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Journal of Tea Science ›› 2003, Vol. 23 ›› Issue (1): 6-10.doi: 10.13305/j.cnki.jts.2003.01.002

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Optimization of Agrobacterium-mediated and Particle Bombardment-mediated Transformation Systems in Tea Plant (Camellia sinensis)

WU Shan, LIANG Yue-rong, LU Jian-liang, KIM Hyesuk, WU Ying   

  1. Tea Research Institute, Zhejiang University;Hangzhou 310029, China
  • Received:2002-10-12 Revised:2003-01-06 Online:2003-05-25 Published:2019-09-16

Abstract: When tea explants were pre-cultured on medium containing PVP 16g/L for 2-3d before infiltrated by Agrobacterium tumefaciens the transformation frequency was increased. Optimum conditions for preparing bombardment micro-particle were as follows: 10βµl tungsten suspension (60βmg/1ml), 1.6βµl plasmid DNA (1βµg/βµl), 4βµl spermidin (0.1βmol/L) and 15βµl CaCl2 (l2.5βmol/L). Ethanol was used to dilute the final volume up to 48βµl. The loading volume for each bombardment was 8-10βµl. The survival rate of resistant calli was 5.0%-12.1%, when the bombarded explants were cultured on selection media with kanamycin or hygromycin.

Key words: Tea science, Tea plant, Transformation, Agrobacterium tumefaciens, Particle bombardment

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