根癌农杆菌介导柞蚕抗菌肽D基因转化柑桔的研究  

Studies on Agrobacterium mediated Antibacterial Peptide D Gene Transfer in Citrus

作　　者：陈善春[1,2,3] 张进仁[1,2,3] 黄自然[1,2,3] 高峰[1,2,3] 陈凤珍[1,2,3] 隆有庆[1,2,3] 

机构地区：[1]中国农业科学院柑桔研究所 [2]华南农业大学遗传工程室 [3]重庆教育学院生物系

出　　处：《中国农业科学》1997年第3期7-13,T001,共8页Scientia Agricultura Sinica

基　　金：中国农业科学院院长基金;重庆市中青年专家基金

摘　　要：以含有双元载体（携带人工合成的柞蚕抗菌肽Ｄ基因、ｎｐｔ－Ⅱ基因和ｎｏｓ基因）的根癌农杆菌菌株ＳＥ转化锦橙、新会橙（ＣｉｔｒｕｓｓｉｎｅｎｉｓＯｓｂｅｃｋ）、沙田柚（Ｃ．ｇｒａｎｄｉｓ）的上胚轴和子叶外植体，经共培养和预培养后在含卡那霉素（Ｋｍ）的诱芽培养基中诱导并筛选Ｋｍ抗性芽。探明了柑桔品种（基因型）、预培养时间和不同的芽伸长培养基对Ｋｍ抗性芽诱导和伸长的影响。抗性芽经伸长培养后在含Ｋｍ的生根培养基中生根或经离体微嫁接，获得了转化植株。Epicotyls and cotyledons from 15 day old in vitro seedlings of Jin orange, Xinhui orange(Citrus sinensis Osbeck) and Shatian pummelo (C.grandis) were infected with Agrobacterium strain SE harboring binary vectors that contained synthetic antibacterial peptide D (AP D) gene of A.pernyi, npt Ⅱ gene and nos gene. The kanamycin resistant (Km r) shoots were directly produced from explants on shoot induction medium supplemented with kanamycin. Effect of citrus varieties (genotyps), duration of pre culture time and various shoot elongation medium on induction and elongation of Km r shoots was studied. The shoots were cultured on rooting medium or were in vitro micrografted onto 15 day old seedlings of Jin orange or Trifoliate orange as rootstocks, that resulted in Km r plantlets production. The evaluation on nos activity and Southern blot analysis verified that the nos gene and AP D gene had already been integrated into the genome of transgenic plants.

关 键 词：柑桔 遗传转化 根癌农杆菌 抗菌肽D基因 

分 类 号：S666.034[农业科学—果树学]