摘要
薄荷是我国传统中草药,具有极大的药用和经济价值.脱落酸(ABA)受体PYLs在植物生长发育及逆境响应方面具有重要作用.克隆薄荷McPYL4 基因,对其蛋白特征、基因表达和蛋白互作进行分析,将为薄荷抗逆遗传改良和分子设计育种提供基因资源.故而通过生物信息学分析、烟草叶片瞬时表达、RT-qPCR、Y2H技术对McPYL4 的蛋白特性、亚细胞定位、基因表达模式和蛋白互作进行分析.结果显示,McPYL4 基因全长 621 bp,编码 206 个氨基酸,其蛋白具有SRPBCC保守结构域,与丹参SmPYL4 最同源;McPYL4 蛋白定位于细胞膜和细胞核;McPYL4 基因在薄荷的各个组织中均有表达,根中表达最高,叶片次之,叶 1 至叶 8 呈现先上后下的表达模式;在叶片和根中,McPYL4 基因均响应外源激素ABA、茉莉酸甲酯(MeJA)以及干旱、AlCl3、NaCl、CdCl2 和CuCl2 处理,并且,McPYL4 在MeJA处理下的叶片和根中,以及AlCl3 胁迫处理 1 h的叶片中上调表达,而其他处理下McPYL4 在叶片和根中均呈现下调的趋势;蛋白互作结果显示,McPYL4 以ABA不依赖的形式与AtABIs蛋白相互作用.该研究表明McPYL4 响应ABA、茉莉酸(JA)以及多个非生物胁迫处理,并且McPYL4 参与薄荷中ABA信号转导,从而参与薄荷的叶片发育及各种非生物胁迫的调控.
Abstract
Mentha canadensis is a traditional Chinese herb with great medicinal and economic value.Abscisic acid(ABA)receptor PYLs have important roles in plant growth and development and response to adversity.The M.canadensis McPYL4 gene was cloned,and its protein characteristics,gene expression,and protein interactions were analyzed,so as to provide genetic resources for genetic improvement and molecular design breeding for M.canadensis resistance.Therefore,the protein characteristics,subcellular localization,gene expression pattern,and protein interactions of McPYL4 were analyzed by bioinformatics analysis,transient expression of tobacco leaves,RT-qPCR,and yeast two-hybrid(Y2H)techniques.The results showed that the McPYL4 gene was 621 bp in length,encoding 206 amino acids,and its protein had the conserved structural domain of SRPBCC and was highly homologous with Salvia miltiorrhiza SmPYL4.McPYL4 protein was localized to the cell membrane and nucleus.The McPYL4 gene was expressed in all tissue of M.canadensis,with the highest expression in roots,followed by leaves,and it showed a pattern of up-regulation followed by down-regulation in leaves 1-8.In both leaves and roots,the McPYL4 gene responded to the exogenous hormones ABA,MeJA,and the treatments of drought,AlCl3,NaCl,CdCl2,and CuCl2.Moreover,McPYL4 was up-regulated for expression in both leaves and roots under the MeJA treatment,as well as in leaves treated with AlCl3 stress for 1 h,whereas McPYL4 showed a tendency to be down-regulated in both leaves and roots under other treatments.Protein interactions showed that McPYL4 interacted with AtABI proteins in an ABA-independent manner.This study demonstrated that McPYL4 responded to ABA,JA,and several abiotic stress treatments,and McPYL4 was involved in ABA signaling in M.canadensis and thus in the regulation of leaf development and various abiotic stresses in M.canadensis.
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