草业科学2017，Vol.34Issue(5)：988-996,9.DOI:10.11829/j.issn.1001-0629.2016-0571
马蔺重金属ATP酶基因HMA2的RNAi载体构建及其遗传转化
Construction of RNAi expression vector of heavy metal ATPases gene HMA2 and genetic transformation in Iris lactea
摘要
Abstract
Heavy metal ATPases HMA2 mediated Cd long-distance transport plays important roles in the distribution and detoxification of Cd2+ in plants.However,the molecular mechanism underlying Cd2+ long-distance transport in Iris lactea remains unknown.Total RNA was extracted from the roots of I.lactea to obtain a fragment of IlHMA2 by RT PCR.Then RNAi expression vector pARTG1G2 with inverted repeats of HMA2 driven by the promoter CaMV35S was constructed based on the intermediate vector pHANNIBAL and the plant expression vector pART27 by restriction enzyme digestion and ligation.Moreover,the recombinant plasmid was transformed into agrobacterium GV3101 by freezing and thawing method,and then was transformed into I.lactea to obtain IlHMA2-RNAi lines.This would be a good basis for clarifying the IlHMA2 mediated Cd2+ long distance transport in the adaptation of Ⅰ.lactea to Cde+ contaminated environments.关键词
马蔺/重金属ATPase酶/镉富集与耐受性/RNAi载体构建Key words
Iris lactea/heavy metal ATPases/Cadmium accumulation and tolerance/RNAi vector construction分类
农业科技引用本文复制引用
张亚楠,蔺银鼎,孟林,毛培春,田小霞,郭强..马蔺重金属ATP酶基因HMA2的RNAi载体构建及其遗传转化[J].草业科学,2017,34(5):988-996,9.基金项目
北京市自然科学基金(6152008) （6152008）
北京市农林科学院科技创新能力建设专项(KJCX20140103、KJCX20170110) （KJCX20140103、KJCX20170110）
