观赏海棠叶片McmiR159a克隆及功能验证

摘要

[Objective] The experimentis to verify the effect of miR159a on anthocyanin biosynthesis in Malus ornamental crabapple leaves.[Methods]Weselected theleaves of ever-red-leafed ‘Royalty’ of Malus ornamental crabapple as experimental material to identify the sequence of miR159a precursor gene and topredictitstarget.The McmiR 159a over-expression vector was constructed and transiently transferred into the ‘ Royalty’ tissue culture seedlings.The content of anthocyanin and the expression of structural genes in the pathway of anthocyanin biosynthesis were analyzed by HPLC and qRT-PCR,respectively.[Results] Over expression of McmiR159a in theleavesof Malus ornamental crabappleincrease the anthocyanin content by downregulating the expression of target and up-regulating the expression of key genes in anthocyanin biosynthesis pathway.[Conclusion] McmiR159amaypositively effect on anthocyanin synthesis in Malus ornamental crabapple leaves.%[目的]为了验证观赏海棠叶片McmiR 159a对花色苷代谢的调控作用.[方法]以观赏海棠红叶品种‘王族’为试验材料,确定McmiR 159a前体基因序列并对其靶基因进行预测,构建McmiR 159a过表达载体并瞬时转入‘王族’组培苗.其次通过高效液相色谱和qRT-PCR分析瞬时侵染植株中花色苷的含量及其合成途径中结构基因的表达量.[结果]结果表明,McmiR15 9a在观赏海棠中过表达下调其靶基因表达,上调花色苷代谢途径关键基因的表达,进而增加花色苷的含量.[结论]研究结果表明,观赏海棠中McmiR 159a对花色苷的合成具有正调控作用.