Factors affecting genetic transformation efficiency for stylo (Stylosanthes guianensis) with Agrobacterium tumefaciens

doi:10.11686/cyxb2015532

Abstract

Abstract: The objective of this study was to investigate effects of Agrobacterium concentration, infection time and co-culture duration on the genetic transformation efficiency for stylo (Stylosanthes guianensis) using a high-efficiency regeneration system that had been developed previously. Hypocotyls from stylo seedlings were used as plant material and the β-glucuronidase (GUS) gene was used as the reporter gene. The callus transformation efficiency was evaluated by GUS staining. The callus transformation efficiency reached 72% under the following conditions: Agrobacterium at a concentration of OD₆₀₀=0.4-0.6, with 15 mins infection and 3 days of co-cultivation. The exogenous gene had been successfully introduced into the stylo genome as indicated by GUS staining and PCR detection after callus selection, shoot differentiation and rooting. The results indicate that this A. tumefaciens-mediated transformation system is suitable for genetic improvement and functional gene analysis of stylo.

CHEN Cai-Hong, CHUAN Xiu-Juan, WANG Hui, JIA Yan-Xing, CHEN Zhi-Jian, LIU Guo-Dao, LUO Li-Juan. Factors affecting genetic transformation efficiency for stylo (Stylosanthes guianensis) with Agrobacterium tumefaciens[J]. Acta Prataculturae Sinica, 2016, 25(6): 102-108.

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