棉铃虫ABCA2作为Cry2Ab受体的功能分析  

Analysis of ABCA2 as a functional receptor for Cry2Ab in cotton bollworm Helicoverpa armigera

作　　者：Ei Thinzar Soe 姚雪 杨朔 段云鹏 魏纪珍 梁革梅[1] Ei Thinzar Soe;Yao Xue;Yang Shuo;Duan Yunpeng;Wei Jizhen;Liang Gemei(State key Laboratory for Biology of Plant Diseases and Insect Pests,Institute of Plant Protection,Chinese Academy of Agricultural Sciences,Beijing 100193,China;College of Plant Protection,Henan Agricultural University,Zhengzhou 450002,Henan Province,China)

机构地区：[1]中国农业科学院植物保护研究所,植物病虫害生物学国家重点实验室,北京100193 [2]河南农业大学植物保护学院,郑州450002

出　　处：《植物保护学报》2021年第5期999-1007,共9页Journal of Plant Protection

基　　金：国家重点研发计划(2016ZX08011-002)。

摘　　要：为明确棉铃虫Helicoverpa armigera ATP结合盒转运蛋白家族A蛋白亚家族2(ATP binding cassette transporter subfamily A member 2,ABCA2)在Cry2Ab杀虫机制中的作用,利用酵母双杂交技术研究棉铃虫ABCA2与Cry2Ab的结合特性,并利用RNA干涉(dsRNA和siRNA)降低ABCA2在细胞和幼虫中的表达,结合细胞毒理学试验分析ABCA2的功能。结果表明,棉铃虫ABCA2能与Cry2Ab特异性结合;棉铃虫的ABCA2序列与美洲棉铃虫H.zea相关序列的相似度为92.86%,利用dsRNA干涉美洲棉铃虫中肠细胞ABCA2的表达能显著降低Cry2Ab对中肠细胞的毒力;利用siRNA干涉技术降低ABCA2在棉铃虫幼虫活体中的表达也显著降低了Cry2Ab对幼虫的毒力。表明棉铃虫ABCA2不仅是Cry2Ab的特异性结合蛋白,而且参与Cry2Ab的杀虫过程,是Cry2Ab的一个重要功能受体。To clarify the role of cotton bollworm Helicoverpa armigera ABCA2(ATP binding cassette subfamily A member 2)in the insecticidal mechanism of Cry2Ab,the binding character between ABCA2 and Cry2Ab was studied using the membrane yeast two-hybrid(MYTH)system,and the function of ABCA2 was further clarified by using RNAi techniques in toxicity tests.The results showed that ABCA2 could specifically bind to Cry2Ab.The amino acid sequence of ABCA2 from H.armigera and H.zea had 92.86%homology,cytotoxicology tests in H.zea midgut cells showed knocking down endogenous ABCA2 using dsRNA significantly reduced the sensitivity of midgut cells to Cry2Ab.Knocking down ABCA2 using siRNA in H.armigera larvae also reduced the sensitivity of the larvae to Cry2Ab.The results demonstrated that ABCA2 was not only a specific binding protein of Cry2Ab,but also a major functional receptor of Cry2Ab in H.armigera,therefore,played an important role in the insecticidal mechanism of Cry2Ab.

关 键 词：ABCA2 CRY2AB 棉铃虫 酵母双杂交 RNA干涉 

分 类 号：S435.622.3[农业科学—农业昆虫与害虫防治]