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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

Targeted mutation of tomato WRKY3 gene based on CRISPR/Cas9 technology

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基于CRISPR/Cas9技术靶向突变番茄WRKY3基因

Journal Information

J. Hefei Univ. Tech. (Nat. Sci.), Jan.2026, Vol.49 No.1: 73-79

DOI: 10.3969/j.issn.1003-5060.2026.01.011

Authors

SONG Qianqian, LIU Zhouyuan, TANG Xiaofeng

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

Abstract and Keywords

Abstract: WRKY transcription factors play important roles in plant growth and development, secondary metabolite synthesis, and response to biotic and abiotic stresses. In order to investigate the function and mechanism of action of WRKY3 in plant adversity response, this study analyzed the structural domains and evolutionary trees of WRKY3 in different species by bioinformatic method and found that WRKY3 was highly conserved. TomExpress database was used to analyze WRKY3 expression level in different tissues of tomato. The result showed that WRKY3 was expressed in various tissues, and the highest in the fruit at the red-fruit stage. The CRISPR knockdown vector of WRKY3 gene was constructed. WRKY3 transgenic tomato plants were obtained by using Agrobacterium-mediated transformation and identified by polymerase chain reaction (PCR) and sequencing. In this study, the stable genetic knockout plants of tomato WRKY3 was obtained, which lays the foundation for the investigation of the function of WRKY3 in plant growth and development and fruit ripening under adversity stress.

Keywords: tomato(Solanum lycopersicum); WRKY3 gene; gene editing; CRISPR/Cas9 technology; vector construction

作者信息

宋茜茜，刘周圆，唐晓凤

（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: WRKY 转录因子在植物生长发育、次生代谢产物合成以及对生物和非生物胁迫的响应等方面发挥重要作用。为探究 WRKY3 在植株逆境应答中的功能及作用机制，文章通过生物信息法对不同物种 WRKY3 的结构域及进化树进行分析，发现 WRKY3 具有较高的保守性；TomExpress 数据库显示 WRKY3 在各种组织中均有表达，且在红果期果实中表达量最高；并构建 WRKY3 基因的 CRISPR 敲除载体，利用农杆菌介导法转化番茄获得 WRKY3 基因的转基因番茄材料，经聚合酶链式反应（polymerase chain reaction，PCR）及测序鉴定发现，已成功获得敲除 WRKY3 基因的转基因番茄植株。该研究获得了番茄 WRKY3 基因敲除突变体稳定遗传株系，为探究 WRKY3 在逆境胁迫下植物的生长发育及果实成熟中所发挥的功能奠定了基础。

关键词: 番茄；WRKY3基因；基因编辑；CRISPR/Cas9技术；载体构建

Funding

安徽省自然科学基金资助项目（2208085MC66）

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