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园艺学报 ›› 2020, Vol. 47 ›› Issue (3): 590-600.doi: 10.16420/j.issn.0513-353x.2019-0432

• 研究报告 • 上一篇    下一篇

杜仲类钙调蛋白基因 EuCML5 的克隆及表达分析

刘 伟 1,3,*,滕 腾 2,3,*,赵懿琛 2,3,**,赵德刚 3,4   

  1. 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
  • 出版日期:2020-03-25 发布日期:2020-03-25
  • 基金资助:
    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)

Cloning and Expression Analysis of EuCML5 Gene in Eucommia ulmoides

LIU Wei1,3,*,TENG Teng2,3,*,ZHAO Yichen2,3,**,and ZHAO Degang3,4   

  1. 1College of Pharmacy,Guizhou University,Guiyang 550025,China;2College of Tea Science,Guizhou University, Guiyang 550025,China;3College of Life Sciences,Academy of Agricultural Bioengineering,The Key Laboratory of Plant Resources Conservation and Germplasm Innovationin Mountainous Region(Ministry of Education),Guizhou University,Guiyang 550025,China;4Guizhou Academy of Agricultural Sciences,Guiyang Branch Center for DUS Testing of New Plant Varieties of the Ministry of Agriculture of China,Guiyang 550006,China
  • Online:2020-03-25 Published:2020-03-25

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。

关键词: 杜仲, 类钙调蛋白, 表达, 胁迫

Abstract: Based on the Eucommia ulmoides genome database,the Calmodulin-like(CML)gene EuCML5 was cloned. The open reading frame of EuCML5 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 EuCML5expression.

Key words: Eucommia ulmoides, CML, expression, stress

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