摘要
【目的】烟碱型乙酰胆碱受体(nicotinic acetylcholine receptors,nAchRs)作为昆虫中枢神经系统中重要的神经递质受体,是乙基多杀菌素的重要靶标。本研究旨在克隆草地贪夜蛾Spodoptera frugiperda nAchR基因,并探析其对乙基多杀菌素胁迫的响应。【方法】基于前期草地贪夜蛾转录组数据,采用RT-PCR和RACE分别克隆草地贪夜蛾nAchRα6和α7亚基基因;乙基多杀菌素(0.400 mg/L)处理草地贪夜蛾3龄幼虫48 h后,采用RT-qPCR技术检测nAchRα6和α7亚基基因的表达量的变化。【结果】nAchRα6亚基基因(GenBank登录号:MT951400)开放阅读框长1506 bp,编码502个氨基酸,并具有跨膜区与信号肽;nAchRsα7亚基基因(GenBank登录号:MW557608)开放阅读框长1524 bp,编码508个氨基酸,并具有跨膜区与信号肽。根据氨基酸多序列比对分析,草地贪夜蛾nAchR亚基α6和α7具有烟碱性乙酰胆碱受体α家族典型特征。0.400 mg/L乙基多杀菌素胁迫48 h后,草地贪夜蛾3龄幼虫nAchRα6亚基基因表达量显著增加,而nAchRα7亚基基因表达量无显著性变化。【结论】乙基多杀菌素可能作用于草地贪夜蛾的nAchRα6亚基。本研究为深入研究草地贪夜蛾对乙基多杀菌素的靶标抗性机制奠定了初步基础。
【Aim】Nicotinic acetylcholine receptors(nAchRs),as the most crucial neurotransmitter receptors in the central nervous system of insects,are the important targets of spinetoram.This study aims to clone the nAchR genes of Spodoptera frugiperda and investigate their response to spinetoram stress.【Methods】Genes of nAchRα6 andα7 subunits of S.frugiperda were cloned via RT-PCR and RACE based on the previous transcriptome database of S.frugiperda.After the 3rd instar larvae of S.frugiperda were exposed to spinetoram(0.400 mg/L)for 48 h,the changes of expression levels of nAchRα6 andα7 subunit genes were assayed by RT-qPCR.【Results】The full-length ORF of nAchRα6 subunit gene(GenBank accession no.:MT951400)is 1506 bp in length,encoding 502 amino acids with transmembrane region and signal peptide.The full-length ORF of nAchRα7 subunit gene(GenBank accession no.:MW557608)is 1524 bp in length,encoding 508 amino acids with transmembrane region and signal peptide.nAchRα6 andα7 subunits of S.fruiperda have the typical characteristic of nAchRαfamily according to amino acid sequence multiple alignment analysis.After the 3rd instar larvae of S.frugiperda were exposed to 0.400 mg/L spinetoram for 48 h,the expression level of their nAchRα6 subunit gene increased significantly,while that of nAchRα7 subunit gene did not change significantly.【Conclusion】Spinetoram may have an impact on the nAchRα6 subunit of S.fruiperda.This study lays a preliminary foundation for future in-depth research on the underlying target resistance mechanisms of S.frugiperda in response to spinetoram.
作者
高祖鹏
静大鹏
黄晓丹
王振营
GAO Zu-Peng;JING Da-Peng;HUANG Xiao-DAN;WANG Zhen-Ying(State Key Laboratory of Biology of Plant Diseases and Insect Pests,Institute of Plant Protection,Chinese Academy of Agricultural Sciences,Beijing 100193,China;Engineering Research Center of Natural Enemy Insects,Institute of Biological Control,Jilin Agricultural University,Changchun 130118,China)
出处
《昆虫学报》
CAS
CSCD
北大核心
2022年第9期1098-1105,共8页
Acta Entomologica Sinica
基金
国家重点研发计划资助(2021YFD1400705)
财政部和农业农村部:国家现代农业产业技术体系建设专项(CARS-02)。
