Cem ERDOĞAN, M. Oktay GÜRKAN, Graham MOORES, A. Sibel VELİOĞLU

Pamuklarda zarar yapan Aphis gossypii Glover (Hemiptera: Aphididae) populasyonlarının biyokimyasal yöntemlerle direnç mekanizmalarının belirlenmesi

Bu araştırma, Akdeniz Bölgesi pamuk ekiliş alanlarından toplanan değişik Aphis gossypii populasyonlarında, insektisitlere direnç ile ilişkili olabilen mekanizmaların biyokimyasal yöntemlerle incelenmesi amacıyla 2000-2003 yılları arasında yürütülmüştür. Denemelerde, Adana'nın dört pamuk alanı ile Antalya'dan toplanan Aphis gossypii populasyonları ve yurtdışından getirtilen standart populasyonlar kullanılmıştır. Direnç ile ilişkili olabilen toplam esteraz aktivitesi kinetik mikroplate assay yöntemi kullanılarak belirlenmiştir. Populasyonlarin esteraz bant dizilişleri poliakrilamid jel elektroforez (PAGE) yöntemi kullanılarak incelenmiştir. Bunlara ilaveten, asetilkolinesteraz (AChE) duyarsızlaşması sonucunda ortaya çıkan insektisitlere direnç durumu mikroplate assay yöntemi kullanılarak araştırılmıştır. Yapılan çalışmalar, toplanan populasyonlardan dördünün yüksek düzeyde esteraz aktivitesine sahip olduğunu göstermiştir. Elektroforez ile ülkemiz populasyonlarının esteraz bant dizilişleri belirlenmiş olup, bu populasyonlarin dirençli 1081K populasyonu ile benzer bant dizilişinde olduğu tespit edilmiştir. Modifiye AChE çalışmasında, inhibitor olarak pirimicarb ile demeton-S-metil kullanılarak populasyonlarin hedef alan duyarlılıkları incelenmiştir. Populasyonlarin pirimicarb ve demeton-S-metil'e değişen oranlarda duyarsız asetilkolinesteraz aktivitesi sergiledikleri belirlenmiştir.

Determination of the insecticide resistance mechanisms using biochemical methods in the Aphis gossypii Glover (Hemiptera: Aphididae) populations Damaging on cotton in Turkey

This research was performed between 2000 and 2003, to explore the role of the mechanisms in conferring insecticide resistance of Aphis gossypii populations collected from the cotton areas of the Mediterranean region of Turkey using biochemical methods. These populations collected from Antalya and four different fields of Adana and standard A. gossypii populations were used throughout the studies. Total esterase activity associated with insecticide resistance was detected using kinetic microplate assay. All populations were characterized using polyacrylamide gel electrophoresis (PAGE) to determine the esterase banding pattern. In addition, insecticide resistance resulted from insensitivity of acetylcholinesterase was investigated using microplate assay. According to results from the biochemical assays, four cotton aphid populations were found to have high esterase activity. Esterase banding patterns were determined with electrophoresis. This demonstrated that all Turkish cotton aphids collected were the same banding pattern as the standard resistant 1081K population in the PAGE. In modified AChE study, target site sensitivities of the populations were investigated using pirimicarb and demeton-S-methyl as inhibitors. All of these populations were identified with differing levels of sensitivity to these chemicals.

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Andrews, M.C., A. Callaghan, L.M. Field, M.S. Williamson and G.D. Moores. 2004. Identification of mutations conferring insecticide-insensitive AchE in the cotton-melon aphid, Aphis gossypii Glover. Insect Mol. Biol., 13 (5): 555-561.
Anonim 2005. Tarımsal Yapı (Üretim, fiyat, değer) 2003. DİE Matbaası. Ankara. 546s.
Anonymous 2007. Arthropod Pesticide Resistance Database. http://www.pesticideresistance.org/ search IV
Benting, J. and R. Nauen. 2004. Biochemical evidence that an S431F mutation in acetylcholinesterase-1 of Aphis gossypii mediates resistance to pirimicarb and omethoate. Pest Manag. Sci. 60: 1051-1055.
Deguine, J.P. 1996. The evolution of insecticide resistance in Aphis gossypii Glover (Hemiptera: Aphididae) in Cameroon, Resistant Pest Management News Letter 8 (1): 13-14.
Delorme, R., D. Auge, M. Bethenod and F. Villate. 1997. Insecticide resistance in a strain of Aphis gossypii from southern France, Pestic. Sci., 49, 90-96.
Devonshire, A.L. and R.M. Sawicki. 1979. Insecticide -resistant Myzus persicae as an example of evolution by gene duplication. Nature 280: 140-141.
Devonshire, A.L. and G.D. Moores. 1982. A carboxylesterase with broad substrate specifity causes organophosphorus, carbamate and pyrethroid resistance in peach-potato aphids (Myzus persicae), Pestic. Biochem. Physiol., 18: 235-246.
Devonshire, A.L. 1989. Resistance of aphids to insecticides, In: Aphids, Their Biology, Natural Enemies and Control, eds: Minks, A. K. and Harrewijn, P., Volume C, Elsevier Science Publishers B. V., Amsterdam, 123-139.
Dong, S.J., M.C. Andrews, F. Li, G.D. Moores, Z.J. Han and M.S. Williamson. 2005. Acetylcholinesterase genes and insecticide resistance in aphids. Chem. Biol. Interact.,157-158:373-374.
Ffrench-Constant, R. H. and A. L. Devonshire. 1987. A multiple homogenizer for rapid sample preparation in immunoassays and electrophoresis. Biochem. Genet., 25: 493- 499.
Furk, C., D. F. Powell and S. Heyd. 1980. Pirimicarb resistance in the mellon and cotton aphid, Aphis gossypii Glover, Plant Pathol., 29: 191-196.
Furk, C. and C.M. Hines. 1993. Aspects of insecticide resistance in the melon and cotton aphid, Aphis gossypii (Hemiptera: Aphididae), Ann. appl. Biol., 123: 9-17.
Han, Z., G.D. Moores, I. Denholm and A.L. Devonshire. 1998. Association between biochemical markers and insecticide resistance in the cotton aphid, Aphis gossypii Glover, Pestic. Biochem. Physiol., 62: 164-171.
Herron, G.A., K. Powis and J. Rophail. 2001. Insecticide resistance in Aphis gossypii Glover (Hemiptera: Aphididae), a serious threat to Australian cotton, Aust. J. Entomol., 40:85-91.
Javed, N., R. Viner, M.S. Williamson, L.M. Field, A.L. Devonshire and G.D. Moores. 2003. Characterization of acetylcholinesterases, and their genes, from the hemipteran species Myzus persicae (Sulzer), Aphis gossypii (Glover), Bemisia tabaci (Gennadius) and Trialeurodes vaporariorum (Westwood). Insect Mol. Biol., 12 (6): 613-620.
Li, F. and Z. Han. 2004. Mutations in acetylcholinesterase associated with insecticide resistance in the cotton aphid, Aphis gossypii Glover. Insect Biochem. Mol. Biol., 34: 397-405.
Metcalf, R. L. 1989. Insect resistance to insecticides, Pestic. Sci., 26, 333-358.
Moores, G. D., I. Denholm, F. J. Byrne, A. L. Kennedy and A. L. Devonshire. 1988. Characterising acetylcholinesterase genotypes in resistant insect populations, Brighton Crop Protection Conference-Pests and Diseases, 451-456.
Moores, G. D., X. Gao, I. Denholm and A. L. Devonshire. 1996a. Characterisation of insensitive acetylcholinesterase in insecticide-resistant cotton aphids, Aphis gossypii Glover (Homoptera: Aphididae), Pestic. Biochem. Physiol., 56: 102-110.
Moores, G. D., Z. J. Han, I. Denholm and A. L. Devonshire. 1996b. Two forms of insecticide insensitive acetylcholinesterase in Aphis gossypii, Brighton Crop Protection Conference-Pests and Diseases, 745-750.
O'Brien, P.J. and J.B. Graves. 1992. Insecticide resistance and reproductive biology of Aphis gossypii Glover, Southwest. Entomol., 17 (2): 115-122.
O'Brien, P.J., Y.A. Abdel-Aal, J.A. Ottea and J.B. Graves. 1992. Relationship of insecticide resistance to carboxylesterases in Aphis gossypii (Homoptera: Aphididae) from midsouth cotton, J. Econ. Entomol., 85 (3): 651-657.
Saito, T. 1993. Insecticide resistance of the cotton aphid, Aphis gossypii Glover (Homoptera: Aphididae) VI. Qualitative variations of aliesterase activity, Appl. Entomol. Zool., 28 (2): 263-265.
Saito, T. and H. Hama. 2000. Carboxylesterase isozymes responsible for organophosphate resistance in the cotton aphid, Aphis gossypii Glover (Homoptera: Aphididae), Appl. Entomol. Zool., 35 (1): 171-175.
Silver, A.R.J., H.F. van Emden and M. Battersby. 1995. Biochemical nature of pirimicarb resistance in two glasshouse clones of Aphis gossypii. Pestic. Sci. 43 (1): 21-29.
Sun, L, X. Zhou, J. Zhang and X. Gao. 2005. Polymorphisms in a Carboxylesterase Gene Between Organophosphate-Resistant and -Susceptible Aphis gossypii (Homoptera: Aphididae). J. Econ. Entomol. 98 (4): 1325-1332.
Suzuki, K., H. Hama, and Y. Konno. 1993. Carboxylesterase of the cotton aphid, Aphis gossypii Glover (Homoptera: Aphididae), responsible for fenitrothion resistance as a sequestering protein, Appl. Entomol. Zool., 28 (4): 439-450.
Takada, H. and Y. Murakami. 1988. Esterase variation and insecticide resistance in Japanese Aphis gossypii, Entomol. Exp. Appl., 48: 37-41.
Toda, S., S. Komazaki, T. Tomida and Y. Kono. 2004. Two aminoacid substitutions in acetylcholinesterase associated with pirimicarb and organophosphorous insecticide resistance in the cotton aphid, Aphis gossypii Glover (Homoptera: Aphididae). Insect Mol. Biol., 13 (5): 549-553.
Tomlin, C. D. S. (ed.) 1997. The Pesticide Manual. 11th edition, The British Crop Protection Council, BCPC Publications, 1606 p., UK.
Williams, D. E. and R. A. Reisfeld. 1964. Disc electrophoresis in polyacrylamide gels: extension to new conditions of pH and buffer, Ann. NY. Acad. Sci., 121: 373-381.