乙烯诱导的油茶酵母cDNA文库构建及CoFAD7上游调控因子筛选  

Construction of yeast cDNA library of Camellia oleifera induced by ethylene and screening of CoFAD7 upstream regulators

作　　者：马晓玲 陈佳昕 柏芮 欧阳翔 MA Xiaoling;CHEN Jiaxin;BAI Rui;OUYANG Xiang(Key Laboratory of Cultivation and Protection for Non-wood Forest Trees of the Ministry of Education,Central South University of Forestry and Technology,Changsha 410004,China;State Key Laboratory of Hybrid Rice,Hunan Hybrid Rice Research Center,Changsha 410125,China)

机构地区：[1]中南林业科技大学,经济林培育与保护教育部重点实验室,长沙410004 [2]湖南杂交水稻研究中心,杂交水稻全国重点实验室,长沙410125

出　　处：《植物生理学报》2024年第8期1301-1309,共9页Plant Physiology Journal

基　　金：湖南省自然科学基金(2023JJ41036);长沙市自然科学基金(kq2208412)。

摘　　要：外施乙烯可促进油茶种仁α-亚麻酸(α-linolenic acid,ALA)积累,FAD7是影响ALA合成的关键基因。为了研究油茶CoFAD7表达的分子调控机制,本研究克隆CoFAD7的启动子,构建乙烯诱导的油茶种仁酵母单杂交cDNA文库,并通过酵母单杂交技术筛选与CoFAD7启动子结合的上游调控因子。结果表明,1395 bp的CoFAD7启动子序列含有多种激素调节与环境胁迫相关的顺式元件及转录因子结合元件。构建的cDNA文库库容为9.5×10^(6) CFU·mL^(-1),平均插入片段长度约1000 bp。通过酵母单杂交筛选获得4个转录因子AP2-3、NFYC-1、F-box和ZFP1,可以与CoFAD7启动子结合。以上结果为进一步探究CoFAD7基因响应乙烯调控ALA含量的分子机理提供参考。Exogenous ethylene could promote the accumulation ofα-linolenic acid(ALA)in Camellia oleifera seeds,FAD7 is a key gene affecting ALA synthesis.In order to elucidate the molecular regulatory mechanism of CoFAD7 expression in C.oleifera,this study cloned the promoter of CoFAD7 and constructed yeast one hybrid cDNA library of C.oleifera seeds induced by ethylene.The upstream regulatory factors that bind to the CoFAD7 promoter were screened through yeast one hybrid technology.The results indicate that the 1395 bp CoFAD7 promoter sequence contains multiple cis-elements and transcription factor binding elements related to hormone regulation and environmental stress.The constructed cDNA library had a capacity of 9.5×10^(6) CFU·mL^(-1) and an average insertion fragment length of approximately 1000 bp.Four transcription factors AP2-3,NFYC-1,F-box and ZFP1 were obtained through yeast one hybrid screening,which can bind to the CoFAD7 promoter.The above results lay the foundation for further exploring the molecular mechanism of CoFAD7 gene responsing to ethylene on regulation of ALA content.

关 键 词：油茶 CoFAD7启动子 酵母单杂交 CDNA文库 转录因子 

分 类 号：S794.4[农业科学—林木遗传育种]