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园艺学报 ›› 2021, Vol. 48 ›› Issue (8): 1437-1445.doi: 10.16420/j.issn.0513-353x.2020-0668

• 研究论文 •    下一篇

‘巨峰’葡萄细胞分裂素响应调节因子VlRR5的克隆与表达分析

边璐, 郭大龙, 于可可, 韦同路, 裴茂松, 刘海楠, 余义和()   

  1. 河南科技大学园艺与植物保护学院,河南洛阳 471023
  • 收稿日期:2021-02-23 修回日期:2021-06-07 出版日期:2021-08-25 发布日期:2021-09-06
  • 通讯作者: 余义和 E-mail:yuyihe@haust.edu.cn
  • 基金资助:
    国家自然科学基金项目(31701893);国家自然科学基金项目(32072517);国家重点研发计划项目(2018YFD1000105);河南省高校科技创新人才项目(21HASTIT035);河南省高等学校青年骨干教师培养计划项目(81);河南省高校科技创新团队支持计划项目(21IRTSTHN021);洛阳市科技发展计划项目(2101102A)

Cloning and Expression Analysis of the Cytokinin Response Regulator VlRR5 in Kyoho Grapevine

BIAN Lu, GUO Dalong, YU Keke, WEI Tonglu, PEI Maosong, LIU Hainan, YU Yihe()   

  1. College of Horticulture and Plant Protection,Henan University of Science and Technology,Luoyang,Henan 471023,China
  • Received:2021-02-23 Revised:2021-06-07 Online:2021-08-25 Published:2021-09-06
  • Contact: YU Yihe E-mail:yuyihe@haust.edu.cn

摘要:

葡萄在生长发育过程中会出现3次落果高峰,‘巨峰’葡萄的落果现象更为严重。从‘巨峰’葡萄中克隆得到1个细胞分裂素响应调节因子VlRR5,该基因有1个磷酸受体结构域(REC)和1个MYB样DNA结合结构域,cDNA全长2 597 bp,编码693个氨基酸,定位在4号染色体上。亚细胞定位在细胞核中。酵母转化试验中,转入pGBKT7-VlRR5的酵母菌可以在SD/-Trp/-Ade/-His/+X-α-Gal缺陷型培养基上正常生长且显蓝色,证明VlRR5具有转录激活活性。荧光定量PCR分析显示,VlRR5的表达具有组织特异性,在叶和茎中表达量较高;在幼果发育阶段呈现先升高后降低的趋势;并且在外源细胞分裂素CPPU处理后表达量上升,在细胞分裂素合成抑制剂洛伐他汀处理后表达量下降。这些结果表明VlRR5可以响应外源细胞分裂素处理,在细胞分裂素介导葡萄坐果过程中发挥重要作用。

关键词: 葡萄, 细胞分裂素, CPPU, 细胞分裂素响应调节因子, VlRR5

Abstract:

There are three fruit-dropping peaks in grape growth and development. Grape fruit-dropping phenomenon is more serious in Kyoho grapevine. In this study,a cytokinin response regulator VlRR5 was cloned from Kyoho grapevine,which has a phosphoric acid receptor domain(REC)and a MYB-like DNA binding domain. Sequence analysis revealed that cDNA of VlRR5 was 2 597 bp in length,encoding 693 amino acids,and located on chromosome 4 of grape. Subcellular localization assay showed that VlRR5 was localized in the nucleus. The yeast transformed into pGBKT7-VlRR5 could grow normally and turn blue on SD/-Trp/-Ade/-His/+X-α-Gal deficient medium,which proved that VlRR5 was a transcription factor with transcriptional activation activity. By quantitative real-time PCR analysis,VlRR5 was characterized by tissue specific expression and showed high expression level in leaf and stem,which was increased and then decreased in the development stage of young fruit. The expression level increased after treatment with exogenous cytokinin,and decreased after treatment with the cytokinin synthesis inhibitor lovastatin. These results indicated that VlRR5 can respond to cytokinin and plays an important role in participating in cytokinin to improve the fruiting rate of grape.

Key words: grapevine, cytokinin, CPPU, cytokinin response regulators, VlRR5

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