Ha 51to be the dom- importantspecies, is the cereal cyst nematode (CCN) H. inatingpathotype of CCN inNorway. Our studiescon- avenae,whichwas Žrst reportedin Norwayin 1925. The Žrm thepresence of H. Ž lipjevi,andindicate the possible dominatingcereals in Norwayare barley, followed by oats presenceof new Heterodera spp.The increasing damage andwheat. Field damage by CCN isshown as patches mayrelate to the lack of resistance to Ha 51in current withuneven growth, which is most clearly seen in oats. cultivars,or changein thepathotype frequencies. The in- Inaddition to cereals, several species of grassare hosts of creasedacreage of wheat,and susceptible cereals as pre- CCN.Analysis of 218samplesfrom theperiod 1995-1998 cropsto oats and wheat could also contribute to the in- demonstratesthe genus Heterodera tobe commonin Nor- creaseddamage. The occurrence of species and patho- wegiancereal Ž elds. Heterodera hasbeen recorded from typesof cystnematodes on cerealsin Norwayneeds to be thesouthern part of Rogaland county, up to a position studiedfurther, and current cultivars need to be screened 65.5°Ninthe county of Nordland.The highest frequen- for resistanceagainst cyst nematodes. The developmentof ciesof occurrencewere observedin oats and wheat.There effectivenematode control strategies is a priority.

Vol.2( 7),2000 763 Symposiumabstract

IdentiŽcation of novel nematicidal compounds by microinjection into syncytia

Roland A. HOLZ 1, Linda VAN DER MEY 1, Marcel BREEDEVELD 1, Amanda CARLILE 2,FlorianM.W .G RUNDLER 3 andStephan A. O HL 1 1 ZENECA MOGEN, P.O.Box628, 2300 AP Leiden,The Netherlands 2 ZENECA Agrochemicals,Jealott’s Hill Research Station, Bracknell, Berkshire, UK 3 Institutfü r Phytopathologie,Hermann-Rodewald Str .9,24118 Kiel, Germany Presentedat the 25th International Nematology Symposium, Herzliya, Israel, 2-7 April 2000

Economicallyimportant plant-parasitic nematode genera, jectpotentially nematicidal compounds into Heterodera such as Meloidogyne , Heterodera and Globodera, are ob- schachtii feedingsites. Some compounds with known ne- ligatefeeders on plants. Attempts to culture them in the maticidalactivity, e.g. proteinaseinhibitors, tested posi- absenceof theirhost plant have been largely unsuccessful tivein this assay. Screening extracts from microbialor- sofar. Screensfor orallyactive nematicidal compounds ganismswe haveidentiŽ ed potentially novel sources of havetherefore mainly relied on bacterial feeders such as nematicidalactivity. Their biochemical characterisation is C. elegans. At ZENECA, we applya methodto microin- inprogress.

Antioxidant proteins of Globodera rostochiensis

John T. JONES 1, Lee ROBERTSON 2, Alison PRIOR 1, Dave KNOX 3 and Mark S. PHILLIPS 1 1 Mycology,Bacteriology and Nematology Unit, Scottish Crop Research Institute, Invergowrie, Dundee, DD2 5DA,UK 2 UniversidadAutonoma de Madrid,Departmento de Biologia, Laboratorio de Žsiologiavegetal, Cantoblanco, 28049 Madrid, Spain 3 MoredunResearch Institute, P entlandsScience P ark,Bush Loan, P enicuik,EH26 0PZ, UK Presentedat the 25th International Nematology Symposium, Herzliya, Israel, 2-7 April 2000

Plantparasitic nematodes are likely to be exposed to re- idase,thioredoxin peroxidase and superoxide dismutase. activeoxygen species generated by theirown metabolism Wehavebiochemically analysed the proteins encoded by andby plant defence mechanisms but almost nothing is thesegenes in orderto investigate their function. Our Žnd- knownabout how they protect themselves from oxida- ingssuggest that thioredoxin peroxidase, superoxide dis- tivestress. W ehavecloned cDNAs encodinga variety mutaseand a secretedfatty acid binding protein (GPSEC- ofsecretedantioxidant proteins of G.rostochiensis which mayplay a rolein protecting these parasites from active 2)mayfunction in the host-parasite interaction but that oxygenspecies generated by their hosts. Genes identi- otherantioxidant proteins such as glutathioneperoxidase Žedto date include secreted forms ofglutathioneperox- havea differentrole.

764 Nematology Symposiumabstract

ESTsfrom potato cyst nematodes —from sequences to functional analysis

John T. JONES 1, Vivian C. BLOK 1, Alison PRIOR 1, Mark S. PHILLIPS 1, Herman POPEIJUS 2, Erin BAKKER 2, Hans HELDER 2 and Geert SMANT 2 1 Mycology,Bacteriology and Nematology Unit, Scottish Crop Research Institute, Invergowrie, Dundee, DD2 5DA,UK 2 NematologyDepartment, University of Wageningen,W ageningen,The Netherlands Presentedat the 25th International Nematology Symposium, Herzliya, Israel, 2-7 April 2000

Expressedsequence tag (EST) projectsoffer arapidroute beencharacterised and approximately 10% of thecDNAs tothe discovery of novelgenes. Genes expressed in awide sequencedare apparently PCN speciŽc. Secretedproteins rangeof parasiticnematodes of medicalor veterinaryim- havebeen identiŽ ed includinga PCNhomologueof cho- portancehave been investigated using EST analysisbut rismatemutase, a cDNA recentlycloned from thegland thesetechniques are only now being applied to plant par- cells of Meloidogynejavanica .Whilelarger scale projects asiticnematodes. W ehaveundertaken a smallscale EST ofthis type will undoubtedly be important in the future, projectusing cDNA librariesmade from thetwo species ourefforts arenow focussed on analysing the function ofpotatocyst nematode Globoderarostochiensis and G. oftheproteins encoded by theidentiŽ ed genes. Function pallida inorderto assessthe utilityof thisapproach and to isbeinganalysed using large-scale approaches including identifycDNAs encodingabundantly expressed secreted differentialscreening and in situ hybridisation.More de- proteins.1000 sequences have been obtained from G. ros- tailedstudies on themost interesting genes are also under- tochiensisand 100 from G. pallida.Avarietyof geneshas way.

Thephloem-speciŽ c sucrose carrierAtSUC2 isactivated insyncytia induced by the beet cyst nematode Heterodera schachtii

Katja JUERGENSEN 1, Joachim SCHOLZ-STARKE 2, Norbert SAUER 2, Paul HESS 3, Aart J.E. VAN BEL 3 andFlorian M.W .G RUNDLER 1 1 Institutfuer Phytopathologie, Universitaet Kiel, Hermann-Rodewald-Strasse 9, 24098 Kiel, Germany 2 LehrstuhlBotanik II, UniversitaetErlangen, Staudtstrasse 5, 91058Erlangen, Germany 3 Institutfü r AllgemeineBotanik und P anzenphysiologie, Senckenbergerstrasse 17, 35390 Giessen, Germany Presentedat the 25th International Nematology Symposium, Herzliya, Israel, 2-7 April 2000

Syncytiaare metabolically highly active cell complexes Itcould be shown that the promoter of the sucrose car- andact as strongsinks within the roots. As thesyncytium rier AtSUC2which is exclusively expressed in the phloem issymplastically isolated from thesurrounding root tis- companioncells of uninfected plants is alsoactive in syn- suethe mechanism of phloem unloading appears to be cytia.T wodays after cyst nematode infection 80% of the apoplastic.Accordingly, the assimilate must be translo- inducedsyncytia showed GUS activity.The speciŽ c ex- cated via speciŽc transportproteins. T oidentifysuch a pressionof the gene product SUC2 was conŽrmed by transporter,transgenic Arabidopsisthaliana plants with RT-PCRand immunolocalisation .Wesuggestthe sucrose promotersof several sugar transporters fused to the re- carrier SUC2to be an important link in the assimilates porter gene b-glucuronidase(GUS) orGFP (green uo- translocationduring the syncytium differentiation and ne- rescentprotein) were infectedwith Heteroderaschachtii . matodedevelopment.

Vol.2( 7),2000 765 Symposiumabstract

Environmental factors affecting sexual differentiation inthe entomopathogenic nematode Heterorhabditis bacteriophora

Hamutal KAHEL-RAIFER and Itamar GLAZER Departmentof Nematology, ARO, VolcaniCenter ,BetDagan, 50250, Israel Presentedat the 25th International Nematology Symposium, Herzliya, Israel, 2-7 April 2000

Thepresent study was aimedat determining the in uence (“Dog-food”agar) resulted in reduction in theproportion ofvarious environmental factors on sex differentiation ofhermaphrodites. Nematodes introduced to the symbi- (SD)inthe entomopathogen icnematode Heterorhabditis oticbacterium obtained from othernematode strains (IS- bacteriophora HP88strain, under in vivo and in vitro cul- 5andIS-33) developedsimilarly to theculture reared on tureconditions. Injection of individualnematodes into last theHP88 bacteria. Rearing the nematodes at a temper- instars of Galleriamellonella resultedin development of a aturerange between 21 °C to 30°Calsodid not have a similarnumber of femalesand hermaphrodites (35-40%) signiŽcant effect on thesexual differentiation among ne- and20-25% males. Increasing the number of nematodes matodesthat were culturedon NGM. Theproportion of thatwere injectedinto the insectdid notchangethese pro- hermaphroditesincreased as starvationperiod of hatching portions.In smaller insects (0.7-1.5 cm long) an increase nematodejuveniles lengthened ( > 6h).The data obtained inthe proportion of hermaphroditeswas recordedas com- paredwith larger size cadavers (2.4-2.7 cm long). When inthepresent study strongly suggest that the main factor individualhermaphrodites were placedon NGM, thepro- affectingsex differentiation in H.bacteriophora is the nu- portionof hermaphrodites,females and male progeny was tritionsource. The practicaland biological implications of 63%,31% and 6%respectively.Rearing on richermedium theresults are discussed.

Survival and infectivity of Meloidogyne hapla, M.fallax and M.chitwoodi at different temperatures

C.J.(Hans) K OK and Ate DE HEIJ PlantResearch International, P .O.Box16, 6700 AA Wageningen,The Netherlands Presentedat the 25th International Nematology Symposium, Herzliya, Israel, 2-7 April 2000

Hatching,survival and infectivity of three temperate At highertemperatures the survival time and infectivity Meloidogyne specieswas comparedat 5, 10, 15, 20 and was stronglyreduced. The energyreserves of thejuveniles 25°C.Eggmasses were incubatedon hatching sieves was correlatedto infectivity. At lowtemperatures hatch- andthe hatched nematodes (J2) were countedover time. ing of M. fallax and M.chitwoodi was fasterthan that of Batchesof J2were inoculatedin containers with sand and M. hapla.Thisshows that M. hapla isless adapted to cold storedat thedifferenttemperatures. On sampling, J2 were temperaturesthan M.chitwoodi or M. fallax. The high ac- extractedfor survivalmeasurements. In other containers tivity of M. fallax and M.chitwoodi atlow temperatures lettuceor tomatowere plantedas bioassayplants to deter- indicatesthat delaying the time of sowingin spring could minethe infectivity of J2.The energy reserves of J2were beeffectiveagainst these nematodes. The dynamics of the determinedwith oil red O. At 5 °Cinfectivejuveniles of all hatchingprocess do notagree with the linear degree-days threespecies were foundeven after 1 yearof incubation. modelused for nematode-temperatureinteractions.

766 Nematology Symposiumabstract

IdentiŽcation and expression of transcription factors in Meloidogyne incognita-induced tomato giant cells

Hinanit KOLTAI,JenniferE. S CHAFF,MurielleG. T HIERY, Ann E. GREENE andDavid M. B IRD PlantNematode Genetics Group, Department of PlantP athology,North Carolina State University, Box 7616, Raleigh, NC 27695,USA Presentedat the 25th International Nematology Symposium, Herzliya, Israel, 2-7 April 2000

Weareinterested in the cascade of host gene expres- tobe spatially and temporally coincident with those of sionnecessary for formationand function of giant cells. Tkn2 KNOX gene,in marked contrast to its expression Onegene expressed in Lycopersiconesculentum giant patternin simple leaf plants. W ealsofound KNOX and cells,deŽ ned by the clone DB#280 ( Le-phan), encodes PHAN tobeco-expressedin giant cells, which might re- aMybtranscription factor that exhibits strongest iden- inforcea generalrole for thesegenes in maintenance of tity to PHANTASTICA (PHAN), from Antirrhinummajus. meristems(implying that giant cells exhibit meristem- PHAN isinvolved in the formation and maintenance of aticcharacteristics). Alternatively, Le-phan might play a meristems,and based on experimentsin maizeand Antir- moregeneral role in theestablishment of cellfate. Trans- rhinum mayact as anepigeneticrepressor of class-I knot- genicplant experiments currently being conducted in our tedhomeobox genes ( KNOX) or vice versa.Weexamined labwill reveal the functional relationship of PHAN and thepattern of Le-phan expressionin tomato in healthyand KNOX, includingthe role of thesegenes in theinduction nematode-infectedtissues, and found Le-phan transcripts andmaintenance of the pathological state.

Root-knot nematodes, Meloidogyne spp., inPoland

Stefan KORNOBIS PlantProtection Institute, Miczurina 20, 60-318 P oznan,´ Poland Presentedat the 25th International Nematology Symposium, Herzliya, Israel, 2-7 April 2000

At present,eight species of the Meloidogyne genus are ofPoland.The mostcommon root-knot nematode species knownin Poland. M.arenaria,M. javanica and M. incog- in Poland is M. hapla. Itoccurs both in open Ž eldand nita occurronly in glasshouses, because in open Ž elds glasshouses.Polish populations of thespecies cannot de- theycannot survive the winter conditions. M.ardenensis velopon cerealsand maize, but in theŽeldtheysurvive on was detectedthree times on birch roots. M. kralli occurrs dicotyledonousweeds.Within the populations there are sometimeson peatymeadows, on plants from the Cypera- cae family. M. duytsi was foundonly once on sandydunes threedistinct morphological groups of the second stage inwestern part of PolishBaltic coast. M. naasi was found juveniles.They signiŽ cantly differ in body length and all atlow population densities in two sites in north-west part relatedmeasurements.

Vol.2( 7),2000 767 Symposiumabstract

Host suitability of some important arable and green manure crops to Meloidogyne chitwoodi

Gerard KORTHALS, Rien NIJBOER andLeendert M OLENDIJK PAV: Appliedresearch for Arable farming and Field production of Vegetables,P .O.Box430, 8200 AK Lelystad,The Netherlands Presentedat the 25th International Nematology Symposium, Herzliya, Israel, 2-7 April 2000

Inthe south east of theNetherlands the root-knot nema- atehosts. Sugar beet and hemp decreased the population todes Meloidogynefallax and M.chitwoodi causeserious stronglyand can be considered as very poor hosts. The problemsin arable farming and the production of Ž eld greenmanure crops T agetesand Englishryegrass declined vegetables.From 1996until spring 1998 a Želdtrial was thepopulation as much as black fallow and are therefore conductedon soil with a naturalinfestation of M. chit- nonhosts. The crops oil radish, Italian ryegrass and white woodi.Duringtwo years the most important arable crops, mustardare poor hosts while rye is a moderatehost leav- Želdvegetables and green manure crops were grownin ingŽ nalpopulation densities on thesame level as maize. plots of 6 ´ 6minat least four replications. The host Furthermorethe results will be comparedwith our present statuswas determined.Each plot was sampledin March (Pi), October(Pf )andagain after winter in thenext March knowledgeon Meloidogyne .Thenewly developed infor- (Pi after). Potatoproved to be averygood host followed mationon host-statuswill be usedto control M.chitwoodi bysummer wheat, summer barley, and maize as moder- withinrotation schemes.

Survey of baited insectparasitic nematodes from the lowland regions of Switzerland

Igor KRAMER 1, Odile HIRSCHY 2 andJuerg M. G RUNDER 1 1 SwissF ederalResearch Station for Fruit-Growing, V iticultureand Horticulture, P .O.Box185, CH-8820 Wä denswil, Switzerland 2 Universityof Applied Sciences ( HSW),CH-8820 Wä denswil, Switzerland Presentedat the 25th International Nematology Symposium, Herzliya, Israel, 2-7 April 2000

About600 soilsamples were baitedfor insectparasitic ne- couldbelong to S. species B, S. speciesE1 and other matodeswith the larvae of thegreater wax moth Galleria notyet described species of Steinernema .Twostrains mellonella.In112 cases insect parasitic nematodes could of Heterorhabditismegidis and one of H.bacteriophora beharvestedfrom the Galleria cadaver.They belonged to couldbe isolated from sandysoils close to river banks the group of Steinernema (104strains), Heterorhabditis inSwitzerland. At leastŽ veadditional species could be (3) andother genera from Rhabditidae(5). Almosthalf recordedfor theŽ rst timein Switzerland. Species rich- of all Steinernema recordswere S.intermedium . Other nessseems to bemuchhigher in the lowlands than in the speciesfound, ranked in decreasing frequency, were S. Swiss Alps(Steiner, 1998). Also improved determination kraussei,S. afŽne, S. feltiae,S. bicornutum,S. carpocap- keysand the consequent checking of the lateral Ž eldre- sae anda singlerecord of a memberof the S. glaseri gionof theinfective juveniles could explain some of the group.Several specimens have still to be conŽ rmed and gainedspecies richness.

768 Nematology Symposiumabstract

Ty3-gypsy group of LTRretrotransposons inplant parasitic nematodes

Amar KUMAR, Julia GROSKI, Ann HOLT, Jane WISHART-LINDSEY GRAY, Vivian C. BLOK and Mark S. PHILLIPS ScottishCrop Research Institute, Invergowrie, Dundee, DD2 5DA,Scotland, UK Presentedat the 25th International Nematology Symposium, Herzliya, Israel, 2-7 April 2000

Thegenomic organisation of the T y3- gypsy groupretro- sesof synonymousand nonsynonymoussubstitutionssug- transposonsin three different plant parasitic nema- gestedthat there is a highpercentage of elements car- todes,namely Globoderapallida, G. rostochiensis and ryingfunctional RT domains in T y3- gypsy populations. Meloidogyneincognita hasbeen investigated using the Southernhybridization analyses are being conducted to partialsequences of the reverse transcriptase genes. A assesswhether T y3- gypsy elementsare present in variable populationof partial sequences of the reverse transcrip- copywithin geographically diverse races of each nema- tasegenes was ampliŽed using a pairof degenerated todespecies. This is the Ž rst reporton the presence of primersderived from theconserved domains of the LTR(longterminal repeat) retrotransposons in the plant gene.Sequence and phylogenetic analyses showed that thereverse transcriptase sequences among three nema- parasiticnematodes. Additionally, these elements have the todespecies are different but they are highly homoge- potentialto beusedas molecular markers for geneticdi- neouswithin each individual species. Moreover, analy- versitystudy within these plant parasitic nematodes.

Distribution patterns of virus symptoms and theirvectors Longidorus arthensis and L.macrosoma intwo cherry orchards

Paul KUNZ andJuerg M. G RUNDER SwissF ederalResearch Station for Fruit-Growing, V iticultureand Horticulture, P .O.Box 185, CH-8820 Wä denswil, Switzerland Presentedat the 25th International Nematology Symposium, Herzliya, Israel, 2-7 April 2000

Inthe central part of Switzerland, close to Arth and inHorgenwent always parallel with the proven presence inHorgen on the Lake of Zü rich, two different cherry of L.arthensis .Inthe orchard in Horgenit could be shown orchardswere sampledintensively for thepresence of thatthe spreading of thenematodeand the infestation with thenematodes Longidorusarthensis and L.macrosoma thevirus always started on a quadrantof thetree that is andtheir virus CR V(CherryRosette V irus)and RR V exposedtowards the focus of the nematode presence. L. (RasperryRingspot Virus), respectively.The analysis of macrosoma was foundonce in Arthbut in Horgen this ne- asequencewith four aerial photographs from thelast 18 matodeis distributed regularly throughout the whole or- yearsresulted in a calculationof thespreading of thevirus chard.The analysis of the nematode distribution pattern symptoms.On average, the speed of spreadingwithin the verticallyand horizontally did not give any evidence of orchardin Arthwas 1.9m peryear .Thevirus infestation competitionbetween L.arthensis and L.macrosoma.

Vol.2( 7),2000 769 Symposiumabstract

Marker-assisted selection( MAS)of the Ma genes for complete root-knot nematode (RKN)resistance in Prunus rootstocks

Anne-ClaireL ECOULS 1,Maria-JoseR UBIO-CABETAS 1,ElisabethD IRLEWANGER 2, Henri DUVAL 3, Georges SALESSES 2 and Daniel ESMENJAUD 1 1 INRA, UnitéSanté Vé gé tale et Environnement, Nematology Group, 06600 Antibes, France 2 INRA, Unitéde Recherchessur les Espè ces Fruitiè res et laVigne,33883 V illenaved’Ornon, France 3 INRA, Unitéde Géné tique et d’Amé lioration des Fruits et Lé gumes, 84140 Montfavet, France Presentedat the 25th International Nematology Symposium, Herzliya, Israel, 2-7 April 2000

TheMyrobalan plum ( Prunuscerasifera ; Prunophora aroundthe gene were identiŽed andtwo of themwere con- subgenus)is a rootstockspecies in which the clones vertedinto more reliable SCAR markers with the nearest P.2175,P .1079and P .2980are highly resistant (R) to (SCAL19) locatedat 3.7cM from Ma1.SCAL19has been alltested RKN speciesincluding Meloidogynearenaria evaluatedfor MASinanEuropeanrootstock programme (MA), M.incognita (MI), M. javanica (MJ)andalso basedon Myrobalanplum ´ Amygdalus (peach,almond anunclassiŽ ed population M. sp.Florida (FL) thatover- oralmond-peach)hybrids and provedto beusablereliably comesthe resistance in other commonly used Prunus inparticular to detect any of the Ma genesin segregat- sourcesfrom the Amygdalus subgenus.All three clones ingprogenies involving the resistance sources mentioned carrya singlemajor dominant gene, respectively desig- above,such as thepeach Nemared, the wildpeach Prunus nated Ma1, Ma2 and Ma3,consideredas allelic or closely davidiana andthe almond Alnem. In the Japanese plum linked,that confers a highand complete-spectrum re- (anotherspecies of the Prunophora subgenus,interfertile sistance(including at least MA, MI,MJ, FLand M. withMyrobalan plum), a newresistance gene ( R ) has mayaguensis ). Bulkedsegregant analysis associated with jap theRAPD orAFLP techniqueswere performedto de- beenevidenced from anintraspeciŽc crossJ13 (H) ´ J222 tectmarkers linkedto the Ma1 geneusing segregating in- (R)andappears to confera resistancesimilar to Ma. One traspeciŽc progeniesfrom P.2175( Ma1ma1) crossed by markerelaborated from SCAL19and located at 10-15cM severalhost (H) parents( ma1ma1). FourRAPD andthree from thisgene has been obtained and suggests that Ma AFLP markerscovering a totaldistance of approx. 27 cM and Rjap genescould be homologous.

Hoplolaimus sp. —an under-estimated parasite of banana roots inthe Jordan Valley,Israel

Yuval LEVY 1, Yair ISRAELI 1, Daniel ORION 2 and Evgeni KOZODOY 3 1 JordanV alleyBanana Experiment Station 15132 2 Departmentof Nematology,ARO, TheV olcaniCenter ,Bet-Dagan50250, Israel 3 PlantProtection and Inspection Services, Bet-Dagan 50250, Israel Presentedat the 25th International Nematology Symposium, Herzliya, Israel, 2-7 April 2000

Inthe past the spiralnematode Helicotylenchusmulticinc- reinfestation.In recent years, growers use tissue culture tus was identiŽed and recognized as the main limiting plants,with the advantage that they are nematode-free. factorto banana production in the Jordan V alley.One of Itwas thereforeassumed that this new planting system themajor problems was thatthe use of traditionalplant- wouldreduce the nematode population in banana plan- ingmaterial to establish a neworchard caused nematode tationsproviding that a cropcycle is alsoapplied. How-

770 Nematology Symposiumabstract everthis assumption has proved wrong. In many cases, ofthespiral nematode and its population dynamics. The plantingtissue culture plants even after proper crop rota- resultsof the survey show that many of the new planta- tionresulted in a rapidbuild up ofnematodepopulations tionsplanted with tissue culture were infestedby Hoplo- causingroot injury in the form oflesions and other vis- laimus sp.,most of them located in the southern part of iblesymptoms of root damage. T oinvestigatethis phe- theSea of Galilee.It was alsofound the Helicotylenchus nomenon,a 3year(1997-99) survey was conductedin multicinctus graduallydisappeared from thesoil after 2- theregion of theJordan V alleyaround the Sea of Galilee, 3yearsof crop rotation. It is concluded that the Hoplo- thatincluded 40 banana plots, with a total480 samples. laimus sp.is apolyphagousectoparasite that can survive Nematodeswere extractedfrom rootsand soil, and root croprotation systems, and a strategyto control it should damagewas assessed.This data clariŽ ed the distribution beconsidered.

Environmental characterisation of three speciesof the genus Aporcelaimellus Heyns, 1965 (Nematoda: Dorylaimida) insoutheastern Spain

Gracia LIÉBANAS 1, Raimundo REAL 2, Ana L. MÁRQUEZ 2 and Reyes PEÑA-SANTIAGO 1 1 Departamentode Biologí a Animal,V egetaly Ecología, Universidadde Jaé n, Paraje“ LasLagunillas s/ n”23071-Jaé n, Spain 2 Departamentode Biologí a Animal,F acultadde Ciencias, Universidad de Málaga, 29071-Má laga, Spain Presentedat the 25th International Nematology Symposium, Herzliya, Israel, 2-7 April 2000

Threespecies of thegenus Aporcelaimellus Heyns, 1965 terns(corotypes) have been detected for A.amylovorus havebeen collected in a recentstudy of dorylaimsfrom and Aporcelaimellus sp.,whereas the same did not occur theSierra Mágina Natural Park (province of Jaén, south- for A.obtusicaudatus. Speciesdistribution with respect to easternSpain): A.amylovorus (Thorne& Swanger,1936) 18abiotic/ bioticenvironmental factors was analysedus- Andrássy, 1959, A.obtusicaudatus (Bastian,1875) Al- inglogistic regression to characterisethe corotypes. Dis- therr,1968 and Aporcelaimellus sp.,the latter probably tributionof A.amylovorus ismainly in uenced by CaCO beingan undescribedspecies. Their distribution has been 3 studiedand analysed. The species were collectedin 30.0, content (y = 0.0280CaCO3 ­ 1.5766),whereas that of 79.8and 29.0%, respectively, of nearly200 soil samples Aporcelaimellus sp.is inuenced by altitudeand claycon- collected;two signiŽ cantly different distributional pat- tent (y = 0.0011Altitude + 0.0592Clay­ 4.3643).

Vol.2( 7),2000 771 Symposiumabstract

How isthe biodiversity of dorylaims inuenced by altitude ina natural area from southeastern Spain?

Gracia LIÉBANAS 1, Ana L. MÁRQUEZ 2, Raimundo REAL 2 and Reyes PEÑA-SANTIAGO 1 1 Departamentode Biologí a Animal,V egetaly Ecología, Universidadde Jaé n, Paraje“ LasLagunillas s/ n”23071-Jaé n, Spain 2 Departamentode Biologí a Animal,F acultadde Ciencias, Universidad de Málaga, 29071-Má laga, Spain Presentedat the 25th International Nematology Symposium, Herzliya, Israel, 2-7 April 2000

Thedistribution of thetaxocoenosis constituted by dory- normaldistribution in the studied area. A univariatelinear laimidnematodes has been studied in the Sierra Mágina regressionanalysis was usedto Ž ndout the relationship NaturalPark (province of Jaén, southeasternSpain). This betweenspecies richness ( y )andaltitude ( x );the follow- studyconsiders several environmental factors, altitude ingregression equation was obtained: y = 0.0066x + beingone of them. Over ninethousand specimens of 3.0749.Thus,the number of dorylaimid species signif- 138species collected in 203 soil samples have been icantlyincreases with altitude in the geographical area studied.Altitudinal range of the region is 600-2100 m. considered, i.e.,analtitudinal distributional pattern is de- Kolmogorov-Smirnovtest showed that altitude presents a tected.

Protein MJE36: astage- and genus-speciŽc putative collagen gene product detected in Meloidogyne javanica

Jia LIU 1, Hinanit KOLTAI 1, Nor CHEJANOVSKY 2 andYitzhak S PIEGEL 1 1 Departmentof Nematology,A.R.O., TheV olcaniCenter ,Bet-Dagan50-250, Israel 2 Departmentof Entomology, A.R.O., TheV olcaniCenter ,Bet-Dagan50-250, Israel Presentedat the 25th International Nematology Symposium, Herzliya, Israel, 2-7 April 2000

Wehavecloned and expressed a fusionprotein contain- tractedby BME, whichsuggests that MJE36 expression inga nonconserved58-amino acid sequence encoded by isstage-speciŽc. Abandof thesame molecular size was aDNA sequencebetween the homology block A andthe detectedwith M.incognita eggsextracted by BME but Žrst (Gly-X-Y)n blockfrom Meloidogynejavanica Mj- notdetected with Heteroderaavenae. TheseŽ ndingsim- col-4gene. Rabbit antiserum, which was raisedagainst plythat the expression is alsogenus-speciŽ c. M. javanica thefusion protein, detected a strongreacting protein band eggsimmunolabelling assays showed that the antiserum (Mw 36kDa) onwestern blots of M. javanica eggs ex- reactedonly withthe broken M. javanicaeggsdensely and tractedby 2-mercaptoethanol (BME). This protein was designatedas MJE36 and was foundto be collagenase irregularly,while no reacting bands were detectedwith sensitive.MJE36 was notdetected on westernblots of M. puriŽ ed M. javanica eggshellsextracted by BME, sug- javanica second-stagejuveniles (J2) oradultfemales ex- gestingthat MJE36 was notin M. javanica eggshells.

772 Nematology Symposiumabstract

Theimprovements of enthomopatogenic nematodes inpest control

Attila LUCSKAI and Miklos NADASY PannonUniversity of AgriculturalSciences Georgikon F aculty,Department of Entomology, Deak F .str.57,8360 Keszthely, Hungary Presentedat the 25th International Nematology Symposium, Herzliya, Israel, 2-7 April 2000

Theentomopathogen icnematode procedure of control ecologicalconditions, so it is preferable to choose a pe- canbe wellŽ ttedinto the integrated plant protection pro- riodto spray nematodes in late afternoon to avoid UV grammes.Nematodes practically occur everywhere in the radiationand warmth. These nematodes can be applied soils,but the density of thepresent population is notsufŽ- beforethe activityof soildwelling insects, and for thisin- cientto restrict the increase of pestsin agriculturalcrops. sectmonitoring is needed. Thus they can be employedas Onlyby multiplyingthem and bringingthem back to their apreventivedefence. The nematode preparations can be areaof origin can the individual number of pests be ef- storedfor alongertime when cooled, so it is advisable to Žcientlydecreased. T oimprovethe effectiveness of en- usethem rapidly. Of course,protection against pests can- tomopathogenicnematodes the following factors are im- notbe solvedby themere useof entomopathogens,even portant.Preliminary insect susceptibility tests and experi- ifanentomopathogenicnematodesuitable for allpests of mentsare needed to know which nematode and howmany acropis found,since e.g. weathercan be apowerfullim- shouldbe used.Effectiveness is greatlyin uenced by the itingfactor .

Meloidogyne hapla on Želdcrops inNorway

Christer MAGNUSSON and Bonsak HAMMERAAS TheNorwegian Crop Research Institute, Plant Protection Centre, Department of Entomologyand Nematology, Hø gskoleveien 7, NO-1432Aas, Norway Presentedat the 25th International Nematology Symposium, Herzliya, Israel, 2-7 April 2000

Inrecent years, the root knot nematode, Meloidogyne Damageon carrots, onion, lettuce and turnip has so far hapla,hasbeen found to damageŽ eldgrown vegetables beenrecorded in the southern counties of Vestfold,Aust- inNorway. In a globalperspective M. hapla is known to Agderand Rogaland. In these areas temperature sums occurin areas with annual mean temperatures of + 6°C reach 1000°C ´ daysin many localities. This would al- orhigher,indicating that Norwegian coastal areas, as far mostallow for thedevelopment of twogenerations of M. northas thecounty of Nord-Trøndelag, would offer pos- hapla,andrecurrent infections during the growing season sibilitiesfor nematodeestablishment. The development areexpected. M. hapla maycause severe losses of carrot of M. hapla isknown to require 554 °C ´ days over a crops.In one Želdinfested with M. hapla,muchof thecar- basaltemperature of + 8°C.InNord-Trø ndelag tempera- rotcrop was lost,and the precedinglettuce crop suffered a turesums, at a soildepth of 10cm,exceed 560 °C ´ days, 50%reduction in yield.Farmers needto takeprecautions whichwould allow for thecompletion of onegeneration. to avoid M. hapla infestations.

Vol.2( 7),2000 773 Symposiumabstract

Effect of Meloidogyne incognita on Želdperformance of cassava ( Manihot esculenta Crantz) genotypes

Nakato N. MAKUMBI-KIDZA 1, Paul R. SPEIJER1,† andRichard A. S IKORA 2 1 InternationalInstitute of Tropical Agriculture, Eastern and Southern Africa Regional Centre, P .O.Box7878, Kampala, Uganda 2 Soil-EcosystemPhytopathology and Nematology, Institut fuer P anzenkrankheiten, University of Bonn,Nussallee 9, 53115Bonn, Germany †Deceased Presentedat the 25th International Nematology Symposium, Herzliya, Israel, 2-7 April 2000

Inresponse to a devastatingoutbreak of cassava mo- plantlets(1000 infective juveniles per plant). The main saicdisease in the early 1990s, breeding programmes in plottreatments were genotypes.Root-knot nematodes re- EastAfrica havebeen concentrating on developing and ducedthe totalstorage root yield by asmuch as 25%.This releasingcassava mosaic genotypes tolerant to the dis- losswas mainlycaused by a reductionin the number of ease.Root-knot nematode problems on cassavahave been storageroots formed, which was signiŽcant ( P < 0.05) reportedfrom anumberof African countries, including for MH95/0324,MH95/ 0349and MH95/ 0196.The num- Uganda.Therefore, the more recently developed cassava berof large storage roots was reduced( P < 0.05) for genotypeswere alsoevaluated for resistanceand or toler- MH95/0324,MH95/ 0372,MH95/ 0161and MH95/ 0067. anceto root-knotnematodes. A Želdexperiment was set Theresults show that a relativelylow initial nematode upinwhich25 genotypeswere tested.Plots were divided pressureis able to cause signiŽ cant production losses in intosubplots of equal size. One plot was plantedwith cassavaand that cassava response to Meloidogyneincog- cleanplantlets, while the other was plantedwith infected nita (race 2)isgenotype dependant.

Integrated control strategies for potato cyst nematodes

Stephen MINNIS 1, Patrick HAYDOCK 1 andKenneth E VANS 2 1 PlantNematology and Soil P estsGroup, Crop and Environment Research Centre, Harper Adams University College, Newport, Shropshire,TF10 8NB, UK 2 IACR-Rothamsted,Harpenden, Herts, AL5 2JQ,UK Presentedat the 25th International Nematology Symposium, Herzliya, Israel, 2-7 April 2000

Inthe UK, themost problematic pests of thepotato crop cationwas signiŽcantly decreased with Santé compared arethe potato cyst nematodes (PCN) Globoderapalli- tothe susceptible variety Estima. 1,3-dichloroprop ene da and G.rostochiensis .Populationsof G.rostochien- andoxamyl signiŽ cantly increased speed of crop emer- sis canbe managedby the integrated use of nematicides genceand percentage ground cover .Thecombination of (granularand fumigant), crop rotation and resistant cul- Santé, 1,3-dichloropropeneand oxamyl at full-rate was tivars.A Želdexperiment was doneto compare the use thehighest yielding treatment. In addition, the use of 1,3- ofthe resistant cultivar Santé with 1,3-dichloroprop ene dichloropropeneincreased the overall number of tubers andthe granular nematicide oxamyl at full and half-rates andimprovedthe tubersize distribution. The resultsof the onan experimental site containing both species of PCN (mainly G.rostochiensis )ataninitialpopulation density experimentshow that an integratedapproach to nematode of 189 eggs g­ 1 ofsoil. Root invasion studies showed controlon heavilyinfested sites can lead to signiŽcant de- signiŽcantly different levels of invasionbetween cultivars creasesin nematode population levels and give economic andbetween fumigation treatments. Nematode multipli- yieldbeneŽ ts.

774 Nematology Symposiumabstract

Anew method to overcome underestimation of plant parasitic nematode populations

LeendertP .G.M OLENDIJK, Rien NIJBOER andGerard W.K ORTHALS PAV: Appliedresearch for Arable farming and Field production of Vegetables,P .O.Box430, 8200 AK Lelystad,The Netherlands Presentedat the 25th International Nematology Symposium, Herzliya, Israel, 2-7 April 2000

Modernagriculture depends more and moreupon a proper phenomenon,a newextraction method has been devel- croprotation or thecultivation of resistantvarieties in or- opedby which soil is incubated to allow egg hatching. derto controlspeciŽ c nematologicalproblems. In contrast SeveralŽ eldstudies were carriedout and gave in total tothe chemical way, this relies more heavily on aproper over3000 samples to estimate the impact of thissoil in- identiŽcation of thenematode species involved and an ac- cubationmethod. From thislarge database it was possible curateestimation of the population densities. Although toinvestigatethe in uence of samplingperiod, preceding manynew molecular tools have been developed to help cropand nematode species. The results clearly demon- theidentiŽ cation, there seems less interest in developing stratedthat, depending on these factors, the most com- newextraction methods to improve the estimation of pop- monlyused extraction methods may lead to dramatic un- ulationdensities. Within our work on different Meloido- derestimationand associated risks within today’s advice gyne species,it was observedthat some populations in- tofarmers. Thereforeit isstrongly recommended to usea creasedduring winter without crops. T oinvestigatethis properperiod of soilincubation.

Molecular mechanism of resistance and virulencein the Meloidogyne- tomato interaction

Sergio MOLINARI Istitutodi Nematologia Agraria, Consiglio Nazionale delle Ricerche, V iaAmendola 165/ A,Bari70126, Italy Presentedat the 25th International Nematology Symposium, Herzliya, Israel, 2-7 April 2000

The Meloidogyne -tomatointeraction has been extensively degradingenzymes causes an increase of oxidative and studied in vivo and in vitro.Different(a)virulent popula- peroxidativeactivity in root cells which ultimately leads tions of Meloidogyne spp.collected in Ž eldsor selected tocelldeath. The consequentformation of alargenecrotic inglasshouse were inoculatedon a numberof resistant areasurrounding the juvenile is responsiblefor thearrest tomatocultivars. Changes in antioxidant enzyme activi- ofnematode feeding and reproduction. Normally aviru- tiesof roots have been monitored from 24h afterjuve- lentpopulations can be selectedfor virulenceby repeated nileinoculation either in pots or in cultures of excised inoculationson resistant tomato cultivars. Near-isogenic roots.Catalase and ascorbate peroxidase activities were populationswere testedfor differencein electrophoreti- foundto belowerin infested roots in every incompatible calpatterns of antioxidantisozymes. More marked cata- interactiontested in vivo and in vitro.Isoelectrofocusing laseand peroxidase bands suggested a higheractivity separationsof rootcatalase isozymes showed that speciŽ c in virulent Meloidogyne females.Furthermore, evidence isozymeswere completelyinactivated due to nematode isemerging of a superoxidedismutase (SOD) isozyme infestation.Nematode-induced inhibition of such H 2O2- speciŽcally found in virulentfemales.

Vol.2( 7),2000 775 Symposiumabstract

Anovel apparatus for extraction of nematodes from soil

Mishael MOR Departmentof Nematology, Agricultural Research Organization, The V olcaniCenter ,P.O.Box6, Bet-Dagan,50250, Israel Presentedat the 25th International Nematology Symposium, Herzliya, Israel, 2-7 April 2000

Existingnematode extraction techniques are generally ter.This arrangement also facilitates in situ cleaning of limitedto certain nematode life stages (free-living or in- thecylinder after completion of theextraction operation ert) andsizes. Most methods are laborious and consist withoutthe need to dismantle the apparatus for quantita- ofseveral phases. The objectives in developing the new tivework. The facility is attached to a standardstainless methodwere toachieve extraction of awiderange of ne- sink(length, 120 cm; width, 40 cm) witha drain,serving matodeforms inasingleapparatus to simplifythe extrac- twoapparatuses. The apparatus is compact, simple and tionprocess and to enhance efŽ ciency and convenience convenientto handle. It is relativelycheap and has a set inoperation. Unlike other elutriation techniques, the new ofPerspexsieves (diameter, 15 cm). Oneperson can com- methodmakes use of an up current of small and dense fortablyoperate several units simultaneously. The method airbubbles in the suspension, rather than a water ow, isparticularlysuitable for separatinglarge or heavynema- for separatingnematodes from soilparticles and keeping todeswhich normally sink quickly. Y ieldsof nematode themin suspension. The basiccomponent of theapparatus suspensionare relatively free ofdebris or soil particles. isa graduatedPerspex scaled cylinder (length, 50 cm;di- Severalyears of experimentation with this method have ameter,10 cm;volume, 3 l)withwater entering through a shownthat from a200g soilsample 70-90% of the ne- perforatedstopper at the bottom, above which a streamof matodesare obtained, depending on soil type, nematode airbubbles is deliveredthrough a Perplextubule by means form, operatingtime, and number of repetitions. These ofapumpand ow-meter.At theupper end of thecolumn, experimentswere conductedboth with infested samples aplastictube which serves as acysttrap is attached;the takenfrom theŽ eldand artiŽ cially infested soil samples otherend of thetube is connected to a vacuumpump. The previouslyinfected in a laboratorywith a widerange of cylinderis mounted in ametalring Ž xedto anaxiswhich nematodespecies. Cyst retrieval with a vacuum-sucked allowsthe apparatusto revolve 180 °,therebyenabling for- streamof wateris 15%more efŽ cient than that obtained mationof awell-mixedsuspension of thesoil in the wa- withthe Fenwick technique.

On the identity of Heterodera latipons Franklin

Mishael MOR 1 and Dieter STURHAN 2 1 Departmentof Nematology, A.R.O., TheV olcaniCenter ,P.O.Box6, Bet-Dagan50250, Israel 2 BiologischeBundesanstalt, Institut fü r Nematologieund Wirbeltierkunde, T oppheideweg88, 48161 Mü nster ,Germany Presentedat the 25th International Nematology Symposium, Herzliya, Israel, 2-7 April 2000

Thecereal cyst nematode, Heteroderalatipons , was de- Re-examinationof H. latipons typespecimens showed scribedfrom Gilat,Israel, in 1969 and has been recorded thateven the type material consists of twomorphological sincethen in many countries, mainly the Mediterranean types,which are mainly distinguished by cystcharacteris- andthe Oriental regions. Comparative biochemical stud- tics,one typeagreeing with that Ž guredin the original de- iesof cystnematode populations of differentgeographical scriptionof H. latipons,theother resembling the related originprovided Ž rst evidencethat the species previously species H.hordecalis whichis mainly known in north- consideredas H. latipons isactually a mixtureof species. ernand central Europe. The H.“hordecalis” type was

776 Nematology Symposiumabstract alsofound among populations identiŽ ed as H. latipons in speciesof grassesand varieties of severalcereals. For ex- Libya,Iran andthe Azores. It could be isolatedeven from ample, Phalarisminor and P. paradoxa inGilatserve as thetype locality Gilat recently, where it occurred together goodhosts, whereas in a nearbylocality they are non- with H. latiponss.str .Populationsof H. latiponss. str. and hosts.There is evidence of an additional non-described H.“hordecalis” showeddifferences in hostrange among specieswithin the H. latipons speciescomplex.

Theeffects of predatory nematodes for harmful insects( Melolonthidae, Noctiudae, Blattidae) inHungary

Miklos NADASY 1, Gyula SARINGER 1, Attila LUCSKAI 1, Szilvia PEKAR 1, Andras FODOR 2, Csaba BUDAI 3 andMichael G. K LEIN 4 1 PannonUniversity of Agricultural Sciences Georgikon F aculty,Department of Entomology,Deak F .str.57,8360 Keszthely, Hungary 2 EotvosUniversity, Department of Genetic,Nuzeum krt. 4/ A,1088Budapest, Hungary 3 CsongradCounty Plant Health and Soil Conservation Station, Biological Control and Quarantine Development Laboratory, P .O. Box99, 6800 Hodmezovasarhely, Hungary 4 JapaneseBeetle Laboratory, O.A.R.D.C. Wooster,OHIO 44691,USA Presentedat the 25th International Nematology Symposium, Herzliya, Israel, 2-7 April 2000

Theentomopathogen icnematodes are potential biological treatedat two concentrations (100, 1000 and 10 000in- controlagents against several insect pests. In the course fectivejuvenile (IJ)/ ml) with4 replications.Mortality of oftheexperiments the following pests were subjectedto larvaewas evaluatedon 5 th and 7th dayafter treatment, examination: Scotiasegetum Schiff, Mammestrabrassi- respectively.Against white grubs, the 100 and 1000 IJ/ ml cae L., Melolonthamelolontha L., Polyphillafullo L., concentrationsgave an average of 25.4%andnearly 100% Blattaorientalis L. and Periplanetaamericana L. The en- mortality.The order of effectivenessagainst grubs was H. tomopathogenicnematodes used in theexperiments were: bacteriophora HH, S. glaseri, S.riobravis and H. bacte- Steinernemafeltiae Filipjev, S.carpocapsae Weiser, S. riophora AZ32.Against Noctuidae larvae H.bacterio- glaseri Steiner, S.scapterisci Nguyenand Smart, S. rio- phora HHwas themost effective, resulting in some50% brave Cabaillas et al., and Heterorhabditisbacteriophora mortality.The mortality of cockroachestreated with 100 Poinar.Theexperiments were carriedout in laboratory and10 000IJ/ mlper dish was 74and100%, respectively, culturepots at room temperature. The insect larvae were after 24 h.

Vol.2( 7),2000 777 Symposiumabstract

Testing inferenceof phylogeographical patterns under specieslevel in Meloidogyne arenaria

Alfonso NAVAS 1, Philippe CASTAGNONE-SERENO 2, Pilar FLORES-ROMERO 3 and Jesus BLAZQUEZ 1 1 MNCN, CSIC,Madrid, Spain 2 Lab.Biol. Invertebres, Antibes, France 3 H.Ramón yCajal,Madrid, Spain Presentedat the 25th International Nematology Symposium, Herzliya, Israel, 2-7 April 2000

Fortyeight isogenetic isolates have been used to de- numberof repeatcopies (n5 to n12) inthe isolates. Allele scribethe pattern of distributionof M. arenaria at popu- frequenciesseem to have a randomdistribution at species lationlevel in arestrictedtobacco monoculture area. Pre- level.However allele frequencies are not randomly dis- viousgenetic characterisation of the isolates resulted in tributedinto MLEE variants showing, in addition,statisti- sixMultilocus Enzyme Electrophoresis (MLEE) variants. calsigniŽ cant differences among them. Nei’s minimum As HighlyPolymorphic T andemRepeat loci seem to be andstandard genetic distances ( Dm, Ds) andstepwise promisingmolecular markers, we havefocused on appli- weightedgenetic distance ( Dsw) arecalculated and com- cationof mitochondrial VNTRs (VariableNumber T an- pared.According to that and stepmatrices analysis, hy- demRepeats) as indicatorsof relatednessamong isolates potheticalphylogenetic relationships among MLEE vari- andMLEE variants. PCR ampliŽcation was performed antshave a highhomoplasy while these relationships are usingDNA from individualfemales centered in the 63 moreconsistent among isolates within each MLEE vari- lengthbp mitochondrial VNTR. This locus showed high ant.The phylogenies of isolatesare geographically repre- heterozygositywith eight different alleles according to the sentedand epidemiological considerations are discussed.

Heat shock genes from the entomopathogenic nematode Heterorhabditis bacteriophora (HP88)

Yael NEVO-CASPI 1, YaŽ t ATIYA-NASAGI 1, Revital BRONSTEIN 1, Itamar GLAZER 2 and Daniel SEGAL 1 1 Dept.Molecular Microbiology & Biotechnology,T el-AvivUniversity, Israel 2 Dept.Nematology, V olcaniCenter ,ARO, BetDagan 50250, Israel Presentedat the 25th International Nematology Symposium, Herzliya, Israel, 2-7 April 2000

Entomopathogenicnematodes,such as Heterorhabditis purposewe havecloned from thecommercial strain of bacteriophora ,arehighly effective bioinsecticides against H.bacteriophora (HP88) cDNAs andgenomic fragments awiderange of pests.However, their sensitivity to envi- correspondingto two genes from the HSP70 family. We ronmentalextremes, especially heat and desiccation, pre- foundthat these proteins are > 90%similar to Hsp70A ventsthem from achievingtheir full biocontrol potential. andto Hsp70C of C. elegans.Northernanalysis showed Heattolerance is achieved in organisms by expressing that the HSP70A and HSP70C genesare overexpressed highlevels of heatshock proteins ( HSP). Transgenicen- (2- and3-fold, respectively) following a 4-hheat shock tomopathogenicnematodes expressing high levels of an at 33°C.Wefoundthat a naturalnon-commercial heat HSP gene from C. elegans havebeen shown to display tolerantisolate of Heterorhabditis (IS5) overexpresses enhancedheat tolerance. W ereasonedthat releasing into HSP70A and HSP70C 5-and2.5-fold, respectively, with theŽ eldtransgenic nematodes overexpressing their own thesame heat treatment. These genes should therefore be heatshock genes may meet less regulatory restrictions usefultools for generatingtransgenic entomopathogen ic thantransforming them with foreign HSP genes.For this nematodesthat will be heattolerant.

778 Nematology Symposiumabstract

Nematode-antagonistic trichothecenes from the fungus Fusarium equiseti

James K. NITAO 1,SusanL.F .M EYER 1, Walter C. SCHMIDT 2, James C. FETTINGER 3 andDavid J. C HITWOOD 1 1 NematologyLaboratory, Agricultural Research Service, U.S. Department of Agriculture, Building 011A, Beltsville, MD 20705,USA 2 EnvironmentalChemistry Laboratory, Agricultural Research Service, U.S. Department of Agriculture,Building 011A, Beltsville, MD 20705, USA 3 Departmentof Chemistryand Biochemistry, University of Maryland,College P ark,MD 20742,USA Presentedat the 25th International Nematology Symposium, Herzliya, Israel, 2-7 April 2000

CultureŽ ltratesof a strainof Fusariumequiseti iso- icacolumns with chloroform-methanol mixtures, and high latedfrom eggsof the soybean cyst nematode (Hetero- performanceliquid chromatography with a gradientof deraglycines )inhibithatching of the root-knot nema- 40-90%acetonitrile in water on a C 18 column.Com- tode Meloidogyneincognita .Inthis study, we isolatedand poundswere identiŽed with infrared spectroscopy, 1H identiŽed two compounds with nematode-antagon isticac- and 13Cnuclearmagnetic resonance spectroscopy, elec- tivity from F.equisetivia bioassay-drivenfractionation tronimpact and chemical ionization mass spectrome- ofthe culture broth. The bioassay consisted of incubat- try,and X-ray crystallography.T woactive compounds ingsurface-sterilized eggs with DMSO-solubilized frac- were discoveredto be responsible for thenematode- tionsand determiningthe percentages of eggsthat hatched antagonisticactivity: 4,15-diacetoxy-12 ,13-epoxy-3,7- andof juveniles that remained mobile. The fractiona- tionprocess consisted of adsorption of compounds in dihydroxytrichothec-9-en-8-one(4,15-diacetyl nivalenol) thebroth onto Amberlite XAD-16 resin,partition of the and4,15-diacetoxy-1 2,13-epoxytrichothec-9-en-3-ol(di- methanoleluate from theXAD-16 betweenethyl ac- acetoxyscirpenol). Thisis the Ž rst publishedreport of etateand 7% methanol in water, chromatography on sil- toxicactivity of thesetwo compounds to nematodes.

Nematicidal activity of essential oilsand theircomponents against the root-knot nematode Meloidogyne javanica

Yuji OKA 1, Sengul NACAR 2, Eli PUTIEVSKY 3, Uzi RAVID 3, Zohara YANIV 4 andYitzhak S PIEGEL 1 1 Departmentof Nematology,Agricultural Research Organization ( ARO), BetDagan 50250, Israel 2 BiologyDepartment, F acultyof Science,KSU University,K. Maras,Turkey 3 Departmentof Aromatic,Medicinal and Spice Crops, ARO, NeweY a’arResearch Center ,RamatY ishay30095, Israel 4 Departmentof NaturalResources, ARO, BetDagan 50250, Israel Presentedat the 25th International Nematology Symposium, Herzliya, Israel, 2-7 April 2000

Nematicidalactivity of essential oils extracted from 27 ducedthe rootgalling of cucumberseedlings in potexper- spicesand aromatic plants were evaluated in vitro and in iments.The main components of theseessential oils were potexperiments. T welveof the 27 essential oils immo- testedfor theirnematicidal activity. Carvacrol, t-anethole, bilizedmore than 80% of juveniles of the root-knot ne- thymol,and (+)-carvone were foundto immobilize the ju- matode Meloidogynejavanica ataconcentrationof 1000 venilesand inhibit hatching at > 125 l/l in vitro. Most of l/l.Most of theseoils also inhibited nematode hatching at thesecomponents, mixed in sandy soil at concentrations thisconcentration. Essential oils of Carumcarvi, F oen- of75 and 150 mg/ kg,reduced root galling of cucumber culumvulgare, Mentha rotundifolia, and Menthaspicata seedlings.In 3 lpotexperiments, nematicidal activity of showedthe highest nematicidal activity among the tested theessential oils and their components was conŽrmed at oils in vitro.Theseoils and those from Origanumvulgare , 200and 150 mg/ kg,respectively. The results suggest that Origanumsyriacum and Coridothymuscapitatus mixed theessential oils and their main components may serve as insandy soil at concentrations of 100and 200 mg/ kgre- nematicides.

Vol.2( 7),2000 779 Symposiumabstract

Fungal eggs parasites of root-knot nematodes on vegetable crops inSpain

Cesar ORNAT 1, Soledad VERDEJO-LUCAS 1 andF .JavierS ORRIBAS 2 1 Institutde Recercai TecnologiaAgroalimentà ries, Crta. Cabrils s/ n08348-Cabrils,Barcelona, Spain 2 EscolaSuperior d’Agricultura de Barcelona,Comte Urgell 187, 08036-Barcelona, Spain Presentedat the 25th International Nematology Symposium, Herzliya, Israel, 2-7 April 2000

Surveyswere conductedto determine fungal egg para- (15isolates) and M. arenaria (2isolates). These fungi sitismof root-knot nematodes in areas of intensive veg- included Verticilliumchlamidosporium (2isolates), V. etableproduction of Barcelona (22 sites) and Almerí a catenulatum (1isolate), Gliocladiumroseum (2isolates), (35sites). Selected sites were currentlyinfested by root- Acremoniumstrictum (4isolates), Cylindrocarpon spp. knotnematodes or had a historyof nematode problems. (2isolates), Fusariumoxysporum (2isolates), Fusarium Thespecies of Meloidogyne identiŽed in Almerí a were solani (2isolates), Fusarium spp.(6 isolates),two sterile M. javanica (66%), and M.incognita (34%),and those mycelium,and Ž veunidentiŽed fungi. The adaptation of inBarcelona were M. javanica (40), M.incognita (38%), thefungi to climate conditions is suggested since A. stric- and M. arenaria (22%).A diversityof fungal parasites was isolatedfrom nematodeeggs in 50 and 43% of the tum was onlydetected in the warmer southernprovince samplescollected in Barcelona and Almerí a, respectively, ofAlmerí a whereas Verticillium spp.were foundin the andthey infected M.incognita (11isolates), M. javanica coolernorthern province of Barcelona.

DNA Žngerprinting of bacterial communities of Meloidogyne fallax egg masses

Artemis PAPERT 1,2 andC.J. (Hans) K OK 1 PlantResearch International P .O.Box16, 6700 AA Wageningen,The Netherlands 2 Presentaddress: ICAPB, Universityof Edinburgh,W estMains Road, Edinburgh, EH9 3JT,UK Presentedat the 25th International Nematology Symposium, Herzliya, Israel, 2-7 April 2000

Thegelatinous matrix in which Meloidogyne eggs are em- potato,growing in pots with Ž eldsoil from twoexperi- beddedis considered to be a protectionagainst physi- mentalŽ eldsin the Netherlands. The DNA Žngerprints calstress on the eggs and against egg parasitic micro- showthat the bacterial community of eggmasses is dif- organisms.The bacterial community of the gelatinous ferentfrom thebacterial community of the rhizoplane. matrixmay in uence the protective action of the ma- Furthermore,site-speciŽ c differencesin the communities trix,by being antagonistic either to the nematode eggs were found.Our conclusionis that the species compo- orto egg parasitic micro-organisms. The bacterial com- sitionof the bacterial community of the egg masses of munity of Meloidogynefallax eggmasses was analysed Meloidogynefallax isdifferent from thecomposition of withDenaturing Gradient Gel Electrophoresis (DGGE) ofPCR-ampliŽ ed 16S DNA fragments.This technique adjacentrhizoplane communities. This stresses the poten- yieldsa DNA-Žngerprint of the major bacterial species tialimportance of the bacterial egg mass communityas inthe community, including the non-culturable species. anecological factor in the reproduction efŽ ciency of the Sampleswere takenfrom eggmasses and rhizoplane of nematodeand the biological control of Meloidogyne.

780 Nematology Symposiumabstract

Thegrowth dynamics of Caenorhabditis elegans and why nematodes moult

Mavji N. PATEL,RicardoB.R. A ZEVEDO andArmand M. L EROI Departmentof Biology, Imperial College of Science,T echnologyand Medicine, Silwood P ark,Ascot, Berkshire SL5 7PY ,UK Presentedat the 25th International Nematology Symposium, Herzliya, Israel, 2-7 April 2000

Thereare 20 000describedspecies of nematode,and each eachlarval stage, the nematodes increased in volumein a hasa cuticlewhich it shedsand regenerates several times strictlylinear fashion and the rate of growthonly changed beforeadulthood. Why they do sohas long been obscure, aftereach moult. If C. elegans were togrow to adult- sinceunlike arthropods, nematodes do notneed to moult hoodat therate set by theŽ rst larvalstage then it would inorder to grow. W emeasuredthe growth of Caenorhab- take173 hours against 43 hoursthat it actuallytakes. Our ditiselegans from egghatch to adulthood at 20 °C. Growth measurementswere madeat about 30 min intervals dur- Žndingsmay provide an explanation for theexistence of ingeach of the four larval stages. W efoundthat during moultsin nematodes.

Contribution to the knowledge of the genus Takamangai Yeates, 1967 inSpain (Nematoda: Dorylaimida)

Reyes PEÑA-SANTIAGO, Gracia LIÉBANAS, Joaquín ABOLAFIA and Pablo GUERRERO Departamentode Biología Animal,V egetaly Ecología, Universidad de Jaén, P araje“ LasLagunillas s/ n”23071-Jaé n, Spain Presentedat the 25th International Nematology Symposium, Herzliya, Israel, 2-7 April 2000

Anematologicalsurvey carried out mainly in natural ar- &Swanger,1936) Andrá ssy, 1991, and two unidenti- easof theSierra Mágina Natural Park (province of Jaén, Žed(probablyundescribed) species. Measurements, illus- southeasternSpain) during the period 1996-1998 yielded trationsand maps of their distribution are provided. T. interestingmaterial belonging to the genus Takamangai eroshenkoi and T.ettersbergensis arethe most widely dis- Yeates,1967 (syn. Thonus Thorne,1974). Seven species tributedspecies in the geographical area studied, having havebeen distinguished: T.eroshenkoi Andrássy, 1991, beenrecorded in 23.1 and 17.2% respectively of thenear T.ettersbergensis (deMan, 1885) Andrá ssy, 1991, T. twohundred soil samples collected. T.ettersbergensis,T . kaszabi (Andrássy, 1959) Andrá ssy, 1991, T. mediana kaszabi and T. mediana arerecorded for theŽ rst timein (Eroshenko,1976) Andrá ssy, 1991, T.cf.nothus (Thorne theIberian Peninsula.

Vol.2( 7),2000 781 Symposiumabstract

An interesting belondiridspecies (Nematoda: Dorylaimida) from southeastern Spain

Reyes PEÑA-SANTIAGO 1 and Manuel PERALTA 2 1 Departamentode Biologí a Animal,V egetaly Ecología, Universidadde Jaé n, Paraje“ LasLagunillas s/ n”23071-Jaé n, Spain 2 I.E.S.“ Auringis”,Avenida de Andalucía s/n,23005-Jaé n, Spain Presentedat the 25th International Nematology Symposium, Herzliya, Israel, 2-7 April 2000

Abundantmaterial belonging to an undescribed be- tailrounded conoid; spicules and lateral guiding pieces londiridnematode has been collected in severalsoil sam- dorylaimoid;and few ventromediansupplements situated plesfrom naturalhabitats in southeasternSpain. It is char- intwo pairs outside the spicules range. This material acterizedby having a thincuticle; numerous lateral and showsa combinationof characters of Belondirinae and ventralglandular bodies; lip region cap-like; circumoral Dorylaimellinae,and could be abridgebetween both sub- refractiveplatelets present; odontostyle relatively short, families.The nature of the stylet (with robust fusiform robustand fusiform,wider than the cuticle at itslevel, and odontostyleand odontophore without  anges)clearly re- withwide aperture; odontophore rod-like; pharynx con- semblesBelondirinae, but the lip region shape (cap-like sistingof aslenderand weaklymuscular anterior part with andwith perioral cuticularized platelets), the thin cuticle, aspindle-shaped,enlarged and valve containing region andthe abundance of lateraland ventral glandular bodies behindthe odontophore base, and expanding gradually conformto amorphologicalpattern more similar to Dory- intothe basal bulb which occupies about half of thetotal lamellinae.However, stylet morphology is here regarded necklength and is surrounded by a thickdextrorse spi- ashaving higher taxonomic weight, which is why it has ralsheath; female genital system didelphic-amphid elphic; beenprovisionally classiŽ ed under Belondirinae.

Resistance and tolerance of Rosa spp. to Pratylenchuspenetrans

Yunliang PENG 1, Jan DE MEUTTER 2,GodelieveG HEYSEN 2 andMaurice M OENS 1 1 AgriculturalResearch Centre, Crop Protection Department, Burg. V anGansberghelaan 96, 9820 Merelbeke, Belgium 2 Universityof Gent, Laboratory of Genetics,K.L. Ledeganckstraat,35, 9000 Gent, Belgium Presentedat the 25th International Nematology Symposium, Herzliya, Israel, 2-7 April 2000

Pratylenchuspenetrans isone of themost damaging ne- tion of six P.penetrans populationswas comparedon 13 matodesof roses. T odevelopalternatives for chemical roserootstocks. A Frenchpopulation (Lavaltrie) repro- control,resistance and tolerance of Rosa spp. to P. pene- ducedsigniŽ cantly less than the others. R.virginiana was trans were studied.T enday-oldseedlings of 21roseroot- resistantto all populations. Differences in pathogenicity stockswere inoculatedwith 250 P.penetrans.R. virgini- to R.dumetorum ‘Laxa’between the six populations were ana was foundto beresistant and R. canina cv. Superba signiŽcant. An increase in guaiacol peroxidase activity was verysusceptible. When 70 day-oldrooted Rosa spp. was observedin infected roots of R.virginiana , whereas andcultivars were inoculatedwith 500 P.penetrans , dif- inrootsof ‘Laxa’and R. canina ‘Pollmers’we observed ferenceswere foundin the nematode multiplication be- tweenthe 140 tested accessions. For toleranceobserva- higherKCN-inhibitable peroxidase-like activities 50 days tion,14 roserootstocks were inoculatedtwice with 1000 afterinoculation. Increased HO-production in the infected P.penetrans . R.rubiginosa ,was themost tolerant with rootsof ‘Laxa’was foundto berelatedto itssusceptibil- theleast reduction of theplant fresh weight.Multiplica- ity.

782 Nematology Symposiumabstract

Lifehistory and development of the mediterranean cerealcyst nematode (MCCN), Heterodera latipons ,under Želdconditions

John PHILIS SpyrosStavrinides Chemicals Ltd, Stassinou 28, Nicosia 1060, Cyprus Presentedat the 25th International Nematology Symposium, Herzliya, Israel, 2-7 April 2000

Juvenilesof H. latipons startedto invade young roots of thata basaltemperature of 7 °Cisrequiredfor thedevel- barley(Hordeumvulgare L.cv.Athenais) thirteen days af- opmentof thenematodethen the accumulated heat, above tersowing, reaching peak soil populations at plantemer- 7°C,from nematodeinvasion to the ‘cortexruptured’, ‘ ap- gence.No juveniles were foundin the soil after 80 days pearanceof cysts’,‘ formationof eggs’and ‘ embryonated from sowing.Newly formed cysts were observedon the eggs’developmental stages reached 130, 215, 277 and roots68 days from sowing,most of them of small size 386 day ´ °Cor21, 54, 70 and 98 days, respectively. (570 ´ 415 m m) whileegg laying began 83 days from Thenematodecompleted only onegenerationduring plant sowing,at a meansoil temperature of 14.5 °C. Assuming growth.

Improving foliar application technologies for entomopathogenic nematodes

Simon J. PIGGOTT 1, John S. CLAYTON 2, Roma L. GWYNN 3, Graham A. MATTHEWS 1, Clare SAMPSON 4 andDenis J. W RIGHT 1 1 Departmentof Biology,Imperial College of Science, T echnologyand Medicine, Silwood P ark,Ascot, Berkshire, SL5 7PY ,UK 2 MicronSprayers Limited, Three Mills, Bromyard, Herefordshire, HR7 4HU,UK 3 MicroBioLimited, Unit 1, HarwoodRoad Industrial Estate, Littlehampton, W estSussex, BN17 7AU, UK 4 HorticultureResearch International, Stockbridge House, Cawood, Selby, North Y orkshire,YO8 0TZ, UK Presentedat the 25th International Nematology Symposium, Herzliya, Israel, 2-7 April 2000

Controlof cryptic pests such as leafminers ( Liriomyza polymer-basedformulation giving the greatest deposition spp.)has proved difŽ cult and a numberof factors have rate (ca 32 IJs cm­ 2). Thedensity of nematodesdeposited beenidentiŽ ed asreducingefŽ cacy. The selection of the usingthe prototype spinning disc was onethird less than applicatorand the formulation can be particularly criti- withthe above combination but was achievedwith only cal,a waterapplication through a hydraulicnozzle being halfthe appliednematodes. The nematodedeposition den- thestandard procedure for bothŽ eldand protected crops. sityshowed a positivecorrelation with the % mortalityof Thepresent study describes the assessment of variousfor- L. bryonae larvaedue to nematode infection, with aver- mulationsand applicators, including a prototypespinning agemortalityof ca 57%beingobtained with the hydraulic disc,against L. bryonae onglasshousetomato. Consider- nozzle-polymercombination. The work is discussedin re- abledifferences in the density of infective juveniles (IJ) lationto further developments in spray application tech- R of Steinernemafeltiae UK76(Nemasys ®)depositedon nologyand the possible integration of entomopathogenic leafsurfaces were found,with hydraulic nozzles and a nematodeswith other biological control agents.

Vol.2( 7),2000 783 Symposiumabstract

Use of Vydate L(oxamyl)indripirrigated crops inSpain: an example of an integrated solution for root-knot nematode control and foliar horticultural pests

Sebastian PONS, Paris MICHAELIDES andStephen I RVING DuPontSpain and DuP ontde Nemours ( France),S.A. European Research and Development Center ,F-68740Nambsheim, France Presentedat the 25th International Nematology Symposium, Herzliya, Israel, 2-7 April 2000

Theintensive horticulture under greenhouse or plasticin Appliedafter transplanting on a regularbasis during the Spainproduces high value crops in shorter cycle crops. cropcycle controls the nematode problem (nemostatic ef- Theuse of plastic covers, chemicals, drip irrigation and fect),and reduces signiŽ cantly leafminer damage ( Liri- selectedplant varieties allows the grower to obtaincrops omyzahuidobrensis ). Theapplicationmethodology of ox- practicallyall year round. At thesame time this situation amylhas been shown to be important to obtain the best beneŽts many pests like nematodes ( Meloidogyne spp.), productperformance. The type of application,by dripir- white ies,leafminers, etc. Oxamyl, the active ingredi- entof Vydate L, isregistered in Spain for dripirriga- rigation,which reduces enormously the risk of applicator tionapplications. Oxamyl has proved crop selective, sys- exposure,and the selectivity to naturalenemies and polli- temicand active on nematodesand several foliar insects. natorsmakes oxamyl Ž tinIPM programs.

Relative importance of plant-parasitic nematodes, the banana weeviland leaf diseases as Musa production constraints inZanzibar

Khadija A. RAJAB 1,2, Paul R. SPEIJER3,† and Arnold VAN HUIS 2 1 PlantProtection Division, P .O.Box1062, Zanzibar ,Tanzania 2 AgriculturalUniversity W ageningen,P .O.Box 8031, 6700 EH Wageningen,The Netherlands 3 InternationalInstitute of Tropical Agriculture, Eastern and Southern Africa Regional Centre, P .O.Box7878, Kampala, Uganda †Deceased Presentedat the 25th International Nematology Symposium, Herzliya, Israel, 2-7 April 2000

Bananasand plantains are important staple food crops in Radopholussimilis, Pratylenchus coffeae andthe leaf dis- Zanzibar,ranking second in production after cassava. A ease MycosphaerellaŽ jiensis were observedboth at Un- surveywas conductedat theend of therainy season May- gujaand Pemba. Banana weevil, Cosmopolitessordidus , July1997, to establishthe relative importance of pestsand damagewas onlyobserved in Unguja and appearedhighly diseasesaffecting Musa productionin the two islands of associatedwith R. similis and P. coffeae incidence.None Zanzibar,Unguja and Pemba. A totalof 12 representa- ofthe biotic constraints in Unguja negatively affected tivesites with Musa cultivarsof thegenome groups AAA, bunchweight. It is mostlikely that, in Unguja, poor soil AAB andABB were selected.Pest incidence and sever- fertility(sandy, coral rag soils) and perhaps low rainfall, itywere assessedand pest impact was measuredby relat- ingdamage levels to plant growth parameters. In Unguja, arethe major production constraints. In Pemba, bunch bunchweights were low,almosthalf of those recorded weightwas affectedby root damage caused by thethree inPemba. The nematodes Helicotylenchusmulticinctus, nematodespecies, followed by leafdiseases.

784 Nematology Symposiumabstract

Geneticpolymorphism and relationships of Meloidogyne spp. from Brazil

Onivaldo RANDIG 1, Frédéric LEROY 1, Michel BONGIOVANNI 1, Regina GOMES CARNEIRO 2 andPhilippe C ASTAGNONE-SERENO 1 1 INRA, UnitéSanté Vé gé tale et Environnement, BP2078, 06606 Antibes cedex, France 2 EMBRAPA,RecursosGeneticos e Biotecnologia,CP02372, CEP70849-970, Brasilia, DF ,Brasil Presentedat the 25th International Nematology Symposium, Herzliya, Israel, 2-7 April 2000

Theroot-knot nematodes (RKN), Meloidogyne spp., are scribedspecies. Genomic DNA was puriŽed from each majorpests of manyimportant crops in Brazil. However, ofthe18 representativeisolates selected for analysis,and thereis little knowledge about the genetic variability and usedas template in RAPD-PCR experiments.AmpliŽ ed therelationships of the main species, and no molecular fragmentswere scoredas 0-1 characters, and the whole dataare currently available on RKNpresentin Brasil. In dataset was analysedusing a parsimonymethod. The orderto investigate the range of genetic polymorphism RAPD markers providedreliable polymorphisms at inter- bothbetween and within RKN species,a collectionwas andintraspeciŽ c levels,and the 18 Meloidogyne isolates built,including isolates from themajor species ( M. are- usedin this study could be clearly identiŽ ed, based on naria, M.incognita , M. javanica and M. hapla), from thepresence/ absenceof diagnostic bands. Clustering of someless damaging species ( M. exigua and M.paranaen- isolatesand relationships inferred from thephylogenetic sis), andalso isolates presumably belonging to unde- analysiswill be presented.

IdentiŽcation and variability for virulenceto Triticeaeof CCN populations from mediterranean regions

Roger RIVOAL 1, Sylvie VALETTE 1, Makram BEL HADJ FRADJ 1, Aïssa MOKABLI 2,Jean-PierreG AUTHIER 1, Joseph JAHIER 1 and Julie NICOL 3 1 INRA, UMR Biologiedes Organismes et des P opulationsappliqué e àlaProtection des Plantes ( Bio3P),BP 29,35653 Le Rheu cedex,France 2 INA, Département de Zoologie Agricole, El-Harrach, DZ, Algeria 3 CIMMYT International,Wheat Program, Lisboa 27, Apdo. P ostal6-641, 06600 Mexico Presentedat the 25th International Nematology Symposium, Herzliya, Israel, 2-7 April 2000

Cerealcyst nematodes (CCN) belongingto thegenus He- havebeen conŽ rmed to belong to either H. avenae or terodera composea groupof speciesof whichthe major H. latipons upontheir restriction patterns. Most of these are H. avenae, H. latipons and H. Ž lipjevi.Thesespecies H. avenae populations,compared to other populations arepresumed to causedamage to bread and durumwheats from SouthEurope, Australia and India, differed signiŽ - incircum-mediterranean countries. This work has twoob- cantlyin their ability to reproduce(Ž tness) on susceptible jectives: i)tocharacterise the species status of popula- hoststested in miniaturised controlled conditions. These tionssampled in this area with molecular tools as restric- populationsalso exhibited variation in (a)virulence spec- tionpolymorphism of ribosomal DNA (PCR/CAPS); ii) to trumtowards the known and unknown resistance genes testthe resistance efŽ ciency of knownor unknowngenes orgenotypes such as the Ae.geniculata accessions.No andgenotypes of cultivatedand wild Triticeae to awider relationshiphas beenshown between the (a)virulencesta- rangeof CCN populations.Attention has been focused tusof thenematode populations and the geographical ori- onthe host reactions of asetof Aegilopsgeniculata ac- ginof theplant genotypes tested. Several of thesegeno- cessionsoriginating from differentlocations around the typespresent an indisputable interest for regionaland/ or mediterraneansea. Populations recently sampled in bread globalutilisation in resistance breeding programmes of anddurum wheat Ž eldsfrom NorthAfrica andW estAsia breadand durum wheats.

Vol.2( 7),2000 785 Symposiumabstract

Analysis of the sensory responses of J2s of Globodera rostochiensis using electrophysiological techniques

Richard N. ROLFE 1, John BARRETT 2 andRoland N. P ERRY 1 1 Entomologyand Nematology Department, IACR-Rothamsted, Harpenden, Herts AL5 2JQ,UK 2 Instituteof BiologicalScience, University of Wales,Aberystwyth, SY23 3DA, W ales,UK Presentedat the 25th International Nematology Symposium, Herzliya, Israel, 2-7 April 2000

Theelectrophysiolog icaltechnique for analysingsensory usinga varietyof semiochemicals provided some in- responsesof livenematodes has been modiŽ ed for work terestingcontrasts. For example,signiŽ cant increases in withsecond-stage juveniles (J2) of G.rostochiensis. To spikeactivity of J2were obtainedon stimulationwith D- illustratethe potential of the technique, concentration- glutamicacid whereas J2 did not respond to the L isomer. dependentresponses of J2were obtainedto acetylcholine IncubatingJ2 for 24hina monoclonalantibody showing andthe delay in response and adaptation were quan- speciŽcity to amphidial secretions resulted in blockingof tiŽed. SigniŽ cant increases in spike activity were ob- thenormalresponse to PRD; the potential of thetechnique tainedon stimulation of J2 with potato root diffusate for quantifyingperturbation of sensoryresponses and de- (PRD), whereasnon-host diffusate and female secretory- terminingthe rate of turnover of sense organ secretions excretoryproducts elicited no response.Comparative tests willbe discussed.

Use of the electropharyngeogram to analyse stylet activation

Richard N. ROLFE andRoland N. P ERRY Entomologyand Nematology Department, IACR-Rothamsted, Harpenden, Herts AL5 2JQ,UK Presentedat the 25th International Nematology Symposium, Herzliya, Israel, 2-7 April 2000

Theelectropharyngeog ram (EPG)techniquewas Žrst tivitywas initiatedby exposure to the neurotransmitter usedto analyse pharyngeal pumping of Caenorhabditis serotoninand the changes in membraneelectrical poten- elegans.Wehaveused the technique to record electrical tialwere recordedcorresponding to the excitatory and re- activityof styletprotractor muscles of fourth-stagejuve- polarisationphases of thestylet protractor muscles. Data niles of Ditylenchusdipsaci andsecond-stage juveniles of willbe presentedon theresponses to a rangeof agonists Globoderarostochiensis .Individualnematodes were cap- turedby the anterior end in a suctionpipette electrode, andantagonists to illustratethe potential of thistechnique from whichthe electrical activity was recorded.Stylet ac- toevaluatestylet activity.

786 Nematology Symposiumabstract

Root-knot nematode pathogenicity factor analysis at the protein level

Marie-Noëlle R OSSO 1, Neil LEDGER 1,Stéphanie J AUBERT 1, Arjen SCHOTS 2,RichardS. H USSEY 3 and Pierre ABAD 1 1 InstitutNational de laRecherche Agronomique, USVE, 123Bd Francis Meilland, 06600 Antibes, France 2 Laboratoryof Nematology,W ageningenAgricultural University, 6709 PD Wageningen,The Netherlands 3 Departmentof Plant P athology,University of Georgia, Athens 30602-7274, USA Presentedat the 25th International Nematology Symposium, Herzliya, Israel, 2-7 April 2000

Inorder to identify and characterise Meloidogyne sp. ofthisscFv led to 40%reductionin thenematodeprogeny pathogenicityfactors two strategies based on protein ontransformedplants as comparedto guscontrols.Analy- analysishave been developed. The Ž rst strategyis in- sisof nematodedevelopment in the transformed roots and direct,and relies on monoclonal antibodies selected for cytologicalobservations will help understanding of the speciŽc bindingto secreted proteins expressed in the oe- roleof the 6D4-secreted antigen in parasitism. The sec- sophagealglands. Such antibodies are used for immunop- ondapproach is thedirect puriŽ cation of styletsecretions uriŽcation of the secreted protein or for itsinhibition in afterlarge scale growing of nematodes.2D electrophore- planta.InAntibes, we focuson inhibitionof styletsecre- sisanalysis allows internal sequencing of theisolated pro- tions in planta byexpressionof single-chainantibodies in teinsand the design of degenerate oligonucleotid esfor infestedroots. A single-chainantibody named scFv 6D4 RT-PCR.The ampliŽ ed products are used for Southern- anddirected to a styletsecretion from Meloidogyne was blotanalysis and in situ hybridisation.The recently ob- expressedin tobacco plants. The cytoplasmic expression tainedresults are discussed.

Consequences of errorsdue to subsampling and other laboratory procedures

Corrie H. SCHOMAKER andH. ThomasB EEN PlantResearch International, P ostbox16, 6700 AA, Wageningen,The Netherlands Presentedat the 25th International Nematology Symposium, Herzliya, Israel, 2-7 April 2000

Inmany nematological studies, in whichsoil sampling is laboratory.Cysts were closeto randomlydistributed in the animportant step in data collection, it is common prac- well-mixedsamples resulting in variancebetween pairs or ticeto collect a bulksample but to investigate only a triplesaccording to a binomialor trinomial distribution, part,taken after mixing. Unfortunately, the rationale be- respectively.The average coefŽ cient of variation associ- hindthe choice of bothbulk sample size and subsample atedwith elutriation ranged from 3.6to 9.6% for thelab- sizeis seldom documented. T oenablethis choice a gen- oratoriesinvolved. However, the estimated 95% quantiles eraltheory is neededabout the relationbetween nematode ofthecoefŽ cients of variationdepended on thenumberof counts,sampling error andsubsampling error anderrors cyststhat were countedin the samplesand rangedfrom 73 dueto laboratory procedures (extraction, counting) on the to42%for laboratory1, from 43to19%for laboratory2 totalsampling error. For thispurpose, Ž eldsinfested with andfrom 10to3% for laboratory3. Theconsequences of potatocyst nematodes were intensivelysampled. Bulk theselaboratory errors for thediscrimination between ne- sampleswere splitinto two or three subsamples. Sub- matodedensities, the detection probability of fociand the samplesof randomly chosen bulksamples were sendto estimationof parametersin distributionpattern of nema- threedifferent laboratories for elutriation.All other lab- todes,as well as methodsto choose optimal sample sizes, oratoryprocedures including counting were donein one arediscussed.

Vol.2( 7),2000 787 Symposiumabstract

Cloning and characterisation of map-1,aputative avirulencegene from the root-knot nematode Meloidogyne incognita

Jean-PhilippeS EMBLAT, Antoine DALMASSO, Pierre ABAD andPhilippe C ASTAGNONE-SERENO INRA, UnitéSanté Vé gé tale et Environnement,BP2078, 06606 Antibes cedex, France Presentedat the 25th International Nematology Symposium, Herzliya, Israel, 2-7 April 2000

Intomato, the Mi resistancegene controls the three Southern-blotexperiments, this fragment was conŽrmed parthenogeneticroot-knot nematode (RKN) species, i.e., tobe differential at the genomic level. Screening of a Meloidogynearenaria , M.incognita and M.javanica. cDNA libraryallowed the isolationof afull-lengthcDNA However,virulent populations able to reproduceon resis- codingfor aproteinof 458amino acids, with no signif- tantplants have been isolated from manytomato-growing icanthomology in databases. The protein, named MAP- areas.T ounderstandthe molecular basis of (a)virulence 1,is characterized by the presence of a signalpeptide, inRKN, we selectedthree independent pairs of avirulent suggestingits secretion, and repetitive motives of 13and and Mi-virulentnear-isogenic M.incognita lines, and a 58amino acids. Using antibodies, MAP-1 was localized differentialanalysis of theirgenome was performedusing aroundthe amphids of second-stage juveniles, thus con- AFLP molecularmarkers. Among the ca 25000ampli- Žrmingit is secreted.The putative role of MAP-1 inthe Žedfragments, one appeared to be polymorphicbetween relationshipsof M.incognita with(resistant) tomatoes avirulentand virulent lines. When used as a probein willbe discussed.

Characterisation of Meloidogyne javanica surface coat

Edna SHARON 1, Yitzhak SPIEGEL 1 and Rosane CURTIS 2 1 Departmentof Nematology,Agricultural Research Organization ( ARO), VolcaniCenter ,P.O.Box6, Bet-Dagan, Israel 2 Departmentof Entomologyand Nematology, IACR-Rothhamsted, Harpenden, AL5 2JQ,Herts, UK Presentedat the 25th International Nematology Symposium, Herzliya, Israel, 2-7 April 2000

Severalpolyclonal and monoclonalantibodies, previously antibodiesPC E2and3F .4labeled spots on thehypoder- raisedagainst Meloidogyneincognita race1, second-stage mis,indicating the possible origin of these surface anti- juvenile(J2) surface-coatand excretory-secretory prod- gens.Antibodies were usedfor immunolocalizationofthe ucts,were testedfor cross-reactivitywith M. javanica live surfaceantigens in planta oncryosections of M. javan- J2,using immuno uorescence. Three of six monoclonal ica infectedtomato roots. PC E2labeled the surface coat antibodies(IACR-Misec 9F .5,7E.1 and 3F .4)and only ofparasitic stages and shedding of the surface was ob- one(PC 373)of six polyclonal antibodies tested, reacted served.Immunolabeling of M. javanica extracts,on West- with M. javanica surface.A polyclonalantibody previ- ernblots with PC 373and E2 revealed different labeling ouslyraised against M. javanica J2(PC E2),was usedas well.None of theantibodieswhich recognized the surface patterns.Bioassays were developedto test the effect of an- reactedwith J2 excretion-secretion products in vitro, fol- tibodieson nematodemotility and invasion of Arabidop- lowingstimulation with the neurotransmitter DMT. Im- sisthaliana roots.All the antibodies tested changed the munolabelingof cryosections of the J2 showed that the movementpattern of theJ2 and inhibited root invasion.

788 Nematology Symposiumabstract

Biological control of Radopholus similis through the ‘biological-enhancement’ of banana tissue culturederived plantlets with mutualistic fungal endophytes

Richard A. SIKORA1,†, Luis POCASANGRE 1, Bjoern NIERE 1 and Paul R. SPEIJER 2 1 Soil-EcosystemPhytopathology and Nematology, Institut fuer P anzenkrankheiten, Universitä tBonn, Nussallee 9, D-53115Bonn, Germany 2 Nematologist,Biological Control Program, ESARC, InternationalInstitute of Tropical Agriculture, Kampala, Uganda †Deceased Presentedat the 25th International Nematology Symposium, Herzliya, Israel, 2-7 April 2000

Alargediversity of fungi have been detected in healthy R. similis densitiesbelow threshold levels in boththe roots rootsand corms of a widespectrum of banana cultivars andrhizosphere. Isolates have the ability to colonize the from Africa, Asiaand South America. These fungi have corticaltissue extensively after inoculation to the plantlets beenshown to be non-pathogenic,causingno visibledam- intheform ofsporesuspensions. SigniŽ cant levels of bio- ageto the plant tissue even at high inoculum level. Re- controlwere obtainedon widelydifferent Musa genomes searchhas clearly demonstrated that many of thesenon- representingboth African Highland as well as commer- pathogenicfungal isolates grow endophytically at high cialdessert cultivars. The results of thesestudies will be densitieswithin banana root tissue. Furthermore, inves- tigationshave shown that some isolates have strong bio- presentedand the concepts of ‘ biological-enhancement’ controlcapabilities when applied externally to tissuecul- oftissue culture planting material for practicalŽ elduse turederived plantlets. The isolates reduced signiŽ cantly willbe outlinedand discussed.

Nematode taxonomists at the beginning of the third Millennium

N. SMOL NematologyCourse, Department of Biology,Ghent University, Gent, Belgium Presentedat the 25th International Nematology Symposium, Herzliya, Israel, 2-7 April 2000

Insome western countries the number of nematode tax- ofnematodesstudied, context of taxonomy,tools used for onomistsis decreasing. An overview of nematologists identiŽcation; experts helping in identiŽ cation, future per- world-wide,with special reference to taxonomists, will spectives,needs, funding agencies, international contacts, bepresented to conclude if this trend is present in other partsof theworld. The following items will be discussed: participationin symposiaand/ orworkshops, etc. The re- numberof taxonomistsper country, per continent; group sultswill be expressedin a north-southratio.

Vol.2( 7),2000 789 Symposiumabstract

Relative importance of plant-parasitic nematodes, banana weeviland leaf diseases as production constraints to plantain ( Musa spp, AAB group) in Southern Nigeria

Paul R. SPEIJER1,†,M.OmolaraR OTIMI 1,2 and Dirk DE WAELE 2 1 InternationalInstitute of TropicalAgriculture ( IITA),Eastern and Southern Africa Regional Centre (ESARC), P.O.Box7878, Kampala,Uganda 2 Laboratoryof TropicalCrop Improvement, Katholieke Universiteit Leuven, Kardinaal Mercierlaan 92, B-3001 Heverlee, Belgium †Deceased Presentedat the 25th International Nematology Symposium, Herzliya, Israel, 2-7 April 2000

Plantainproduction in W estand Central Africa accounts 50and 46% of the sites, while J2 stages of the genus for 50%of the world output. Nigeria is considered the Meloidogyne occurredat 68% of the sites. W eevildam- largestproducer. However, various biotic constraints, in- agewas observedin 71% of the sites. Black Leaf Streak cludingplant-parasitic nematodes, the banana weevil and diseasewas observedat all sites. Factors, derived from leafdiseases, affect Nigeria’s plantain production. T oes- aPrincipalComponent Analyses of damageobservations tablisha rankingamong these constraints, a surveywas for nematodes,banana weevil and leaf diseases, were re- conductedby theInternational Institute of TropicalAgri- latedto plantaingrowth. The results suggest that the ne- culture(IIT A)inthe plantain growing regions of South- matodespecies P. coffeae,followedby R. similis are the ernNigeria. The predominant nematode species found in rootsof plantain was Helicotylenchusmulticinctus , oc- majorbiotic production constraints to plantain in Nigeria. curringat all69 sites. Hoplolaimuspararobustus , Praty- Higherlosses are anticipated from thesespecies than from lenchuscoffeae and Radopholussimilis were foundin 64, thebanana weevil or leafdiseases.

Cadusafos: amost effective nematicide

Charles A. STAETZ FMC ResearchCenter ,P.O.Box 8, Princeton, NJ 08543,USA Presentedat the 25th International Nematology Symposium, Herzliya, Israel, 2-7 April 2000

R Cadusafos,Rugby ®,isa veryeffective compound with actionof cadusafos on nematodes as well as its impact activityagainst a widerange of nematodes and also oninsectsand plants will be presented.In addition, other againstmany soil-dwelling insect pests. Cadusafos is cur- topicssuch as soil movement in different soil types and rentlyutilized on a widerange of crops, including ba- theperformance of several formulations under different nanas,vegetables, citrus, tobacco, ornamentals, potatoes Želdconditions will be reviewed. Lastly, the impact of andspice plants, grown around the world under different resistanceand the phenomenon of accelerated microbial soil,climatic and crop grouping conditions. The mode of degradationupon cadusafos will be discussed.

790 Nematology Symposiumabstract

Phasmids inT ylenchidae and Anguinidae?

Dieter STURHAN 1 and Karin KIONTKE 2 1 BiologischeBundesanstalt, Institut fü r Nematologieund Wirbeltierkunde, T oppheideweg88, 48161 Mü nster ,Germany 2 FreieUniversitä tBerlin, Institut fü r Biologie/Zoologie,Kö nigin-Luise-Str .1-3,14195 Berlin, Germany Presentedat the 25th International Nematology Symposium, Herzliya, Israel, 2-7 April 2000

Phasmid-likestructures located in thepostmedian region ever,commonly observed in many members of Rhabditi- ofthe body in a dorsosublateralposition have been re- dae,Cephalobidae, Panagrolaimidae, Diplogasteridae and portedfor manymembers of T ylenchidae,several mem- otherfamilies of Rhabditida, where they are called pos- bersof Anguinidaeand for Aphelenchusavenae in Aphe- teriordeirids. W eobservedsuch structures also in males lenchidae.Our recentlight-microscopical studies revealed andjuveniles of many Steinernema species.In these taxa, thepresence of suchstructures also in several Eutylenchus inaddition to posterior deirids, phasmids are present in speciesin T ylenchidae,in Halenchus and Pseudhalenchus acaudalposition. The ‘ phasmid-likestructures’ in post- speciesin Anguinidae and in Paraphelenchus species in medianportion of the body in T ylenchidaeare likely to Aphelenchidae,but nothing could be detectedin, e.g., Psi- behomologous with the posterior deirids in Rhabditida lenchushilarulus, Macrotrophurus arbusticola, Pleuroty- ratherthan being homologous with phasmids. This is in- lenchussachsi, Boleodorus spp.,members of the fami- liesDolichodoridae, Belonolaimidae, Pratylenchidae, Ho- dicatedby theirpresence in taxa with phasmids in thetail plolaimidae,Heteroderidae and Aphelenchoididae ,most regionand by the obvious absence of an external pore. ofwhich have phasmids in a caudalposition. Struc- Truephasmids are obviously absent in Anguinidae and turesin a similardorsosublateral position were, how- mostgenera of Tylenchidae.

Studies on the interaction and Pasteuria penetrans and Glomus sp. as bio-management agents against Meloidogyne incognita on tomato

Miguel TALAVERA andT akayukiM IZUKUBO CropNematode Lab. National Agriculture Research Center ,Kannondai3-1-1, Tsukuba, Ibaraki, 305-8666 Japan Presentedat the 25th International Nematology Symposium, Herzliya, Israel, 2-7 April 2000

Theeffects of thetriple interaction M.incognita ´ P. pen- ducedby 42.0%in mycorrhizal plants, by 43.3%after P. etrans ´ Glomus sp.on tomatogrowth, nematode repro- penetrans applicationand by 60.7%when both treatments duction,bacterial infectivity and mycorrhizal colonisa- were appliedtogether .Infestationby M. incognita did not tionof rootswere testedin agreenhouse-potexperiment. affectroot colonisation by Glomus sp.when mycorrhizal Tomatoshoot weight and length were reducedin alltreat- inoculumwas establishedtwo weeks before nematode in- ments when M. incognita was present,but an improve- mentin plant growth could be observed when tomato oculum.No signiŽ cant effects of the mycorrhizal treat- plantswere treatedjointly with Glomus sp. and P. pen- ment on P.penetrans attachmenton M. incognita juve- etrans.Finaldensities of M.incognita J2in soilwere re- nilesor numberof femalesinfected were observed.

Vol.2( 7),2000 791 Symposiumabstract

Radopholus similis :apossible trigger of the decliningproduction spiralof highland cooking bananas ( Musa AAA) inUganda

HerbertA.L. T ALWANA 1, Paul R. SPEIJER1,† and Dirk DE WAELE 2 1 InternationalInstitute of TropicalAgriculture ( IITA),East and Southern Africa Regional Centre ( ESARC), Namulonge,P .O.Box 7878Kampala, Uganda 2 Laboratoryof TropicalCrop Improvement, Katholieke Universiteit Leuven, Kardinaal Mercierlaan 92, 3001 Heverlee, Belgium †Deceased Presentedat the 25th International Nematology Symposium, Herzliya, Israel, 2-7 April 2000

Twonematode species Radopholussimilis and Praty- out R. similis.Inaddition, bunch weight was 30%lower lenchusgoodeyi co-existon highlandbanana ( Musa spp., andthe number of standingleaves was lowerfrom plants AAA type)in Central and Southern Uganda. The rela- where R. similis was presentcompared to those where it tiveimportance of these two species to banana produc- was absent.Soil temperature recorded in between adja- tiondecline was investigatedin Mbarara (South W estern centmats and close to themats was higherwhere R. sim- Uganda,altitude 1430 m) bysampling2-y old individual ilis was presentthan where it was not.The increase in matsdiffering in nematode species proŽ les. Small suckers soiltemperature induced by Radopholussimilis may trig- were detachedfrom plantsat harvest and the nematode gertheunrecoverable production decline observed in East populationsin the roots were estimated.Suckers from mats with R. similis were smallerin girth,had a reduced Africanhighland bananas. High soil temperature favours rootsystem, and supported lower populations of P. good- R. similis reproductionand accelerates soil organic matter eyi comparedto those suckers detached from matswith- decomposition.

Characterisation of resistance to root-knot nematodes inpepper ( Capsicum spp.)

Judy A. THIES 1 andRichard L. F ERY 1 1 U.S.V egetableLaboratory, USDA, ARS, 2875Savannah Highway, Charleston, SC 29414,USA Presentedat the 25th International Nematology Symposium, Herzliya, Israel, 2-7 April 2000

Root-knotnematodes ( Meloidogyne spp.)are major pests comparedto 24 °C.However,at 32 °C,reproductionon ofpeppersworldwide. The N geneconditions M. incog- resistantcultivars was only20% of that of susceptible nita resistancein therecently released bell pepper ( C. an- cultivarsand root gall indices were withinthe moder- nuum)cultivarsCharleston Belle and Carolina W onder, atelyresistant range. At ambientgreenhouse temperatures andan allele of the N genecontrols the resistance in (26 3°C), these M.incognita -resistantgenotypes exhib- C. chinense § theScotch Bonnet ( )cultigensP A-353,P A- itedresistance to M. arenaria races1 and2 and M. javan- 398,and P A-426.Peppers are often grown in hot cli- ica, but not M. hapla.Althougha partialloss of resistance mateswhere several Meloidogyne spp.occur .Therefore, to M.incognita occurredat high temperatures, resistant we characterizedthe heat stability and speciŽ city of re- sistancein these M.incognita -resistant C. annuum and C. peppercultivars should be ausefulcomponent of cropping chinense genotypes.All Ž vegenotypes exhibited a par- systemsdesigned to manage multiple species of root-knot tialloss of resistance to M.incognita at 28 and 32°C nematodes.

792 Nematology Symposiumabstract

Morphometrics of threespecies of Hirschmanniella (Nemata: Pratylenchinae) parasitising rice,with remarks on the genus

Alberto TROCCOLI Istitutodi Nematologia Agraria C.N.R., viaAmendola, 165/ A,70126Bari, Italy Presentedat the 25th International Nematology Symposium, Herzliya, Israel, 2-7 April 2000

The genus Hirschmanniella Luc& Goodey,1964 contains monlyoccurring on ricecrops) allowed an accuratecom- mainlytropical species, most of which are recorded as parativestudy of their morphology, with remarks on in- parasitesof rice ( Oryzasativa L.).They are easily dis- traspeciŽc variabilityfor keycharacters within the genus. tinguishableamong members of thefamily by their very Morphometryof thethree species is presented,with par- longand slender body, massive cephalic sclerotization, ticularemphasis on cephalicregion, stylet morphometry, andelongate oesophageal gland lobe, but their identiŽ - shapeof female and male tail, cuticular annulation, and cationat species level is rather difŽ cult, owing to astrong othercharacters with diagnostic value, making identiŽ - resemblanceto eachother, and thehabitof feedingprefer- cationeasier. A newgeographic record is also given for ablyon thesame host. The Ž ndingof threepopulations of Hirschmanniella from differentgeographic origins (iden- oneof thespecies ( H. gracilis), whichwas foundfor the tiŽ ed as H. gracilis from Greece, H. oryzae from China Žrst timein a naturalhabitat of anorthwesternprovince and H.spinicaudata from Venezuela,all of them com- of Greece.

Measures to control TRVtransmission by trichodorid nematodes in ornamental bulb crops

AnneSophie V AN BRUGGEN 1, Frans C. ZOON 2, Cees J. ASJES 1, Ate DE HEIJ 2 andF .AstridD E BOER 1 1 BulbResearch Centre, P .O.Box85, 2160 AB Lisse,The Netherlands 2 PlantResearch International, P .O.Box 16, 6700AA W ageningen,The Netherlands Presentedat the 25th International Nematology Symposium, Herzliya, Israel, 2-7 April 2000

Tobaccorattle virus (TR V)mayinfect several ornamental effectof severalcrops on the multiplication of T. similis bulbcrops. TR Vistransmittedby trichodoridnematodes. andthe associated TR Vserotypewas studied.A selection Inprevious research Paratrichodorusteres appeared to ofthese crops is being tested in a Želdexperiment with bethemost important vector species in bulb crops in the anaturalinfection of viruliferous T. similis.Theinfection Netherlands.Effective cultural measures were developed potentialof thesoil(IPS) afterculture of thetest crops was tocontrol virus transmitted by P. teres.Theyinclude cul- determinedby aMostProbable Number (MPN) method. tivationof fodderradish ( Raphanussativus )duringsum- Cultivationof fodderradish resulted in areductionof the mer priorto a gladiolusor tulipcrop. Trichodorussimilis IPS, whereasother crops had no effect.This preliminary isanother important vector .Inthis study, we investigated resultindicates that a cultureof fodder radish may be a whethercultivation of fodder radish reduces TR Vtrans- feasiblecontrol measure for varioustrichodorid-TR Vdis- missionby this species as well. In a potexperiment the easecomplexes.

Vol.2( 7),2000 793 Symposiumabstract

Hatching behaviour of the Northern root-knot nematode Meloidogyne hapla

Jan VAN DE HAAR RZResearchBV ,P.O.Box 2, NL-9123ZR, Metslawier,TheNetherlands Presentedat the 25th International Nematology Symposium, Herzliya, Israel, 2-7 April 2000

Large-scalescreening of plants for nematoderesistance applicationof low salinity treatment (0.3 M NaCl)pro- usingmonoxenic tests requires the aseptic production of hibitedthe hatching of free eggs,but did not adversely af- hugeamounts of juveniles.The Northern root-knot nema- fectthe embryonic development of theseeggs. It not only tode M. hapla was rearedin tissue culture containers. At seemedto synchronise the hatchafter transferring the eggs 20°Cthehighest amounts of juvenileswere yieldedafter towater,but also increased the number of hatchedeggs up anincubation period of 10 weeks after inoculation. Af- tothe level of thatfrom egg-masses.These observations terthe liberation of the eggs from egg-massesby using donotsupport the hypothesis that hypochloride has a di- hypochloride,the hatching usually stops after 3 weeks, rectnegative effect on the hatching of eggs.Probably the resultingin a relativelylow percentage of hatching. Pri- marilythe immature eggs do nothatch. Incubation of un- compositionof thematrix within the egg-mass facilitates treatedegg-masses prolongs the hatching period and in- thedevelopment of theyoung eggs. It seemed to bepossi- creasesthe Ž nalnumbers of juveniles. The well-known bleto imitatethese conditions by lowsalinity.

Pathogenic variation in Meloidogyne chitwoodi on Solanum bulbocastanum

J.G.(Hans) V AN DER BEEK and Leo M. POLEIJ PlantResearch International, W ageningenUniversity and Research Centre, P .O.Box 16, NL-6700 AA Wageningen,The Netherlands Presentedat the 25th International Nematology Symposium, Herzliya, Israel, 2-7 April 2000

Resistancein potatoto thequarantineroot-knot nematode vealed4 pathotypesand 3 resistancefactors. Experiments Meloidogynechitwoodi isone of themost promising and withvirulent selections of theseisolates, derived from S. environmentallysafe controlmeasures for thispest organ- bulbocastanum genotypes,showed that at leastsome iso- ism.Adequate resistance must be effective to the widest lateswere mixturesof pathotypes.Breeding programmes possiblegenetic range of the parasite. Testing of 18 ne- inpotatofor resistanceto M.chitwoodi arehampered by matodeisolates to 6 genotypesof S.bulbocastanum re- thisgenetic variation in pathogenicity.

794 Nematology Symposiumabstract

Resistance of hybrid citrusrootstocks to the citrusnematode

Soledad VERDEJO-LUCAS 1, F. Javier SORRIBAS 2, Cesar ORNAT 1, Magda GALEANO 1, Juan B. FORNER 3 andAntonio A LCAIDE 3 1 Institutde Recercai TecnologiaAgroalimentà ries, Crta. Cabrils s/ n08348-Cabrils,Barcelona, Spain 2 EscolaSuperior d’Agricultura de Barcelona,Comte Urgell, 187. 08036 Barcelona, Spain 3 InstitutoV alencianode Investigaciones Agrarias, Apartado oŽ cial. 46113 Moncada, V alencia,Spain Presentedat the 25th International Nematology Symposium, Herzliya, Israel, 2-7 April 2000

Tylenchulussemipenetrans isthe most important plant- resistantstandards to the citrus nematode, respectively. parasiticnematode that affects citrus orchards in Spain. Fifteenselections of Cleopatra mandarin ´ P.trifoliata , Theuse of nematode resistant rootstocks can be an ef- three of Citrusvolkameriana ´ P.trifoliata , and one of fective,economic and environmentally safe controlman- Kingmandarin ´ P. trifoliata were resistantto the cit- agement.The response of 66 citrus hybrid rootstocks to rusnematode. The rootstockswith Troyer citrange in their aMediterraneanbiotype of T.semipenetrans was deter- parentagesupported nematode reproduction but showed minedin greenhouse tests. One-year old trees were in- differentlevels of susceptibility.The resistance response oculatedwith approximately 1 ´ 104 eggs + juveniles ofsome selected citrus hybrids to increasing inoculum perplant, and nematode infectivity (females/ groot)and reproductivepotential (eggs + juveniles/groot)were as- pressurehas been conŽ rmed in microplots. Some of these sessed6 monthsafter infestation. Sour orange and Pon- hybridspossess other characteristics that are of interestfor cirustrifoliata Rubidouxwere includedas susceptible and citrusproduction in Spainand elsewhere.

Parasite speciŽc genes from Meloidogyne artiellia

Pasqua VERONICO 1, John T. JONES 2,FrancescaD E LUCA 3, Mauro DI VITO 3,MariaRosaria C ORTESE 1, Franco LAMBERTI 3 and Carla DE GIORGI 1 1 Dipartimentodi Biochimicae BiologiaMolecolare, University of Bari, Bari, Italy 2 ScottishCrop Research Institute, Invergowrie, Dundee, Scotland, UK 3 Istitutodi Nematologia Agraria, CNR, Bari,Italy Presentedat the 25th International Nematology Symposium, Herzliya, Israel, 2-7 April 2000

Wehavecharacterised three Meloidogyneartiellia genes, genesisolated is unclear. One gene is similar to bacter- twoof whichmay be speciŽc toplantparasitic nematodes. ialgenes involved in capsularpolyglutamate biosynthesis. TheŽ rst geneshows similarity to chitin synthases from Thisgene hasnohomologsin the C. elegans genome or in Caenorhabditiselegans andfungi. The M.artiellia chitin thelargeanimal parasite EST dataset.It is expressedin all synthaseis present as a singlecopy in the genome and M.artiellia stagesexamined. The third gene is smalland isexpressed in unhatchednematodes. It seems likely that istranscribedin alllife cycle stages tested. No homologs nematodechitin synthases are involved in production of theeggshell chitin layer. W eareexamining the temporal ofthisgene are present in any of thedatabases, although andspatial expression patterns of the C. elegans and M. proteinswith similar Mr andpredicted pI arepresent in C. artiellia genesin more detail. The function of the other elegans.

Vol.2( 7),2000 795 Symposiumabstract

Belgiannematologists prepare for battle against potato cyst nematodes

Nicole VIAENE 1 and Tina MAHIEU 1 1 AgriculturalResearch Centre, Department of CropProtection, Burg. V anGansberghelaan96, 9820 Merelbeke, Belgium Presentedat the 25th International Nematology Symposium, Herzliya, Israel, 2-7 April 2000

Sincethe detection of Globoderarostochiensis in Bel- rostochiensis ,threecontained G. pallida andone Želdwas giumin 1946, few studieshave been conducted on the infestedwith both species. The survey will be expandedto spreadof potatocyst nematodes (PCN). Somefarmers use otherregions. A testfor choiceof cultivars,performed in croprotation and resistant varieties for themanagement of closedcontainers, was optimisedfor pathotypesof G. ros- PCN, butlack information about the degree of Želdinfes- tochiensis andmade available for farmers. Thetest will be tationand the pathotype present in their Ž eld.In a recent adapted for G. pallida.Detectionof cystsin Želdsneeds surveyin thenorth of Belgium,PCN was foundin 8.6% improvement.Several infested Ž eldswere sampledinten- of1320Ž eldsthat were sampledby taking60 soil cores sivelyto measurethe nematode distribution. The datawill perhectare (about 2 kgof soil).Species identiŽ cation of beusedto simulate a typicaldistribution pattern for Bel- cystsof 60 Ž eldsshowed that 56 were infestedwith G. gianŽ eldsand develop an adequatesampling scheme.

Diagnostic value of morphological characters insome genera of Actinolaimidae (Nematoda: Dorylaimida) with notes on the phylogeny of the family

MariaTeresa V INCIGUERRA andMirella C LAUSI Dipartimentodi BiologiaAnimale, Università di Catania, via Androne 81, 95124 Catania, Italy Presentedat the 25th International Nematology Symposium, Herzliya, Israel, 2-7 April 2000

Thefamily Actinolaimidae, among Dorylaimida, is char- Brittonematinae.Recently the identity of some of these acterisedby having four large onchia surrounding the generahas been discussed in various papers by different odontostyle.Many genera have been described as belong- authorsand the diagnostic value of somemorphological ingto thisfamily, mainly differing from eachother by one charactershas beenquestioned. Our study,conducted both orveryfew morphologicalcharacters. Still, some genera bylight and scanning electron microscopy, reviews the ofthis family differ from allthe others in severalcharac- variabilityand the consequent diagnostic value of those tersshared by allof them:a peculiarstructure of pharynx, charactersand allows a betterdeŽ nition of thegenera of moreslender odontostyle, higher lip region and a cuticle Brittonematinaeconsidered valid. The main evolutionary longitudinallyridged. This clearly monophyletic group trendsof the family and their phylogenetic implications ofgenerais attributed by some authors to the subfamily arealso discussed.

796 Nematology Symposiumabstract

Parasitism on roots and rhizomesof anthurium ( Anthuriumandraenum ) by Radopholus similis in Madeira

Nicola VOVLAS 1, Isabel M. DE O. ABRANTES 2, Alberto TROCCOLI 1,MargaritaP ESTANA 3 andM. Susana DE A. SANTOS 2 1 Istitutodi NematologiaAgraria, C.N.R., ViaAmendola165/ A,70126Bari, Italy 2 Departamentode Zoologia, Universidade de Coimbra, Coimbra, P ortugal 3 Laboratorioagricola da Madeira,Dir .Serv.Investigação Agricola,PT -9135Camacha, Madeira, P ortugal Presentedat the 25th International Nematology Symposium, Herzliya, Israel, 2-7 April 2000

Extensivesample collections were conductedin many an- zometissues showed that the burrowing nematode is an thuriumgrowing areas of Madeira, Portugal, since the exclusivelycortical feeder when it parasitises roots, but Žrst reportof occurrence of the burrowing nematode feedscommonly on meristematicperipheral tissues of rhi- Radopholussimilis (Cobb,1893), Thorne, 1949. The sur- zomes.Occasionally, vascular elements can be damaged veywas conductedon anthurium plants ( Anthuriuman- byitsfeeding activity. Newly laid and embryonated eggs, draeanum Lindenand Andre) imported and grown in ar- juvenilestages, mature females and males were recov- tiŽcial substrates in shade-houses to determine the level eredfrom necrotictissues; the males/ femalesratio, in well ofparasitism, population dynamics, parasitic habits, and establishedpopulations, varied from 1:20to 1:50. Ac- comparativemorphology of thedetected population with cordingly,this nematode reproduced efŽ ciently in thean- thosefrom otherhosts and origins. Radopholussimilis thuriumrhizome. Morphometric and morphological char- was foundparasitising roots (28-52 specimens/ gofroot actersof one population of R. similis aredescribed, il- tissues)and sometimes the rhizome, with induction of lustratedand compared with populations from otherhosts necroticlesions. Infected plants showed signs of losing andothergeographic origins and carrot-disk cultures. The vigour,with slightly yellowed leaves, and undersized, damageby thisphytoparasitic species to ower cultivation distorted owers.Histological analysis of root and rhi- andproduction is discussed.

PCR identiŽcation of singleindividuals of the dagger nematodes, Xiphinema index, X.diversicaudatum and X.vuittenezi using speciŽc primersfrom ribosomal genes

Xirong WANG 1, Chantal CASTAGNONE 2, Roger VOISIN 2, Pierre ABAD 2 and Daniel ESMENJAUD 2 1 NematologyLaboratory, South China Agricultural University, GuangZhou, P .R.C. 2 INRA-USVE, Nematologygroup, B.P .2078,06606 Antibes, France Presentedat the 25th International Nematology Symposium, Herzliya, Israel, 2-7 April 2000

The species Xiphinemaindex (XI), X.diversicaudatum suspectedas theputative vector of grapevinechrome mo- (XD) and X.vuittenezi (XV)areoften sympatric in North- saicvirus (GCMV) inCentral and Eastern Europe. All ernEuropean vineyards. For grapevine, X. index and threespecies may be mixedin theŽ eldand are difŽ cult to X.diversicaudatum arethe vectors of grapevine fanleaf identifyreliably when only single or few adultor juvenile virus(GFL V)andarabis mosaic virus (AMV), respec- individualsare available. With the objective of developing tively,the Žrst onebeingthe economically most important adiagnosticmethod answering this concern, an approx. 1 nepovirusesworldwide. Xiphinemavuittenezi has been kbITS regionspanning the 18s and 5.8s ribosomal genes

Vol.2( 7),2000 797 Symposiumabstract was sequencedin one population of each species, using (adultor juveniles)for eachspecies, illustrated by i) the twoconserved primers. Internal sense primers speciŽ c to presenceand ii)theexpected size of thediagnostic frag- eachspecies were designed.A singleampliŽ cation prod- ment.The study is in progressto conŽrm thespeciŽcity of uctwas obtainedfor eachspecies ( i.e. approx.330 bp theprimers in otherpopulations of thethree Xiphinema for XI, 810bp for XDand600 bp for XV). Thena re- speciesand the possibility to usethem in amultiplextest liableidentiŽ cation was obtainedfrom singleindividuals for mixedspecies.

Comparison of various combinations of ethylene dibromideand metham sodium with methyl bromide for control of weeds and nematodes intobacco seedbeds

Jennifer WAY andZibusiso S IBANDA TobaccoResearch Board, P .O.Box1909, Harare, Zimbabwe Presentedat the 25th International Nematology Symposium, Herzliya, Israel, 2-7 April 2000

TheefŽ cacy of the standard recommendation of methyl ofbroad leafed weeds in all treated plots but only the bromideapplied at 50g/m 2 for nematodeand weed con- methylbromide treatment gave good control of grasses. trolin tobacco seedbeds was comparedwith metham Althoughthe pressure of nematodes in the experiments 2 sodiumat 50ml/ m alone,and in combination with eth- was verylow, metham sodium on its own was notable ylenedibromide (EDB) 41%at 35, 28 and 21 ml/ m 2. toprovidesatisfactory control. A suitablecombination of Similarcomparisons were madekeeping the rate of EDB methamsodium and EDB, which provides good nema- 41%constant at 35 ml/m 2 whilevarying rates of metham sodiumfrom 15,25 to and 35 ml/ m 2.Alltreatments re- todeand weed control, could be agoodcandidate for the sultedin betterand similar germination of tobaccocom- replacementof methylbromide for seedbedfumigation in paredto the untreated control. There were lowernumbers tobaccoproduction in Zimbabwe.

798 Nematology Symposiumabstract

Development of diagnostics for distinguishing Meloidogyne chitwoodi and M. fallax from other root-knot nematodes

Jane WISHART, Vivian C. BLOK, John T. JONES and Mark S. PHILLIPS Departmentof Mycology, Bacteriology and Nematology, Scottish Crop Research Institute, Invergowrie, DD2 5DA Scotland,UK Presentedat the 25th International Nematology Symposium, Herzliya, Israel, 2-7 April 2000

Meloidogynechitwoodi and M. fallax haverecently be- widehost range they are difŽ cult to control.W ehavede- comequarantine organisms in the European community. velopeda sensitivePCR-based diagnostic which can be Theyare thought to have been introduced into The Nether- usedto distinguish single juvenile or female nematodes landsfrom theUnited States, probably by infestedtubers, of M.chitwoodi and M. fallax from other Meloidogyne ornamentalor root-stock material. Initially, these nema- spp. (M. hapla, M.incognita , M. javanica, M. arenaria todeswere misidentiŽed as M. hapla butnow two species and M.mayaguensis ), followinga simpleprocedure in- arerecognized. They are serious pathogens of severalEu- volvingdisruption of thenematodein water. W ehavealso ropeancrops particularly potato, but also infect alfalfa, producedantisera to thesenematodes which can be used thecereals, maize, beet, carrot etc.,andbecause of this todetectinfected plant material.

Material balance of 14C-oxamyl after the translocation inthe plant and the consequence for root-knot nematode control strategies

Albert E. WISSING 1,2,RichardA. S IKORA 2 andStephen N. I RVING 1 1 DuP ontde Nemours(France) S. A.,European Research Development Center ,24,Rue des Moulin, F-68740 Nambsheim, France 2 Institutfü r Panzenkrankheiten, Phytopathology in Soil-Ecosystems& Nematology,Universitaet Bonn, Nussallee 9, D-53115 Bonn, Germany Presentedat the 25th International Nematology Symposium, Herzliya, Israel, 2-7 April 2000

R Oxamyl(Vydate ®)hasunique plant systemic properties. cantuptake of oxamylby theplant prior to anydegrada- Itcontrols effectively most nematode species and a wide tion.Up to 81%of thesoil applied oxamyl was takenup rangeof sucking and piercing insects as well as miteson bytheplant within 10 days.The results show further that manyŽ eldcrops, vegetables, fruits and ornamentals. T o thecrop itself exerts a majorrole in preventing leaching effectivelyintegrate oxamyl into a givenpest management ofoxamylout of therooting zone into the sub-soil water. programme,the systemic action, which includes both up- Themultiple application via adripirrigation system per- ward acropetaland downward basipetal translocation of activematerial in the plant, needs to bequantiŽed in order mitsthe utilisation of smaller amounts of oxamyl. This tooptimise efŽ cacy and minimise environmental impact. prolongsthe time of uptakeby the plant and thereby re- Thisstudy with radiolabelled 14C-oxamylshows signiŽ - ducesthe chance of groundwater contamination.

Vol.2( 7),2000 799 Symposiumabstract

Planting depth and nematicideincorporation depth affect control of the potato cyst nematode and yieldof potatoes

Simon WOODS andPatrick H AYDOCK PlantNematology and Soil P estsGroup, Crop and Environment Research Centre, Harper Adams University College, Newport, Shropshire,TF10 8NB, UK Presentedat the 25th International Nematology Symposium, Herzliya, Israel, 2-7 April 2000

AŽeldexperiment was doneat Harper Adamsto investi- prox.10 cm), medium(approx. 15 cm) anddeep (approx. gatethe effect of nematicideincorporation and seed tuber 25cm). Resultsshowed that the medium depth nematicide plantingdepth on the yield of thepotato cultivar Estima incorporation,when tubers were plantedat a shallowor andthe population control of the potato cyst nematode mediumdepth, reduced root invasion compared with the Globoderarostochiensis .Nematicidewas appliedat com- othertreatments. Medium depth nematicide incorporation mercialŽ eldrate and incorporated to three depths: shal- alsogave consistently larger ware yieldsand better nema- low,which involved the broadcast of thenematicide on to todecontrol than the other incorporation methods, which thesoil surface prior to planting,medium incorporation to were notsigniŽcantly different from thecontrol.However, 20cmanda deepincorporation down to 35cm.Potatoes yield and Pf/Pi ratioswere notsigniŽ cantly affected by were mechanicallyplanted to three depths: shallow (ap- plantingdepth.

Incorporating granular nematicidesinto potato seedbeds: the current UK situation

Simon WOODS andPatrick H AYDOCK PlantNematology and Soil P estsGroup, Crop and Environment Research Center ,HarperAdams University College, Newport, Shropshire,TF10 8NB, UK Presentedat the 25th International Nematology Symposium, Herzliya, Israel, 2-7 April 2000

Theuse of granular nematicides for managinginfesta- areincorporated into the seedbed. Soil is cultivated to a tionsof potatocyst nematodes remains Ž nanciallyattrac- greaterdepth (over30 cm) andplanting/ ridgingis donein tiveto UK growersas partof anintegrated management onepass with tubers often planted at depthsgreater than strategy.Nematicide granules were traditionallybroad- 20cm.Many of thecultivation practices will not place the castonto the soil surface and rotavated or harrowed to nematicidegranules and seed tubers together in the soil. adepthof 15 cm. Potatoes would then be planted 5- Thishas consequences for nematicideefŽ cacy. The ma- 10cm deep and ridging up would take place after the chinerycombinations that can be potentially used in the crophad emerged. This method followed the industryrec- ommendationsand growers still using this approach are Želdfor incorporatingnematicide granules and the conse- unlikelyto have problems with nematicide efŽ cacy. The quencesof thesetechniques for cropgrowth and nematode adoptionof potatobeds has changed the way nematicides controlare discussed.

800 Nematology Symposiumabstract

Interactions between plant parasitic nematodes and the banana weevilon smallbanana plants

Nathalie WUYTS 1, Carine DOCHEZ 2, Paul R. SPEIJER2,† and Dirk DE WAELE 1 1 Laboratoryof Tropical Crop Improvement, Catholic University of Leuven( KUL), KardinaalMercierlaan 92, 3001 Heverlee, Belgium 2 InternationalInstitute of TropicalAgriculture ( IITA)— Easternand Southern African Regional Centre (ESARC), P.O.Box7878, Kampala,Uganda †Deceased Presentedat the 25th International Nematology Symposium, Herzliya, Israel, 2-7 April 2000

Cookingand beer bananas are primary staples in East- similis alone, Pratylenchusgoodeyi alone,or notinfested. ernAfrica. Productionis threatened by a pestcomplex Fourweeks after inoculation, the plantswere infestedwith consistingof thebanana weevil, Cosmopolitessordidus, thebananaweevil or not(control).Presence of thebanana plant-parasiticnematodes and leaf diseases. Surveys con- weevilsigniŽ cantly increased percentage coefŽ cient of in- ductedin East Africa suggestan interaction between festation(PCI) andperipheral damage, particularly in R. thebanana weevil and nematodes, whereby nematode- similis infestedplants. Moreover, the banana weevil had infestedplants suffer ahigherattack by weevils. T oin- astrongpreference for plantsinfested by R. similis, when vestigatethis interaction, 4-week old plants of the com- compared to P. goodeyi infestedor controlplants. On the monlygrown cultivars, Mbwazirume ( Musa AAA, high- otherhand, penetration of weevillarvae into the corm is landcooking banana type) and Pisang A wak( Musa ABB, less for R. similis infestedplants and higher for P. goodeyi brewingbanana type), were infestedwith Radopholus infestedplants, when compared to thecontrol.

Adaptation of apopulation dynamics model for prediction of potato cyst nematode distribution within Želds

Jingyi YANG 1,2, Joseph PERRY 2, Patrick HAYDOCK 1 andKenneth E VANS 2 1 PlantNematology and Soil P estsGroup, Crop and Environment Research Center ,HarperAdams University College, Newport, Shropshire,TF10 8NB, UK 2 IACR-Rothamsted,Harpenden, Herts, AL5 2JQ,UK Presentedat the 25th International Nematology Symposium, Herzliya, Israel, 2-7 April 2000

Thepopulation dynamics equation of Jonesand Perry has putscreenhas beendesigned to allow input parameter val- beenused for thedevelopment of amodelthat will permit uestobe speciŽed for inputof eitheraverage Ž eldpopula- simulationsto be made of within-Ž eld changes in popu- tiondensities or datasetsof within-Želd distributions, and lationdensities of potatocyst nematodes. Detailed infor- examplesof thelatter are given. Other purposes for which mationfrom oneŽ eldhas allowed us toestimatethe most themodel can be used,such as modellingnematode pop- appropriatevalues for thatŽ eldof the input parameters requiredby the model. Analyses of thesensitivity of the ulationchanges through rotations or followingnematicide modelto theseparameters are presented. A convenientin- treatments,are also illustrated.

Vol.2( 7),2000 801 Symposiumabstract

IdentiŽcation of Meloidogyne incognita, M.javanica and M. arenaria using sequencecharacterised ampliŽed region (SCAR)based PCR assays

Carolien ZIJLSTRA 1,DorineT.H.M. D ONKERS-VENNE 1 andMireille F ARGETTE 2 1 PlantResearch International, P .O.Box 16, NL-6700 AA Wageningen,The Netherlands 2 CBGP (Centrede Biologie et de Gestion des P opulations)— IRD(ex-ORSTOM),BP 5045,34032 Montpellier Cedex 1, France Presentedat the 25th International Nematology Symposium, Herzliya, Israel, 2-7 April 2000

Threerandomly ampliŽ ed polymorphic DNA (RAPD) identify M.incognita , M. javanica and M.arenaria. The markers,species speciŽ c tothe root-knot nematode lengthvariant SCAR markers can be ampliŽed from DNA species Meloidogynearenaria , M.incognita and M. ja- from eggmasses, second stage juveniles and females. This vanica,were identiŽed. After sequencingthese RAPD- speciesidentiŽ cation technique is therefore independent PCRproducts, longer primers of 18 to 23 nucleotides ofthe nematode life cycle stage. Moreover, the SCAR- were designedto complement the terminal DNA se- PCRassaywas successfullyapplied using DNA extracts quencesof the DNA fragments.This resulted in three from infestedplant material. The method has potential to pairsof speciesspeciŽ c primersthat were usedto amplify thesequence characterised ampliŽ ed regions (SCARs). beoptimised for routinepractical diagnostic tests facil- Thedeveloped sets of SCAR primers were successfully itatingthe control of these economically important pest usedin straightforward, fast and reliable PCR assays to organisms.

Isolation and characterisation of glycogen synthase gene from the insect parasitic nematode Steinernema feltiae IS6

Tali ZITMAN GAL 1, Ahron SOLOMON 2, Itamar GLAZER 1 andHinanit K OLTAI 1 1 Departmentof Nematology,ARO, TheV olcaniCenter ,BetDagan 50250, Israel 2 Departmentof AnimalSciences, F acultyof Agriculture of theHebrew University of Jerusalem, Rehovot 76100, Israel Presentedat the 25th International Nematology Symposium, Herzliya, Israel, 2-7 April 2000

Steinernemafeltiae isused as abiologicalcontrol agent. encodea 694-residueprotein. Sf-gsy-1mRNA istrans- S. feltiae canpartially tolerate desiccation by entering a splicedby the nematodetrans-spliced leader SL-1. South- shallowdormancy. A new,higherdesiccation tolerant S. ernblot analysis suggested that Sf -gsy-1is a singlecopy feltiae strain,IS-6, was recentlyisolated from theNegev gene.W ealsoestablished the changes in thesteady state desertin Israel.W eareinterested in the metabolicchanges level of Sf -gsy-1transcripts upon dehydration, and ob- thatenable the high desiccation tolerance of IS-6 strain. serveda signiŽcant decline after 24 hofdesiccationstress. Byusing differential display we identiŽed novel individ- Thereduction in the steady state level of glycogen syn- ualgenes whose transcript level is modiŽed uponinduc- thasetranscripts correlated with reduction in glycogen and tionof anhydrobiosis.One of thegenes is glycogensyn- increasein trehaloselevels. Understanding the genetic and thase,the rate-limiting enzyme in the synthesis of glyco- biochemicalnet that results in highdesiccation tolerance gen.W eisolatedthe entire open reading frame ofIS6 maycontribute to theimprovement of nematodesas bio- glycogensynthase ( Sf -gsy-1)and found it to potentially logicalcontrol agents.

802 Nematology Symposiumabstract

Quantitative detection of tobacco rattle virus (TRV)intrichodorid vector nematodes by Taqman RT-PCR

Frans C. ZOON, Paul C. MARIS, Orhan BICAKCI and Cor D. SCHOEN PlantResearch International, PO Box16, 6700 AA Wageningen,The Netherlands Presentedat the 25th International Nematology Symposium, Herzliya, Israel, 2-7 April 2000

Amolecularassay was developedto detecttobacco rattle tionof thenumber of TRV-particlesin individual nema- virus(TR V)intrichodorid vector nematodes. It is based todescould be made by using real time measurement of onselectiveampliŽ cation of cDNA synthesizedfrom vi- uorescenceduring the PCR anddilution series of RNA ralRNA, anddetection of thePCR product by auores- extractsfrom puriŽed TRV.Therange found was consis- centT aqman-probe.The target RNA-sequence is a part tentwith estimations based on thesize and virus packing ofthe RNA1 genome common in TR Visolatesstudied ofthenematode oesophagus. The method provides a sen- sofar .TRVwas detectedin plant samples infected with sitivetool for quantitativestudies on virus transmission. differentTR Vserotypes,in individual viruliferous Para- Thepossible use of this quantitative RT -PCRfor quick trichodorusteres andin nematode fauna extracted from measurementof theTR Vinfectionpotential in soil sam- soilscontaining various trichodorid species. An estima- plesis under study.

Image Digitisation inNematology —Digital Databases for Research, Teaching, and Extension, NemaPix, Vol. 2(1999)

Ulrich ZUNKE 1 andJonathan D. E ISENBACK 2 1 Universityof Hamburg, F acultyof Biology, Plant Protection, Institute of AppliedBotany, D-20355 Hamburg, Germany 2 VirginiaT ech.,Dept. PPWS, Blacksburg,VA 24060,USA Presentedat the 25th International Nematology Symposium, Herzliya, Israel, 2-7 April 2000

Therapid development of image digitization for storage images.The images are useful for observationdirectly anddisplay on personal computers offers togreatly en- onthe computer screen or they can be printed on pa- hanceentomological research and teaching through the per,overhead transparencies, or even photographic Ž lm. developmentof digital databases. One example of digi- Theimages can be used for presentationswith a digital tisedimages stored electronically on a CD-ROM is A projectoror overhead projector panel connected directly Journalof Nematological Images editedby J.D. Eisen- toa computeror powerbook. The equipment and steps back& U.Zunke.NemaPix, V ol.1 was publishedin for thedevelopment and use of digitisedimages are pre- 1997/98.Similar to V ol.1 NemaPix,V ol.2 (Dec. 1999) sentedin a diagram.On V ol.2 thereare 11 short digi- isnow published. There are 1064 images from 26contrib- tizedvideos, a slideshow(200 Images for Presentation utorsworldwide stored on this2nd V olumeon CD-ROM etc.)andreferences to books which are already out of for usewithin research, teaching, and advisory services. Thephotographs and micrographs are easily searched print (e.g.,Cobb).For moreinformation please contact: andaccessed for easeof use. The pictures are stable [email protected] or [email protected] or homepages: andnot subject to degradation such as fading, scratch- http://dreamwater.com/biz.mactodeor http://www.zunke- ing,staining, etc.,thatare common with photographic photography.com.

Vol.2( 7),2000 803