青稞酵母双杂文库的构建及HVUL0H33228.2互作蛋白的筛选  

Construction of Highland Barley Yeast Double Miscellaneous Library and Screening of HVUL0H33228.2 Interacting Proteins

作　　者：羊海珍 顿珠加布 徐齐君[1,2] Yang Haizhen;Dunzhu Jiabu;Xu Qijun(State Key Laboratory of Hulless Barley and Yak Germplasm Resources and Genetic Improvement,Tibet Academy of Agriculture and Animal Hus-bandry Sciences,Lhasa,850002;Research Institute of Agriculture,Tibet Academy of Agriculture and Animal Husbandry Sciences,Lhasa,850002)

机构地区：[1]西藏自治区农牧科学院,省部共建青稞和牦牛种质资源与遗传改良国家重点实验室,拉萨850002 [2]西藏自治区农牧科学院农业研究所,拉萨850002

出　　处：《分子植物育种》2023年第1期185-191,共7页Molecular Plant Breeding

基　　金：西藏重点研发及转化计划项目(XZ202001ZY0016N);财政预算内项目(2015ZX001)共同资助

摘　　要：白粉病是青稞生产中危害最为严重的病害之一,会造成产量严重下降。本实验室前期筛选到一个白粉菌诱导后上调表达的NF-YB转录因子HVUL0H33228.2。为了进一步明确该基因的作用机制,本研究构建了青稞受白粉菌诱导的酵母双杂交cDNA文库,以HVUL0H33228.2为诱饵进行酵母双杂交筛选。结果表明:所获得的cDNA文库滴度为5.6×108CFU/mL,插入片段长500~2 000 bp,重组率为94%,符合酵母双杂文库质量要求。利用酵母双杂交系统在该文库中筛选到6个与HVUL0H33228.2相互作用的蛋白,分别为铁氧还蛋白-NADP+氧化还原酶、核酮糖二磷酸羧化酶、叶绿素a/b结合蛋白2和6、光合系统Ⅱ反应中心蛋白CP43和醛缩酶型TIM桶家族蛋白。本研究为进一步揭示HVUL0H33228.2应答白粉菌侵染的分子机制提供了参考依据。Powdery mildew disease,one of the major destructive diseases of Tibetan hulless barley,caused heavy loss in grain yield.A NF-YB transcription factor HVUL0H33228.2 with up-regulated expression induced by powdery mildew was identified in previous research.To further explore the biological mechanisms of HVUL0H33228.2,cDNA library of Tibetan hulless barley under powdery mildew stress was constructed,and interacting proteins were screened using HVUL0H33228.2 as a bait.The results showed that library titer was 5.6×10~8CFU/mL,with the inserted fragment of 500~2 000 bp in length and 94%recombination rate,which could reach the quality requirements of yeast Two-hybrid library.Six proteins interacted with HVUL0H33228.2,including ferredoxin-NADP reductase,ribulose bisphosphate carboxylase,photosystemⅡCP43 reaction center protein,chlorophyll a/b binding protein 2,chlorophyll a/b binding protein 6,aldolase-type TIM barrel family protein were identified by yeast two hybrid system.This study provides a reference for further exploring the molecular mechanisms of HVUL0H33228.2in response to powdery mildew.

关 键 词：青稞 白粉病 CDNA文库 HVUL0H33228.2 互作蛋白 

分 类 号：S435.123[农业科学—农业昆虫与害虫防治]