鉴定和表征菜粉蝶 17 个谷胱甘肽 S-转移酶基因。
Identification and characterisation of seventeen glutathione S-transferase genes from the cabbage white butterfly Pieris rapae.
机构信息
College of Plant Protection, Anhui Agricultural University, Hefei, Anhui 230036, China.
College of Plant Protection, Anhui Agricultural University, Hefei, Anhui 230036, China.
出版信息
Pestic Biochem Physiol. 2017 Nov;143:102-110. doi: 10.1016/j.pestbp.2017.09.001. Epub 2017 Sep 8.
Insect glutathione S-transferases (GSTs) play essential roles in the detoxification of insecticides and other xenobiotic compounds. The cabbage white butterfly, Pieris rapae, is an economically important agricultural pest. In this study, 17 cDNA sequences encoding putative GSTs were identified in P. rapae. All cDNAs include a complete open reading frame and were designated PrGSTd1-PrGSTz2. Based on phylogenetic analysis, PrGSTs were divided into six classes (delta, epsilon, omega, sigma, theta and zeta). The exon-intron organizations of these PrGSTs were also analysed. Recombinant proteins of eight PrGSTs (PrGSTD1, PrGSTD2, PrGSTE1, PrGSTE2, PrGSTO1, PrGSTS1, PrGSTT1 and PrGSTZ1) were heterologously expressed in Escherichia coli, and all of these proteins displayed glutathione-conjugating activity towards 1-chloro-2,4-dinitrobenzene (CDNB). Expression patterns in various larval tissues, at different life stages, and following exposure to sublethal doses of abamectin, chlorantraniliprole or lambda-cyhalothrin were determined by reverse transcription-quantitative PCR. The results showed that PrGSTe3, PrGSTs1, PrGSTs2, and PrGSTs4 were mainly transcribed in the fat body, while PrGSTe2 was expressed predominantly in the Malpighian tubules. Four genes (PrGSTe2, PrGSTo4, PrGSTs4 and PrGSTt1) were mainly expressed in fourth-instar larvae, while others were ubiquitously expressed in egg, larval, pupa and/or adult stages. Abamectin treatment significantly upregulated ten genes (PrGSTd1, PrGSTd3, PrGSTe1, PrGSTe2, PrGSTo1, PrGSTo3, PrGSTs1, PrGSTs3, PrGSTs4 and PrGSTt1). Chlorantraniliprole and lambda-cyhalothrin treatment significantly upregulated nine genes (PrGSTd1, PrGSTd2, PrGSTe1, PrGSTe2, PrGSTe3, PrGSTs1, PrGSTs3, PrGSTs4 and PrGSTz1) and ten genes (PrGSTd1, PrGSTd3, PrGSTe1, PrGSTe2, PrGSTo1, PrGSTo2, PrGSTs1, PrGSTs2, PrGSTs3 and PrGSTz2), respectively. These GSTs are potentially involved in the detoxification of insecticides.
昆虫谷胱甘肽 S-转移酶（GSTs）在昆虫杀虫剂和其他外来化合物的解毒中发挥着重要作用。菜粉蝶（Pieris rapae）是一种重要的农业害虫。本研究在菜粉蝶中鉴定了 17 个编码假定 GSTs 的 cDNA 序列。所有 cDNA 均包含完整的开放阅读框，并被命名为 PrGSTd1-PrGSTz2。根据系统发育分析，PrGSTs 分为六个类（delta、epsilon、omega、sigma、theta 和 zeta）。还分析了这些 PrGSTs 的内含子-外显子组织。通过大肠杆菌异源表达了 8 种 PrGSTs（PrGSTD1、PrGSTD2、PrGSTE1、PrGSTE2、PrGSTO1、PrGSTS1、PrGSTT1 和 PrGSTZ1）的重组蛋白，所有这些蛋白均对 1-氯-2,4-二硝基苯（CDNB）表现出谷胱甘肽共轭活性。通过反转录定量 PCR 测定了在不同幼虫组织、不同生命阶段以及亚致死剂量的阿维菌素、氯虫苯甲酰胺或氯氰菊酯暴露后的表达模式。结果表明，PrGSTe3、PrGSTs1、PrGSTs2 和 PrGSTs4 主要在脂肪体中转录，而 PrGSTe2 主要在马氏管中表达。四个基因（PrGSTe2、PrGSTo4、PrGSTs4 和 PrGSTt1）主要在第四龄幼虫中表达，而其他基因则在卵、幼虫、蛹和/或成虫阶段广泛表达。阿维菌素处理显著上调了 10 个基因（PrGSTd1、PrGSTd3、PrGSTe1、PrGSTe2、PrGSTo1、PrGSTo3、PrGSTs1、PrGSTs3、PrGSTs4 和 PrGSTt1）。氯虫苯甲酰胺和氯氰菊酯处理分别显著上调了 9 个基因（PrGSTd1、PrGSTd2、PrGSTe1、PrGSTe2、PrGSTe3、PrGSTs1、PrGSTs3、PrGSTs4 和 PrGSTz1）和 10 个基因（PrGSTd1、PrGSTd3、PrGSTe1、PrGSTe2、PrGSTo1、PrGSTo2、PrGSTs1、PrGSTs2、PrGSTs3 和 PrGSTz2）。这些 GSTs 可能参与了杀虫剂的解毒作用。
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