薄荷茉莉酸受体McCOI1a基因的克隆与表达模式分析

Cloning and Expression Pattern Analysis of Jasmonic Acid Receptor Gene McCOI1a in Mentha canadensis L.

【目的】茉莉酸(jasmonic acid,JA)受体COI1在调节植物生长发育、逆境响应方面具有重要作用。克隆薄荷McCOI1a基因,并分析其蛋白特征与表达模式,为薄荷分子育种提供基因资源。【方法】基于转录组数据从薄荷叶片中克隆McCOI1a,并通过生物信息学分析、烟草叶片瞬时表达、实时荧光定量PCR技术对McCOI1a的蛋白特性、亚细胞定位、基因表达模式进行分析。【结果】McCOI1a基因全长1842 bp,编码613个氨基酸;McCOI1a蛋白具有保守的F-box和LRR结构域,与丹参SmCOI1蛋白同源性最高;亚细胞定位结果显示McCOI1a蛋白定位于细胞核;McCOI1a在不同组织中均有表达,在根中表达量最高,在叶序中表达呈逐渐上升的趋势;在叶片中,McCOI1a在MeJA、干旱、NaCl、AlCl3、CdCl2、CuCl2处理下的表达呈不同程度的上调,并且AlCl3处理下其上调最明显,表达量最高可达1206倍;在根中,McCOI1a的表达在CuCl2处理下呈先下调而后上调的模式,在其余处理下,McCOI1a的表达都呈现出不同程度的下调。【结论】McCOI1a响应MeJA和非生物逆境胁迫,可能在调控薄荷生长发育、逆境响应方面发挥重要作用。

【Objective】The jasmonic acid(JA)receptor COI1 plays a vital role in regulating plant growth and development,and stress response in plants.Cloning and analysis of the protein characterization,and analyzing expressions of McCOI1a from Mentha canadensis L.may provide the gene resources for its molecular breeding.【Method】The McCOI1a gene was cloned from the leaves of M.canadensis based on transcriptome data,and then the characteristics,subcellular localization,and expression profiles of McCOI1a were analyzed through bioinformatics analysis,transient expression in the leaves of tobacco,and real-time quantitative PCR.【Result】The full-length of McCOI1a gene was 1842 bp and it encoded 613 amino acids.McCOI1a protein possessed the conserved F-box and LRR domains and that was highly homologous with Salvia miltiorrhiza SmCOI1.Subcellular localization results showed that McCOI1a protein was located in the nucleus.The qRT-PCR results showed that McCOI1a was broadly expressed in different tissues,with the highest expression in the root;McCOI1a expression gradually increased during leaf development.In addition,the expressions of McCOI1a in the leaves were up-regulated to varying degrees under MeJA,drought,NaCl,AlCl3,CdCl2,and CuCl2 treatments,and the up-regulation was the most obvious under AlCl3 treatment,with the highest expression reaching 1206-fold.And McCOI1a expression in the root was firstly down-regulated and then up-regulated under CuCl2 treatment,while the expression of McCOI1a showed varying degrees of down-regulation under other treatments.【Conclusion】McCOI1a responds to MeJA and abiotic stress,and may play an important role in regulating the growth and development,and stress response in M.canadensis.