香蕉长链非编码RNA Malnc2310 启动子区域分析及上游调控因子筛选  

Analysis of long noncoding RNA Malnc2310 promoter from banana(Musa spp.)and identification of its regulators

作　　者：李文彬[1] 于晓玲[1] 闫羿辰 孙建波[1] LI Wenbin;YU Xiaoling;YAN Yichen;SUN Jianbo(Institute of Tropical Bioscience and Biotechnology/Key Laboratory of Biology and Genetic Resources of Tropical Crops,Ministry of Agriculture and Rural Affairs/Hainan Academy of Tropical Agricultural Resource,Chinese Academy of Tropical Agricultural Sciences,Haikou 571101,China;Qiongtai Normal University,Haikou 571127,China)

机构地区：[1]中国热带农业科学院热带生物技术研究所/农业农村部热带作物生物学与遗传资源利用重点实验室/海南省热带农业资源研究院,海南海口571101 [2]琼台师范学院,海南海口571127

出　　处：《南京农业大学学报》2023年第2期287-296,共10页Journal of Nanjing Agricultural University

基　　金：海南省基础与应用基础研究计划(自然科学领域)高层次人才项目(2019RC292);国家重点研发计划项目(2019YFD1000204)。

摘　　要：[目的]为了深入研究香蕉长链非编码RNA Malnc2310的功能和特性,对Malnc2310的启动子顺式作用元件、病原菌响应模式及结合因子进行了初步的研究。[方法]将Malnc2310全长启动子P1(1625 bp)及从5′端到3′端序列互补的启动子片段P2(400 bp)、P3(800 bp)和P4(425 bp)转入拟南芥,研究其在高盐和尖孢镰刀菌粗毒素胁迫下的响应模式;以Malnc2310启动子P1为诱饵,利用酵母单杂交技术筛选获得与该启动子结合的转录因子,并确定启动子的核心功能区。[结果]Malnc2310的启动子包含多个与光照、病原菌、激素响应相关的顺式作用元件;所有启动子片段在尖孢镰刀菌粗毒素处理的GUS活性比高盐处理后更高,且P3驱动下GUS的活性与全长P1驱动下的活性相似;通过酵母单杂文库筛选到128个潜在的结合因子序列,其中双C2H2结构的锌指蛋白ZFP-WIP2经点对点酵母单杂交验证可与P1和P3互相结合。[结论]Malnc2310的启动子对尖孢镰刀菌粗毒素比对高盐处理更敏感,筛选到1个锌指蛋白ZFP-WIP2可与启动子核心功能区(425~1225 bp)结合。[Objectives]To understand the characteristic and the function of Malnc2310,the cis-acting elements on the promoter,the response to Fusarium crude toxin and the potential binding factors were investigated through genetic methods.[Methods]The response of promoter P1(1625 bp)and three complement fragments including P2(400 bp),P3(800 bp)and P4(425 bp),to high salinity and fusarium crude toxin were investigated through the Arabidopsis transformation.The potential binding factors on the promoter P1 and the core binding domain were studied through yeast one-hybrid screening system.[Results]The promoter of Malnc2310 contained many cis-elements,mainly including constitutive regulatory elements,phytohormone response elements,pathogen and salicylic regulatory elements,and light responsive elements through online database analysis.The average activities of GUS in all promoter fragments were higher under the Fusarium crude toxin than high salinity treatment.Moreover,the activities of GUS under P3 were similar with those under P1 during different treatments.One zinc finger protein with two C2H2 domains called ZFP-WIP2 was verified to bind to P1 and P3 through one-hybridization system.[Conclusions]This research verifies the promoter of Malnc2310 is more responsive to Fusarium crude toxin than high salinity,and one zinc finger protein ZFP-WIP2 can bind to the promoter core domain(425-1225 bp).

关 键 词：香蕉 Malnc2310 启动子 顺式作用元件 调控因子 

分 类 号：S668.1[农业科学—果树学] Q943.2[农业科学—园艺学]