Cloning, Sequence Analysis and Expression of Helicoverpa armigera CYP9G5 Gene under Stress of Deltamethrin

doi:10.16409/j.cnki.2095-039x.2018.04.004

Abstract

Abstract: Cytochrome P450 is a supergene family, which belongs to one of the three important detoxification enzymes in vivo and participates in the synthesis and oxidation metabolism of endogenous and exogenous substances. In this paper, a P450 gene CYP9G5 (GenBank:KM016757.1) was cloned through data analysis of the transcription group of H. armigera, encoding 493 amino acids with a molecular weight of about 57.930 kDa, and theoretical isoelectric point pI 7.16. Using the technology of real-time quantitative analysis the CYP9G5 genes in different groups of the Helicoverpa armigera and deltamethrin treatment after expressing quantity change, found CYP9G5 highest expression quantity of the head, after treated with deltamethrin CYP9G5 express volume also increased significantly. It is speculated that CYP9G5 is very likely to be involved in the metabolism of H. armigera to cypermethrin, and may play a role in the resistance of H. armigera. This study lays a foundation for further exploring the resistance of H. armigera and the development of new pesticides.

Key words: Cytochrome P450, Helicoverpa armigera, deltamethrin, Real-time quantitative PCR

CLC Number:

S435.622

ZHANG Kun, DENG Denghui, GUO Jianglong, HAN Weili, WU Shaoying. Cloning, Sequence Analysis and Expression of Helicoverpa armigera CYP9G5 Gene under Stress of Deltamethrin[J]. journal1, 2018, 34(4): 520-526.

References

[1] King A B S. Heliothis/Helicoverpa (Lepidoptera:Noctuidae)[M]//Matthews G A, Tunstall J P eds. Insect Pests of Cotton. Wallingford, United Kingdom:CAB International, 1994, 39-106.
[2] McCaffery A R. Resistance to insecticides in Heliothine Lepidoptera:a global view[J]. Philosophical Transaction of the Royal Society of London. Series B:Biological Sciences, 1998, 353(1376):1735-1750.
[3] Shen J, Wu Y. Resistance of Helicoverpa armigera to insecticides and its management[M]. Beijing:China Agricultural Press, 1995, 35-38.
[4] Forrester N W, Cahill M, Bird L T, et al. Management of pyrethroid and endosulfan resistance in Helicoverpa armigera (Lepidopteran:Noctuidae) in Australia[J]. Bulletin of Entomological Research-Supplement, 1993, 5(1):21-27.
[5] Keeling C, Bohlmann J. Genes, enzymes and chemicals of terpenoid diversity in the constitutive and induced defence of conifers against insects and pathogens[J]. New Phytologiet, 2006, 170(5):657-675.
[6] Nelson D R. Progress in tracing the evolutionary paths of cytochrome P450[J]. Biochimica et Biophysica Acta, 2011, 1814(1):14-18.
[7] Nelson D R, Strobel H W. Evolution of cytochrome P-450 proteins[J]. Molecular Biology Evolution, 1987, 4(6):572-593.
[8] Feyereisen R. Insect P450 enzymes[J]. Annual Review Entomo1ogy, 1999, 44(1):507-533.
[9] Scott J G, Wen Z. Cytochromes P450 of insects:the tip of the iceberg[J]. Pest Management Science, 2001, 57(1):958-967.
[10] 郭亭亭, 姜辉, 高希武. 昆虫细胞色素P450基因的多样性、进化及表达调控[J]. 昆虫学报, 2009, 52(3):301-311.
[11] Bautista M A, Miyata T, Miura K, et al. RNA interference- mediated knockdown of a cytochrome P450, CYP6BG1, from the diamondback moth, Plutella xylostella, reduces larval resistance to permethrin[J]. Insect Biochemistry and Molecular Bio1ogy, 2008, 39(1):38-46.
[12] Wang R L, Liu S W, Scott R B, et al. Identification and functional analysis of a novel cytochrome P450 gene CYP9A105 associated with pyrethroid detoxification in Spodoptera exigua Hübner[J]. Molecular Science, 2018, 19(3):737.
[13] John V, Linda G, John M, et al. Rapid selection of a pyrethroid metabolic enzyme CYP9K1 by operational malaria control activities[J]. Proceedings of the National Academic of Science, 2018, 115(18):4619-4624.
[14] Livak K J, Schmittgen T D. Analysis of relative gene expression data using real-time quantitative PCR and the 2^-ΔΔCTmethod[J]. Methods, 2001, 25(4):402-408.
[15] Strode C, Wondji C S, David J P, et al. Genomic analysis of dctoxification genes in the mosquito Aedes aegypti[J]. Insect Biochemistry Molecular Biology, 2008, 38(1):113-123.
[16] Huber D P W, Erickson M L, Leutenegger C M, et al. Isolation and extreme sex-specific expression of cytochrome P450 genes in the bark beetle, Ips paraconfusus, following feeding on the phloem of host ponderosa pine, Pinus ponderosa[J]. Insect Molecular Biology, 2007, 16(3):335-349.
[17] Aw T, Schlauch K, Keeling C I, et al. Functional genomics of mountain pine beetle (Dendroctonus ponderosae) midguts and fat bodies[J]. BMC Genomics, 2010, 11(1):1-12.
[18] Cano-Ramírez C, López M F, Cesar-Ayala A K, et al. Isolation and expression of cytochrome P450 genes in the antennae and gut of pine beetle Dendroctonus rhizophagus (Curculionidae:Scolytinae) following exposure to host monoterpenes[J]. Gene, 2012, 11(1):59.
[19] Keeling C I, Henderson H, Li M, et al. Transcriptome and full-length cDNA resources for the mountain pine beetle, Dendroctonus ponderosae Hopkins, a major insect pest of pine forests[J]. Insect Biochemistry Molecular Biology, 2012, 42(1):525-536.
[20] 王燕红. 野桑蚕细胞色素P450基因的克隆、组织表达及其功能研究[D]. 苏州:苏州大学, 2009.
[21] Zhu F, Parthasarathy R, Bai H, et al. A brain-specific cytochrome P450 responsible for the majority of deltamethrin resistance in the QTC279 strain of Tribolium castaneum[J]. Proceedings of the National Academic of Science, 2010, 107(19):8557-8562.
[22] Pilar M, Carmen L, Marian M, et al. Response of last instar Helicoverpa armígera larvae to Bt toxin ingestion:changes in the development and in the CYP6AE14, CYP6B2 and CYP9A12 gene expression[J]. PLoS ONE, 2014, 9(6):e99229.
[23] Wu S Y, Shi X Y, Wang Y, et al. Response of cytochrome P450 expression to maize volatiles in Helicoverpa armigera[J]. Journal of Integrative Agriculture, 2013, 12(4):646-652.
[24] Zhang L, Lu Y, Xiang M, et al. The retardant effect of 2-Tridecanone, mediated by cytochrome P450, on the development of cotton bollworm, Helicoverpa armigera[J]. BMC Genomics, 2016, 17(1):954-970.
[25] Xu L, Li D Z, Qin J Y, et al. Over-expression of multiple cytochrome P450 genes in fenvalerate-resistent field strains of Helicoverpa armigera from north of China[J]. Pesticide Biochemistry and Physiology, 2016, 84(1):127-134.