月季品种‘绿萼’<em>RcAG</em>基因启动子克隆及分析

doi:10.11913/PSJ.2095-0837.2021.40407

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摘要

月季品种‘绿萼’（Rosa chinensis ‘Viridiflora’）是中国古老月季最宝贵资源之一，其花瓣、雄蕊及雌蕊均萼片化。本研究以花器官正常发育的月季品种‘月月粉’（R.chinensis ‘Old Blush’）为对照，利用实时荧光定量PCR技术对RcAG基因在‘绿萼’花器官中的表达情况进行分析，阐明RcAG基因在‘绿萼’花器官发育中的作用。结果显示，RcAG基因在‘绿萼’中的表达明显下调。进一步克隆‘绿萼’和‘月月粉’的RcAG基因启动子，序列分析结果表明两个启动子均含有TATA、TATC-box和MBS等顺式作用元件，但在‘绿萼’RcAG基因启动子中发现了光响应元件MRE和昼夜节律调控元件Circadian，而光响应元件TCT-motif只在‘月月粉’RcAG基因启动子中被发现。同时，本研究采用重亚硫酸盐测序技术分析了‘绿萼’和‘月月粉’RcAG基因启动子甲基化的情况，发现‘绿萼’RcAG基因启动子区域中有4个CpG位点均发生甲基化，其甲基化程度远高于‘月月粉’。研究结果表明RcAG基因在‘绿萼’花器官中的下调表达可能与其启动子的顺式作用元件及甲基化修饰相关。

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	李蓉
	眭梦洁
	王其刚
	余春霞
	杜光辉
	晏慧君

关键词 ：月季品种&lsquo, 绿萼&rsquo, 花器官发育, 荧光定量PCR, 启动子克隆, 甲基化分析

Abstract：

Rosa chinensis ‘Viridiflora’ is a valued Chinese garden rose cultivar and represents an important rose germplasm resource, in which the petals, stamens, and pistils are converted into sepal-like organs (i.e., sepalody). Expression of the RcAG gene in flower tissues of the R. chinensis ‘Viridiflora’ and ‘Old Blush’ varieties was analyzed by real-time quantitative polymerase chain reaction (PCR). Results showed that the expression level of RcAG was down-regulated in R. chinensis ‘Viridiflora’. To explore the reason for its down-regulation, the promoter sequences of RcAG were cloned in ‘Viridiflora’ and ‘Old Blush’, respectively. Results showed that the two promoter sequences contained TATA-box, TATC-box, and MBS cis-elements. Notably, MRE and Circadian were specifically found in the RcAG promoter in ‘Viridiflora’, while the TCT motif was only found in ‘Old Blush’. Furthermore, double sulfite sequencing technology was used to analyze promoter methylation. Results showed that all four CpG sites in the promoter region of ‘Viridiflora’ were methylated, and the degree of methylation was much higher than that of ‘Old Blush’. Our findings suggest that the down-regulation of the RcAG gene in ‘Viridiflora’ may be related to its promoter cis-elements and methylation.

Key words： Rosa chinensis ‘Viridiflora’ Flower organ development Real-time quantitative PCR Promoter cloning Methylation analysis

收稿日期: 2020-12-11

ZTFLH:

Q943.2

基金资助:

国家自然科学基金项目（32060693，31660579）。

通讯作者: 杜光辉, 晏慧君 E-mail: dgh2012@ynu.edu.cn;hjyan8203@126.com

作者简介: 李蓉(1997-),女,硕士研究生,研究方向为月季花发育(E-mail:443618756@qq.com)。

引用本文:

李蓉, 眭梦洁, 王其刚, 余春霞, 杜光辉, 晏慧君. 月季品种‘绿萼’RcAG基因启动子克隆及分析[J]. 植物科学学报, 2021, 39(4): 407-414. Li Rong, Sui Meng-Jie, Wang Qi-Gang, Yu Chun-Xia, Du Guang-Hui, Yan Hui-Jun. Cloning and analysis of RcAG promoter in Rosa chinensis ‘Viridiflora’. Plant Science Journal, 2021, 39(4): 407-414.

链接本文:

http://www.plantscience.cn/CN/10.11913/PSJ.2095-0837.2021.40407 或 http://www.plantscience.cn/CN/Y2021/V39/I4/407

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