ptc-miR801人工microRNA表达载体构建及功能初步研究  

Construction of Expression Vector of ptc-miR801 Gene and Function Analysis

作　　者：杨惠琴[1,2] 蒋晶[1] 刘明英[1] 乔桂荣[1] 姜彦成[2] 卓仁英[1] 

机构地区：[1]中国林业科学研究院亚热带林业研究所,富阳311400 [2]新疆大学生命科学与技术学院,乌鲁木齐830046

出　　处：《植物研究》2013年第5期599-604,共6页Bulletin of Botanical Research

基　　金：浙江省科技厅重大专项(2010C12010)"重要生态经济树种转基因平台建立及耐盐转基因研究"

摘　　要：在构建盐胁迫下青杨microRNA文库中发现了ptc-miR801,为探索植物在盐胁迫条件下ptc-miR801参与胁迫应答的机制,本实验构建了植物表达载体pCAM2300-ami801,经根癌农杆菌EHA105介导、花序侵染法获得拟南芥转基因植株。RT-PCR半定量结果显示ptc-miR801可以在转基因拟南芥中超表达且NaCl胁迫下ptc-miRNA801转基因植株种子萌发率和根长显著高于野生型,说明ptc-miR801超表达增强了转基因拟南芥耐盐性。该试验为进一步研究miR801在杨树胁迫应答机制中的作用奠定基础。ptc-miR801 was identified from P. cathayana by microRNA library sequence, and it had been confirmed that ptc-miRS01 was induced by salt stress. The putative targets were NAC-domain protein and Scarecrow-like gene, which were shown to play important roles in response to salt stress. To investigate the function of ptc-miR801, the artificial miR801 plant expression vector was constructed and transformed into Arabidopsis thaliana. Transformed plants were confirmed by PCR and RT-PCR. The results showed that amiRNA transcript level was markedly improved in transgenic plant compared with non-transgenic plants and the overexpression of ptc-miR801 could enhance the salt tolerance of transgenic plants. Our results would help to clarify the molecular mechanism of P. cathayana under salt stress.

关 键 词：MIRNA 表达载体 拟南芥 耐盐 

分 类 号：S287[农业科学—农业水土工程]