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[Factors affecting Agrobacterium-mediated genetic transformation of Poncirus trifoliata (L.) Raf]
1Institute of Chinese Materia Medica, University of Traditional Chinese Medicine and Pharmaceutical Sciences, Guangzhou 510407.
Objective:
Establishing an effective system for the Agrobacterium-mediated genetic transformation of Poncirus trifoliata, so as to lay a foundation for improving breeds and introducing foreign genes.
Method:
The explants used for the genetic transformation were the epicotyls from Poncirus trifoliata. The Agrobacterium tumefaciens strain was EHA101, containing vector plasmid PGA482GG. The GUS gene and NPT II gene were introduced into the transformation plasmid.
Results:
20-day epicotyls were suitable for transformation. 2-day precultivation enhanced shoot regeneration. Shoot regeneration frequency was high when the cocultivation lasted 2-3 days. With the presence of acetosyringone during cocultivation, the efficiency of transformation could be greatly enhanced, and the frequency of shoot regeneration rose as high as 27.5%. After the explants were cocultivated with Agrobacterium, 2-day delayed selection could stimulate shoot regeneration.
Conclusion:
An effective genetic transformation system has been established for Poncirus trifoliata on the basis of factors affecting Agrobacterium-mediated genetic transformation.