Exploration on the Functions of IlNRAMP5 Mediating the Cd2+ uptake in Iris lactea

doi:10.11733/j.issn.1007-0435.2023.06.005

Acta Agrestia Sinica ›› 2023, Vol. 31 ›› Issue (6): 1640-1647.DOI: 10.11733/j.issn.1007-0435.2023.06.005

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Exploration on the Functions of IlNRAMP5 Mediating the Cd²⁺ uptake in Iris lactea

LI Qi-dong^1,2, LI Cui², CHEN Zhi-min², ZHANG Li-xiang¹, GUO Qiang²

1. College of Agricultural Resources and Environment, Heilongjiang University, Harbin, Heilongjiang Province 150080, China;
2. Institute of Grassland, Flowers, and Ecology, Beijing Academy of Agriculture and Forestry Sciences, Beijing 100097, China

Received:2023-01-05 Revised:2023-03-08 Online:2023-06-15 Published:2023-06-30

马蔺IlNRAMP5介导Cd²⁺吸收功能初探

李启东^1,2, 李翠², 陈治民², 张李香¹, 郭强²

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

通讯作者: 张李香,E-mail:zhanglixiang@hlju.edu.cn;郭强,E-mail:guoqiang@baafs.net.cn
作者简介:李启东(1996-)，男，朝鲜族，黑龙江延寿人，硕士研究生，主要从事草类植物逆境生理研究，E-mail：1148135691@qq.com
基金资助:
国家自然科学基金项目(31971752)资助

Abstract

Abstract: NRAMP5 is a highly conserved transmembrane heavy metal transporter,which plays an important role in mediating Cd²⁺ uptake. However,it is unclear whether IlNRAMP5 is involved in the process of Cd²⁺ 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 Cd²⁺ 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 Cd²⁺ uptake.

Key words: Iris lactea, IlNRAMP5, Genetic transformation, RNA interference, Cadmium stress

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Cd²⁺吸收中扮演着重要的角色。然而，有关IlNRAMP5在马蔺Cd²⁺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Cd²⁺浓度较野生型显著降低。为进一步解析IlNRAMP5在马蔺Cd²⁺吸收中的作用机制奠定了工作基础。

关键词: 马蔺, NRAMP5, 遗传转化, RNA干扰, 镉胁迫

CLC Number:

Q945.79

LI Qi-dong, LI Cui, CHEN Zhi-min, ZHANG Li-xiang, GUO Qiang. Exploration on the Functions of IlNRAMP5 Mediating the Cd²⁺ uptake in Iris lactea[J]. Acta Agrestia Sinica, 2023, 31(6): 1640-1647.

李启东, 李翠, 陈治民, 张李香, 郭强. 马蔺IlNRAMP5介导Cd²⁺吸收功能初探[J]. 草地学报, 2023, 31(6): 1640-1647.

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Exploration on the Functions of IlNRAMP5 Mediating the Cd²⁺ uptake in Iris lactea

马蔺IlNRAMP5介导Cd²⁺吸收功能初探

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