【摘要】 大麦黄花叶病是我国长江中下游地区大麦种植区域的主要病毒病害,由大麦黄花叶病毒(BaYMV,Barley yellow mosaic virus)及大麦和性花叶病毒(BaMMV,Barley mild mosaic virus)引起,自然条件下病毒寄生于禾谷多黏菌中,通过感染大麦根部导致病害发生。本研究通过分析RNA-seq数据,获得感染BaMMV后上调表达的基因HORVU1Hr1G069640(WRKY55)。荧光定量PCR分析发现WRKY55在感病品种中的表达水平极显著高于抗病材料,说明WRKY55参与到大麦感染黄花叶病的过程。WRKY55全长870 bp,在415～594位核苷酸之间含有WRKY保守结构域,系统进化分析显示该基因位于独立的进化分枝。组织表达分析发现该基因主要在根部和幼嫩组织中表达,而其他成熟部位表达较少甚至没有。农杆菌介导的烟草亚细胞定位实验发现WRKY55位于整个细胞。酵母转录因子活性分析实验未检测到转录激活活性,酵母双杂交实验未检测到WRKY55与感病基因编码蛋白eIF4E和PDIL5-1存在物理相互作用。这些结果显示WRKY55参与到大麦黄花叶病感染。更多还原

【Abstract】 Barley yellow mosaic virus disease,caused by Barley yellow mosaic virus(BaYMV)and Barley mild mosaic virus(BaMMV),is a major viral disease in barley planting regions of Europe and East Asia countries.In the field,this disease is transmitted y the soil-borne fungus Polymyxa graminis via inoculating the roots.Here,by unlocking the transcriptome datasets,we identified a differentially expressed gene HORVU1Hr1G069640(referred WRKY55)that was significantly up-regulated after infection with b BaMMV.Consistently,by analyzing its expression in a group of naturally-inoculated genotypes,a significantly higher expression of WRKY55 was observed in susceptible relative to resistant genotypes.This gene contains an 870 bp of the coding sequence with a conserved WRKY domain between nucleotides 415′-594′.Phylogenetic analysis revealed an independent branch of this gene,differedto the sequence homologous genes.This gene was highly expressed in roots and young tissues,but rarely expressed in other tissues.By Agrobacterium-mediated transformation into Nicotiana benthamiana Domin,the WRKY55-GFP fusion protein was found in the whole cell.In yeast,no trans-activation activity of WRKY55 was detected and no physical interaction between WRKY55 and either susceptibility factors eIF4 E or PDIL5-1 were observed.This work identified a BaMMV-infected upregulated gene WRKY55,which possibly was involved in infection of BaYMV/BaMMV.更多还原