《植物生理学报》 2021, 57(1): 85-93

葡萄VlCKX8基因克隆与植物生长调节剂响应分析

于可可^1,2，杨英军¹，郭大龙^1,2，刘海楠^1,2，裴茂松^1,2，韦同路^1,2，边璐^1,2，余义和^1,2,*

¹河南科技大学园艺与植物保护学院, 河南洛阳471023；²河南园艺植物质量调控工程技术研究中心, 河南洛阳471023

通信作者：余义和；E-mail: yuyihe@haust.edu.cn

摘　要：

细胞分裂素氧化酶/脱氢酶(cytokinin oxidase/dehydrogenase, CKX)作为不可逆降解细胞分裂素的酶, 对维持细胞分裂素稳态发挥重要作用。本研究以‘巨峰’ (Vitis vinifera × Vitis labrusca)葡萄为试材克隆了细胞分裂素氧化酶/脱氢酶8 (VlCKX8)的基因及其启动子, 并进行了表达特性分析和启动子活性分析。结果表明: VlCKX8基因cDNA序列全长为1 278 bp, 包含一个1 212 bp的开放阅读框, 编码403个氨基酸。RT-qPCR结果显示VlCKX8的表达水平在细胞分裂素生物合成抑制剂洛伐他汀处理之后先上调后下调。启动子活性分析表明VlCKX8的启动子活性在氯吡苯脲、吲哚丁酸、茉莉酸甲酯处理后均增强, 在赤霉素、脱落酸处理之后减弱。上述结果为研究细胞分裂素调控坐果的分子机理提供参考依据, 同时也为生产实践中科学使用植物生长调节剂调控果实坐果提供理论指导。

关键词：葡萄; VlCKX8; 启动子活性; 坐果; 细胞分裂素

收稿：2020-09-11   修定：2020-11-27

资助：国家自然科学基金(31701893)、河南省高校科技创新人才计划(21HASTIT035)、河南省高等学校青年骨干教师培养计划(No. 81)、中原科技创新领军人才项目(194200510007)和河南省高校科技创新团队支持计划(21IRTSTHN021)。

loning and plant growth regulator response analysis of VlCKX8 in grape

YU Keke^1,2, YANG Yingjun¹, GUO Dalong^1,2, LIU Hainan^1,2, PEI Maosong^1,2, WEI Tonglu^1,2, BIAN Lu^1,2, YU Yihe^1,2,*

¹College of Horticulture and Plant Protection, Henan University of Science and Technology, Luoyang, Henan 471023, China; ²Henan Engineering Technology Research Center of Quality Regulation and Controlling of Horticultural Plants, Luoyang, Henan 471023, China

Corresponding author: YU Yihe; E-mail: yuyihe@haust.edu.cn

Abstract:

Cytokinin oxidase/dehydrogenase (CKX), as an enzyme that irreversibly degrades cytokinin, plays an important role in maintaining cytokinin homeostasis. In this study, the VlCKX8 gene and its promoter were cloned from ‘Kyoho’ (Vitis vinifera × Vitis labrusca), and expression characteristics and promoter activity were analyzed. The results showed that the cDNA sequence of VlCKX8 was 1 287 bp, and the open reading frame (ORF) length was 1 212 bp, encoding 403 amino acids. Real-time quantitative PCR (RT-qPCR) results showed that the expression level of VlCKX8 was up-regulated and then down-regulated after cytokinin synthesis inhibitor lovastatin treatment. The promoter activity analysis showed that the VlCKX8 promoter activity was enhanced after forchlorfenuron, indobutyric acid and methyl jasmonate treatments and decreased after gibberellin, abscisic acid treatment. Taken together, these results provide a reference for the molecular mechanisms underlying cytokinin regulated fruit setting, and also provide theoretical guidance for the scientifc using plant growth regulators to regulate fruit setting in grape production.

Key words: grape; VlCKX8; promoter activity; fruit setting; cytokinin