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园艺学报 ›› 2017, Vol. 44 ›› Issue (8): 1463-1476.doi: 10.16420/j.issn.0513-353x.2016-0778

• 研究论文 • 上一篇    下一篇

欧洲葡萄中CIPK基因的克隆及表达分析

闫朝辉,李桂荣,扈岩松,周瑞金,扈惠灵,苗卫东,朱自果*   

  1. (河南科技学院园艺园林学院,河南新乡 453003)
  • 出版日期:2017-08-25 发布日期:2017-08-25

Cloning and Expression Analysis of CIPK Genes in Grapevine

YAN Chaohui,LI Guirong,HU Yansong,ZHOU Ruijin,HU Huiling,MIAO Weidong,and ZHU Ziguo*   

  1. (College of Horticulture and Landscape Architecture,Henan Institute of Science and Technology,Xinxiang,Henan 453003,China)
  • Online:2017-08-25 Published:2017-08-25

摘要:

以欧洲葡萄‘佳丽酿’(Carinena)为材料,利用RT-PCR方法获得两个CIPK基因的全长cDNA序列,分别为VvCIPK13和VvCIPK14。VvCIPK13全长为1 501 bp,包括一个1 395 bp的ORF,编码465个氨基酸;VvCIPK14全长为1 616 bp,包括一个1 404 bp的ORF,编码468个氨基酸。生物信息学分析表明两个蛋白均含有两个保守结构域:丝氨酸/苏氨酸激酶结构域和NAF结构域,两个结构域之间有连接域,但VvCIPK13和VvCIPK14的同源性较低,仅有40.33%。利用荧光定量PCR技术研究VvCIPK13和VvCIPK14在葡萄不同组织、植物生长调节剂诱导和逆境胁迫下的表达模式,结果表明:VvCIPK13和VvCIPK14表达具有组织特异性,均在卷须中高表达;VvCIPK13对6-BA诱导有响应,而VvCIPK14对6-BA、ABA、IAA、SA、MeJA和GA3等诱导均有不同程度的响应;VvCIPK13和VvCIPK14均响应高盐和干旱,但不响应低温,其中VvCIPK14的响应最为强烈。推测VvCIPK13和VvCIPK14在葡萄的非生物胁迫过程中发挥重要的作用。

关键词: 葡萄, CIPK, 基因表达, 非生物逆境

Abstract: Two CIPK genes,VvCIPK13 and VvCIPK14,were isolated from‘Carinena’grapevine(Vitis vinifera). The full-length of VvCIPK13 was 1 501 bp containing an open reading frame(1 395 bp),which encoded 465 amino acids;the full-length of VvCIPK14 was 1 616 bp containing an open reading frame(1 404 bp),which encoded 468 amino acids. They had the typical kinase domain,NAF regulatory domain and the junction domain,but their similarity was only 40.33%. Real-time PCR analysis indicated that the expressions of VvCIPK13 and VvCIPK14 were higher in tendril than other tissues. Mean- while,VvCIPK13 was induced only by 6-BA,different from that VvCIPK14 was induced by 6-BA,ABA,IAA,SA,MeJA and GA3. The expressions of VvCIPK13 and VvCIPK14 were both induced by salt and drought stresses,but weren’t induced by cold. And the expression of VvCIPK14 was stronger than VvCIPK13. These results showed that VvCIPK13 and VvCIPK14 may play a key role in abiotic stress in grapevine.

Key words: Vitis vinifera, CIPK, gene expression, abiotic stress

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