Agrobacterium-mediated genetic transformation of Camptotheca acuminata

Abstract

UGPase gene related with wood cellulose synthesis was transferred into C. acuminata using the method of Agrobacterium-mediated genetic transformation, and an efficient transformation system was developed for C. acuminata on the basis of evaluations of several factors affecting Agrobacterium-mediated DNA transfer rate. The highest transformation rate was achieved when pre-cultured leaf explants were infected with an Agrobacterium culture corresponding to OD600 (0.5) for 10 min, and cultured on explant regeneration medium for three days. The results of Southern hybridization showed that genomic DNA of the kanamycin-resistant shoots to an UGPase gene probe substantiated the integration of the transgene. Transformation efficiency (6%) was achieved under the optimized transformation procedure. This system should facilitate the introduction of important useful genes into C. acuminata.

Keywords

Genetic transformation / Camptotheca acuminata / UGpase gene