Plant Regeneration from Agobacterium-mediated CTV-cp Gene Transformation of Poncirus trifoliata Raf.
Objective:To lay a foundation for the resistant breding, the anti-virus CTV-cp gene was transformed into the epicotyles mediated by Agrobacterium tumefaciens in Poncirus trifoliata. Method:The explants used for the genetic transformation were the epicotyls from P.trifoliata.The Agrobacterum tumefaciens strain was EHA101 containing vector plasmid pGA482GG.The coat protein gene (CTV-cp gene),GUS gene and NPT II gene were introduced into the transformation plasmid.Results:Ceftaxime used as antibiotics was better than carbenicillin. The concentration was 300 mg*L-1;The selection pressure for kanamycin was 50 mg*L-1;70.0% of the resistant plants were GUS-positive; extra gene was proved to be in P.trifoliata plant by southern blot examination. Conclusion:An effective genetic transformation mediated by Agrobacterium tumefacines,which harbours a CTV-cp gene, has been developed in P.trifoliata.Transgenic CTV-cp plants were obtained.