碰柑愈伤组织的遗传转化及转化体的再生

Base on multiplieation of callus，induetion of somatic embryo and plant regeneration in Ponkan mandarin(Citrus reticulata Blanco cv.‘Ponkan’), the AP1 and AP一BD genes medieated by Agrobacterium tumefaciens were transformed into the callus wounded by carborundum, transformic plantlets were obtained from the callus through seleetion. A. tumefaciens strain LBA4404 harboring pCDB一Ⅱ or p208一AP1 was used in this transfonnation researeh. pCDB一Ⅱ contains both AP一D and AP一B gene，and p208一API contains API gene.More KanR embryoids were obtained through carborundum wounding than without treatment, their differences is significant. The integratation the AP一D, AP一B and API genes into Ponkan genome was confirmed by PCR, among 138 transfromant tested, there being some plantlets containing AP一D, AP一B or API gene, their pecrcent being 6.5%, 7.2% and 8.7%, respcetively; and some plantlets containing both AP一D and API，or AP一D and AP一D，moreover，4 transformant containing AP一D，AP一B and API. Southern blotting detection showed that the target genes were integrated into plantlets in single copy; this verified the results of PCR.