转GFP基因国庆1号温州蜜柑愈伤组织原生质的再生

Protoplast isolation,culture and embryoid induction were carried out for transgenic GFP(pBIN-mGFP5-ER)Guoqing No.1 embryogenic callus.All purified transgenic GFPCitrus unshiuMarc.cv.Guoqing No.1 protoplasts expressed strong GFP,and the viability reached about 87%.First cell division was observed about 12~14 d after protoplast cuture,and visible colonies were formed after 40 d of protoplast culture and many callus clusters with a diameter of 1~2 mm occurred after 50~60 d culture.Some calli were transferred to embryoid-induction medium,and all calli still remained in a callus stage,surrounded with abundant regenerated fresh calli even after 2 months of culture. Ploidy analysis by flow cytometry showed that all recovered products were diploid. The applications of transgenic GFP Guoqing No.1 in protoplast fusion are discussed