玉米抗病相关基因ZmEDS1的克隆及表达分析

从玉米中克隆ZmEDS1基因的cDNA序列,全长2 164 bp,开放阅读框(Open reading frame,ORF)长1 860 bp,编码619个氨基酸。生物信息学分析表明,该基因编码的蛋白等电点为6.05,分子量为68.74 kDa,内部无信号肽结构,N端有一个酯酶结构域。系统进化树分析表明,玉米ZmEDS1蛋白和高粱EDS1L蛋白的亲缘关系最近,同源性高达94%。采用实时荧光定量PCR分析病毒侵染下该基因在感性材料郑58和抗性材料D863F中的表达模式,在两种材料中,ZmEDS1基因均在病毒侵染48 h的表达量最高,分别达到0 h对照组感、抗材料的1.56、3.47倍,在抗病材料D863F中的表达水平高于感病材料郑58。初步判断,ZmEDS1基因应答病毒的侵染过程,且在感病材料和抗病材料中的响应模式存在差异。

Cloning and Expression Analysis of the Disease-related Gene ZmEDS1 in Maize

The disease-related protein EDS1 plays a key role in the resistance of pathogens during plant growth and development. In this study, the cDNA of ZmEDS1 was cloned from maize, which is 2164 bp in length and encodes a protein with 619 amino acids, 68.74 kDa and theoretical isoelectric point of 6.05. Phylogenetic analysis showed that ZmEDS1 was 94% homology with EDS1L in sorghum. Using rice black-streaked dwarf virus(RBSDV) inoculation method, the expression patterns of ZmEDS1 in D863F(resistant) and Zheng 58 (susceptible) cultivars were respectively studied by RT-qPCR, which revealed that ZmEDS1 gene showed the highest expression levels after viral infected for 48 h in both materials, reaching 1.56 times in Zheng 58 and 3.47 times in D863F when compared with that of control (infected for 0 h), namely, the expression of ZmEDS1 in resistant material was higher than in susceptible cultivar. In all, the ZmEDS1 gene in maize responds to RBSDV infection, while it showed difference expression levels in the resistant and susceptible materials.