农杆菌法将抗菌肽D基因导入蕉柑胚性细胞的研究  

Agrobacterium tumefaciens -Mediated Transformation of Citrus Embryonic Cells

作　　者：王声斌[1] 邹伟权[1] 余让才[1] 房师松[2] 黄自然[2] 

机构地区：[1]华南农业大学生命科学学院,广东广州510642 [2]华南农业大学艺术设计学院,广东广州510642

出　　处：《华南农业大学学报》2002年第2期47-50,共4页Journal of South China Agricultural University

摘　　要：用农杆菌介导的方法成功地将抗菌肽D基因导入到蕉柑胚性悬浮细胞 .再生芽经PCR、Southern杂交证明抗菌肽D基因已整合到其基因组DNA中 .在农杆菌介导的蕉柑胚性悬浮细胞转化过程中 ,共培养介质中的乙酰丁香酮 (As)能显著提高其转化效率 ,与对照相比 ,每克悬浮细胞转化后形成的抗卡那霉素细胞团数从 0提高到 0 .75 .悬浮细胞与农杆菌共培养时间对转化效率也具有显著影响 ,共培养 1、2、3d后 ,每克悬浮细胞转化后形成的抗卡那霉素细胞团数分别为 0 .2 0、0 75、0 .Two clones resistant to kanamycine were obtained in MT+Km 90 mg·L -1 medium from citrus embryonic cell suspensions after Agrobacterium tumefanciens -mediated transformation. Several shoots regenerated from these clones resistant to kanamycine(90 mg·L -1 ). Integration of cecropin D gene into these shoots genome was confirmed by PCR and Southern blot. The efficiency of transformation was markedly increased by co-cultivation of citrus embryonic cells with Agrobacterium tumefanciens that was induced with 200 μmol·L -1 Acetosyringone(As). More clones resistant to kanamycine(90 mg·L -1 ) were obtained by the treatment of 2 days co-cultivation of citrus embryonic cells with Agrobacterium tumefanciens .

关 键 词：抗菌肽D基因 胚性细胞 遗传转化 抗病育种 蕉柑 农杆菌法 

分 类 号：S666.103.6[农业科学—果树学]