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园艺学报 ›› 2021, Vol. 48 ›› Issue (6): 1079-1093.doi: 10.16420/j.issn.0513-353x.2020-0902

• 研究论文 • 上一篇    下一篇

山葡萄VaCBL6参与非生物胁迫和ABA途径的响应

马俊杰1, 宋丽娜1, 李乐1, 马晓春1, 靳磊1, 徐伟荣2,3,4,*()   

  1. 1宁夏大学农学院,银川 750021
    2宁夏大学食品与葡萄酒学院,银川 750021
    3中国葡萄酒产业技术研究院,银川 750021
    4宁夏优势特色作物现代分子育种重点实验室,银川 750021
  • 收稿日期:2021-03-16 修回日期:2021-05-11 出版日期:2021-06-25 发布日期:2021-07-07
  • 通讯作者: 徐伟荣 E-mail:xuwr@nxu.edu.cn
  • 基金资助:
    国家重点研发计划项目(2019YFD1002500);国家自然科学(31560550);国家自然科学(31860542);国家科技重大专项(106001000000150012)

VaCBL6 from Vitis amurensis Involved in Abiotic Stress Response and ABA-mediated Pathway

MA Junjie1, SONG Lina1, LI Le1, MA Xiaochun1, JIN Lei1, XU Weirong2,3,4,*()   

  1. 1College of Agronomy,Ningxia University,Yinchuan 750021,China
    2School of Food & Wine,Ningxia University,Yinchuan 750021,China
    3China Wine Industry Technology Institute,Yinchuan 750021,China
    4Key Laboratory of Modern Molecular Breeding of Dominant and Special Crops in Ningxia,Yinchuan 750021,China
  • Received:2021-03-16 Revised:2021-05-11 Online:2021-06-25 Published:2021-07-07
  • Contact: XU Weirong E-mail:xuwr@nxu.edu.cn

摘要:

对山葡萄(Vitis amurensis)‘左山-1'中的1个类钙调素磷酸酶B亚基蛋白基因VaCBL6进行克隆及功能分析。VaCBL6位于第4条染色体,开放阅读框为777 bp,编码258个氨基酸;结构域分析表明,该蛋白包含4个EF-hand与1个跨膜结构域;蛋白进化树分析表明,VaCBL6与欧洲葡萄VvCBL9和VvCBL6相似性较高,分别为98.84%、94.96%。VaCBL6编码蛋白定位于细胞核与细胞膜。VaCBL6在根与卷须中有较高的表达,且其表达受低温、干旱、NaCl和ABA的诱导。基于酵母体系的转录激活分析表明,VaCBL6无转录激活活性。100 ~ 125 μmol · L -1 NaCl和0.25 ~ 1.25 μmol · L -1 ABA处理均抑制了VaCBL6转基因拟南芥种子的萌发,且转化拟南芥幼苗对NaCl与ABA的敏感性提高。以上结果说明VaCBL6在植物的逆境胁迫应答中具有重要作用。

关键词: 山葡萄, 类钙调素磷酸酶B亚基蛋白, 基因克隆, 非生物胁迫, 功能分析

Abstract:

A calcineurin B-like phosphatase subunit protein gene VaCBL6 from Vitis amurensis ‘Zuoshan-1'was cloned and functionally analyzed. VaCBL6is located on chromosome 4 and has an open reading frame of 777 bp,encoding 258 aa;protein domain analysis shows that the protein contains four EF-hands and one transmembrane domain. Phylogenetic analysis using amino acid sequence showed that high similarities between VaCBL6 and VvCBL9,VaCBL6 and VvCBL6 are 98.84% and 94.96%,respectively. VaCBL6 is located in the nucleus and cell membrane,and is highly expressed in roots and tendrils,and its expression is induced by low temperature,drought,NaCl and ABA. Analysis of transcriptional activation based on the yeast system displayed that VaCBL6 had no transcriptional activation activity. Experiments on the growth stage ofArabidopsis seedlings overexpressingVaCBL6with different treatments found that 100-125 μmol · L -1 NaCl and 0.25-1.25 μmol · L -1 ABA treatments inhibited the germination of VaCBL6 transgenic Arabidopsis seeds,and the transgenic Arabidopsis seedlings increased the sensitivity to NaCl and ABA. The results indicated that VaCBL6 plays an important role in plant stress response.

Key words: Vitis amurensis, calcineurin B-like phosphatase subunit protein, gene cloning, abiotic stress, functional analysis

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