桂花色泽关键基因OfCCD4互作转录因子的鉴定分析  

Identification of transcriptome-based Osmanthus fragrans OfCCD 4 interaction transcription factors

作　　者：史德静 邬俊媛 韩柳青 张瑞珍 林楠 尚富德 刘燕培 SHI Dejing;WU Junyuan;HAN Liuqing;ZHANG Ruizhen;LIN Nan;SHANG Fude;LIU Yanpei(College of Life Sciences,Henan Agricultural University,Zhengzhou 450046,China)

机构地区：[1]河南农业大学生命科学学院,河南郑州450046

出　　处：《河南农业大学学报》2024年第5期757-764,共8页Journal of Henan Agricultural University

基　　金：河南省重大公益专项项目(201300110900);全国重点实验室开放课题资助项目(Horti-KF-2023-10);河南省青年科学基金项目(222300420183);河南农业大学科技创新基金项目(KJCX2021A17);河南省留学人员科研择优资助项目(30602425)。

摘　　要：【目的】明确调控桂花类胡萝卜素代谢关键基因OfCCD4的转录因子,探究转录因子与OfCCD4结合并调控其表达的分子机制,为解析桂花类胡萝卜素代谢的分子机制及桂花花色的基因工程育种提供理论依据。【方法】选取杭州临安太湖源区桂花花瓣为研究材料,对前期筛选的类胡萝卜素代谢关键基因OfCCD4进行转录因子预测,并利用实时荧光定量PCR(RT-qPCR)、酵母单杂交等分子生物学试验初步鉴定OfCCD4的转录因子。【结果】转录组分析发现,有10个转录因子与OfCCD4表达水平呈现高相关性;Y1H酵母转录激活活性结果显示,其中有8个转录因子具有一定的转录自激活能力;酵母单杂交结果表明,3个转录因子与OfCCD4启动子有强相互作用,3个转录因子与OfCCD4启动子存在弱相互作用。【结论】筛选到3个具有自激活活性,且与OfCCD4存在强互作关系的转录因子,推测其可能通过调控桂花OfCCD4基因表达影响花色差异。【Objective】Defining the transcription factors of OfCCD 4,a key gene that regulates carotenoid metabolism in Osmanthus fragrans,as well as exploring the molecular mechanism of transcription factors binding to OfCCD 4 and regulating OfCCD 4 expression,which provide a theoretical basis for revealing the molecular mechanism of carotenoids metabolism as well as for O.fragrans color genetic engineering breeding.【Method】The petals of Osmanthus flower were obtained in Taihuyuan,which located in Lin’an,Hangzhou City.The transcription factors of OfCCD 4 were initially predicted through the online websites,and further identified by real-time fluorescence quantitative(RT-qPCR)and yeast-one-hybrid assays.【Result】Transcriptome analysis were that 10 transcription factors were highly correlated with the expression level of OfCCD 4.Transactivation activity in Y1H yeast showed that 8 transcription factors of them had transactivation activity.Finally,3 transcription factors displayed strong interaction and 3 transcription factors exhibited weak interaction with OfCCD 4 promoter in yeast-one-hybrid assay.【Conclusion】In this study,three transcription factors with transactivation activity and strong interaction with OfCCD 4 were screened,indicating they might affect flower color by regulating OfCCD 4 expression in Osmanthus fragrans.

关 键 词：桂花 类胡萝卜素 OfCCD4基因 转录因子 酵母单杂交 

分 类 号：S685.13[农业科学—观赏园艺]