Biotechnology Bulletin ›› 2022, Vol. 38 ›› Issue (2): 263-268.doi: 10.13560/j.cnki.biotech.bull.1985.2021-0635

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Establishment of an Efficient in planta Transformation Method for Camellia sinensis

ZHOU Cheng-zhe1,2,3(), CHANG Xiao-jun1,3, ZHU Chen1,2,3, CHENG Chun-zhen1,3, CHEN Yu-kun1,3, LAI Zhong-xiong1,3, LIN Yu-ling1,3, GUO Yu-qiong1,2()   

  1. 1. College of Horticulture,Fujian Agriculture and Forestry University,Fuzhou 350002
    2. Key Laboratory of Tea Science in Universities of Fujian Province,Institute of Tea Industry Research,Fujian Agriculture and Forestry University,Fuzhou 350002
    3. Institute of Horticultural Biotechnology,Fujian Agriculture and Forestry University,Fuzhou 350002
  • Received:2021-05-14 Online:2022-02-26 Published:2022-03-09
  • Contact: GUO Yu-qiong E-mail:chengzhechou@foxmail.com;guoyq828@163.com

Abstract:

To address the lack of an efficient and stable genetic transformation system for tea trees,an in planta transformation transgenic method based on Agrobacterium tumefaciens-mediated transformation of tea trees was established,laying a solid foundation for gene function research and germplasm innovation of tea tree. The seedlings of tea cultivar ‘Tieguanyin’,‘Baiye1’,and ‘Longjing43’ were used as receptor plants,and the apical bud and axillary bud were removed. Using the A. tumefaciens broth infecting the wounds of the seedling,regenerative buds were obtained through resistance screening. After molecular biological identification,transgenic plants were obtained by short-ear cutting method. The results demonstrated that the GUS(-glucuronidase)and HYG(hyglymycin)labeled genes were tested to be positive with multiple PCR,and the PCR product sequence was consistent with the labeled gene sequence. The transformation efficiency of ‘Tieguanyin’,‘Baiye1’,and ‘Longjing43’ was 8.14%,2.99%,and 2.53%,respectively. This study establishes an in planta transformation method of tea plant that does not rely on tissue culture,and the transformation protocol characterizes of simple operation,high transformation rate,low cost,and short experiment cycle.

Key words: Camellia sinensis, in planta transformation, agrobacterium tumefaciens