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J. Hefei Univ. Tech. (Nat. Sci.)

合肥工业大学学报(自然科学版)

JOURNAL OF HEFEI UNIVERSITY OF TECHNOLOGY (NATURAL SCIENCE)

Competent Authorities: Ministry of Education of the People's Republic of China

Sponsor by: Hefei University of Technology

ISSN 1000-1646, CN 34-1001/N

Construction of pXB94 vector of Arabidopsis thaliana NHX4 gene and identification of transgenic plants

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拟南芥 NHX4 基因 pXB94 载体的构建及转基因植株的筛选

Journal Information

J. Hefei Univ. Tech. (Nat. Sci.), Oct.2025, Vol.48 No.10: 1378-1382

DOI: 10.3969/j.issn.1003-5060.2025.10.012

Authors

WANG Tong, FAN Tingting, CAO Junxuan

(School of Food and Biological Engineering, Hefei University of Technology, Hefei 230601, China)

Abstract and Keywords

Abstract: Soil salinization is a crucial factor restricting crop yields. To alleviate the inhibitory effect of high-salt environments on crop growth and development, clarifying the gene regulatory mechanisms in response to salt stress is of great significance for agricultural production. Given that the vacuolar membrane transporter NHX4 can enhance plant salt tolerance by storing $ Na^{+} $, and the transcription factor MYBX may positively regulate the expression of NHX4 at the molecular level, this study aims to further verify the regulatory relationship between MYBX and NHX4. The laboratory constructed an overexpression vector of the NHX4 and introduced the recombinant plasmid into Agrobacterium using electroporation. Subsequently, Agrobacterium containing the recombinant plasmid was used to infect the Arabidopsis thaliana mutant myba-1 via the floral dip method. Positive plants were obtained through resistance screening, and homozygous transgenic plants myba-1/35S;NHX4 were further screened out by polymerase chain reaction (PCR) identification, providing experimental materials for subsequent verification of the relationship between MYBX and NHX4. This study establishes a foundation for further clarifying the molecular interaction mechanism between MYBX and NHX4.

Keywords: Arabidopsis thaliana; NHX4 gene; vector construction; transgenic plants; salt stress

作者信息

王彤，樊婷婷，曹君璇

（合肥工业大学食品与生物工程学院，安徽合肥230601）

摘要和关键词

摘要: 土壤盐碱化是制约农作物产量的重要因素,高盐环境对作物生长发育有抑制作用,因此研究响应盐胁迫的基因调控机制具有重要意义。鉴于液泡膜转运蛋白NHX4可通过储存 $ Na^{+} $增强植物耐盐性和转录因子MYBX可能在分子水平正向调控NHX4的表达,为了验证MYBX与NHX4之间的调控关系,文章构建NHX4基因的过表达载体,采用电击法将重组质粒导入农杆菌;利用浸花法将含重组质粒的农杆菌侵染拟南芥突变体myba-1,经抗性筛选获得阳性植株,通过聚合酶链式反应(polymerase chain reaction,PCR)鉴定筛选出纯合的转基因植株myba-1/35S;NHX4,为后续验证MYBX与NHX4的关系提供实验材料。该研究为进一步明确MYBX与NHX4的分子互作机制奠定了基础。

关键词: 拟南芥；NHX4基因；载体构建；转基因植株；盐胁迫

Funding

国家自然科学基金资助项目（31872803）；中央高校基本科研业务费专项资金资助项目（JZ2018HGTB0248）

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