文献类型：期刊文献

中文题名：牡丹花青素苷合成关键基因PsDFR启动子活性分析

英文题名：Promoter Functional Analysis of the Key Gene PsDFR Involved in Paeonia suffruticosa Anthocyanin Biosynthesis

作者：周琳[1] 袁梦[1] 齐宇[1,2] 张梦杰[1] 王雁[1]

第一作者：周琳

机构：[1]中国林业科学研究院林业研究所,国家林业和草原局林木培育重点实验室,北京100091;[2]山东省农业科学研究院,山东济南250100

年份：2024

卷号：37

期号：2

起止页码：16-26

中文期刊名：林业科学研究

外文期刊名：Forest Research

收录：CSTPCD;;Scopus;北大核心:【北大核心2023】；CSCD:【CSCD2023_2024】；

基金：国家自然科学基金项目(31972456);国家重点研发计划项目(2018YFD1000405)。

语种：中文

中文关键词：牡丹;花青素苷;PsDFR;启动子;顺式作用元件;启动子活性

外文关键词：Paeonia suffruticosa;anthocyanin;PsDFR;promotor;cis-acting elements;promoter activity

分类号：S685.11

摘要：[目的]分析牡丹花青素苷合成关键基因PsDFR启动子的顺式作用元件及活性,为进一步研究PsDFR启动子的功能及其在牡丹花色形成中的调控机制奠定基础。[方法]以‘黑花魁’牡丹花瓣中提取的基因组DNA为模板,通过染色体步移法克隆PsDFR的启动子序列。利用生物信息在线软件预测分析启动子序列的顺式作用元件。构建5个不同长度缺失启动子与GUS基因融合的表达载体,并瞬时转化烟草叶片,通过GUS组织化学染色和酶活性检测分析缺失启动子的活性,及其对脱落酸(ABA)、茉莉酸甲酯(MeJA)、光照等不同胁迫处理的响应。[结果]克隆得到PsDFR上游1687 bp的启动子序列。生物信息学分析发现PsDFR启动子含有多个光信号、激素响应、逆境响应及组织特异表达等顺式作用元件,这暗示PsDFR的表达可能受光信号、激素和胁迫等多种信号的共同调节。GUS组织化学染色和酶活性检测表明,随启动子片段的缩短,启动子活性逐渐下降,-1623至-916区段对于启动子活性具有重要作用。MeJA和黑暗处理对各启动子片段活性均有显著抑制作用,恢复光照可促使启动子活性明显回升,而参与ABA响应的核心调控区域位于-443至-76 bp。[结论]PsDFR启动子含有多个光信号、激素响应、逆境响应及组织特异表达等顺式作用元件,其活性受光的正调控以及MeJA的负调控;-1623至-916 bp间的区域对于启动子活性具有重要作用,-443至-76 bp是响应ABA处理的核心区域。该研究为进一步揭示PsDFR应答环境信号参与牡丹花色形成的分子调控机制提供参考。
[Objective]To analyze the cis-acting element and activity of the tree peony anthocyanin biosynthetic key gene PsDFR promoter for further research on its function and regulation mechanism involved in tree peony flower coloration.[Method]The genomic DNA extracted from the petals of tree peony‘Hei Hua Kui’was used as a template.PsDFR promoter was isolated by genomic walking method.The cis-acting elements of promoter were analyzed and predicted through Bioinformatics online software.Five different length of deletion fragments were fused with GUS gene to construct promoter-reporter vectors,and then were transiently expressed in tobacco leaves.The activities of deletion promoters and their response to different stress treatments such as Abscisic acid(ABA),Methyl jasmonate(MeJA)and light were analyzed by GUS staining and GUS activity assay.[Result]A 1687 bp sequence of PsDFR promoter was isolated.The results of bioinformatics analysis showed that the promoter contains multiple cis-acting elements associated with light signals,hormone response,stress response,and tissue-specific expression,indicating that the expression of PsDFR may be regulated by various signals such as light signaling,plant hormone and stress.GUS staining and quantitative analysis of GUS activity showed that GUS activities decreased gradually with decrease of the length of PsDFR promoters,and the region of-1623 bp to-916 bp played an important role on the activity of the PsDFR promoter.The GUS activities were inhibited significantly by MeJA or dark treatment,and were induced obviously after light restoration.And core regulation region involved in ABA-response might be located between-443 and-76 bp.[Conclusion]PsDFR promoter contains multiple cis-acting elements associated with light signals,hormone response,stress response,and tissue-specific expression.Its activity is positively regulated by light and negatively regulated by MeJA.The region of-1623 bp to-916 bp is important for the activity of the PsDFR promoter,and-443 and-76 bp is the core region in response to ABA treatment.This study provides a reference for further revealing the regulatory mechanism of PsDFR response to environmental signals involved in tree peony flower coloration.