摘要
乙酰胆碱酯酶(acetylcholinesterase,AChE)是生物体内神经传导中的一种关键酶,它能够快速水解神经递质乙酰胆碱,保证神经信号在生物体内正常传递。本研究鉴定并克隆了一条黏虫乙酰胆碱酯酶基因MsAChE(GenBank登录号:PP473664),其cDNA全长5430 bp,包含一个长度为1917 bp的开放阅读框,编码638个氨基酸。qRT-PCR分析显示,MsAChE基因在黏虫不同发育阶段和不同组织中均有表达。为探究其生物学功能,利用RNAi技术将MsAChE基因沉默,以研究该基因对黏虫生长发育的影响。当MsAChE基因被沉默后,在3~24 h内基因的表达量被显著抑制,其酶活性先升高后降低,在RNA干扰6 h时,基因沉默效果最显著,基因表达量下降78.84%;用毒死蜱和西维因两种杀虫剂处理干扰后的黏虫,死亡率显著上升,处理后24 h,死亡率分别达到71.1%和58.9%;转录组测序数据分析表明黏虫体内许多与信号传导和代谢相关重要差异基因表达量发生变化。本研究结果表明MsAChE基因参与黏虫体内新陈代谢、信号传导等重要途径,为进一步解析AChE的生物学功能提供了重要参考,为靶向杀虫剂的开发提供重要理论依据。
Acetylcholinestrase(AChE)is one of the key enzymes in cholinergic nervous system.One of the neurotransmitters,named acetylcholine,can be hydrolyzed rapidly by AChE.In this study,an acetylcholinesterase gene MsAChE(GenBank accession number:PP473664)of Mythimna separata was successfully identified and cloned.The length sequence of MsAChE was 5430 bp with an open reading frame(ORF)of 1917 bp,encoding 638 amino acids.The qRT-PCR results showed that MsAChE was expressed in all tested developmental stages and tissues.In order to explore its biological function,the MsAChE was silenced by RNAi to study the effect on the growth and development of M.separata.The interference efficiency experiment demonstrated successful silencing of MsAChE,resulting in significant inhibition of gene expression within 3 to 24 hours.When RNA interfered for 6 h,the gene silencing effect was the most significant,and the gene expression decreased by 78.84%.Enzyme activity itially increased before decreasing,and the mortality of RNA-interfered M.separata treated with chlorpyrifos and carbaryl also increased significantly,reaching 71.1%and 58.9%respectively at 24 h after treatment.Transcriptome data revealed alterations in the expression levels of numerous key differential genes associated with signal transduction and metabolism in M.separata.These findings suggest that the MsAChE gene plays a crucial role in essential pathways,including metabolism and signal transduction.It provides a foundation for further investigations of the biological function of AChE,and laid a theoretical basis for development of the AchE-targeted insecticides.
作者
张维佳
杨洪佳
王艺晓
威力斯
吕佳炜
单志勐
张鑫鑫
樊东
ZHANG Weijia;YANG Hongjia;WANG Yixiao;WEI Lisi;LÜJiawei;SHAN Zhimeng;ZHANG Xinxin;FAN Dong(College of Plant Protection,Northeast Agricultural University,Harbin 150030,China)
出处
《中国生物防治学报》
北大核心
2025年第2期309-320,共12页
Chinese Journal of Biological Control
基金
国家重点研发计划(2023YFD1401000)。
关键词
黏虫
乙酰胆碱酯酶
RNA干扰
转录组分析
Mythimna separata
acetylcholinestrase
RNA interference
transcriptome analysis
