grains. Key words:Bioinsecticide, maize weevil, peumo,stored showed repellenteffect, butnotfumigant activity. was notaffected. Both,thepowder andtheoiltreatments wheat seedstreatedwithC.albapowderandessentialoil grain and40mL essentialoilkg levels wereobtainedat80gpowderkg mortality higher The replicates. 10 and treatments five with seed germination. A completelyrandomizeddesignwasused repellent effect, andimpactonwheat (Triticum L.) aestivum by contactandfumigantactivity, adultemergence (F against ( properties ofthepowderandessentialoilpeumo to evaluate,underlaboratoryconditions,theinsecticidal such asbotanicalinsecticides. The aimofthisresearchwas has led tothesearch for morefriendly control alternatives insecticides butproblemsastoxicresiduesandresistance insect speciesisusuallycontrolledbymeansofsynthetic asthe pests insect maize weevil(SitophiluszeamaisMotschulsky).This is crops of these problems significant animal diet.Understorageconditions,oneofthemost Cereals constitutearelevantpartofhumananddomestic ABSTRACT essential oilkg S. zeamais(F of adultS.zeamaissurpassed80%. The emergence ofadults doi:10.4067/S0718-58392016000100007 Accepted: 6October2015. Received: 17 April 2015. México-Texcoco, Montecillo, EstadodeMéxico,México. 2 * 595, Casilla537,Chillán,Chile. 1 Looser against Motschulsky J. Concepción Rodríguez Juan J.Pinto essential oilof Insecticidal activityofpowderand [Molina] Looser; ) alba[Molina]Looser;Lauraceae)leaves Colegio dePostgraduados,Campus Montecillo, km 38,5Carr. Corresponding author([email protected]). Universidad de Concepción, Facultad de Agronomía, Vicente Méndez CHILEAN JOURNAL OF AGRICULTURALCHILEAN JOURNAL OF AGRICULTURAL RESEARCH 76(1) JANUARY RESEARCH 76(1)JANUARY S. zeamais. The variablesassessedweretoxicity 1 ) wasreducedby100%at80gpowderkg -1 ; in both cases, the mortality grainofC.alba;inbothcases,themortality 1 , GonzaloSilva CHILEAN JOURNAL OF AGRICULTURAL RESEARCH 76(1)JANUARY 2 , andAngelLagunes -1 grain. Germination of grain.Germinationof 1* , InésFigueroa Sitophilus zeamais Sitophilus Cryptocarya alba Cryptocarya -1 grainand40mL 1 , MaritzaTapia 1 2 ), ), -1 - -

MARCH 2016 MARCH 2016 oil from conditions, insecticidal properties ofthe powder andthe essential others. The aim of thisresearchwas to assess,underlaboratory (Coleoptera: Tenebrionidae) (Tapondjou etal.,2005),among (Kordali etal.,2006)andTribolium confusum Jacquelin du Val et al.,2007),SitophilusgranariusL.(Coleoptera: Curculionidae) of storedproductsasS.zeamais(Tapondjou etal.,2005;Kouninki components, have shown insecticidal activity against insects pest that contain terpinen-4-ol, 1,8-cineole or cymol as the main 1,8-cineole and terpinen-4-ol, α-pinene, (Avello et al., 2012). Studies of essential oils extracted from other components cymol, main β-terpinene, the being β-pinene, oil, essential of 0.3% foliage, reaching 15-20 m tall (Hoffmann, 1983). Its leaves contain to LosLagosRegion.Itisaperennial tree withdenseanddark is a tree native from Chile that grows from southern Coquimbo important toexploreotherChileanplantspecies. & G. Forst.(Winteraceae) (Andrade et al., 2009). Hence, it is (Monimiaceae) (Ortiz et al., 2012), and (Torres et al., 2014), 2010), as boldo(PeumusboldusMolina;Monimiaceae) (Betancur etal., derivates is verylimited and focusesonareduced number ofspecies the essentialoilisabetteroption. mixing it with the grain. However, if fumigant effect is required, against stored grain pest consistondryingthe foliage and then (Silva et al., 2003). The easiest way tousebotanical insecticides contact, fumigant, antifeedant, and repellent activity against insects and essential oils for many years. These pesticides have shown an alternative. Hence, the use ofbotanical insecticides should be considered as L.), maize (Zea maysL.),andsorghum (Sorghum bicolorL.) different insecticides usedagainst thispestonrice (Oryza sativa State University, S.zeamaishas32reportsofresistance to nine Resistance Database (www.pesticideresistance.org) ofMichigan synthetic insecticides but accordingtothe Arthropod Pesticide worldwide. This insect species is usually controlled by means of weevil (Sitophilus zeamais Motschulsky)is a key pest of cereals Because of the grain damage on stored conditions, the maize INTRODUCTION The peumo (Cryptocarya alba The Chileanresearchaboutinsecticidal activity of native plants The botanical insecticides have beenusedaspowders,extracts, 1 Laurelia sempervirens (Ruiz & Pav.) Tul. (Monimiaceae) , AngélicaUrbina C. albafoliageagainst (Molina) - MARCH 2016

Laureliopsis phillippiana 1 , S. zeamais. [Molina] Looser; Lauraceae), Drimys winteri (Looser) Schodde J.R.Forst.

48 RESEARCH CHILEAN JOURNAL OF AGRICULTURALCHILEAN JOURNAL OF AGRICULTURAL RESEARCH 76(1) JANUARY RESEARCH 76(1)JANUARY mA; scanrate1s as follows: ionization voltage 70 eV; emission current 40 with asplitratio1:20.Massspectrometry conditions were University of Concepcion, Campus Chillan, and they were cultures maintained in the Laboratory of Entomology of The insects used in those study were obtained from stock Insects andgrain mL 1 rate flow (helium), min gas carrier conditions wereasfollow:injector temperature, 250 °C; Operation µm). 0.25 thickness film m, 30 length mm, 0.25 diphenyl 95% dimethylsiloxane bonded phase column (i.d. California, USA). Separation was achieved using a 5% poly (HPGC-MS series II 5890; Hewlett Packard, Palo Alto, high performance gas chromatography mass spectrometry using a coupled tomassspectrometrydetection (GC-MS), with sodiumsulfatetoeliminatewatertraces. the essentialoilwastreated by gaschromatography(GC), containers at 4.5 °C. Previous tothe bioassays andanalysis Subsequently, theoilwasstoredinamber-colored glass water inaClevengertypeapparatus(Kouninkietal.,2007). were obtainedbysteamdistillation during4husingdistilled has the highest level of grain adherence. The essential oils USA). According toSilvaetal.(2003)thisgranulometry a20 throughout was sieved mesh (0.841mm)(DualManufacturing, Chicago, Illinois, which powder fine a obtain coffee grinder(A5052HF, Moulinex, Aleçon, France)to at 40°C. After that,thefoliage was groundedinanelectric a stove (UNB500,Memmert Gmbh, Schwabach,Germany) in h 48 during out dried were leaves the field-collected, Agronomy ofUniversity of Concepcion at Chillan. Once of tovoucher identification CONC-CH 5488and deposited in herbarium of Faculty of according taxonomic verified The was foliage criteria. field-collected (1997) al. et Ñuble Province, Biobío Region, Chile, using the Vogel 2012 fromPintocounty(36°42’ S,71°54’ W, 286ma.s.l.), December in field-collected was Cryptocarya albafoliage Essential oilextraction andanalysis MATERIALS ANDMETHODS rate of 10°Cmin isothermal at 60 °C and then increased to 210 °C,at the range was35-300Da. The oventemperature was 2min were calculatedusingamixtureofn-alkanes(C analysis. Intheseconditionsthelinearretention indices (RI) in the same experimental conditions described for GC-MS detector ionization flame using analysis GC from obtained literature. The relative amounts of each component were and Technology [NIST], Gaithersburg,or Maryland,USA) the library NIST NBS54K(National Institute of Standards available in our laboratory, and by matching the MS data with MS fragmentation pattern was checked with the standards Samples (1 µL) were dissolved with CH Chemical analysisofessentialoilwasassessedbyGC -1 , andto260°C,atrate of 10°Cmin -1 ; sourcetemperature 285 °C.Mass 2 -1 Cl andsplit injection 2 (1:100v/v). The 8 -C 28 - - MARCH 2016 MARCH 2016 ). -1 . then frozenat-4±1°Cfor48hpriortoitsuse. in a stove (UNB 500, Memmert Gmbh) at 25 °C for 12 h and healthy grain it was washed with drinkable water and dried from the and vegetable market of city of Chillan. Only determined usingthecriteriaproposedbyHalstead(1963). 749, Memmert Gmbh, Schwabach, Germany). The sex was (IPS chamber bioclimatic a in darkness total and RH 5% ± 60 °C, 1 ± 30 at wheat containing flasks glass L 1 in reared oil kg powder kg previously described. as assessed was °C, Mortality darkness. total 1 and RH ± 5% ± 30 60 at chamber bioclimatic a in stored were units infested with 20 insect couples, 48 h-old. The experimental were flasks evaporate. the to that, acetone After the allow to °C) 5 ± (20 temperature room at h 2 for left and uncovered hand shaken and covered during 15stoproperlycoverthegrainswithoil;thenthey were flasks the Then, grains. wheat g 200 containing flasks mLglass 400 to applied were Reichmuth (1997). Solutionsof1mL essential oil in acetone carried out usingthemethodology ofObeng-Oferiand environmental conditions. to abioclimatic chamber under previouslydescribed returned flasks respective the and removed were insects all a needleduring30s. After recordingthepercent mortality, when theyfailedtomoveafterbeingproddedgentlywith was discardedandrepeated. An insectwasconsidereddead exceeded 10%mortality, thewholerespectivereplicate means of the Abbott’s formula (Abbott, 1925), but if control assessed at7dafterinfestation (DAI) and wascorrected by 5% RHand total darkness). The percentage of mortality was ± 60 °C, 1 ± (30 chamber bioclimatic a to transferred were with 20pairsofinsects. After insect infestation, containers control consisted inajarwith100gwheatgrainsinfested After that,20insectcouples,48h-oldwereadded.Untreated grains. Then thecontainer was handshakenduring1min. was addedinto 250 mL glassjarscontaining 100 gofwheat methodology of Silvaetal.(2005). The respective treatment The bioassaywithpowderwascarriedoutusingthe Bioassays ofcontact toxicity the medianlethal concentration (LC emergence oftheF the after 36(week3),505)and557)DAI Cary, NorthCarolina,USA). Statistical Analysis System (SAS) software(SASInstitute, to ProbitanalysisusingthePROCPROBIT procedure of seeds per replicate were selected, then they were laid down on seeds per replicate were selected, then they were laid down on evaluated 55 DAI. To set up the experiment, 20 healthy wheat considering valuesoftheuntreated controlas100%. were recorded and percent emergence was calculated, Wheat forinsectrearingandbioassayswasobtained The evaluated treatments were 5.0,10,20, 40,and80g The contacttoxicity bioassays withessentialoilwere For both, thepowderandessentialoilbioassays, For The impact of the treatments on wheat germination was The impactofthetreatments onwheatgerminationwas -1 grain.Everytreatment had 10replicates. To obtain -1 grain and 2.5, 5.0, 10, 20, and 40 mL essential 1 was assessed. was The amountofF 50 ), dataweresubjected 1 adults

49 CHILEAN JOURNAL OF AGRICULTURALCHILEAN JOURNAL OF AGRICULTURAL RESEARCH 76(1) JANUARY RESEARCH 76(1)JANUARY Bioassays offumigant andrepellent effect considering untreatedcontrolas100%wasestimated. germination arelative hence certified, not were bioassays in conditions inabioclimaticchamberduring7d. The grainsused a wet filter paper and kept under of 24 ± 2 °C and 60 ± 5% RH design. The variable percentage was transformedtothe The experimental designwasacompletely randomized analysis Experimental design andstatistical kg treatments assessed were 5.0, 10, 20, 40, and 80 g powder assessment by the fumigant effect was made 5 DAI. The with 20insectswithoutpowderinthetube. The mortality consisted in plastic containers with 20gwheatgrainsinfested and infestedwith20unsexedinsects. The untreated control grains, wheat g 20 with filled was tube the and container the the environment. Then, the rest oftheinnerspacebetween powder, butallowing the release of volatile compounds into organza fabric to prevent direct contact of insects with the fine of piece a with covered was tube PVC of end the Then, respective treatment, was stuck vertically with silicone. tube of5cmlongand2.5indiameter, containingthe (2005). At thebottomof200mL plasticcontainers,aPVC alba powderwasadapted from Tavares and Vendramim The methodology to evaluate the fumigant effect of the and Vendramim (2003), who classified the treatment as treatment the neutral iftheindex=1,attracting>1andrepellent< 1. classified who (2003), Vendramim and was calculated with theseresultsaccording to Mazzonetto possible interference of external factors. The repellent index replicate, the treatments were randomlyrotatedtoavoidthe was recorded.Each treatment had 10replicates and ineach for 24 hand the number of insects that moved to each treatment darkness), total and RH 5% ± 60 °C, 1 ± (30 chamber et al.,2003). The choicearenaswerekeptinabioclimatic grain and the other two 20gofuntreated grain (Procopio and 0.5cmdiameter), two dishescontained20gtreated dish was connected to four dishes using tubes (10 cm long 48 h-oldofS.zeamaiswithoutsexing. 20 adults, This Petri was achoice arena consistinginacentral Petri dishwith (2003) wasusedbyrepellent effect. The experimental unit each treatment. Mortality was assessed5DAIand10replicates were runfor infested with10adultinsects,48-holdwithoutsexing. equivalent to 0.5L)and200gwheatgrainwasadded of screwcapsa500-mL glasscontainer (air volume UK). Then, the treated paper was attached to undersurface Kent, Maidstone, Whatman, diameter; cm (5.5 paper filter 10 nr Whatman circular a on oil essential of μL 35 and 30, (2006), whichconsistedofapplying 0 (control), 15, 20,25, of essential oil, weusedthe methodology of Pireset al. -1 The methodologyproposedbyMazzonettoand Vendramim To carryoutthebioassaytoevaluate the fumigant toxicity grainandeachtreatmentconsistedof10replicates. - - MARCH 2016 MARCH 2016 C. fiducially limits did not overlap at a given mortality level (50%) (RobertsonandPreisler, 1992). givenmortality ata not overlap did limits fiducially responses wereconsidereddifferent whentheirrespective analysis Probit 0.05). ≤ (p used was test comparison means one treatment wasdifferenta fromtherest.Ifso, Tukey = (α ANOVA out 0.05) testwiththeSASprogramtodetermine if atleast carry to prior function arcsine √x/100 and as a male pheromone of ectoparasitoid plants inresponsetoinsectherbivory(Zhuangetal.,2012) a sesquiterpene identified as a volatile compound released by insecticidal properties is the(E)-beta-bergamotene which is 71°54’ W), ÑubleProvincefoothills. December (summer season) in Pinto county (36°42’ S; in Nonguen Valley wood(36º49’ S;73º01’ W) andwein Region but Avello etal.(2012)inNovember(springseason) and Avello etal.(2012)andwecollected foliage inBiobío Ocoa (32°54.957’ S;71°02.7080’ W), Valparaiso Region, Teillier (2007) collected foliage on April (fall season) in and zone Niemeyer field-collected. was the geographical material the on where and date field-collection the in attribute these differences to the fact that there are differences (21.4%) and 1-terpinen-4-ol (28.19%), respectively. We 1,8-cineole identified mainly who (2012), al. et Avelloand Our resultsdonotagreewithNiemeyerand Teillier (2007) (1.53), jasmoline 1 (1.47%),and safrole (1.1%) (Table 1). β-cubebene (1.85%), (2.93%), 1,8-cineole (2.74%), α-farnesene β-himachelene (3.38%), propanone cyclopentane β-apo13carotenone(5.3%), linalool (4.4%),(-)-terpinen-4-ol(3.45%),2-methyl- (7.65%), (8.5%), germacrene-D viridiflorol (15.6%), (E)-beta-bergamotene were The principal constituents of the essential oil of RESULTS ANDDISCUSSION kg powder wasobtainedwiththe treatment of 80gpowder The highestmortality reached bytheuseofC.alba Contact toxicity activity against Huang etal.(2002)indicated that safrole hasinsecticidal Dahms (Hymenoptera: Eulophidae) (Consoli et al., 2002). S. granarius, Oferi andReichmuth (1997)thiscompound alsoaffects (Coleoptera: Tenebrionidae) and according to Obeng- (Hemiptera: Psyllidae)(Liuetal.,2014),amongothers. et al., 2011), andCacopsylla chinensis (Yang & Li, 1981) Sitophilus oryzae L.(Coleoptera: Curculionidae) (Prieto insecticidal activity against S.zeamais(Chuetal.,2012), of theessential oils inotherplantspecies,andhaveshown (-)-terpinen-4-ol and 1,8-cineole,which arecomponents identified compounds are viridiflorol, germacrene- Horn) (Coleoptera: Bostrichidae). truncatus (G.H. The other -1 The main secondary metabolite identified with identified metabolite secondary main The grainwhichcaused 85% ofmortality (Table 2), being S. zemais, S. zeamais and T. castaneum, Tribolium castaneum (Herbst) Melittobia digitata and Prostephanus d, linalool, C. alba

50 CHILEAN JOURNAL OF AGRICULTURALCHILEAN JOURNAL OF AGRICULTURAL RESEARCH 76(1) JANUARY RESEARCH 76(1)JANUARY showed amortality greaterthan80%,our resultsexhibit 80 gpowderkg to controlS.zeamaisonlyatconcentrations higher than can inferthatthepowderofC.alba foliagehasthepotential (p ≤ 0.05). The rest of treatments (5.0 to 40 g powder kg powder g 40 to (5.0 treatments of rest The 0.05). ≤ (p treatments other the with comparison in higher significantly Dehydro-aromadendrene Veridiflorol 2-Methyl-cyclopentane propanone α-Cubebene 3,9-Dodecadiine β-Phenylethyl butyrate β-Himachelene 3-Decine Jasmoline I β-Apo-13-carotenone -cyclohexanol methylene-but-3-enylidene)-1 1,3,3-Trimethyl-2-(3-methyl-2- Unknown α-Farnesene 2-(1,3-Butadienyl) mesitylene Cubinene β-Cubebene Bicyclosesquifelandrene ∂-Elemene α-Bulnesene α-Cadinene Germacrene- Phenethyl isovalerate ∂-Cadinene Isoledene α-Caryophyllene β-Bisabolene (-)-Aristolene E-β-Bergamotene (-)-Germacrene β-Phenylethyl butyrate α-Cubebene Safrole 2-Nonanone 7-Octen-2-one Linalyl formate Terpineole (-)-Terpinen-4-ol (3Z)-2,2,5,5-Tetra methylhex-3-eno Pinocarveol cis-p-2-Menthen-1-ol Unknown 3-Isopentyl isovalerate Linalool Isoterpinolene 1-8-Cineole 1,5-Dimethyl-1,5-cyclooctadiene 1,3,8-p-Menthatriene β-Terpinene α-Felandrene Sabinene Isoterpinolene Heptanal Cryptocarya alba Table 1.Chemicalcompositionoftheessentialoilfrom was 50gpowderkg grain) showedalevel of mortality lower than30%. The LC database, Kovatsretentionindices(IK),andpurestandards(S). †† † Compound Retention time. Compounds identified by comparison with mass spectra (MS) from mass spectra with comparison by identified Compounds d . -1 grain. Although, 80gpowderkg -1 grainandaccordingtothisresults, we 16.105 15.570 15.470 15.410 15.340 15.280 15.220 15.060 14.866 14.710 14.660 14.540 14.330 14.216 14.070 13.944 13.850 13.66 13.55 13.49 13.47 13.44 13.43 12.22 13.13 13.00 12.95 12. 81 12.27 12.25 10.87 10.82 10.60 10.27 11.69 9.40 9.22 8.86 8.63 8.33 8.18 8.13 7.93 7.79 6.89 6.83 6.76 6.63 6.43 6.17 5.55 4.73 RT min % †

Relative content 15.60 0.45 8.50 3.38 0.84 0.28 0.17 2.74 0.25 1.47 5.30 0.15 0.52 2.93 0.25 0.90 1.53 0.81 0.29 0.25 0.16 7.65 0.76 0.18 0.79 0.23 0.22 0.28 0.44 0.32 0.73 1.01 0.24 0.24 0.22 0.87 3.45 0.58 1.84 0.22 0.26 2.20 4.40 0.45 1.85 0.43 0.15 1.64 0.77 0.50 0.27 0.18

Identification MS, IK,S MS, IK,S MS, IK,S MS, IK,S MS, IK,S MS, IK,S MS, IK,S MS, IK,S MS, IK,S MS, IK,S S IK, MS, MS, IK,S MS, IK,S MS, IK,S MS, IK,S MS, IK,S MS, IK,S MS, IK,S MS, IK,S MS, IK,S MS, IK,S MS, IK,S MS, IK,S MS, IK,S MS, IK,S MS, IK,S MS, IK,S MS, IK,S MS, IK,S MS, IK,S MS, IK,S MS, IK,S MS, IK,S MS, IK,S MS, IK,S MS, IK,S MS, IK,S S IK, MS, MS, IK,S MS, IK,S MS, IK,S MS, IK,S MS, IK,S MS, IK,S MS, IK,S MS, IK,S MS, IK,S MS, IK,S MS, IK,S MS, IK,S MS, IK,S MS, IK,S - - MARCH 2016 MARCH 2016 -1 grain †† 50 -1

(40 g powder kg mortality with concentrations similar or lowerthan 4.0% and against a lowertoxicitythanothervegetablepowdersassessed 94% deadinsects. This treatment had an estimated LC essential oil kg with concentrations equal orhigherthan2.0%(20mL using plants assessed against lower toxicity than essential oils fromotherChilean native which didnotsurpass60%mortality. concentrations Ourresultsshowa assessed others than 0.05) ≤ (p toxic more 14.6 mL essential oilkg insect mortality was 80 mL essential oil kg 5% ofdeadinsects. Molina (Rosaceae)(Silvaetal.,2005)thatdidnotexceed (Myrtaceae) (Silva etal.,2003)andQuillajasaponaria better resultsthat EucalyptusglobulusLabill. C. albashows that we requiredabout 20 kgofC.albafoliage. However it will be required 8 kgofpeumopowder; so weestimate of required powder. Forexample, to protect 100 kg of cereal should beconsideredimpractical becauseofthehighamount essential oil kg (Hydrangeaceae) that at the concentration of 4.0% (40 mL species as occurred with Salvia hydrangea DC.exBenth than other essential oils evaluated against the same insect is impractically high. However, C.albaismoretoxic to LC n Essential oil LC n Powder 5.0 lethal concentration50%(LC of Tablepowder 2.Mortalitybycontacteffectofessentialoiland R R R according to Tukey’s test (p≤0.05). different nonsignificantly are letter same the with column a Valueswithin & ¶ † concentration: g powderkg Treatments Probit adjustment slope (b)andstandarderrorof(SE); g kg Confidence limitsat95%. 2 : Coefficientofdetermination. In case of the essential oil, the treatment with the highest Cryptocarya albaagainstadultsofSitophiluszeamaisand C. albain the sense that the required oil concentration -1 P. boldus(Silvaetal.,2003),whichhave100% grain; P. boldusessential oil obtained mortality over 80% S. zeamaisasD.winteri(Andradeetal.,2009) †† mL kg -1 -1 -1 grain). In thiscase, the situation is similar grain) led to 68.3% mortality (Kotan et -1 grain). Moreover, a concentration of 8.0% grain; b ±SE b ±SE Concentration ††† -1 S. zeamais. Betancur et al. (2010), -1 grain (Table 2), being significantly (95% CL) (95% CL) § § grain or mL essential oil kg Total numberofinsectstreated. 50 14.6 40.0 94.0a 20.0 59.5b 10.0 23.0c 50 80.0 84.6a 40.0 22.9b 20.0 15.4b 10.0 10.0b ††† 5.0 13.0c 2.5 2 † 2 ¶ ¶ 0.99 200 0.99 200 50 †† † ) and95%(LC 11.2b 15.5c & & 12.8-16.2 44.2-56.7 -1 grainandcaused 95 ). 1.33 ±0.46 1.70 ±0.55 Mortality (%) -1 grain; § * Lethal 50 of

51 CHILEAN JOURNAL OF AGRICULTURALCHILEAN JOURNAL OF AGRICULTURAL RESEARCH 76(1) JANUARY RESEARCH 76(1)JANUARY 5 (50DAI). The treatment of 80gpowderkg effect against (2008) indicated that 1,8-cineole and terpineole have toxic capitata L. (Lamiaceae) documented 90% of mortality in 1,8-cineole isolated from essential oil ofLavandulaspica T. confusum.Furthermore, Clemente et al. (2007), using and caused of 100%mortality against oils oftwospeciesthegenusAchillea activity. According to Calmasur et al. (2006), the essential compounds existinotherplantspecieswithinsecticidal compounds suchas1,8-cineole and terpineole. These alba against 10% ofdeadinsects,respectively(Tapondjou etal.,2005). 7, thetreatments of 5,10,and20gpowder kg treatments. week other the all At with 0.05) ≤ differences(p Cryptocarya alba to different concentrations ofpowder andessentialoil of treatments which caused an emergence of F treatments of 40and80gpowderkg Powder 0.0 63.2%, and70.4%ofadultF 80.3%, with control, untreated the from significantly differ essential oil kg Eucalyptus saligna Sm. (Myrtaceae) that at 5.0%(50mL al., 2008), Essential oil Table 3.Emergence(F an F the oneobservedwith40and80gpowderkg of 10.5%and0%,respectively. The F at concentrations of2.0%(20gpowderkg quinoa such asE.globulus(Silva emergence inhibitionthanotherplantsassessedaspowder evaluations, 20 and 40 mL essential oil kg the emergence ofinsectsstartedintheweek5,butall 74% and82%ofS.zeamaisemergence. These treatments showedanF and F † according to Tukey’s test(p≤0.05). different significantly not are letter same the with column a Valueswithin Emergence ofcontroltreatmentwasconsideredas100%. Treatments g kg The emergence ofadultinsectswasobservedfromweek The toxicity of thepowderandessentialoilC. The essentialoilshowedasimilartrendvaluesbecause -1 1 grain; mrec o 0 (al 3 ehbtn significant exhibiting 3) (Table 0% of emergence 1 lower than 1.0%. These treatments values were Willd. (Chenopodiaceae) (Silva et al., 2005), which (Wiedemann) (Diptera: Tephritidae). Kotanetal. †† Cupressus sempervirens L. (Cupressaceae) and mL kg S. zeamais,ismaybe due to the bioactivity S. granariusandT. confusum. Concentration -1 . grain) of essential oil reached a 25% and -1 40.0 20.0 10.0 80.0 40.0 20.0 10.0 5.0 2.5 0.0 5.0 grain. †† †

1 ) of adults of Sitophiluszeamais ) ofadults et al.,2003)andChenopodium Week 3 0.0a 0.0a 0.0a 0.0a 0.0a 0.0a 0.0a 0.0a 0.0a 0.0a 0.0a 0.0a 1 emergence, respectively. 1 emergence higher than Insect emergence (%) -1 1 valuesobtainedwith grain show a higher grainshow 0.0a 0.0a 8.2ab 15.9b 20.7b 0.0a 8.6b 51.4c 48.6c 74.1d 100.0d 100.0c 100.0b Week 5 S. granariusand have1,8-cineole 1 -1 adult -1 -1 grain)showed grain showed -1 grain showed graindidnot S. zeamais Ceratitis - - 54.6b 55.7b 46.1b 10.5b 0.0a 0.9a 0.0a 70.4c 63.2c 80.3c MARCH 2016 MARCH 2016 Week 7 -1 grain Powder 0.0 therefore anovicidaleffect isassumed. we didnotdetected any deadlarvaeinsideoroutsidegrains, essential oil), all wheat grains per replicate were checked larvicidal toxicity. However, inbothbioassays(powderand and Stamopoulos(2002),theessential oils haveovicidal and concentrations ofpowder andessentialoilofCryptocaryaalba Tabledifferentmixed with 4.Germinationofwheat and isosafrolagainst al. (2002) obtained a significant fumigant toxicity with safrol against insectpestcontrolinstoredproducts,andHuanget of themostpromissorymonoterpeneswithfumigantactivity (1.01%). According toLee et al.(2004)1,8-cineole is one lower concentration of 1,8-cineole (1.85%) andsafrole showed 0%mortality. These results aremaybe due tothe All assessedtreatment of powderoressential oil ofC.alba Fumigant andrepellent effect Essential oil of essentialoilshowedalowerF emergence begins inthe week 5andthe higher concentrations L. sempervirens (Torres et al., 2014).Inbothcases,theinsect Chilean native plants as P. boldus(Betancur et al., 2010) and those observed in the evaluation ofother essential oils from between 46% and56% (Table 4). Ourresultsagree with and did not differ with those of the control with emergences significantly lower than the observed in the rest of treatments treatments toprotectgrainandseed. thus increasingthepossibilityofusingmoreeffective grain oilcomplywiththerequirementtoexportseeds; 90%) in treatments of 20 and 40 mL essential oil kg germination. The highergerminationvaluesobtained(> according Pérezetal.(2007),P. boldusreducesmaize oils thatnegativelyaffect germination. For example, alba has advantages over other plant powder or essential untreated control(Table 4). According theseresults,C. showed agerminationpercentagesimilar(p>0.05)tothe were registered in treatments with higher concentrations of were registeredintreatmentswithhigherconcentrationsof repellent activity. Although theindexeswithlowervalues(>repellency) as classifies (2003) Vendramim and Mazzonetto showed arepellenceindex<1(Table 5),whichaccording † (Tukey, p≤0.05). Values withinacolumn withthesameletterarenotsignificantlydifferent Treatments g powderkg All treatmentswithpowderandessentialoilofC.alba All treatmentswithpowderandessentialoilofC.alba -1 grain, †† mL essentialoilkg S. zeamaisandT. castaneum. Concentration 40.0 20.0 10.0 80.0 40.0 20.0 10.0 5.0 2.5 0.0 5.0 †† † 100.0a 1 100.0a . According toPapachristos -1 grain. Germination (%) 96.0a 90.0a 86.5a 93.5a 95.0a 88.3a 86.7a 86.3a 77.9a 82.1a -1 .

52 CHILEAN JOURNAL OF AGRICULTURALCHILEAN JOURNAL OF AGRICULTURAL RESEARCH 76(1) JANUARY RESEARCH 76(1)JANUARY contact insecticidetocontrolinsectsofstoredproducts. alba mayhave potential to bedeveloped as anewnatural without affecting wheat germination. The essential oil C. and repellence effect against adults of Sitophilus zeamais under laboratoryconditionshavecontact insecticidal activity The powderandessential oil offoliage of Cryptocaryaalba CONCLUSIONS in essentialoils,thelowerwillberepellency. concentration islower, usuallythemostabundantcomponents with monoterpenesconcentration. Thus ifthemonoterpenes According to Conti et al. (2010) this effect is directly related powder oressentialoil,therepellenteffect isdetectable. Essential oil Cryptocarya alba Table 5. Repellence index of powder andessential oil of Consoli, F.L., H.J. Williams, S.B. Vinson, R.W. Matthews, andM. Clemente, S., G.Mareggiani, A. Broussalis,yG.Ferraro.2007. Calmasur, O.,S.Kordali, O. Kaya, andI. Aslan. 2006. Toxicity Betancur, J., G.Silva, J.C. Rodríguez, S. Fischer, and N. Zapata. Avello, M.,C.López, Gatica,E.Bustos, A. Brieva,E.Pastene,et Andrade, Y., G.Silva,N.Zapata, M. Tapia, J. Rodríguez, y A. Abbott, W.S. 1925. A method of computing the effectiveness of an REFERENCES Powder 5.0 † repellent (IR<1),attractive>1). Treatments g powderkg 28:1675-1689. ectoparasitoid Cooperband. 2002. trans -Bergamotenes-male pheromone of the 23:29-34. Ceratitis capitata Actividad insecticida de1,8-cineol sobre moscadelosfrutos Val). JournalofPlantDiseases andProtection113:37-41. Sitophilus granaries (L.)andTribolium confusum(Jacquelin du of essentialoilsvapoursobtained from Achillea spp. to Agricultural Research70:399-407. oil against 2010. Insecticidal activity of PeumusboldusMolinaessential de PlantasMedicinales17(1):73-83. de lasfamilias Lauraceae y Atherospermataceae. Revista Cubana al. 2012. Efectos antimicrobianos de extractos de plantas chilenas 25(1):33-40. (Coleoptera: Curculionidae) en laboratorio. Agro-Ciencia J.R. etG.Forster)contraSitophiluszeamaisMotschulsky Lagunes. 2009. Toxicidad de polvoscanelo (Drimys winteri insecticide. JournalofEconomicEntomology18:265-267. -1 grain, Sitophilus zeamais Motschulsky. Chilean Journal of Concentration Melittobia digitata againstadultsofSitophiluszeamais. Wied.(Diptera: Tephritidae). Dominguezia †† 40.0 20.0 10.0 80.0 40.0 20.0 10.0 mL essential oil kg 5.0 2.5 †† † 0.98 0.28 . Journal of Chemical Ecology Repellence index (IR) 0.19 Repellent 0.21 Repellent 0.27 Repellent 0.19 Repellent 0.82 Repellent 0.91 Repellent 0.92 Repellent 0.92 Repellent -1 grain,

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53 CHILEAN JOURNAL OF AGRICULTURALCHILEAN JOURNAL OF AGRICULTURAL RESEARCH 76(1) JANUARY RESEARCH 76(1)JANUARY Tapondjou, A.L., C. Adler, D.A.Fontem,H.Bouda,andC. Silva, G.,D.Pizarro,P. Casals,yM.Berti.2003.Evaluación Silva, G., R. Kiger, R. Hepp, y M. Tapia. 2005. Control de Sitophilus Robertson, J.L.,andH.K.Preisler. 1992.Pesticide bioassays with J. Procopio, S., Vendramin,Santos. 2003. Ribeiro, eJ. J. Journal ofStoredProductsResearch41:91-102. Sitophilus zeamais Motschulsky and of Reichmuth. 2005. Bioactivities of cymol and essential oils Agrociência 9(4):383-388. zeamais Motschulskyenmaíz almacenado. Revista Brasileira de de plantas medicinales en polvoparaelcontrol de Sitophilus 40(10):953-960. del géneroChenopodium.Pesquisa Agropecuária Brasileira zeamais arthropods. 224p.CRCPress,BocaRaton,Florida,USA. Agrotecnologia 27:1231-1236. Sitophilus zeamais Mots.(Coleoptera: Curculionidae). Ciencia e Bioatividade de diversospósorigenvegetal em relaçao a Cupressus sempervirens and Motschulsky con polvosvegetalesdetresespecies Eucalyptus saligna against Triboliumdu Val. confusum - - MARCH 2016 MARCH 2016 Zhuang, X., T.G. Köllner, N.Zhao,G.Li, Y. Jiang,L.Zhu,etal. Vogel, H., I.Razmilic, y U.Doll. 1997. Contenido de aceite esencial Torres, C.,G.Silva,M. Tapia, J.C.Rodríguez,I.Figueroa, Tavares, M.,eJ. Vendramim. 2005. Bioatividade da erva-de-santa- Journal 69:70-80. biosynthesis insorghum andrelated grass crops. The Plant 2012. Dynamic evolution of herbivore-induced sesquiterpene Mol.) CienciaeInvestigación Agraria 24:1-6. y alcaloides en diferentes poblaciones de boldo(Peumusboldus 74:421-426. zeamais Motschulsky. ChileanJournalof Agricultural Research sempervirens (Ruiz&Pav.) Tul. essentialoilagainstSitophilus A. Lagunes,etal.2014.Insecticidal activity of Laurelia 34:319-323. Mots. (Coleoptera: Curculionidae). Neotropical Entomology maria, Chenopodium ambrosioides L.,sobreSitophiluszeamais

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