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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 and identification of AtRBOHD gene expression vector in Arabidopsis thaliana

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拟南芥 AtRBOHD 基因表达载体构建及鉴定分析

Journal Information

J. Hefei Univ. Tech. (Nat. Sci.), Sept.2025, Vol.48 No.9: 1218-1221, 1242

DOI: 10.3969/j.issn.1003-5060.2025.09.010

Authors

YE Min, FAN Tingting, SONG Hui, ZHANG Jing, CHEN Yifan

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

Abstract and Keywords

Abstract: As an important NADPH oxidase gene in plants, AtRBOHD gene can generate reactive oxygen species by catalyzing oxygen in response to adversity stress, but whether it participates in the regulation of plant heavy metal cadmium stress is unclear. In this study, using Arabidopsis thaliana as experimental material, the promoter region of AtRBOHD gene was cloned by polymerase chain reaction (PCR) technology, and the ProAtRBOHD:GUS fusion expression vector was successfully constructed. The fusion expression vector was transformed into wild-type Arabidopsis thaliana by floral dip method, and homozygous and positive transgenic plants were screened out by resistance screening, PCR identification and resistance isolation identification. Furthermore, cadmium stress was performed on the obtained transgenic materials, and it was found that the GUS activity of the treated transgenic materials increased. This study provides an important basis for the preliminary research on the function and transcription level of AtRBOHD gene under cadmium stress.

Keywords: Arabidopsis thaliana; cadmium stress; AtRBOHD gene; vector construction; GUS staining

作者信息

叶敏，樊婷婷，宋慧，张景，陈逸凡

（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: AtRBOHD 基因作为植物体内重要的 NADPH 氧化酶基因, 可将氧催化生成活性氧以响应逆境胁迫, 但其是否参与植物重金属镉胁迫的调控尚不清楚。文章以拟南芥为实验材料, 利用聚合酶链式反应(polymerase chain reaction, PCR)技术克隆出 AtRBOHD 基因的启动子区域, 成功构建 ProAtRBOHD:GUS 融合表达载体; 并通过浸花法将该融合表达载体转化到野生型拟南芥中, 通过抗性筛选、PCR 鉴定、抗性分离鉴定筛选出纯合阳性转基因植株。进一步对获得的转基因材料进行镉胁迫处理, 发现经处理后的转基因材料 GUS 活性提高。为初步研究镉胁迫下 AtRBOHD 基因功能和转录水平变化提供重要依据。

关键词: 拟南芥；镉胁迫；AtRBOHD基因；载体构建；GUS染色

Funding

安徽省自然科学基金资助项目（1508085QC50）；中央高校基本科研业务费专项资金资助项目（JZ2023HGQB0167）

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