沙田柚遗传转化的探讨

采用含ｎｏｓ基因和人工合成柞蚕抗菌肽Ｄ基因ＡＰ－Ｄ基因的根癌农杆菌对沙田柚外植体进行转基因研究，与根癌农杆菌共培养产生的芽，苗经Ｋｍ敏感性筛选，获得了若干转基因植株，并对这些植株进行了报告基因ｎｏｓ表达的电泳检测，同位素标记、ＡＰ－Ｄ基因为探针的点印迹杂交，外源ＡＰ－Ｄ基因的ＰＣＲ扩增和Ｓｏｕｔｈｅｎ杂交。结果显示，报告基因ｎｏｓ、抗菌肽Ｄ基因已转入沙田柚植株细胞中。

Research on the Genetic Transformation of Shatian Pummelo

Some transgenic plantlets of shatian pummelo (Citrus grandis Osb.) have been achievely obtained by infection of Agrobacterium tumefaciena,which containing NPTII gene, nos gene,and AP-D gene (the promoter, CaMV 35s is included).The infected tissues and buds were selected on the MS medium containing kanamycin (Km) of different concentration. The gene transformation was confirmed on Kmr plantlets by analysis of nopaline detection electrophores,DNA dot blot,PCR and southern bolt.The results shown that the reporter-nos,and the goal gene, Antibacterial peptide D gene has been integrated into the genome of tramsgenic plant cells of Citrus grandis cv. shatian pummelo.