Acta Agrestia Sinica ›› 2022, Vol. 30 ›› Issue (6): 1388-1395.DOI: 10.11733/j.issn.1007-0435.2022.06.009

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Cloning and Expression Analysis of SgNramp1 and SgNramp2 in Stylosanthes guianensis

ZOU Xiao-yan1, WANG Lin-jie1, HUANG Jie1, LUO Li-juan1, LIU Guo-dao2, CHEN Zhi-jian1,2   

  1. 1. College of Tropical Crops, Hainan University, Haikou, Hainan Province 570228, China;
    2. Institute of Tropical Crop Genetic Resources, Chinese Academy of Tropical Agriculture Sciences and Key Laboratory of Crop Gene Resources and Germplasm Enhancement in Southern China, Ministry of Agriculture and Rural Affair, Haikou, Hainan Province 571101, China
  • Received:2021-11-17 Revised:2022-01-04 Published:2022-07-05

柱花草SgNramp1SgNramp2基因克隆与表达分析

邹晓燕1, 王林杰1, 黄杰1, 罗丽娟1, 刘国道2, 陈志坚1,2   

  1. 1. 海南大学热带作物学院, 海南 海口 570228;
    2. 中国热带农业科学院热带作物品种资源研究所/农业农村部华南作物基因资源与种质创制重点实验室, 海南 海口 571101
  • 通讯作者: 陈志坚,E-mail:jchen@scau.edu.cn
  • 作者简介:邹晓燕(1994-),女,甘肃兰州人,硕士研究生,主要从事热带牧草抗逆研究,E-mail:243710275@qq.com
  • 基金资助:
    国家自然科学基金项目(31861143013);财政部和农业农村部:国家现代农业产业技术体系(CARS-34)资助

Abstract: To explore the natural resistance associated macrophage proteins (Nramps) function as metal ion transporters that involve in the transportation,translocation,and distribution of metal ions in plants. In this study,SgNramp1 and SgNramp2 genes were cloned in Stylosanthes guianensis (stylo) using PCR method,and the expressions of SgNramp1 and SgNramp2 were further detected. The result showed that the full lengths of SgNramp1 and SgNramp2 were 1 654 and 1 716 bp,respectively. Both SgNramp1 and SgNramp2 contained 12 transmembrane domains. Tissue expression analysis showed that SgNramp1 was mainly expressed in roots,while SgNramp2 displayed specific expression in leaves. qRT-PCR analysis showed that SgNramp1 and SgNramp2 exhibited different responses to deficient or excess metal stresses. Among them,manganese deficiency (-Mn) inhibited the expressions of SgNramp1 and SgNramp2 in both leaves and roots. Iron deficiency (-Fe) enhanced and inhibited the expressions of SgNramp2 in leaves and roots,respectively. Furthermore,the expressions of SgNramp1 and SgNramp2 were regulated by at least one excess metal stress in leaves or roots. In addition,both SgNramp1 and SgNramp2 were up-regulated by cadmium (Cd) treatment in roots. Both aluminum (Al) and lanthanum (La) treatments inhibited SgNramp1 but increased SgNramp2 transcripts in roots. These results together suggest that SgNramp1 and SgNramp2 may be involved in metal ion transport.

Key words: Stylosanthes guianensis, Nramp, Gene expression, Metal transporter, Element stress

摘要: 为了探究植物天然抗性相关巨噬蛋白(Nramp)在金属元素吸收、转运和分配等方面的重要作用,本研究以柱花草(Stylosanthes guianensis)为材料,采用PCR方法克隆了柱花草SgNramp1/2基因,并分析了该基因的表达模式。结果表明:SgNramp1SgNramp2基因全长分别为1 654 bp和1 716 bp,且均含有12个跨膜结构域。基因表达分析发现,SgNramp1在柱花草根部的表达高于叶部,而SgNramp2则在叶中特异性表达;金属元素缺乏和过量胁迫对SgNramp1/2基因表达的影响有所差异,缺锰(-Mn)处理抑制了SgNramp1/2基因在根和叶中的表达,而缺铁(-Fe)处理则增强了SgNramp2基因在叶中的表达,但抑制了其在根中的表达,SgNramp1/2基因在根和叶中的表达至少受到一种元素过量胁迫的影响。此外,金属镉(Cd)处理均增强了SgNramp1/2基因在柱花草根中的表达,而铝(Al)和镧(La)处理抑制了SgNramp1基因的表达,但增强了SgNramp2基因的表达。本研究结果表明SgNramp1/2基因可能参金属离子的转运。

关键词: 柱花草, Nramp, 基因表达, 金属转运蛋白, 元素胁迫

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