桂花番茄红素β-环化酶基因LCYB上游B2亚组ERF转录因子的筛选和鉴定  

Screening and identification of ERF transcription factors of B2 subgroup involved in regulating lycopeneβ-cyclase gene LCYB in Osmanthus fragrans

作　　者：李莉 庞天虹[1,2,3] 付建新 张超[1,2,3] LI Li;PANG Tianhong;FU Jianxin;ZHANG Chao(College of Landscape and Architecture,Zhejiang A&F University,Hangzhou 311300,Zhejiang,China;Zhejiang Provincial Key Laboratory of Germplasm Innovation and Utilization for Garden Plants,Zhejiang A&F University,Hangzhou 311300,Zhejiang,China;Key Laboratory of National Forestry and Grassland Administration on Germplasm Innovation and Utilization for Southern Garden Plants,Zhejiang A&F University,Hangzhou 311300,Zhejiang,China)

机构地区：[1]浙江农林大学风景园林与建筑学院,浙江杭州311300 [2]浙江农林大学浙江省园林植物种质创新与利用重点实验室,浙江杭州311300 [3]浙江农林大学南方园林植物种质创新与利用国家林业和草原局重点实验室,浙江杭州311300

出　　处：《浙江农林大学学报》2025年第1期86-93,共8页Journal of Zhejiang A&F University

基　　金：浙江省基础公益研究计划项目(LY19C160002)。

摘　　要：【目的】筛选并鉴定参与调控桂花Osmanthus fragrans番茄红素β-环化酶OfLCYB基因的B2亚组ERF转录因子。【方法】以桂花品种‘堰虹桂’O. fragrans ‘Yanhong Gui’为材料,从桂花转录组数据库筛选B2亚组OfERF基因,通过生物信息学分析、实时荧光定量PCR (RT-qPCR)以及酵母单杂交技术,对OfERF基因序列和表达特性及其对Of LCYB基因启动子结合情况进行分析。【结果】OfLCYB基因启动子序列含有2个ATCTA顺式作用元件;基于桂花转录组数据库筛选出4个B2亚组ERF基因,均包含1个AP2保守结构域;RT-qPCR结果表明:OfERF72a与OfERF72b基因表达量均随着开花进程逐渐下降,与OfLCYB基因表达显著负相关,P分别为0.033 8、0.029 6;酵母单杂交结果证明:OfERF72b与OfLCYB启动子之间存在物理结合。【结论】OfERF72b可能通过调控OfLCYB的转录参与桂花类胡萝卜素的代谢。[Objective]This study aims to screen and identify ERF transcription factors of B2 subgroup involved in regulating lycopeneβ-cyclase OfLCYB gene of Osmanthus fragrans.[Method]‘Yanhong Gui’,a cultivar of O.fragrans,was used as the material to screen OfERF genes of B2 subgroup from the O.fragrans transcriptome database.Bioinformatic analysis,real-time quantitative PCR(RT-qPCR)and yeast one-hybridization were used to analyze the sequence and expression characteristics of the OfERF gene and its binding to the OfLCYB gene promoter.[Result]The promoter of the OfLCYB gene contained two ATCTA cis-acting elements.Four OfERF genes of B2 subgroup were screened based on the O.fragrans transcriptome database,all of which contained an AP2 conserved structural domain.The RT-qPCR results showed that the expression levels of OfERF72a and OfERF72b genes gradually decreased with the flowering process,and were significantly negatively correlated with the expression of OfLCYB gene,with P values of 0.0338 and 0.0296,respectively.The results of yeast one-hybridization proved that there was a physical binding between OfERF72b and the OfLCYB promoter.[Conclusion]OfERF72b may participate in the metabolism of carotenoid in O.fragrans by regulating the transcription of OfLCYB.

关 键 词：桂花 类胡萝卜素 ERF转录因子 基因功能 

分 类 号：S722.3[农业科学—林木遗传育种]