香蕉MaAQP1启动子诱饵载体及干旱胁迫酵母单杂交cDNA文库的构建  

Constructions of Banana MaAQP1 Bait Vector and Drought-resistance cDNA Library

作　　者：许奕[1,2] 李羽佳 魏卿[1] 王安邦[1] 王笑一[1] 宋顺 李敬阳[1] XU Yi;LI Yujia;WEI Qing;WANG Anbang;WANG Xiaoyi;SONG Shun;LI Jingyang(Haikou Experimental Station,Chinese Academy of Tropical Agricultural Sciences/Hainan Key Laboratory of Banana Genetic Improvement,Haikou,Hainan 571101,China;Hainan Key Laboratory for Biosafety Monitoring and Molecular Breeding in Off-Season Reproduction Regions,Sanya,Hainan 572000,China)

机构地区：[1]中国热带农业科学院海口实验站/海南省香蕉遗传改良重点实验室,海南海口571101 [2]海南省南繁生物安全与分子育种重点实验室,海南三亚572000

出　　处：《福建农业学报》2020年第10期1078-1085,共8页Fujian Journal of Agricultural Sciences

基　　金：中国热带农业科学院基本科研业务费专项项目(1630092020002);国家自然科学基金项目(31501371);中央级公益性科研院所基本科研业务专项项目(1630092020007)

摘　　要：【目的】香蕉MaAQP1能够提高植物的耐旱性,研究香蕉MaAQP1的相关特性可为了解其干旱胁迫响应机制奠定基础。【方法】通过克隆MaAQP1的启动子,将启动子构建到pHIS2诱饵质粒上,转化酵母菌构建诱饵表达载体,同时构建干旱胁迫的香蕉(Musa acuminat L. AAA group cv. Brazilian)cDNA文库。【结果】克隆获得1 362 bp的启动子序列,通过分析其顺式作用元件,结果显示启动子序列中共有72个顺式作用元件,包括了TATA-box和CAAT-box核心元件,ABA响应元件、MYB元件、MYC元件、ERE元件、MeJA响应元件、光响应元件以及分生组织响应元件等;成功构建了MaAQP1诱饵载体和干旱胁迫条件下的cDNA文库,文库库容为1.25×107 CFU,插入片段平均在1 200 bp左右。【结论】本研究克隆获得了香蕉MaAQP1启动子并构建了干旱胁迫酵母单杂交cDNA文库,为下一步运用酵母单杂交筛选MaAQP1互作的转录因子、解析MaAQP1响应干旱胁迫的作用机制奠定了基础。【Objective】 To clone MaAQP1 from a drought stressed banana plant using a constructed bait vector, and establish a cDNA library of the transformed drought-resistance gene from single-hybrids yeast cells. 【 Method】 The promoter of MaAQP1 from a banana plant(Musa acuminat L. AAA group cv. Brazilian) under draught-stress was constructed on the pHIS2 plasmid as the bait to be transformed into yeast cells for the gene cloning. A cDNA library of MaAQP1 in the yeast was subsequently established for the study. 【Result】 The MaAQP1 bait vector was successfully constructed. The sequence of the1 362 bp promoter was cloned and analyzed to show 72 cis-acting TATA-box and CAAT-box core elements as well as elements of MYB, MYC, ERE, and MeJA as well as those of ABA, light, and meristem responses, etc. A drought-resistance cDNA library of 1.25×10~7 CFU in capacity with an average insert size of about 1 200 bp was established. 【Conclusion】 The results provided a basis for screening transcription factors of the cloned MaAQP1 in the yeast single-hybrids to decipher the mechanism of MaAQP1 in response to drought for improving the stress resistance of plants.

关 键 词：香蕉 干旱 MaAQP1 诱饵载体 CDNA文库 

分 类 号：S668.1[农业科学—果树学]