Optimization of Agrobacterium tumefaciens-mediated transformation systems in tea plant (Camellia sinensis (L.) O. Kuntze)

Q.R. Lv, C.S. Chen, Y.J. Xu, S.K. Hu, L. Wang, K. Sun, X. Chen, X.H. Li
The cotyledon callus of Camellia sinensis were used as the receptors for transformation by Agrobacterium tumefaciens EHA105 containing PS1aG-3. Some factors which affected the result of Agrobacterium-mediated transformation of C. sinensis were studied on the basis of GUS transient expression system. The optimum system of Agrobacterium-mediated transformation was that the cotyledon callus were pre-cultured for 3 d, and then infected for 15 min by EHA105 followed by 3 d co-culture in the dark on the YEB medium containing 150 μmol L-1 AS. The transient expression rate of GUS gene was 62.6%. An appropriate time of delayed selective culture could improve the conversion rate of the tea callus. After delaying the selective culture for 3 d, infected callus were transferred into the differentiation medium and the root induction medium, both of which were supplemented with 100 mg L-1 Spectinomycin, in the end resistant seedlings of C. sinensis were obtained. The conversion rate was about 3.6%.
Lv, Q.R., Chen, C.S., Xu, Y.J., Hu, S.K., Wang, L., Sun, K., Chen, X. and Li, X.H. (2018). Optimization of Agrobacterium tumefaciens-mediated transformation systems in tea plant (Camellia sinensis (L.) O. Kuntze). Acta Hortic. 1208, 199-206
DOI: 10.17660/ActaHortic.2018.1208.26
https://doi.org/10.17660/ActaHortic.2018.1208.26
Camellia sinensis, cotyledon callus, Agrobacterium tumefaciens, GUS transient expression
English

Acta Horticulturae