Insecticidal effects of essential oils of eleven plant species from Lamiaceae on Sitophilus granarius (L.) (Coleoptera: Curculionidae)

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1 Romanian Biotechnological Letters Vol. 16, No. 6, 2011 Copyright 2011 University of Bucharest Printed in Romania. All rights reserved ORIGINAL PAPER Insecticidal effects of essential oils of eleven plant species from Lamiaceae on Sitophilus granarius (L.) (Coleoptera: Curculionidae) Abstract 6702 Received for publication, March 7, 2011 Accepted, May 28, 2011 EROL YILDIRIM 1, SABAN KORDALI, GULTEN YAZICI Atatürk University, aculty of Agriculture, Department of Plant Protection, 250, Erzurum- TURKEY 1 Corresponding author: Tel.: ; fax: , address: yildirimerol@hotmail.com our different concentrations of the essential oils obtained from eleven plant species (Origanum onites L., Origanum rotundifolium Boiss., Rosmarinus officinalis L., Salvia hydrangea Dc., Salvia numerosa L., Salvia multicaulis Vahl., Salvia sclarea L., Satureja hortensis L., Satureja spicigera (C. Koch) Boiss., Thymus fallax isch. & Mey. and Thymus sipyleus Boiss.(Lamiaceae) were tested against adults of Sitophilus granarius (L.) on Petri dishes. After exposure, mortality of the adults was determined at, and h. The results show that essential oils of O. onites, O. rotundifolium, R. officinalis, S. hydrangea, S. hortensis, S. spicigera, T. fallax and T. sipyleus have an insecticidal effect on adults of S. granarius in comparison with controls. The insecticidal effect was influenced by the doses of the essential oils and the exposure time. Higher doses and longer exposure time resulted in maximum toxicity on S. granarius. Treatment with essentials oils of O. onites, O. rotundifolium, R. officinalis, S. hydrangea, S. hortensis, S. spicigera, T. fallax and T. sipyleus leads to a higher and S. multicaulis to less mortality of adults of S. granarius but essential oils of S. numerosa and S. sclarea had no effect. The mortality rates after h of treatment with the maximum dose (20 µl) of essential oils of S. hortensis, O. rotundifolium, O. onites, S. spicigera, R. officinalis, T. fallax T. sipyleus, S. hydrangea, S. multicaulis, S. sclarea and S. numerosa was determined as 100,.97, 95.27, 94.27, 93.93, 88.88, 82.82, 67.33, 55.21, and % for S. granarius, respectively. Separately, the mortality rates after h of treatment with the 1 µl in dose of essential oils of S. hortensis was established to the highest mortality rate in S. granarius with %, and 5, 10 and 20 µl in dose of S. hortensis with 100 %. Keywords: Sitophilus granarius, Lamiaceae, Essential oils, Insecticidal effects Introduction Since the middle ages, essential oils have been widely used for bactericidal, virucidal, fungicidal, antiparasitical, insecticidal, medicinal and cosmetic applications, especially nowadays in pharmaceutical, sanitary, cosmetic, and agricultural and food industries. Because of the mode of extraction, mostly by distillation from aromatic and medicinal plants, they contain a variety of volatile molecules such as terpenes and terpenoids, phenolderived aromatic components and aliphatic components [1]. The essential oils of a large number of plant species have been found to have toxic and/or repellent effects against different insects [2, 3]. Recent investigations in several countries confirm that some plant essential oils not only repel insects, but also have contact and fumigant insecticidal actions against species pests [4]. Recent research has focused on natural product alternatives for pest control in developing countries. They could lead to the development of new classes of safer insectcontrol agents. Recently, there has been a growing interest in research concerning the possible use of plant extracts as alternatives to synthetic insecticides. The toxicity of a large number of

2 EROL YILDIRIM, SABAN KORDALI, GULTEN YAZICI essential oils and their constituents has been evaluated against a number of stored-product insects. Some essential oils were found to have potential for the control of stored product insect pests. The insecticidal activity of many plant derivates against several stored-product pests has been demonstrated [5-30]. The granary weevil, Sitophilus granarius (L.) (Coleoptera: Curculionidae) is a well-known pest causing economically important yield loss in stored products in Turkey and many other countries. Various publications have previously documented the insecticidal activity of the essential oils from medicinal plant species against storage-food pests [31-33]. But, the antimicrobial and insecticidal activities of the plant essential oils can change according to their harvest period, ecological and climatic conditions. The aim of the present study was to evaluate the insecticidal effect of hydrodistilled essential oils of Origanum onites L., Origanum rotundifolium Boiss., Rosmarinus officinalis L., Salvia hydrangea DC. ex Benth, Salvia numerosa L., Salvia multicaulis Vahl., Salvia sclarea L., Satureja hortensis L., Satureja spicigera (C. Koch) Boiss., Thymus fallax isch. & Mey. and Thymus sipyleus Boiss. from Lamiaceae against adults of Sitophilus granarius (L.) in in-vivo conditions. Materials and Methods Insects and rearing conditions. The insects were collected from Erzurum storage house. Wheat grains were purchased from local market and stored in a freezer at -20 C. Wheat for S. granarius was washed in tap water, dried and heated, to prevent pre infestation before using it for the experiments. S. granarius adults were reared in laboratory at 25 ± 1 C, 64 ± 5 relative humidity and L:D = 12 h:12 h in the Department of Plant Protection, at Atatürk University. The adults obtained from laboratory cultures were stored in separate insect cages, including wheat. Tests were also carried out under the same conditions in the same laboratory. Plant material and isolation of essential oil. Origanum onites L., Origanum rotundifolium Boiss., Salvia hydrangea DC. ex Benth, Salvia numerosa L., Salvia multicaulis Vahl., Salvia sclarea L., Satureja hortensis L., Satureja spicigera (C. Koch) Boiss., Thymus fallax isch. & Mey. and Thymus sipyleus Boiss. (Lamiaceae) were collected at the flowering stage in August in Turkey. Separately, the aerial parts of Rosmarinus officinalis L. were purchased at vegetative stage from a local market. Collected plant materials were dried in shadow and ground in a grinder. The dried plants samples (500 g) were subjected to hydrodistillation (plant material in boiling water) using a Clevenger-type apparatus for 4 hours. Hydrodistillation of O. onites, O. rotundifolium, R. officinalis, S. hydrangea, S. numerosa, S. multicaulis, S. sclarea, S. hortensis, S. spicigera, T. fallax and T. sipyleus yielded 3.67, 0.87, 1.46, 0.53, 0.38, 0.80, 1.35, 2.30, 1.56, 1.95 and 0.98 % (w/w) of essential oils, respectively. The yields were based on dry materials of plant samples. Determination of the age of the adults. or S. granarius adults test, 4-6 day-old adults were used as the test material. In order to make test in the adult of the same age, some more larvae and pupa with wheat grain were placed in separate Petri dishes. After adult emergence pupa, in the same time emergence adults were collected and used in the same tests. Bioassays. Romanian Biotechnological Letters, Vol. 16, No. 6,

3 Insecticidal effects of essential oils of eleven plant species from Lamiaceae on Sitophilus granarius (L.) (Coleoptera: Curculionidae) In order to test the toxicity of the oils against to the pest adults, 33 adults of the insects with 33 grains of wheat were placed to Petri dishes (9 cm). 1, 5, 10 and 20 microliters of the oils were applied with an automatic pipette on Whatman No. 1 filter papers (2 cm 2 cm) and were attached to the upside of the Petri dish, corresponding to 7.70, 38.47, and μl/l air concentrations. After exposure, mortality of the adults was determined at, and h. Petri dish applied with only sterile water was used as control group. Three replicates were used for each dose and exposure time combination and insecticidal activity of the oil was expressed as % mean mortality of the adults. Statistical analysis. The differences among the fumigant activities of essential oils tested were determined according to analysis of variance (ANOVA) test by using SPSS 13.0 software package. Differences between means were tested through Duncan tests and values with p<0.01 and p<0.05 were considered significantly different. Results and Discussions The toxicity effects of the essential oils extracted from Origanum onites, O. rotundifolium, Rosmarinus officinalis, Salvia hydrangea, S. numerosa, S. multicaulis, S. sclarea, Satureja hortensis, S. spicigera, Thymus fallax and T. sipyleus (Lamiaceae) on adults of Sitophilus granarius are summarized in Table 1 and 2. The results show that essential oils of O. onites, O. rotundifolium, R. officinalis, S. hydrangea, S. hortensis, S. spicigera, T. fallax and T. sipyleus have an insecticidal effect on adults of S. granarius in comparison with controls (ig. 1, 2, 3). The analysis of variance demonstrates that the effects on the mortality rate of adults of S. granarius are highly significant on the basis of doses and exposure time (Table 1). Higher doses and longer exposure times result in maximum toxicity on S. granarius. Treatment with essentials oils of O. onites, O. rotundifolium, R. officinalis, S. hydrangea, S. hortensis, S. spicigera, T. fallax and T. sipyleus leads to a higher and S. multicaulis less mortality of adults of S. granarius but essential oils of S. numerosa and S. sclarea have no effect (Table 1, 2). The mortality rates after h of treatment with the maximum dose (20 µl) of essential oils of O. onites, O. rotundifolium, R. officinalis, S. hydrangea, S. numerosa, S. multicaulis, S. sclarea, S. hortensis, S. spicigera, T. fallax and T. sipyleus were determined as 95.27,.97, 93.93, 67.33, 39.73, 55.21, 41.76, 100, 94.27, and % for S. granarius, respectively (ig. 1, 2, 3). However, there was no mortality in the controls of each species Romanian Biotechnological Letters, Vol. 16, No. 6, 2011

4 Source Time (h) Dose (µl) EROL YILDIRIM, SABAN KORDALI, GULTEN YAZICI Table 1. The result of ANOVA belonging to exposure dose and exposure time of eleven plant species essential oils on Sitophilus granarius adults O. onites O. rotundifolium R. officinalis S. hydrangea S. numerosa S.multicaulis S. sclarea S. hortensis S. spicigera T. fallax T.sipyleus Df ** ** ** ** ** ** ** ** ** ** ** ** ** * ** ** ** * ** ** ** Time x Dose * ** ** ** ** ** ** ** ** Error Total 36 **Values followed by different letters in the same column differ significantly at p <0.01. *Values followed by different letters in the same column differ significantly at p <0.05. Table 2. Mortality effects of eleven plant species essential oils on Sitophilus granarius adults. Doses (µi) Control Exposure Time (h) O. onites O.rotundifolium R. officinalis S. hydrangea S. numerosa S. multicaulis S. sclarea S. hortensis S. spicigera T. fallax T. sipyleus 21.00b± a± a± b± b± a± b± b± a± b± b± a± c± b± a± b± b± a± a± b± b± a± b± b± a± c± b± a± b 1± a 1± c 1± d 1± c 1± c 1± d 1± b± c 1± d 1± c 1± b± a± b± b± a± c± b± a± b± b± a± c± a± a± c± b± a± c± b± b± b± a± c± b± a 1± a 1± b 1± a 1± b 1± b 1c 1± c 1± b 1± c 1± b 1± b± a± b± b± c± b± c± b± a± c± b± a± c± b± a± b± a± b± b± a± c± b± a 1± a 1± ab 1± a 1± b 1± a 1b 1± b 1± b 1± b 1± a 1± b± a± a± b± b± c± b± c± b± a± c± b± c± b± a± a± b± b± a± b± a± a 1± a 1± a 1± a 1± a 1± a 1± a 1± a 1± a 1± a 1±7.45 a,b,c: in the same column at each dose the exposure times differ (P<0.01). a 1,b 1,c 1 d 1 : in the same column the doses differ (P<0.01). ±S X :of three replicates, each set-up with 33 adults. Romanian Biotechnological Letters, Vol. 16, No. 6,

5 Insecticidal effects of essential oils of eleven plant species from Lamiaceae on Sitophilus granarius (L.) (Coleoptera: Curculionidae) Separately, the mortality rates after h of treatment with the 1 µl in dose of essential oils of S. hortensis was established as the highest mortality rate in S. granarius with %, and 5, 10 and 20 µl in dose of S. hortensis with 100 % (ig. 1, 2, 3). The present results show that the essential oils of O. onites, O. rotundifolium, R. officinalis, S. hydrangea, S. multicaulis, S. hortensis, S. spicigera, T. fallax and T. sipyleus have varying degrees of insecticidal activity against S. granarius adults. The insecticidal activity increased with increasing dose and exposure times. The essential oils caused significant mortality (ig. 1, 2). igure 1. Mortality of Sitophilus granarius (L.) exposed to eleven essential oils at different doses. igure 2. Mortality of Sitophilus granarius (L.) according to treatment times of essential oils of eleven plant species Romanian Biotechnological Letters, Vol. 16, No. 6, 2011

6 EROL YILDIRIM, SABAN KORDALI, GULTEN YAZICI igure 3. Mortality of Sitophilus granarius (L.) regression analysis to eight essential oils at different doses. Romanian Biotechnological Letters, Vol. 16, No. 6,

7 Insecticidal effects of essential oils of eleven plant species from Lamiaceae on Sitophilus granarius (L.) (Coleoptera: Curculionidae) Some previous studies demonstrated that, in general, the toxicity of essential oils isolated from plant samples against stored-product pests is related to their major components [6-9, 11-13, 16-30]. These results suggested that the essential oils isolated from different plant species might have different toxicity, which can be attributed to their different chemical composition and different major and/or minor components. The observed fumigant activity shows that essential oils are sources of biologically active vapours which are potentially efficient insecticides. Consequently, the possibility of employing these natural fumigants to control insects in stored products may be worthy of further investigation [13]. A large number of essential oils extracted from different species and herb plants were screened for the insecticidal activity [14]. Some essential oils were found to be the most potential fumigants of all the oils tested against some major stored- product insect pests [26, 27]. In this study, h of treatment with the maximum dose (20 µl) of essential oils of O. onites, O. rotundifolium, R. officinalis, S. hydrangea, S. hortensis, S. spicigera, T. fallax and T. sipyleuswas determined the highest mortality rate in S. granarius (ig. 1, 2). These compounds may be used as potential insecticidal agents for controlling of S. granarius adults in stored food products. Our results and those reported earlier clearly indicate variations in the activity of essential oils in regard to the stage of the insect, the species of insect and plant origin of the essential oils. Not all the essential oils tested showed satisfactory activity, but the essential oils of O. onites, O. rotundifolium, R. officinalis, S. hydrangea, S. hortensis, S. spicigera, T. fallax and T. sipyleus proved to be promising as control agents against stored - product insects, especially S. granarius. The existence of naturally occurring insecticidal plant components has been known for centuries. However, relatively few of these compounds are actually used in crop protection today [3]. Increasing problems concerning the use of modem synthetic insecticides, such as persistence of residues, resistance, and damage to the environment and human health have generated interest in naturally occurring compounds. It should be noted that biologically active compounds of food plants are assumed to be environmentally more acceptable and less hazardous than others to humans. The results presented in this study suggest that some plant essential oils or their major constituents could be efficient fumigants and also could be integrated with other pest management procedures. References 1. BAKKALI., AVERBECK S., AVERBECK D., IDAOMAR, M., Biological effects of essential oils A review. ood and Chemical Toxicology, 46, (2008). 2. REGNAULT-ROGER, C., The potential of botanical essential oils for insect pest control. International Journal of Pest Management, 2, (1997). 3. YILDIRIM, E., OZBEK, H., ASLAN, I., Pests of Stored Product. Ataturk University Agricultural aculty Press, No.191, Erzurum, pp 117 (2001). 4. ISMAN, M. B., Plant essential oils for pest and disease management. Crop Protection, 19, (2000). 5. SU, H. C.., SPEERS, R. D., PATRIC, C. M., Toxicity of citrus oils to several stored product insects: Laboratory evaluation. Journal of Economic Entomology, 65, (1972). 6. KLINGAUER,., BESTMANN, H. J., VOSTROWSKY, O., MICHAELIS, K., Wirkung von aterischen Olen auf Schadinsekten. Mitt. Deut. Ges. fuer Allg. Angew Entomology, 4, (1983). 7. SU, H.C.., Laboratory study on effects of Anethum graveolens seeds onfour species of stored-product insects. Journal of Economic Entomology, 78, (1985). 8. SINGH, D., SIDDIQUI, M.S., SHARMA, S., Reproduction retar-dant and fumigant properties in essential oils against rice weevil (Coleoptera: Curculionidae) in stored wheat. Journal of Economic Entomology, 82, (1989). 9. EL NAHAL, G.H., SCHIMIDT, A.K.M., RISHA, E.M., Vapours of Acarus calamus oil-a space treatment for stored-product insects. Journal of Stored Products Research, 25, (1989) Romanian Biotechnological Letters, Vol. 16, No. 6, 2011

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