杜仲类钙调蛋白基因EuCML5的克隆及表达分析  

Cloning and Expression Analysis of EuCML5 Gene in Eucommia ulmoides

作　　者：刘伟 滕腾 赵懿琛 赵德刚 LIU Wei;TENG Teng;ZHAO Yichen;ZHAO Degang(College of Pharmacy,Guizhou University,Guiyang 550025,China;College of Tea Science,Guizhou University,Guiyang 550025,China;College of Life Sciences,Academy ofAgricultural Bioengineering,The Key Laboratory of Plant Resources Conservation and Germplasm Innovationin Mountainous Region(Ministry of Education),Guizhou University,Guiyang 550025,China;Guizhou Academy of Agricultural Sciences,Guiyang Branch Center for DUS Testing of New Plant Varieties of the Ministry of Agriculture of China,Guiyang 550006,China)

机构地区：[1]贵州大学药学院,贵阳550025 [2]贵州大学茶学院,贵阳550025 [3]贵州大学生命科学学院,农业生物工程研究院,山地植物资源保护与种质创新省部共建教育部重点实验室,贵阳550025 [4]贵州省农业科学院,国家农业农村部植物新品种DUS测试贵阳分中心,贵阳550006

出　　处：《园艺学报》2020年第3期590-600,共11页Acta Horticulturae Sinica

基　　金：国家自然科学基金项目(31660076);贵州省教育厅青年科技人才成长项目(黔教科KY[2016]126);贵州省生物学一流学科建设项目(GNYL[2017]009)。

摘　　要：在前期构建的杜仲基因组数据库的基础上,克隆得到了类钙调蛋白(Calmodulin-like protein,CML)基因EuCML5。分析发现EuCML5开放阅读框长度为282 bp,编码93个氨基酸,不含跨膜结构和信号肽,属于胞内蛋白,氨基酸序列中含有EF-Hand motif结构,在进化关系上与一些木本植物类钙调蛋白较近。实时荧光定量PCR分析发现,EuCML5在杜仲根中表达较高,茎和叶中表达较低。杜仲苗经过模拟干旱、高盐、茉莉酸甲酯、生长素处理后,EuCML5表达量有不同程度提高,表明EuCML5能够被非生物胁迫和植物生长调节剂诱导表达,并且受钙离子通道抑制剂LaCl3和钙调蛋白拮抗剂TFP抑制。Based on the Eucommia ulmoides genome database,the Calmodulin-like(CML)gene EuCML5 was cloned.The open reading frame of Eu CML5 was 282 bp in length,encoding a total of 93 amino acids.It contained no transmembrane structure and signal peptide,belonging to intracellular protein.The amino acid sequence contained the EF-Hand motif,close to calmodulin-like protein of some woody plants in evolutionary relationship.Quantitative real-time PCR analysis revealed that the expression of EuCML5 was higher in Eucommia ulmoides roots and lower in stems and leaves.After treatment with drough(Mannitol),salt,methyl jasmonate and auxin,the expression of EuCML5 in Eucommia ulmoides was increased,indicating that EuCML5 can be induced by abiotic stress and phytohormone.In addition,calcium channel inhibitors LaCl3 and calmodulin antagonist TFP inhibited EuCML5 expression.

关 键 词：杜仲 类钙调蛋白 表达 胁迫 

分 类 号：S567[农业科学—中草药栽培]