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The P450 enzyme Shade mediates the hydroxylation of ecdysone to 20-hydroxyecdysone in the Colorado potato beetle, Leptinotarsa decemlineata

文献类型： 外文期刊

作者： Kong, Y. 1 ; Liu, X-P. 1 ; Wan, P-J. 1 ; Shi, X-Q. 1 ; Guo, W-C. 2 ; Li, G-Q. 1 ;

作者机构： 1.Nanjing Agr Univ, Coll Plant Protect, Key Lab Integrated Management Crop Dis & Pests, Educ Minist, Nanjing 210095, Jiangsu, Peoples R China

2.Xinjiang Acad Agr Sci, Dept Plant Protect, Urumqi, Peoples R China

关键词： Shade;ecdysteroidogenesis;ecdysone 20-monooxygenase;RNA interference;performance

期刊名称：INSECT MOLECULAR BIOLOGY （ 2020影响因子：3.585； 五年影响因子：3.215 ）

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收录情况： SCI

摘要： Ecdysone 20-monooxygenase (E20MO), a cytochrome P450 monooxygenase (CYP314A1), catalyses the conversion of ecdysone (E) to 20-hydroxyecdysone (20E). We report here the cloning and characterization of the Halloween gene Shade (Shd) encoding E20MO in the Colorado potato beetle, Leptinotarsa decemlineata. LdSHD has five conserved motifs typical of insect P450s, ie the Helix-C, Helix-I, Helix-K, PxxFxPE/DRF (PERF) and heme-binding motifs. LdShd was expressed in developing eggs, the first to fourth instars, wandering larvae, pupae and adults, with statistically significant fluctuations. Its mRNA was ubiquitously distributed in the head, thorax and abdomen. The recombinant LdSHD protein expressed in Spodoptera frugiperda 9 (Sf9) cells catalysed the conversion of E to 20E. Dietary introduction of double-stranded RNA (dsRNA) of LdShd into the second instar larvae successfully knocked down the LdShd expression level, decreased the mRNA level of the ecdysone receptor (LdEcR) gene, caused larval lethality, delayed development and affected pupation. Moreover, ingestion of LdShd-dsRNA by the fourth instars also down-regulated LdShd and LdEcR expression, reduced the 20E titre, and negatively influenced pupation. Introduction of 20E and a nonsteroidal ecdysteroid agonist halofenozide into the LdShd-dsRNA-ingested second instars, and of halofenozide into the LdShd-dsRNA-ingested fourth instars almost completely relieved the negative effects on larval performance. Thus, LdSHD functions to regulate metamorphotic processes by converting E to 20E in a coleopteran insect species Le. decemlineata.

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[1]Knockdown of juvenile hormone acid methyl transferase severely affects the performance of Leptinotarsa decemlineata (Say) larvae and adults. Fu, Kai-Yun,Li, Qian,Zhou, Li-Tao,Meng, Qing-Wei,Lu, Feng-Gong,Li, Guo-Qing,Guo, Wen-Chao.

[2]CHARACTERIZATION AND FUNCTIONAL STUDY OF A PUTATIVE JUVENILE HORMONE DIOL KINASE IN THE COLORADO POTATO BEETLE Leptinotarsa decemlineata (Say). Fu, Kai-Yun,Lu, Feng-Gong,Li, Guo-Qing,Guo, Wen-Chao.

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[8]Molecular cloning and characterization of a putative proline dehydrogenase gene in the Colorado potato beetle, Leptinotarsa decemlineata. Wan, Pin-Jun,Lu, Dong,Yang, Lu,Mu, Li-Li,Li, Guo-Qing,Guo, Wen-Chao,Ahmat, Tursun. 2014

[9]RNA interference of chitin synthase genes inhibits chitin biosynthesis and affects larval performance in Leptinotarsa decemlineata (Say). Shi, Ji-Feng,Mu, Li-Li,Chen, Xu,Li, Guo-Qing,Guo, Wen-Chao. 2016

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[11]Knocking down a putative Delta(1)-pyrroline-5-carboxylate dehydrogenase gene by RNA interference inhibits flight and causes adult lethality in the Colorado potato beetle Leptinotarsa decemlineata (Say). Wan, Pin-Jun,Fu, Kai-Yun,Lu, Feng-Gong,Wang, Xin-Xin,Li, Guo-Qing,Wan, Pin-Jun,Guo, Wen-Chao.

[12]Knockdown of a putative alanine aminotransferase gene affects amino acid content and flight capacity in the Colorado potato beetle Leptinotarsa decemlineata. Wan, Pin-Jun,Fu, Kai-Yun,Lu, Feng-Gong,Li, Guo-Qing,Guo, Wen-Chao,Wan, Pin-Jun.

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