Combination of Particle Bombardment-mediated and Agrobacterium-mediated Transformation Methods in Tea Plant

吴姗，梁月荣，陆建良，黎昊雁

doi:10.13305/j.cnki.jts.2005.04.004

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Journal of Tea Science ›› 2005, Vol. 25 ›› Issue (4) : 255-264. DOI: 10.13305/j.cnki.jts.2005.04.004

Combination of Particle Bombardment-mediated and Agrobacterium-mediated Transformation Methods in Tea Plant

WU Shan¹, LIANG Yue-rong², LU Jian-liang²，LI Hao-yan¹

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Abstract

A microprojection protocol using particle delivery system and GUS report gene was developed for tea plant (Camellia sinensis) callus. Comparison of several pre-bombardment culture media showed the highest transient gene expression in the media with PVP. Instead of making contribution to improving GUS expression, osmotic pre-treatment would cause the target tissue having difficulty in regeneration. High transformation rate was obtained using 125 μg tungsten coated with 0.25 μg DNA per shot. Bombarding tea callus at 7 MPa with a target distance of 5 cm and one time was suitable for both GUS activity and regeneration. Several transformation methods, including Agrobacterium-mediated transformation(AGR), particle bombardment (BOM), and combination of the two methods (BTA, BPA and BOA), have been tested and compared with each other. Based on the results of GUS expression and survival rates of the resistant calli, the method combining Agrobacterium and particle bombardment systems is suitable to apply for tea plant transfortion.

Key words

Agrobacterium tumefaciens / Genetic transformation / Particle bombardment / Tea plant (Camellia sinensis)

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WU Shan, LIANG Yue-rong, LU Jian-liang, LI Hao-yan. Combination of Particle Bombardment-mediated and Agrobacterium-mediated Transformation Methods in Tea Plant[J]. Journal of Tea Science. 2005, 25(4): 255-264 https://doi.org/10.13305/j.cnki.jts.2005.04.004

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