Fast,simple,efficient Agrobacterium rhizogenes-mediated transformation system to non-heading Chinese cabbage with transgenic roots

作     者：Huiyu Wang Yushan Zheng Qian Zhou Ying Li Tongkun Liu Xilin Hou Huiyu Wang;Yushan Zheng;Qian Zhou;Ying Li;Tongkun Liu;Xilin Hou

作者机构：State Key Laboratory of Crop Genetics&Germplasm EnhancementKey Laboratory of Biology and Genetic Improvement of Horticultural Crops(East China)Ministry of Agriculture and Rural Affairs of ChinaEngineering Research Center of Germplasm Enhancement and Utilization of Horticultural CropsMinistry of Education of ChinaNanjing Agricultural UniversityNanjingJiangsu 210095China Nanjing Suman Plasma Engineering Research InstituteNanjing Agricultural UniversityNanjingJiangsu 210095China 

出 版 物：《Horticultural Plant Journal》 (园艺学报（英文版）)

年 卷 期：2024年第10卷第2期

页      面：450-460页

核心收录：

学科分类：09[农学] 0902[农学-园艺学] 090202[农学-蔬菜学] 

基　　金：funded by National Natural Science Foundation of China (Grant No.32072575) Postgraduate Research & Practice Innovation Program of Jiangsu Province (Grant No.KYCX20_0588) National Vegetable Industry Technology System (Grant No.CARS-23-A16) 

主　　题：Agrobacterium rhizogenes Non-heading Chinese cabbage Transgenic roots Composite plant Transformation efficiency 

摘      要：Non-heading Chinese cabbage, a variety of Brassica campestris, is an important vegetable crop in the Yangtze River Basin of China. However,the immaturity of its stable transformation system and its low transformation efficiency limit gene function research on non-heading Chinese cabbage. Agrobacterium rhizogenes-mediated(ARM) transgenic technology is a rapid and effective transformation method that has not yet been established for non-heading Chinese cabbage plants. Here, we optimized conventional ARM approaches(one-step and two-step transformation methods) suitable for living non-heading Chinese cabbage plants in nonsterile environments. Transgenic roots in composite non-heading Chinese cabbage plants were identified using phenotypic detection, fluorescence observation, and PCR analysis. The transformation efficiency of a two-step method on four five-day-old non-heading Chinese cabbage seedlings(Suzhouqing, Huangmeigui, Wuyueman, and Sijiu Caixin) was 43.33%-51.09%, whereas using the stout hypocotyl resulted in a transformation efficiency of 54.88% for the 30-day-old Sijiu *** one-step method outperformed the two-step method;the transformation efficiency of different varieties was above 60%, and both methods can be used to obtain transgenic roots for functional studies within one month. Finally, optimized ARM transformation methods can easily,quickly, and effectively produce composite non-heading Chinese cabbage plants with transgenic roots, providing a reliable foundation for gene function research and non-heading Chinese cabbage genetic improvement breeding.