马蔺IlNRAMP5介导Cd^(2+)吸收功能初探  

Exploration on the Functions of IlNRAMP5 Mediating the Cd^(2+)uptake in Iris lactea

作　　者：李启东 李翠[2] 陈治民 张李香[1] 郭强[2] LI Qi-dong;LI Cui;CHEN Zhi-min;ZHANG Li-xiang;GUO Qiang(College of Agricultural Resources and Environment,Heilongjiang University,Harbin,Heilongjiang Province 150080,China;Institute of Grassland,Flowers,and Ecology,Beijing Academy of Agriculture and Forestry Sciences,Beijing 100097,China)

机构地区：[1]黑龙江大学,现代农业与生态环境学院,黑龙江哈尔滨150080 [2]北京市农林科学院,草业花卉与景观生态研究所,北京100097

出　　处：《草地学报》2023年第6期1640-1647,共8页Acta Agrestia Sinica

基　　金：国家自然科学基金项目(31971752)资助。

摘　　要：天然抗性相关巨噬细胞蛋白NRAMP5是一类高度保守的跨膜重金属转运蛋白,在介导Cd^(2+)吸收中扮演着重要的角色。然而,有关IlNRAMP5在马蔺Cd^(2+)吸收过程中是否起重要作用至今仍不清楚。本研究以马蔺(Iris lactea)根总RNA为模板,采用RT-PCR方法,获得了IlNRAMP5基因片段。采用In-Fusion无缝连接方法,构建了CaMV35 s启动子驱动的含有内含子发卡结构正反向重复序列的RNA干扰植物表达载体pART27-pHANNIBAL-IlNRAMP5-RNAi(PPIR)。并通过农杆菌介导将PPIR转入马蔺,经抗性筛选及分子检测获得IlNRAMP5-RNAi遗传转化株系。初步发现,镉胁迫下其IlNRAMP5-RNAi株系Cd^(2+)浓度较野生型显著降低。为进一步解析IlNRAMP5在马蔺Cd^(2+)吸收中的作用机制奠定了工作基础。NRAMP5 is a highly conserved transmembrane heavy metal transporter,which plays an important role in mediating Cd^(2+)uptake.However,it is unclear whether IlNRAMP5 is involved in the process of Cd^(2+)uptake.Here,the total RNA of Iris lactea root was used as a template,and IlNRAMP5 gene fragment was obtained by RT-PCR,RNA interference plant expression vector pART27-pHANNIBAL-IlNRAMP5-RNAi(PPIR)with intron hairpin structure driven by CaMV35 s promoter was constructed using In-Fusion method.Subsequently,PPIR was transferred into I.lactea by Agrobacterium,and IlNRAMP5-RNAi transformed lines were obtained by resistance screening and molecular detection.The preliminary finding is that contents of Cd^(2+)in IlNRAMP5-RNAi lines were significantly lower than that of wild type under cadmium stress.This further lays a working foundation for dissecting the role of IlNRAMP5 in Cd^(2+)uptake.

关 键 词：马蔺 NRAMP5 遗传转化 RNA干扰 镉胁迫 

分 类 号：Q945.79[生物学—植物学]