菜用豌豆酵母单杂交cDNA文库及糖转运蛋白PsSWEET15诱饵载体构建  

Construction of Vegetable Pea Yeast One-hybrid cDNA Library and Sugar Transporter PsSWEET15 Bait Vector

作　　者：冯志娟[1] 刘娜 张古文[1] 卜远鹏 王斌 龚亚明[1] Feng Zhijuan;Liu Na;Zhang Guwen;Bu Yuanpeng;Wang Bin;Gong Yaming(Key Laboratory of Vegetable Legumes Germplasm Enhancement and Molecular Breeding in Southern China of Ministry of Agriculture and Rural Affairs,State Key Laboratory for Managing Biotic and Chemical Threats to the Quality and Safety of Agro-products,Institute of Vegetables,Zhejiang Academy of Agricultural Sciences,Hangzhou,310021)

机构地区：[1]浙江省农业科学院蔬菜研究所农业农村部南方豆类蔬菜种质创新与分子育种重点实验室(部省共建),省部共建农产品质量安全危害因子与风险防控国家重点实验室,杭州310021

出　　处：《分子植物育种》2023年第19期6390-6397,共8页Molecular Plant Breeding

基　　金：浙江省公益项目(LGN21C150007,LGN20C150006);国家自然科学基金项目(31601767);浙江省科技计划项目(2022C02016-1,2021C02052-2);浙江省蔬菜新品种选育重大科技专项(2021C02065-6)共同资助。

摘　　要：SWEETs(Sugars will eventually be exported transporter)是近年来新发现的一类重要的糖转运蛋白,调控植物体内的糖转运和分配过程。在菜用豌豆中,有关PsSWEETs基因功能及作用机制的报道甚少。本研究克隆获得了PsSWEET15启动子,长度为1182 bp。通过启动子作用元件分析,发现其含有种子特异表达元件GCN4 motif、光、无氧、乙烯、赤霉素等环境胁迫应答元件以及ERF、DOF、MYB、WRKY、NAC、NF-Y、TCP等多个转录因子结合位点。进一步构建了PsSWEET15启动子酵母单杂诱饵载体pPsSWEET15::HIS3以及菜用豌豆cDNA文库,库容为3.056×10^(8)CFU,插入片段平均长度约1000 bp。研究结果为后续筛选PsSWEET15基因上游转录调控因子并探究PsSWEET15基因调控机制提供了基础。SWEETs(Sugars will eventually be exported transporters)are one class of important sugar transporters discovered in recent years,which play important roles in regulating the transport and distribution of sugar in plants.In vegetable pea,the reports focusing on the gene function and regulatory mechanism of PsSWEETs are stil rare.In present study,the 1182 bp promoter region of PsSWEET15 was cloned.Acting element analyses revealed that the promoter of PsSWEET15 harbored the seed specific expression element of GCN4 motif as well as the light,anaerobic induction,ethylene and gibberellin environmental stress responsive elements and the transcription factor binding sites of ERF,DOF,MYB,WRKY,NAC,NF-Y and TCP.Further,the yeast one-hybrid bait vector of pPsSWEET15::HIS3 and vegetable pea cDNA library of 3.056×10^(8)CFU in capacity with an average insert size of about 1000 bp were constructed.These results provided the foundation for screening the upstream transcriptional regulators and deciphering the regulation mechanism of PsSWEET15 gene.

关 键 词：菜用豌豆 糖转运蛋白 PsSWEET15启动子 酵母单杂 CDNA文库 

分 类 号：S643.3[农业科学—蔬菜学]