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谷胱甘肽-S-转移酶(glutathione-S-transferases,GSTs)是对一种机体的解毒代谢起重要作用并可以被诱导的酶系。为了从分子水平上探究家蚕Bombyx mori GSTs基因与植物次生物质芸香苷代谢的关系,本实验采用双跟踪标定定量PCR(dual-spike-in qPCR)方法,用5×10~(-1),5×10~(-2),5×10~(-3) ng/μL 3个浓度芸香苷溶液处理家蚕5龄幼虫,并对各组织中GSTs Epsilon家族不同基因的转录水平进行检测。结果显示,浓度为5×10~(-2) ng/μL的芸香苷溶液能诱导GSTs基因的转录水平发生显著变化。在中肠中,BmGSTe1,BmGSTe2和BmGSTe6的诱导转录水平较高,在诱导后24 h达到最大值;在脂肪体中BmGSTe1,BmGSTe6和BmGSTe7的转录水平相对较高且分别在诱导后2 h,2 h和4 h达到最大值;而GSTs基因在马氏管中表达量均很低或者检测不到表达。与5×10~(-2) ng/μL浓度相比,5×10~(-1)ng/μL的芸香苷溶液诱导后各基因的转录水平上升的幅度较小并且达到峰值的时间不同,而5×10~(-3) ng/μL浓度的芸香苷溶液并不能使GSTs基因的诱导转录水平发生变化。结果提示,家蚕GSTs Epsilon家族基因对芸香苷的代谢具有重要作用。
Glutathione-S-transferases (GSTs) are enzymes that play an important role in the detoxification metabolism of an organism and can be induced. In order to explore the relationship between the Bombyx mori GSTs gene and the rutin metabolism of plant secondary metabolites at the molecular level, a dual-spike-in qPCR method was used in this experiment. The effects of 5 × 10 ~ (-1) The 5th instar larvae of silkworm (Bombyx mori) were treated with rutin at 5 × 10 -2 and 5 × 10 -3 ng / μL, respectively. The transcript levels of different genes of GSTs Epsilon family were detected in each tissue. The results showed that the concentration of 5 × 10 -2 ng / μL rutin solution can induce a significant change in the transcription level of GSTs gene. In the midgut, BmGSTe1, BmGSTe2 and BmGSTe6 had higher transcriptional transcript levels and reached the maximum at 24 h after induction. The transcript levels of BmGSTe1, BmGSTe6 and BmGSTe7 in adipose tissue were relatively high and reached the peak at 2 h, 2 h and 4 h reached the maximum value; while GSTs gene expression in the Markov tube were low or undetectable expression. Compared with the concentration of 5 × 10 -2 ng / μL, the transcription level of each gene increased slightly and peaked at different times after being induced by 5 × 10 -1 ng / μL rutin. However, rutin at concentrations of 5 × 10 ~ (-3) ng / μL did not change the transcriptional level of GSTs. The results suggest that the silkworm GSTs Epsilon family of genes on rutin metabolism has an important role.