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比较RNA测序分析揭示了赤拟谷盗中新型δ类谷胱甘肽S-转移酶相关基因的表达模式。

Comparative RNA-sequencing profiling reveals novel Delta-class glutathione S-transferases relative genes expression patterns in Tribolium castaneum.

作者信息

Chen Xuhong, Xiong Wenfeng, Li Chengjun, Gao Shanshan, Song Xiaowen, Wu Wei, Li Bin

机构信息

Jiangsu Key Laboratory for Biodiversity and Biotechnology, College of Life Sciences, Nanjing Normal University, Nanjing 210023, China.

Jiangsu Key Laboratory for Biodiversity and Biotechnology, College of Life Sciences, Nanjing Normal University, Nanjing 210023, China.

出版信息

Gene. 2016 Nov 15;593(1):13-20. doi: 10.1016/j.gene.2016.08.013. Epub 2016 Aug 7.

Abstract

Glutathione S-transferases (GSTs) are a large group of enzymes having both detoxification roles conferring insecticide resistance and specialist metabolic functions. Tribolium castaneum GST Delta 1 (TcGSTd1) has been found playing crucial role in insecticide resistance and biological processes in insect species. However, the regulatory system of TcGSTd1 has still rarely been known. Comparing the transcriptome profile of RNAi treated larvae (ds-TcGSTd1) and control larvae of T. canstaneum by using RNA-sequencing, we obtained 14,284,085 sequence reads aligned with 13,275 genes. And 512 differentially expressed genes (DEGs) were identified from ds-TcGSTd1 treated group. Est/CCE, CYP, MRPs were significantly down-regulated in ds-TcGSTd1 group when compared with control group, which illustrated that they cooperated with TcGSTd1 to reduce the activity of cellular metabolism system. While, SNO was up-regulated in ds-TcGSTd1 insects suggested it may also involve in detoxifying alkaloid of insect metabolism system. These results established that TcGSTd1 not only acts as a vital gene for phase II cellular detoxification but also participates in phase 0, I, and III cellular detoxification by cooperating with CSPs, OBPs, CYP9, ESTB1, CCE6, MRPs and other detoxification genes. Knockdown of TcGSTd1 also suppressed several genes encoding antioxidant enzymes, e.g. CuZnSOD, Duox, Prx, HPX, CPO, and MCORP. Suggested that they may modulate the function of TcGSTd1 on lifespan, immune, development and reproduction. All these results shed the new insights into the regulatory mechanism of TcGSTd1 involved in insect physiology and could further facilitate the research of suitable and sustainable managements for the pest control.

摘要

谷胱甘肽S-转移酶(GSTs)是一大类酶,兼具赋予抗杀虫剂能力的解毒作用和特定的代谢功能。已发现赤拟谷盗谷胱甘肽S-转移酶δ1(TcGSTd1)在昆虫抗杀虫剂能力及昆虫物种的生物学过程中发挥关键作用。然而,TcGSTd1的调控系统仍鲜为人知。通过RNA测序比较RNA干扰处理的幼虫(ds-TcGSTd1)和赤拟谷盗对照幼虫的转录组图谱,我们获得了与13275个基因比对的14284085条序列读数。并且从ds-TcGSTd1处理组中鉴定出512个差异表达基因(DEGs)。与对照组相比,酯酶/羧酸酯酶(Est/CCE)、细胞色素P450(CYP)、多药耐药相关蛋白(MRPs)在ds-TcGSTd1组中显著下调,这表明它们与TcGSTd1协同作用以降低细胞代谢系统的活性。而在ds-TcGSTd1处理的昆虫中嗅觉神经元表面蛋白(SNO)上调,表明它可能也参与昆虫代谢系统中生物碱的解毒。这些结果表明,TcGSTd1不仅是II期细胞解毒的关键基因,还通过与化学感受蛋白(CSPs)、气味结合蛋白(OBPs)、CYP9、酯酶B1(ESTB1)、羧酸酯酶6(CCE6)、多药耐药相关蛋白及其他解毒基因协同作用,参与0期、I期和III期细胞解毒。敲低TcGSTd1也抑制了几个编码抗氧化酶的基因,如铜锌超氧化物歧化酶(CuZnSOD)、双氧化酶(Duox)、过氧化物酶(Prx)、六聚体过氧化物酶(HPX)、氯过氧化物酶(CPO)和中肠氧化还原蛋白(MCORP)。这表明它们可能调节TcGSTd1在寿命、免疫、发育和繁殖方面的功能。所有这些结果为TcGSTd1参与昆虫生理学的调控机制提供了新见解,并可进一步促进害虫防治适宜且可持续管理方法的研究。

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