棉花学报2025，Vol.37Issue(1)：38-49,12.DOI:10.11963/cs20240046
5个解毒基因在棉蓟马生长发育中及甲维盐胁迫下的表达模式分析
Expression pattern analysis of five detoxification genes in the growth and development of Thrips tabaci and under emamectin benzoate stress
摘要
Abstract
[Objective]This study aims to clarify the sequence structure of cytochrome P450(CYP450)gene and glutathione S-transferase(GST)gene of Thrips tabaci,and the expression of these genes at the different growth and development stages of T.tabaci and emamectin benzoate stress.[Methods]Based on the transcriptome data of different growth and development stages of T.tabaci,CYP450 genes and GST genes were mined,specific primers were designed,and polymerase chain reaction(PCR)was used to amplify the cDNA of these genes.Bioinformatics software was used to predict the structural characteristics of CYP450 and GST proteins.The indoor toxicity of emamectin benzoate to adult T.tabaci was determined by leaf dipping method.Quantitative real time PCR(qRT-PCR)was used to analyze the expression patterns of CYP450 and GST genes at the different development stages of T.tabaci and under the stress of emamectin benzoate.[Results]Three CYP450 genes(CYP4C101,CYP4C102,CYP6K1)and two GST genes(GST1,GSTX1)were cloned.The results of physicochemical analysis showed that CYP4C101,CYP4C102,CYP6K1,GST1,and GSTX1 were composed of 507,528,513,215 and 207 amino acid residues,respectively,all of which were hydrophilic proteins.Phylogenetic analysis showed that CYP4C101 had the highest homology with CYP4C1 of Frankliniella fusca.CYP4C102,CYP6K1 and GST1 of T.tabaci had the highest relationships with the homologous proteins from F.occidentalis and F.fusca.GSTX1 had the highest homology with F.occidentalis.Domain prediction showed that CYP4C101,CYP4C102 and CYP6K1 had conserved domains of CYP450,and GST1 and GSTX1 had conserved domains of GST.The results of indoor toxicity test showed that the LC20 value of emamectin benzoate was 4.01 mg·L-1 at 48 h.The results of qRT-PCR showed that CYP4C101,CYP4C102,CYP6K1,GST1,and GSTX1 genes were expressed at all development stages,and the expression levels were the highest on the 9th day of adult emergence.The expression levels of the above-mentioned genes were significantly up-regulated under the stress of emamectin benzoate LC20 for 24 h.Among them,CYP4C101,CYP4C102,and CYP6K1 were significantly up-regulated to 4.43,22.91 and 8.48 times,respectively,and GST1 and GSTX1 were significantly up-regulated to 9.06 and 5.26 times,respectively.At 48 h after emamectin benzoate LC20 treatment,the expression levels of CYP4C102 and CYP6K1 were significantly up-regulated by 3.84 and 1.43 times,respectively.The expression levels of CYP4C101,GSTX1,and GST1 were up-regulated but did not reach a significant difference level.[Conclusion]Three CYP450 genes and two GST genes of T.tabaci were cloned,and the expression levels of the five genes were the highest on the 9th day of adult emergence.Under the stress of emamectin benzoate LC20,although these five detoxification genes were induced to express at different times,they may be involved in the response of T.tabaci to emamectin benzoate,providing clues for subsequent functional studies of CYP450 and GST genes.关键词
棉蓟马/解毒基因/甲维盐/基因表达/亚致死浓度Key words
Thrips tabaci/detoxification gene/emamectin benzoate/gene expression/sublethal dose引用本文复制引用
辛渺渺,陈亮亮,帕提玛·乌木尔汗,崔金杰,王晓云,姬继超,高越,雒珺瑜,张银宝,刘军,张文斌,王冬..5个解毒基因在棉蓟马生长发育中及甲维盐胁迫下的表达模式分析[J].棉花学报,2025,37(1):38-49,12.基金项目
安徽荃银高科种业股份有限公司项目(GMLM2023-06) （GMLM2023-06）
中央级公益性科研院所基本科研业务费专项(1610162023010) （1610162023010）
新疆农业大学与中国农业科学院棉花研究所合作项目(2523HXKT1) （2523HXKT1）
