垂丝海棠MhMYB114-Like的克隆及其在苹果愈伤组织中的抗缺铁功能鉴定

从苹果砧木垂丝海棠(Malus halliana Koehne)中克隆得到了Mh MYB114-Like基因(序列号:LOC103405832)。该基因包含全长为602 bp的完整开放阅读框,编码200个氨基酸,蛋白质等电点为9.38。系统进化树分析表明,Mh MYB114-Like与野草莓该家族蛋白亲缘关系最近。分析Mh MYB114-Like的启动子区域发现包含干旱、光、生长素和黄化响应等顺式作用元件。荧光定量PCR分析表明,在缺铁胁迫下Mh MYB114-Like在垂丝海棠叶和根中表达水平显著上升,Mh MYB114-Like的表达明显受缺铁胁迫的诱导。将Mh MYB114-Like遗传转化‘王林’苹果愈伤组织,通过测定相关生理指标进行抗性分析发现,过量表达Mh MYB114-Like可明显提高转基因愈伤组织对缺铁胁迫的抗性。结果表明Mh MYB114-Like可响应缺铁胁迫,在抵抗缺铁胁迫中具有重要作用。

Cloning of MhMYB114-Like from Malus halliana and Its Functional Identification of Iron Deficiency Tolerance in Apple Callus

The Mh MYB114-Like gene(Gen Bank accession number:LOC103405832)was cloned from apple rootstock Malus halliana. The gene contains a full open reading frame of 602 bp,which encodes 200 amino acids,and the protein has an isoelectric point of 9.38. The phylogenetic tree analysis showed that apple Mh MYB114-Like was the closest relative to wild strawberry. Promoter cis-element prediction of apple Mh MYB114-Like showed that there were drought,light,auxin,yellowing and other responsive regulatory elements. Fluorescence quantitative PCR analysis demonstrated that the expression level of Mh MYB114-Like gene was greatly increased in the leaves and roots of Malus halliana under iron deficiency condition,and the expression was obviously induced by iron deficiency stress. In addition,The genetic transformation of Mh MYB114-Like into apple‘Orin’callus and resistance analysis by measuring related physiological indexes showed that the overexpression of Mh MYB114-Like could significantly improve the resistance of transgenic callus to iron deficiency stress. Therefore,Mh MYB114-Like is involved in the response to iron deficiency and play a crucial role in resisting iron deficiency stress.