葡萄VvMSA基因的克隆及表达特性分析

为了解VvMSA基因的功能,以抗性葡萄品种Vidal Blanc组培苗为试材,克隆得到VvMSA基因,对其序列进行了生物信息学分析,并利用实时定量PCR分析了其组织表达特性和多种非生物胁迫和信号应答表达特性。结果表明,VvMSA基因片段大小为450 bp,编码149个氨基酸序列。生物信息学分析结果显示,VvMSA蛋白分子量约为16.703 k Da,等电点为5.68,不稳定系数为41.71,推测为不稳定蛋白。VvMSA含有65个氨基酸组成的ABA/WDS保守结构域。在进化上属于单独一个分支,它和已报道过的番茄Le ASR1分别属于同一个祖先进化出的2个分支。实时荧光定量PCR分析显示,VvMSA在葡萄不同组织中均有表达,花中表达量最高。多种逆境胁迫因子如盐、干旱和低温等能诱导VvMSA不同程度的上调表达,盐胁迫3 h时诱导表达量最高。同时,VvMSA受逆境胁迫信号分子NO和H_2S不同程度诱导表达上调,而SA则抑制VvMSA表达。激素ABA、GA_3、IAA和ET均能不同程度诱导VvMSA表达上调,并且VvMSA盐处理下的表达模式与ABA诱导的类似。综合以上结果推断VvMSA可能通过调控ABA信号途径来调节葡萄对盐胁迫应答。

Gene Cloning and Expression Analysis of VvMSA in Vitis vinifera

To investigate the function of VvMSA,the full-length cDNA of VvMSA was cloned from Vitis vinifera cultivar Vidal Blanc tissue culture seedlings.Bioinformatic analysis and quantitative Real-time PCR was used to an-alyze its characteristics.The results showed that VvMSA amplified fragment size of 450 bp encoding 149 amino acids with molecular weight 16.703 kDa,isoelectric point 5.68.VvMSA was an unstable hydrophilic protein and con-tained a conserved domain ABA /WDS encoding by 65 amino acids.VvMSA and the known homolog LeASR1 in to-mato belongs to different sub-families.Real-time PCR analysis showed that the VvMSA was expressed in all tested tissues with the highest expression in flower.VvMSA was induced by salt,drought and cold stress in different levels, and highest expression level was showed after 3 h salt treatment.In addition,VvMSA was also induced by nitric oxide (NO),hydrogen sulfide(H2 S),abscisicacid(ABA)gibberellin(GA3 ),auxin(IAA)and ethylene(ET),while sali-cylic acid(SA)inhibited its expression.The expression pattern of VvMSA by salt treatment was similar with the ex-pression pattern by ABA treatment.In total,these results suggested that the VvMSA was involved in salt resistance by ABA signaling.