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短角外斑腿蝗GST基因的鉴定及对杀虫剂敏感性
马国强1, 高俊仙2, 廉梅霞1
1.山西林业职业技术学院 林学系;2.山西省林业科学研究院
摘要:
【目的】鉴定短角外斑腿蝗(Xenocatantops humilis brachycerus)谷胱甘肽S-转移酶(glutathione S-transferase,GST)基因,并对其进行系统发育分析;比较GST基因在短角外斑腿蝗不同组织部位的表达水平,探究其对杀虫剂的敏感性。【方法】基于短角外斑腿蝗转录组数据库,通过Blast软件筛选GST基因;采用邻接法构建代表性昆虫GST家族系统进化树;利用实时荧光定量PCR技术,检测XbGSTd7、XbGSTd8、XbGSTe2XbGSTe4在短角外斑腿蝗10个组织部位的表达水平。通过注射dsRNA沉默XbGSTd8和XbGSTe4,结合杀虫剂敏感性试验,比较GST基因沉默后短角外斑腿蝗对4种杀虫剂敏感性的变化。【结果】从短角外斑腿蝗转录组数据库筛选得到15条GST全长序列和12条GST片段序列。系统发育分析表明,15条短角外斑腿蝗GST全长序列分别属于Delta、Epsilon、Theta和Omega家族。实时定量PCR结果显示,XbGSTd8在短角外斑腿蝗胃毛囊、中肠和马氏管的表达量相对较高,XbGSTe4在短角外斑腿蝗胃毛囊和中肠的表达量相对较高。沉默XbGSTe4后,短角外斑腿蝗对4种杀虫剂的敏感性无明显变化;沉默XbGSTd8后,短角外斑腿蝗对毒死蜱的敏感性显著提高。【结论】XbGSTd8可能参与短角外斑腿蝗对毒死蜱的代谢解毒过程。
关键词:  短角外斑腿蝗  谷胱甘肽S-转移酶  组织表达  杀虫剂敏感性
DOI:
分类号:
基金项目:山西省自然科学基金项目(2012011033-4)
Identification of glutathione S-transferase (GST) gene family from Xenocatantops humilis brachycerus and its insecticide sensitivity
MA Guoqiang,GAO Junxian,LIAN Meixia
Abstract:
【Objective】The glutathione S-transferase (GST) genes from Xenocatantops humilis brachycerus were identified and analyzed by phylogenetic analysis,expression levels at different tissues were compared among,and the sensitivity to insecticides was explored.【Method】Based on the transcriptome database of X. humilis brachycerus,GST genes were screened by Blast software,and the phylogenetic tree was constructed by neighboring method.RT-qPCR was used to detect the expression levels of four GST genes among ten tissues of X. humilis brachycerus.XbGSTd8 and XbGSTe4 were silenced by injection of dsRNA.Combined with insecticide bioassay experiments, the sensitivity of X. humilis brachycerus against four insecticides was also determined after GST gene silencing.【Result】Fifteen full-length sequences and twelve fragment sequences of GST were screened by blast software from the transcriptome database of X. humilis brachycerus.Phylogenetic analysis indicated that these full-length sequences belonged to the Delta,Epsilon,Theta and Omega families,respectively.RT-qPCR showed that XbGSTd8 was highly expressed in gastric caecum,midgut and Malpighian tubules,and XbGSTe4 was highly expressed in gastric caecum and midgut.After silencing XbGSTe4,the sensitivity of X. humilis brachycerus against four insecticides changed insignificantly,while silencing XbGSTd8 significantly increased sensitivity against chlorpyrifos.【Conclusion】XbGSTd8 may be involved in the metabolic detoxification process of chlorpyrifos in X.humilis brachycerus.
Key words:  Xenocatantops humilis brachycerus  glutathione S-transferase  tissue-specific expression  insecticide sensitivity