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Construction of 35S:SOS2/WRKY12-OE5 transgenic line of Arabidopsis thaliana

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拟南芥 35S:SOS2/WRKY12-OE5 转基因株系的构建

Journal Information

J. Hefei Univ. Tech. (Nat. Sci.), Sept.2025, Vol.48 No.9: 1213-1217

DOI: 10.3969/j.issn.1003-5060.2025.09.009

Authors

MA Qian, FAN Tingting, WU Xi, JIA Yafeng, ZHANG Zhen, SONG Hui

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

Abstract and Keywords

Abstract: To investigate the genetic relationship between SOS2 and WRKY12 in response to salt stress in Arabidopsis thaliana, the cDNA produced by reverse transcription of wild-type Arabidopsis thaliana RNA was used as a template to amplify the SOS2 fragment, then pART27 and SOS2 fragment were double digested using Kpn I and Xho I, and pART27 was ligated to SOS2 fragment using T4 ligase. The ligated products were transformed into the prepared Escherichia coli receptor state and coated onto resistance plates, and single colonies were picked for polymerase chain reaction (PCR) identification and sequencing of positive strains. Fragments of recombinant plasmids were extracted from the correctly sequenced positive monoclonal strains, transformed into the prepared Agrobacterium receptor state GV3101, and coated onto the double antibody plates. Monoclonal colonies were selected for PCR identification and sequencing, thereby obtaining positive monoclonal colonies. Floral dip transformation was performed on wild-type Arabidopsis thaliana, followed by screening and identification of positive plants to obtain homozygous 35S:SOS2/WT. The resulting 35S:SOS2 and WRKY12-OE5 transgenic lines were then crossed, and through screening and identification, the 35S:SOS2/WRKY12-OE5 transgenic line was obtained, laying the foundation for further investigation into the genetic relationship between SOS2 gene and WRKY12 gene.

Keywords: Arabidopsis thaliana; WRKY12 gene; SOS2 gene; recombinant plasmids; transgenic plants

作者信息

马倩，樊婷婷，吴席，贾亚峰，张震，宋慧

（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: 为探究 SOS2 与 WRKY12 在拟南芥响应盐胁迫时的遗传关系,文章以野生型拟南芥 RNA 反转录产生的 cDNA 为模板扩增出 SOS2 片段,将质粒 pART27 和 SOS2 片段利用 Kpn I 和 Xho I 进行双酶切,使用 T4 连接酶将 pART27 与 SOS2 片段连接。连接产物转化至制备好的大肠杆菌感受态中,涂布至抗性板上,挑取单菌落进行阳性菌株的聚合酶链式反应(polymerase chain reaction, PCR)鉴定与测序。从测序正确的单克隆阳性菌株中提取重组质粒,转化至制备好的农杆菌感受态 GV3101 中,并涂布至双抗平板上,挑取单克隆菌株进行 PCR 鉴定与测序,进而获得阳性单克隆菌落。浸花侵染拟南芥野生型,筛选鉴定阳性植株以获得纯合的 35S:SOS2/WT,再将获得的 35S:SOS2 和 WRKY12-OE5 转基因株系进行杂交,通过筛选鉴定获得 35S:SOS2/WRKY12-OE5 转基因株系,为进一步研究 SOS2 基因与 WRKY12 基因之间的遗传关系奠定基础。

关键词: 拟南芥；WRKY12基因；SOS2基因；重组质粒；转基因植株

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（JZ2023HGQB0167）

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