Construction of Transgenic Drosophila Expressing LdCYP6B53 and Its Susceptibility to Insecticides

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Chinese Agricultural Science Bulletin ›› 2016, Vol. 32 ›› Issue (19) : 97-101. DOI: 10.11924/j.issn.1000-6850.casb16030222

Construction of Transgenic Drosophila Expressing LdCYP6B53 and Its Susceptibility to Insecticides

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Abstract

The objective was to investigate the susceptibility of CYP6B53 to four insecticides including cyfluthrin, deltamethrin, phoxim and chlorantraniliprole. The attP40＞CYP6B53 and VK5＞CYP6B53 transgenic Drosophila were constructed by transforming LdCYP6B53 into Drosophila. The toxicity of insecticides to transgenic Drosophila was measured using bioassay. The CYP6B53 gene was confirmed to stably express into Drosophila by restriction endonuclease digestion, RT-PCR and sequencing. Compared with attP40 and VK5 control Drosophila, the attP40＞CYP6B53 and VK5＞CYP6B53 transgenic Drosophila significantly decreased susceptibility to cyfluthrin with 1.83- fold and 1.61- fold of the control Drosophila for LC50, respectively. However, the attP40＞CYP6B53 and VK5＞CYP6B53 transgenic Drosophila significantly increased susceptibility to deltamethrin with 0.52- fold and 0.21- fold of the control Drosophila for LC50, respectively. No significant difference of susceptibility to phoxim and chlorantraniliprole was detected between attP40＞CYP6B53 and VK5＞CYP6B53 transgenic Drosophila and the control Drosophila. The transgenic Drosophila lines attP40＞CYP6B53 and VK5＞CYP6B53 were successfully constructed by using transgenic technology, and these transgenic Drosophila lines could be used for screening insecticides and provide a method for studying LdCYP6B53 functions.

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transgenic Drosophila; Lymantria dispar; CYP6B53 gene; insecticide; susceptibility

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Construction of Transgenic Drosophila Expressing LdCYP6B53 and Its Susceptibility to Insecticides. Chinese Agricultural Science Bulletin. 2016, 32(19): 97-101 https://doi.org/10.11924/j.issn.1000-6850.casb16030222

References

[1]候雅芹,南楠,李镇宇. 舞毒蛾研究进展[J].河北林果研究, 2009, 24(4):439-444
[2]倪鸣,柴汝松,张鹏,等.不同药剂对舞毒蛾和春尺蠖的毒力测定 [J].东北林业大学学报,2009,37(5):121-122.
[3]Daborm P J, Lumb C, Harrop T W R, Set al. UsingSDrosophila melanogasterSto validate metabolism-based insecticide resistance from insect pests [J]. Insect Biochem Molec Biol, 2012, 42(12):918-924.
ffrench-Constant, Roush R T, Cari?o F A. Drosophila as a Tool for investigating the molecular genetics of insecticide resistance. Molecular approaches to fundamental and applied entomology: Part of the series Springer Series in Experimental Entomology, 1993, 1-37.
[5]Morton RA. Evolution ofSDrosophilaSinsecticide resistance [J].Genome, 1993, 36:1-7.
[6]Daborn P J, Yen J L, Bogwitz M R, et al. ASsingle P450 allele associated with insecticide resistance in Drosophila [J]. Science, 2002, 297(5590):2253-2256.
[7]Bogwitz M R, Chung H, Magoc L, et al. Cyp12a4 confers lufenuron resistance in a natural population of Drosophila melanogaster [J]. P. Natl. Acad. Sci. USA., 2005, 102(36):12807-12812.
[8]Perry T, McKenzie J A, Batterham P. ASDalpha6Sknockout strain ofSDrosophila melanogasterSconfers a high level of resistance to spinosad [J]. Insect Biochem Mol. Biol, 2007, 37(2): 184-188.
[9]Low W Y, Feil S C, Ng H L, et al. Recognition and detoxification of the insecticide DDT bySDrosophila melanogasterSglutathione S-transferase D1 [J]. J. Mol. Biol, 2010, 99(3):358-366.
[10]Zhu F, Parthasarathy R, Bai H, et al. A brain-speci?c cytochrome P450 responsible for the majority of deltamethrin resistance in the QTC279 strain of Tribolium castaneum [J]. P Natl Acad Sci USA., 2010, 107(19):8557-8562.
[11]Riveron J M,SIrving H,SNdula M, et al. Directionally selected cytochrome P450 alleles are driving the spread of pyrethroid resistance in the major malaria vector Anopheles funestus [J]. P Natl Acad Sci USA, 2013, 110(1): 252-257.
[12]Riveron J M,SIbrahim S S,S Chanda E, et al. The highly polymorphicSCYP6M7Scytochrome P450 gene partners with the directionally selectedSCYP6P9aSand CYP6P9bSgenes to expand the pyrethroid resistance front in the malaria vectorSAnopheles funestusSin Africa [J]. BMC Genomics, 2014, 15: 817.
[13]Daborn P, Boundy S, Yen J, et al. DDT resistance in Drosophila correlates with Cyp6g1 over-expression and confers cross-resistance to the neonicotinoid imidacloprid [J]. Mol Genet Genomics, 2001, 266(4):556-563.
[14]耿俊华.人Tau蛋白截短片断过表达转基因果蝇模型的建立及鉴定[D],南京：东南大学,2009,6-17.
[15]王晓军, 张宗俭, 姜辉, 等. NY/T1154.7-2006,农药室内生物测定试验准则[S].北京：中国标准出版社,2006.
[16]Lvaylo S. Cytochrome P450s: coupling development and environment [J]. Trends Genet, 2001, 17(11):629-632.
[17]Zhao Y, Hull A K, Gupta N R, Set al. Trp-dependent auxin biosynthesis inSArabidopsis: involvement of cytochrome P450s CYP79B2 and CYP79B3 [J]. Genes Dev, 2002, 16(23): 3100–3112.
[18]Li X C, Baudry J, Berenhaum M R, et al. Structural and functional divergence of insect CYP6B proteins: From specialist to generalist cytochrome P450 [J]. P Natl Acad Sci USA, 2004, 101(9):2939-2944.
[19]Daborn P J, Lumb C, Boey A, et al. Evaluating the insecticide resistance potential of eightSDrosophila melanogasterScytochrome P450 genes by transgenic over-expression [J]. Insect Biochem Mol Biol, 2007, 37(5): 512–519.
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