Postharvest Control of the Date Moth Ectomyelois ceratoniae ... - Iresa

stored dates and causes weight loss anddowngrading of the commercial value ofthe fruit (2, 14).Dates intended for export undergothe process of conditioning to preserveand improve their quality. Chemicalcontrol using fumigation is the mosteconomical tool for managing this storedpest in postharvest treatment (3). Methylbromide is still the primary insecticide ofpostharvest insect control for dates inTunisia and in several other countries (19,53). Nevertheless, the use of thisfumigant is being restricted because of itshealth and environmental damages (6).Moreover, the Montreal Protocol of theUnited Nations Environment Program(49) recommends the phasing out ofmethyl bromide by 2005 in developedcountries and by 2015 in developingcountries (33).Consequently, research of effectivealternative methods is needed. In recentyears, essential oils have received muchattention as pest control agents because oftheir insecticidal properties (42). Essentialoils are volatile and can act like fumigantsoffering the prospect for use in storedproductprotection (7, 15, 41, 44, 46).Eucalyptus (Myrtaceae) is one of themost cultivated genera in the worldincluding more than 700 species. Variousbiological properties have already beenattributed to the genus Eucalyptus, amongthem insecticidal activity against beetles(8), repellent action against Phlebotomuspapatasi (51) and larvicidal activity onculicids (10). Besides, eucalyptusessential oils are used for medicinal andpharmaceutical purposes (11, 12, 18, 26,37, 43). In Tunisia, 177 Eucalyptusspecies were introduced and acclimatizedinto more than 30 arboretums since 1957(23).The objectives of this study were todetermine the chemical composition ofthe essential oils of Eucalyptuscamaldulensis and E. rudis and toevaluate their fumigant toxicity againstnew emerged adults of E. ceratoniae.MATERIALS AND METHODSInsect rearing. A laboratory rearingcolony was established in the Laboratoryof Plant Protection at the Institut Nationalde la Recherche Agronomique de Tunisie(INRAT) from infested field-collecteddates. The moth was reared on anartificial diet based on wheat bran (60%),sucrose (12%), salt mixture (2%), yeast(1.3%), lysine (1.23%), methyl paraben(0.13%), vitamin C (0.67%), aureomycine(0.67%), glycerine (150 ml) and distilledwater (150 ml) (for details see 34).Rearing was conduced in plastic boxes(20 × 15 × 10 cm) placed in a rearingroom. The rearing conditions were:temperature of 25 ± 1°C, photoperiod of15: 9 (L: D) and 65 ± 5% relativehumidity.Plant material. E. camaldulensisand E. rudis leaves were collected fromnatural populations at the flowering stageon August 25, 2009 from the arboretumof Korbus (North Tunisia) of the InstitutNational des Recherches en GénieRurale, Eaux et Forêts (INRGREF). Theharvested material was air-dried at roomtemperature (20-25°C) for one week andthen stored in cloth bags.Essential oil extraction andanalysis. The essential oils wereextracted by hydrodistillation of driedplant material (100 g of each sample in500 ml of distilled water) using aClevenger-type apparatus for 4 h. The oilswere dried over anhydrous sodiumTunisian Journal of Plant Protection 202 Vol. 5, No. 2, 2010

times (LT 50 ) to cause mortality of 50% oftested insects at the lowest doses (13.16and 26.31 µl/l air). The mortality wasassessed by direct observation of insectsevery hour until all the insects were dead.Data were analyzed using Finney’smethod (16).RESULTSEssential oils composition. GC andGC-MS analysis of E. camaldulensis andE. rudis essential oils were respectivelyreported in Tables 1 and 2. The oil yieldsbased on dry matter weight wererespectively 1.32% for E. camaldulensisand 2.09% for E. rudis.Table 1. Chemical fractions, others constituents and total identifiedcompounds of the essential oil obtained from leaves of E.camaldulensis collected at flowering stage (%)N° Compound Percent. (%) RT KIMonoterpene hydrocarbons 22.521 α-pinène 16.49 9.11 9392 γ-terpinene 4.8 14.13 10753 Dehydro-p-cymene 1.23 14.74 1090Oxygenated monoterpenes 33.824 1,8-cinéole 20.62 12.55 10845 Terpinéne-4-ol 4.45 17.36 12176 Isothymol 7.3 20.54 13367 Piperitone 1.45 19.52 1297Sesquiterpene hydrocarbons 3.938 Aromadendrene 3.93 23.97 1468Oxygenated sesquiterpenes 6.659 Spathulenol 2.36 27.5 161810 Viridiflorol 1.36 27.66 162411 t-muurolol 2.93 29.35 1697Other compounds 23.59Total 94.02RI, KI were respectively Retention Index and Kováts Index calculatedon a HP-5MS capillary column (30 m × 0.25 mm × 0.25 µm).A total of 94.02% from theconstituents of E. camaldulensis leafessential oil were identified (Table 1).Among them, 22.52% were monoterpenehydrocarbons, 33.82% oxygenatedmonoterpenes, 6.65% oxygenatedsesquiterpenes and 3.93% sesquiterpenehydrocarbons. The major compoundswere α-pinene (16.49%), 1,8-cineole(20.62%), p-cymene (21.69%), isothymol(7.3%), γ-terpinene (4.8%), andterpinene-4-ol (4.45%).Tunisian Journal of Plant Protection 204 Vol. 5, No. 2, 2010

Table 2. Chemical fractions, others constituents and total identifiedcompounds of the essential oil obtained from leaves of E. rudiscollected at flowering stage (%)N° Compound Percent. (%) RT KIMonoterpene hydrocarbons 24.441 α-pinene 14.49 9.32 9512 β-pinene 3.91 10.45 9903 γ-terpinene 6.04 13.75 1068Oxygenated monoterpenes 28.854 1,8-cineole 19.87 12.79 10465 Terpinene-4-ol 4.46 17.1 12076 α-terpineol 4.32 17.43 1219Sesquiterpene hydrocarbons 9.229 Aromadendrene 6.37 23.64 144810 Bicyclogermacrene 2.85 25.19 1517Oxygenated sesquiterpenes 5.4712 Isospathulenol 3.09 29.29 169513 t-muurolol 2.34 29.71 1715Others compounds 10.13Total 78.11RI, KI were respectively Retention Index and Kováts Index calculatedon a HP-5MS capillary column (30 m × 0.25 mm × 0.25 µm).For E. rudis leaf essential oil, a totalof 78.11% of the constituents wasidentified (Table 2). The oxygenatedmonoterpenes represented 28.85%followed by the monoterpenehydrocarbons 24.44%, the sesquiterpenehydrocarbons 9.22% and the oxygenatedsesquiterpenes 5.47%. The mainconstituents were α-pinene (14.49%), β-pinene (3.91%), 1,8-cineole (19.87%), γ-terpinene (6.04%), terpinene-4-ol(4.46%), α-terpineol (4.32%) andaromadendrene (6.37%).ceratoniae adults exposed for variousperiods of time to essential oils from E.rudis and E. camaldulensis.Results showed that E.camaldulensis and E. rudis essential oilswere toxic to E. ceratoniae adults (Fig.1). E. camaldulensis was more toxic thanE. rudis for the concentrations exceeding52.63 µl/l air.At the lowest concentration (13.16µl/l air), E. rudis achieved 25% ofmortality after 24 h of exposure comparedwith 10% of mortality for E.camaldulensis. However, at the highestconcentration (131.58 µl/l air), 100%mortality was recorded for E.camaldulensis and E. rudis essential oilsFumigant toxicity. Mortality wascorrected for control mortality by usingAbbott’s formula (1). Results werepresented as percentage of mortality of E. after only 12 h of exposure (Fig. 1).Tunisian Journal of Plant Protection 205 Vol. 5, No. 2, 2010

10080Dose 13.16 µl/ l airE. rudisE. camaldulensisControl10080Dose 26.31 µl/l airE. rudisE. camaldulensisControlMortality (%)6040Mortality (%)6040202000 1 6 12 18 24Exposure time (h)00 1 6 12 18 24Exposure time (h)10080Dose 52.63 µl/l airE. rudisE. camadulensisControl10080Dose 78.94 µl/l airE. rudisE. camaldulensisControlMortality (%)6040Mortality (%)6040202000 1 6 12 18 24Exposure time (h)00 1 6 12 18 24Exposure time (h)Mortality (%)100806040Dose 105.26 µl/l airE. rudisE. camaldulensisControlMortality (%)100806040Dose 131.58 µl/l airE. rudisE. camaldulensisControl202000 1 6 12 18 24Exposure time (h)Fig. 1. Percentage of mortality of E. ceratoniae adults exposed for various periods of time todifferent doses of essential oils from E. rudis and E. camaldulensis00 1 6 12 18 24Exposure time (h)Probit analysis showed that E.ceratoniae was more sensitive to E. rudisessential oil than E. camaldulensis one.The corresponding LC 50 values were31.37 and 34.08 µl/l air, respectively(Table 3).The LT 50 values ranged from 39.50h for the concentration 13.16 µl/l air to18.27 h for the concentration 26.31 µl/lair for E. rudis essential oil. Nevertheless,for E. camaldulensis essential oil, theLT 50 values were 87.73 and 38.31 h forthe concentrations 13.16 and 26.31 µl/lair, respectively (Table 4).Tunisian Journal of Plant Protection 206 Vol. 5, No. 2, 2010

Table 3. LC 50 values calculated for mortality within 24 h ofexposure of E. ceratoniae adults to E. rudis and E. camaldulensisessential oilsPlant species E. rudis E. camaldulensisLC a,b 50 (LD 50)31.3734.37(24.9 - 70.21) (21.07 - 48.26)Slope ± SEM2.17 ± 0.71 4.49 ± 0.88(0.18 - 4.16) (2.03 - 6.95)Degree of freedom 10 10χ2 20.27 10.35aUnits LC 50 = µl/ air, applied for 24 h at 25 °C.b95% lower and upper confidence limits are shown in parenthesis.Table 4. LT 50 values calculated for E. ceratoniae adults exposed to E. rudis and E. camaldulensisessential oilsEssential oilConcentrationLT(µl/l air)50 (h) aDegree ofSlope ± SEfreedomχ2E. rudis13.1639.50 3.0 ± 0.47(32.73 - 47.04) (2.14 - 4.02)10 0.2526.3118.27 2.33 ± 0.31(14.53 - 22.26) (1.72 - 2.95)10 4.6287.73 3.43 ± 0.5013.16E.(73.73 - 102.97) (2.45 - 4.41)10 5.25camaldulensis38.31 4.34 ± 0.5826.31(32.58 - 44.23) (3.19 - 5.50)10 5.10a95% lower and upper confidence limits are shown in parenthesis.DISCUSSIONcineole content in eucalyptus essentialThis study showed that the essential oils and their toxicity to insects.oils of E. camaldulensis and E. rudis Furthermore, this compound whencollected from Korbus were rich in present in eucalyptus essential oilsmonoterpenoids. Moreover, the major showed promising fumigant toxicity (36,common compounds of these two oils 38, 40). Additionally, Yang et al. (52)were 1,8-cineole (20.62- 19.87%), α- attributed the toxicity of E. globuluspinene (16.49- 14.49%), γ-terpinene (4.8- essential oil against the eggs and adults of6.04%) and terpinene-4-ol (4.45- 4.46%). Pediculus humanus capitis (Anoplura:Among various compounds of Pediculidae) to the action of 1,8 cineole.eucalyptus essential oils, 1,8-cineole is Besides, Lee et al. (24) reported thatthe most important. This compound is essential oils rich in 1,8-cineole presentedcharacteristic of the Eucalyptus genus and a strong toxicity against Sitophilusis mainly responsible of its insecticidal oryzae, Tribolium castaneum andproprieties (5). Indeed, 1,8 cineole which Rhyzopertha dominica.is the major compound of E.Moreover, the pesticidal activity ofcamaldulensis and E. rudis essential oils eucalyptus oils has been due to thewas recognized to be toxic on several components such as α-pinene and γ-insect species. Lucia et al. (31) indicated terpinene (4, 27, 28, 29, 39, 47). In thisthe presence of correlation between 1,8-Tunisian Journal of Plant Protection 207 Vol. 5, No. 2, 2010

context, several works reported that α- and Pistacia lentiscus essential oilspinene is listed among major constituents against E. ceratoniae. The authorassociated with pesticidal activities (9, 30, indicated that 100% of mortality was45). Besides, Su et al. (47) indicated that obtained for these two oils atγ-terpinene is involved in biological concentrations 90.91 and 113.64 µl/l airactivities of eucalyptus essential oils.after 18 and 36 h of exposure,On the other hand, terpinen-4-olwhich also represents a major compoundof these two essential oils was reported torespectively.On the other hand, E. camaldulensisessential oil was reported to be toxicbe toxic on several insect species. Indeed, against stored-product beetles (35).Kordali et al. (21) indicated that terpinen-4-ol was the most toxic against Sitophilusgranaries adults after 12 h of exposure.Furthermore, this compound showedpromising fumigant toxicity on the riceMoreover, Tunç et al. (48) indicated thatE. camaldulensis essential oil inducedhigh mortality to E. kuehniella andTribolium confusum. Besides, Winters etal. (50) demonstrated that E. rudis and E.weevil Sitophilus oryzae (25). camaldulensis essential oils wereAdditionally, terpinen-4-ol showed characterized by similar emission ofrelatively strong toxicity against the monoterpenes with respective values oflarvae and adults of Leptinotarsa 370.71 and 210.51µg C m -2 h -1 .decemlineata (22).The present work reported firstThis work demonstrated that investigations on the fumigant activity ofessential oils of E. rudis and E. E. camaldulensis and E. rudis essentialcamaldulensis displayed strong fumigant oils against E. ceratoniae in storage.toxicity against new emerged adults of Results clearly demonstrate thethe date moth E. ceratoniae. The insecticidal activity of these essential oilsinsecticidal activity depends on oil against this major pest of stored dates.species, concentration and exposure time.Few works have been reported onthe fumigant activity of essential oilsagainst the date moth E. ceratoniae.Mansouri (32) first investigated thefumigant toxicity of Lavandula officinalisThus, they could be used as an alternativeto the synthetic fumigant in postharvesttreatment programs. Accordingly, there isan interest to further investigate theessential oil toxicity on other storeddate’s pests.__________________________________________________________________________RESUMEHaouel S., Mediouni-Ben Jemâa J. et Khouja M.L. 2010. Lutte en post-récolte contre la pyraledes dates Ectomyelois ceratoniae moyennant la fumigation aux huiles essentielles d'eucalyptus.Tunisian Journal of Plant Protection 5: 201-212.Ce travail a pour objectifs d’étudier la composition chimique et d’évaluer pour la première fois l’effetfumigène des huiles essentielles de deux espèces d’eucalyptus (Eucalyptus camaldulensus et E. rudis)contre la pyrale des dattes Ectomyelois ceratoniae en entrepôts comme alternative au bromure deméthyle. Les analyses CPG-SM ont montré que l’huile essentielle d’E. rudis présente α-pinène(14.49%), 1,8-cinéole (19.87%), β-pinène (3.91%), γ-terpinène (6.04%), terpinéne-4-ol (4.46%) et α-terpinéol (4.32%) comme constituants majeurs alors que celle d’E. camaldulensis présente α-pinène(16.49%), 1,8-cineole (20.62%), γ-terpinène (4.08%), terpinène-4-ol (4.45%), aromadendrene (3.93%)Tunisian Journal of Plant Protection 208 Vol. 5, No. 2, 2010

et isothymol (7.3%) comme composés majeurs. Les résultats ont montré que l’activité fumigènedépend de l’espèce d’huile, de la concentration et du temps d’exposition. Le potentiel fumigène del’huile essentielle d’E. rudis (CL 50 = 31.37 µl/l air) était supérieur à celui de l’huile essentielle d’E.camaldulensis (CL 50 = 34.08 µl/l air). En outre, les valeurs respectives des temps létaux médians (TL 50 )étaient de 36.10 et 48.84 h. Les résultats suggèrent que les huiles essentielles d’E. rudis et E.camaldulensis peuvent être utilisées comme alternatives aux fumigants de synthèse en traitement postrécoltepour la lutte contre E. ceratoniae.Mots clés: Concentration létale CL 50 , CPG-SM, Ectomyelois ceratoniae, Eucalyptus camaldulensis,Eucalyptus rudis, huile essentielle, temps létal LT 50__________________________________________________________________________ملخصحوال،‏ سمية وجودة مديوني-بن جماعة ومحمد العربي خوجة.‏.2010باستعمال التبخير بالزيوت العطرية للأوآالبتوس.‏Tunisian Journal of Plant Protection 5: 201-212المكافحة في مرحلة ما بعد الجني لعثة التمرEctomyelois ceratoniaeيهدف هذا العمل إلى دراسة الترآيب الكيميائي وتقيم الفعالية السمية بالاختناق للزيوت العطرية/الأساسية لنوعين منلمكافحة عثة التمر ،Ectomyelois ceratoniae لأول مرة فيو الأوآليبتوسأن الزيت الأساسي لتونس،‏ في المخازن آطريقة بديلة لبروميد الميثيل.‏ بينت التحاليل الكروماتوغرافية وويحتوي علي المكونات الأساسية التالية:‏(4.32%)، بينما احتوى الزيت الأساسي لو ووعلى العناصر الأساسية التالية:‏بينت النتائج أن الفعاليةوووالسمية بالاختناق للزيوت الأساسية تختلف حسب نوع الأوآليبتوس وترآيز الزيت ومدة التعرض.‏ آانت الفعالية السميةأفضل من الفعالية السمية للزيت الروحي لللزيت الأساسي لساعة بالنسبة لهذينعلى التوالي 36.10 إضافة إلي ذلك،‏ بلغ الوقت القاتل آطريقة بديلةو لالزيتين وأظهرت النتائج أنه يمكن استعمال الزيوت الأساسية ل للمبخرات الصناعية في برنامج معاملة ما بعد القطاف لمكافحة عثة التمرE. rudis Eucalyptus camaldulensisE.(GC-MS)β-pinene (3.91%) (19.87%) 1,8-cineole (14.49%) α-pinenerudisE.α-terpineol (4.46%) terpinéne-4-ol و (6.04%) γ-terpineneγ-terpinene (20.62%) و 1,8-cineole (16.49%) α-pinenecamaldulensis.(7.3%) isothymol (3.93%) aromadendrene (4.45%) terpinene-4-ol (4.08%)E. camaldulensisو 48.84(TL 50 )E. camaldulensis E. rudis.E. ceratoniae(LC 50 = 31.37 µl/l air) E. rudis.(LC 50 = 34.08 µl/l air)Eucalyptuscamaldulensis, E. rudis__________________________________________________________________________آلمات مفتاحية:‏ التحاليل الكرومتغرافية،‏ الترآيز القاتل،‏ الزيوت الروحية،‏ عثة التمر،‏ الوقت القاتل،‏LITERATURE CITED1. Abbott, W.S. 1925. A method of computing theeffectiveness of an insecticide. J. Econ. Entomol.18: 265-267.2. Al-Azawi, A.F., El-Haidari, H.S., Aziz, F.M.,Murad, A.K., and Al-Saud, H.M. 1984. Theeffect of high temperatures on the dried fruitbeetle Carpophilus hemipterus L., a pest ofstored dates in Iraq. Date Palm Journal 3: 327-336.3. Azelmat, K., El Garrouj, D., Mouhib M., andSayah F. 2006. Irradiation of Bouffegous dates:Effects on chemical composition during storage.Postharvest Biology and Technology 39: 217-222.4. Batish, D.R., Singh, H.P., Setia, N., Kaur, S., andKohli, R.K. 2006. Chemical composition andphytotoxicity of volatile essential oils from intactand fallen leaves of Eucalyptus citriodora. Z.Naturforsch. 61: 465-471.5. Batish, D.R., Sing, P.H., Kohli, K.R., and Kaur,S. 2008. Eucalyptus essential oil as a naturalpesticide. Forest Ecology Management 256:2166-2174.6. Bell, C.H. 2000. Fumigation in the 21 st century.Crop Prot. 19: 563-569.7. Bignell, C.M., Dunlop, P.J., and Brophy, J.J.1997. Volatile leaf oils of some Queensland andnorthern Australian species of the genusEucalyptus (series II) part II. Subgenera (a)Tunisian Journal of Plant Protection 209 Vol. 5, No. 2, 2010

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