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、AlCl_(3)、NaCl、CdCl_(2)和CuCl_(2)处理,并且,McPYL4在MeJA处理下的叶片和根中,以及AlCl_(3)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。
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,AlCl_(3),NaCl,CdCl_(2),and CuCl_(2).Moreover,McPYL4 was up⁃regulated for expression in both leaves and roots under the MeJA treatment,as well as in leaves treated with AlCl_(3)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.
作者
唐伟林
汪霞
康琴
王棵
彭丹丹
孙义凯
吴卫
侯凯
冯冬菊
徐东北
TANG Wei-lin;WANG Xia;KANG Qin;WANG Ke;PENG Dan-dan;SUN Yi-kai;WU Wei;HOU Kai;FENG Dong-ju;XU Dong-bei(College of Agronomy,Sichuan Agricultural University,Chengdu 611100,China;College of Prataculture Technology,Sichuan Agricultural University,Chengdu 611100,China)
出处
《中国中药杂志》
CAS
CSCD
北大核心
2024年第6期1494-1505,共12页
China Journal of Chinese Materia Medica
基金
国家自然科学基金青年基金项目(31900256)
四川省科技厅应用基础项目(2020YJ0406)
天府峨眉计划项目(1649)
四川农业大学双支计划项目(P202108)。
关键词
薄荷
McPYL4基因
蛋白特征
表达分析
蛋白互作
Mentha canadensis
McPYL4 gene
protein characteristics
expression analysis
protein interaction
