狄斯瓦螨唾液毒性蛋白VTP互作蛋白FABP的鉴定与功能分析  

Identification and functional analysis of the Apis cerana protein FABP,which interacts with the varroa toxic protein VTP from salivary glands of Varroa destructor

作　　者：丁兆润 李家豪 陈晓文 李文峰 韩日畴 张祎 DING Zhao-Run;LI Jia-Hao;CHEN Xiao-Wen;LI Wen-Feng;HAN Ri-Chou;ZHANG Yi(Institute of Zoology,Guangdong Academy of Sciences,Guangzhou 510260,China;South China Agricultural University,Guangzhou 510642,China;South China Normal University,Guangzhou 510631,China;Qinghai Normal University,Xi’ning 810008,China;Guangdong Pharmaceutical University,Guangzhou 527527,China)

机构地区：[1]广东省科学院动物研究所,广州510260 [2]华南农业大学,广州510642 [3]华南师范大学,广州510631 [4]青海师范大学,西宁810008 [5]广东药科大学,广州527527

出　　处：《应用昆虫学报》2023年第3期755-763,共9页Chinese Journal of Applied Entomology

基　　金：2022年省级乡村振兴战略专项资金种业振兴项目(2022-440000-4301030211-9465)。

摘　　要：【目的】阐明狄斯瓦螨Varroa destructor唾液毒性蛋白(Varroa toxic protein,VTP)致死中华蜜蜂(以下简称中蜂)Apis cerana工蜂预蛹的机制,从而了解狄斯瓦螨与中蜂的互作机制,为开发蜂螨害防治新技术拓宽思路。【方法】通过酵母双杂交筛选狄斯瓦螨唾液蛋白VTP的互作蛋白;利用RNAi技术和qRT-PCR验证互作蛋白基因功能并检测相关基因表达情况。【结果】利用酵母双杂交筛选到11个候选蛋白基因,进一步通过“回复”验证,脂肪酸结合蛋白(Fatty acid binding protein,FABP)是唯一阳性蛋白;为了进一步验证FABP在体内与VTP结合,通过注射dsRNA-fabp干扰中蜂幼虫fabp的表达,结果发现,注射dsRNA-fabp幼虫组化蛹率(58%)与仅注射VTP组(40%)相比显著提高。qRT-PCR检测发现,fabp沉默后,幼虫的蘑菇体大型凯恩细胞特异性蛋白1基因(Mushroombodylarge-typeKenyon cell-specificprotein1,mblk-1)基因和蜕皮激素受体基因(Ecdysonereceptor,ecr)基因的相对表达量发生变化。【结论】狄斯瓦螨唾液毒性蛋白VTP可能通过与中蜂体内FABP蛋白结合,导致5龄幼虫化蛹失败,从而致死中蜂。[Objectives]To investigate the mechanism by which varroa toxic protein(VTP),found in Varroa destructor saliva,kills Apis cerana,and thereby facilitate the development of new methods for controlling V.destructor.[Methods]A two-hybrid yeast was used to screen candidate proteins that bind to VTP after which RNAi technology and qRT-PCR were used to verify the function of the interacting protein gene and detect the expression of related genes.[Results]Eleven candidate protein genes were identified,however,a"return experiment"revealed that the fatty acid binding protein(FABP)was the only positive protein.In order to further verify the binding of FABP and VTP in vivo,the expression of fabp in bee larvae was suppressed by an injection of dsRNA-fabp.The pupation rate of larvae injected with dsRNA-fabp was significantly higher(58%)than that of those injected with VTP alone(40%).QRT-PCR indicated that the relative expression of the Mushroom body large-type kenyon cell-specific protein 1(mblk-1)gene and ecdysone receptor(ecr)gene in the mushroom body of larvae,changed after fabp was silenced.[Conclusion]The VTP protein in the saliva of V.destructor may cause the death of 5th instar A.cerana larvae by binding to the FABP protein.

关 键 词：中华蜜蜂 狄斯瓦螨 VTP 脂肪酸结合蛋白 

分 类 号：S895.32[农业科学—特种经济动物饲养]