小菜蛾V-ATP酶A亚基表达丰度差异  

Expression abundance analysis of the V-ATPase subunit A gene in Plutella xylostella

作　　者：卢莉娜 朱庆 高美静 谢雅晶 刘贤金[1,2] 张志勇 LU Lina;ZHU Qing;GAO Meijing;XIE Yajing;LIU Xianjin;ZHANG Zhiyong(Jiangsu Key Laboratory for Food Quality and Safety-State Key Laboratory Cultivation Base,Ministry of Science and Technology,Nanjing 210014,China;Institute of Food Safety and Nutrition,Jiangsu Academy of Agricultural Sciences,Nanjing 210014,China)

机构地区：[1]省部共建国家重点实验室培育基地—江苏省食品质量安全重点实验室,江苏南京210014 [2]江苏省农业科学院农产品质量安全与营养研究所,江苏南京210014

出　　处：《浙江农业学报》2025年第4期839-846,共8页Acta Agriculturae Zhejiangensis

基　　金：国家自然科学基金(31901919,31630061)。

摘　　要：V型ATP酶(V-ATPase)在昆虫体内和Cry毒素作用机制中发挥重要作用。本研究采用分子克隆和实时荧光定量PCR(qRT-PCR)技术探究小菜蛾(Plutella xylostella)幼虫体内V-ATP酶A亚基基因在不同生长发育阶段,以及取食Cry1Ac毒素后的表达量丰度差异和变化情况。结果表明,小菜蛾V-ATP酶A亚基基因CDS区为1848 bp,编码615个氨基酸(GenBank登录号:ON108624),与亚洲玉米螟(Ostrinia furnacalis)、二化螟(Chilo suppressalis)和棉铃虫(Helicoverpa armigera)的同源性约为94%。V-ATP酶A亚基基因在小菜蛾卵、幼虫、蛹和成虫期均有表达,在卵和蛹期表达量较低,在幼虫2龄期时表达量达到最高值,为卵期的4.16倍,3~4龄期表达量下降。小菜蛾幼虫取食含有Cry1Ac毒素的饲料后,体内V-ATP酶A亚基基因表达量整体呈显著下降趋势,取食48 h后表达量恢复至正常水平。在小菜蛾幼虫体内,V-ATP酶A亚基基因的表达量与生长发育和代谢变化密切相关,并且与Cry1Ac毒素的作用有一定关系。V-ATPase plays important role in insects and mechanism of Cry toxins.In this study,the molecular cloning and real time fluorogenic quantitative PCR(qRT-PCR)was conducted to explore the expression of Plutella xylostella V-ATPase subunit A gene in different growth and developmental stages as well as before/after feeding Cry1Ac toxin.The results showed that the CDS of V-ATPase subunit A gene was 1848 bp,which encoded 615 amino acids(GenBank No.ON108624)and had approximately 94%homology with V-ATPase subunit A of Ostrinia furnacalis,Chilo suppressalis and Helicoverpa armigera.The qRT-PCR results indicated that V-ATPase subunit A gene was expressed in the whole larval stages,eggs,pupa and adults.In eggs and pupa,the expression level of V-ATPase subunit A gene was low,while in the 2nd instar larvae the expression reached the highest to be 4.16 times as in eggs,and decreased in the 3rd and 4th instar larvae.After feeding the artificial diet with Cry1Ac toxins,the express level of V-ATPase subunit A gene declined and then reached to the normal after feeding 48 h.These results indicated that the express level of V-ATPase subunit A gene in P.xylostella was closely related with development and metabolic changes,and might be involved in the mode action of Cry1Ac toxin.

关 键 词：小菜蛾 V-ATP酶A亚基 Cry毒素 表达丰度 

分 类 号：S433.4[农业科学—农业昆虫与害虫防治]