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园艺学报 ›› 2021, Vol. 48 ›› Issue (2): 265-275.doi: 10.16420/j.issn.0513-353x.2020-0343

• 研究论文 • 上一篇    下一篇

中国野生葡萄抗霜霉病相关基因VpPR4b及其启动子的克隆和功能分析

刘兵*, 李梦媛*, 张娜, 尚博兴, 刘国甜**(), 徐炎**()   

  1. 西北农林科技大学园艺学院,农业农村部西北地区园艺作物生物学与种质创制重点实验室,陕西杨凌 712100
  • 收稿日期:2020-10-15 修回日期:2021-01-09 出版日期:2021-02-25 发布日期:2021-03-09
  • 通讯作者: 刘国甜,徐炎 E-mail:liuguotian555@163.com;yan.xu@nwsuaf.edu.cn
  • 基金资助:
    陕西省自然科学基础研究计划一般项目(2018JQ3012);国家自然科学基金项目(31601716,31972374)

Cloning and Functional Analysis of the CDS and Promoter of VpPR4b Gene Response to Downy Mildew in Chinese Wild Grape

LIU Bing*, LI Mengyuan*, ZHANG Na, SHANG Boxing, LIU Guotian**(), XU Yan**()   

  1. College of Horticulture,Northwest A & F University,Key Laboratory of Horticultural Plant Biology and Germplasm Innovation in Northwest China,Ministry of Agriculture and Rural Affairs,Yangling,Shaanxi 712100,China
  • Received:2020-10-15 Revised:2021-01-09 Online:2021-02-25 Published:2021-03-09
  • Contact: LIU Guotian,XU Yan E-mail:liuguotian555@163.com;yan.xu@nwsuaf.edu.cn

摘要:

以中国野生葡萄(Vitis piasezkii)‘留坝-8’(抗霜霉病)和欧亚种葡萄(V. vinifera)‘黑比诺’(易感霜霉病)为材料,克隆得到抗霜霉病相关基因VpPR4bVvPR4b。序列分析发现VpPR4bVvPR4b核苷酸序列相似性为98.61%。VpPR4b编码区为432 bp,编码143个氨基酸,含有BARWIN保守结构域,属于典型的Ⅱ型PR4蛋白。VpPR4bVvPR4b均可在霜霉菌侵染后诱导表达。亚细胞定位表明VpPR4b定位于细胞膜上。分别从‘留坝-8’和‘黑比诺’中克隆得到了VpPR4bVvPR4b的启动子,其序列相似性高达95.65%,并且含有相似的顺式作用元件。通过酵母单杂交验证了转录因子WRKY40和WRKY75可以结合VpPR4b启动子。本研究的结果表明PR4b可能在葡萄抵御霜霉病侵染过程中发挥一定作用。

关键词: 葡萄, 霜霉病, PR4b, 启动子

Abstract:

VpPR4b and VvPR4b were cloned form Vitis piasezkii‘Liuba-8’(resistant to downy mildew)and V. vinifera‘Pinot Noir’(susceptible to downy mildew). Sequence analysis showed that the identity between VpPR4b and VvPR4b nucleotide was 98.61%. The coding region of VpPR4b is 432 bp,which encodes 143 amino acids and contains the BARWIN conserved domain,indicating it is a typical typeⅡPR4 protein. Both VpPR4b and VvPR4b are highly expressed after inoculation with Plasmopara viticola. Subcellular localization analysis indicated that VpPR4b was localized in the membrane. The promoters of VpPR4b and VvPR4b were cloned from‘Liuba-8’and‘Pinot Noir’,respectively. It showed that VpPR4b and VvPR4b promoter sequence similarity is as high as 95.65%,and contains similar cis-acting elements. It has been verified that the transcription factors WRKY40 and WRKY75 can bind the VpPR4b promoter in yeast. This study showed that PR4b may play a role in the resistance to downy mildew in grape.

Key words: grape, downy mildew, PR4b, promoter

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