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园艺学报 ›› 2015, Vol. 42 ›› Issue (2): 221-232.doi: 10.16420/j.issn.0513-353x.2014-0875

• 果树 • 上一篇    下一篇

野生毛葡萄水通道蛋白基因VhPIP1的克隆及其在干旱胁迫下的表达分析

颜培玲,潘学军,张文娥   

  1. 1贵州大学农学院,贵阳 550025;2贵州省果树工程技术研究中心,贵阳 550025
  • 出版日期:2015-02-25 发布日期:2015-02-25
  • 基金资助:
    贵州省优秀青年科技人才培养专项[黔科合人字(2009)02号];国家科技支撑计划课题(2014BAD23B03)

Cloning of Aquaporion Gene VhPIP1 in Vitis heyneana and Its Expression Under Drought Stress

YAN Pei-ling1,2,PAN Xue-jun1,2,*,and ZHANG Wen-e   

  1. 1 College of Agriculture,Guizhou University,Guiyang 550025,China;2Guizhou Engineering Research Center for Fruit Crops,Guiyang 550025,China
  • Online:2015-02-25 Published:2015-02-25

摘要: 采用RT-PCR和RACE技术从中国野生毛葡萄(Vitis heyneana)‘花溪–9’根中克隆得到1个质膜内在蛋白(plasma membrane intrinsic proteins,PIPs)类的水通道蛋白基因,命名为VhPIP1(登录号为KM026528)。该基因cDNA全长为1 087 bp,包含1个858 bp的完整开放阅读框,编码286个氨基酸。氨基酸序列分析表明,VhPIP1含有水通道蛋白家族高度保守的两个NPA(Asn-Pro-Ala)单元和6个跨膜区,其氨基酸残基与MIP家族蛋白保守区序列完全一致。实时荧光定量RT-PCR分析表明,VhPIP1在野生毛葡萄根、茎、叶中均有表达,在根中的表达量最高,叶片中最少。干旱胁迫下,抗旱性强的毛葡萄‘花溪–9’VhPIP1的表达量随着胁迫时间的延长先增加后降低,而不抗旱的欧亚种‘红地球’VvPIP1的表达量呈下降的趋势。与不抗旱的‘红地球’相比,干旱胁迫下极抗旱的‘花溪–9’较高的VhPIP1转录水平对应较高的叶片相对含水量,同时对应较低的根系细胞膜相对透性。VhPIP1的表达丰度与毛葡萄抗旱性密切相关。

关键词: 毛葡萄, 水通道蛋白, 干旱胁迫, 克隆, 表达分析

Abstract: A plasma membrane intrinsic proteins(PIPs)gene,designated VhPIP1,was cloned from root on wild grapevine(Vitis heyneana‘Huaxi-9’)native to China by RT-PCR and RACE(Rapid amplication of cDNA ends). The full cDNA sequence of VhPIP1 was 1 087 bp,containing an open reading frame(858 bp)and encoding a putative VhPIP1 protein with 286 amino acids. Bioinformatics analysis demonstrated that VhPIP1 exhibited an internal symmetry showing two highly conserved NPA(Asn-Pro-Ala)motifs and a typical structure with six membrane-spanning domains and possessing the MIP family signal consensus sequence. Quantitative real-time PCR results showed that the VhPIP1 expressed in roots,stems and leaves,and the expression level was the highest in roots,and the lowest inleaves. Under drought stress,the expression level of VhPIP1 of Huaxi-9 changed with time extension,which was first rised and then dropped,but the expression level of VvPIP1 of Red Globe was decreased. Compared with drought sensitive genotype Red Globe,the relatively higher expression level of VhPIP1 in drought resistance genotype corresponds to the higher leaves relative water content and the lower ion leakage of plasma membrane. These results suggested that the expression quantity of VhPIP1 closely related to drought resistance.

Key words: Vitis heyneana, aquaporin, drought stress, clone, expression

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