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Research Article | Open Access

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

Huiyu Wanga, Yushan Zhenga, Qian Zhoua, Ying Lia,b, Tongkun Liua( ), Xilin Houa,b( )
State Key Laboratory of Crop Genetics & Germplasm Enhancement, Key Laboratory of Biology and Genetic Improvement of Horticultural Crops (East China), Ministry of Agriculture and Rural Affairs of China, Engineering Research Center of Germplasm Enhancement and Utilization of Horticultural Crops, Ministry of Education of China, Nanjing Agricultural University, Nanjing, Jiangsu 210095, China
Nanjing Suman Plasma Engineering Research Institute, Nanjing Agricultural University, Nanjing, Jiangsu 210095, China

Peer review under responsibility of Chinese Society of Horticultural Science (CSHS) and Institute of Vegetables and Flowers (IVF), Chinese Academy of Agricultural Sciences (CAAS)

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Abstract

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 Caixin. The 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.

Keywords

Transformation efficiencyAgrobacterium rhizogenesNon-heading Chinese cabbageTransgenic rootsComposite plant

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Horticultural Plant Journal
Volume 10 Issue 2,
March 2024
Pages 450-460
DOI: 10.1016/j.hpj.2023.03.018
Cite this article:
Wang H, Zheng Y, Zhou Q, et al. Fast, simple, efficient Agrobacterium rhizogenes-mediated transformation system to non-heading Chinese cabbage with transgenic roots. Horticultural Plant Journal, 2024, 10(2): 450-460. https://doi.org/10.1016/j.hpj.2023.03.018

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Received: 16 March 2023
Revised: 17 May 2023
Accepted: 20 July 2023
Published: 26 September 2023
© 2024 Chinese Society for Horticultural Science (CSHS) and Institute of Vegetables and Flowers (IVF), Chinese Academy of Agricultural Sciences (CAAS).

This is an open access article under the CC BY-NC-ND license (http://creativecommons.org/licenses/by-nc-nd/4.0/).
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