利用酵母双杂交筛选与小麦黄花叶病毒P1互作的寄主蛋白  

Screening of the host factors interacting with P1 of Wheat yellowmosaic virus by yeast two-hybrid system

作　　者：刘丽娟[1] 梁乐乐[1] 戚文平 孙炳剑[1] 施艳[1] 陈琳琳[1] 李洪连[1] LIU Li-juan LIANG Le-le QI Wen-ping SUN Bing-jian SHI Yan CHEN Lin-lin LI Hong-lian(College of Plant Protection, Henan Agricultural University, Zhengzhou 450002, Chin)

机构地区：[1]河南农业大学植物保护学院,郑州450002

出　　处：《植物病理学报》2017年第4期563-567,共5页Acta Phytopathologica Sinica

基　　金：NSFC-河南人才培养联合基金项目(U1304322);公益性行业(农业)科研专项(201303021)

摘　　要：小麦黄花叶病是我国冬小麦上重要的病毒病害之一,自20世纪90年代以来,每年发生面积都在66万hm^2以上,一般减产10%-30%,严重的达70%以上。该病害在我国河南、江苏、山东、安徽、陕西、湖北、四川等省均有分布,Wheat yellow mosaic virus （WYMV） is an important plant pathogen that causes yellow mosaic di- sease in winter wheat worldwide. In order to study the interactions between host plant and WYMV, we investiga- ted the host factors that directly interact with WYMV P1 using MatchmakerTM Gold Yeast Two-Hybrid System. WYMV P1 gene was cloned into the bait vector （pGBKT7） and transformed into yeast strain Y2H GOLD. The transformed yeast was grown on the selective medium lacking histidine, leucine, tryptophan and adenine and supplemented with X-α-Gal to examine the self-activating effect and virulence of WYMV P1. Western blot anal- ysis confirmed WYMV P1 expression in the yeast. The cDNA library was generated from Triticum aestivum L. cv. Chinese spring. The positive clones were sequenced and further analyzed by BLAST, ontology （GO） and CELLO v2.5. Our screening yielded 15 potential Pl-interacting proteins from the library. These proteins could be divided into various functional categories, including photosynthesis Calvin cycle, protein synthesis, posttransla- tional modification, post-translational assembly and cofactor ligation, RNA regulation of transcription, signa- ling, cell division, biodegradation, amino acid and minor carbohydrate （CHO） metabolism, cell wall precursor synthesis, acid and other phosphatases, and not assigned proteins. Our results provide a new insight into the mo- lecular interactions between WYMV and wheat host.

关 键 词：小麦黄花叶病毒 P1 寄主植物 病毒病害 酵母双杂交筛选 大麦黄花叶病毒 自激活 TRITICUM 感受态 黄花叶 

分 类 号：S432.41[农业科学—植物病理学]