陆地棉体细胞胚胎的发生和遗传转化条件初探  

Prime Studies on Somatic Embryogenesis and Genetic Factors Affecting Genetic Transformation of Gossypium Hirsutum L.

作　　者：苟萍[1] 计巧灵[1] 李冠[1] 马东建[1] 

机构地区：[1]新疆大学生命科学与技术学院,新疆乌鲁木齐830046

出　　处：《新疆大学学报（自然科学版）》2002年第3期324-329,共6页Journal of Xinjiang University(Natural Science Edition)

摘　　要：以陆地棉 (冀棉 2 0 )下胚轴为外植体 ,在含 KT0 .3 mg/ L和 2 ,4-D0 .0 8mg/ L的 MS培养基上 ,诱导产生了胚性愈伤组织 .继代培养不需要添加激素 ,胚性愈伤组织即可分化为胚状体 ,并萌发出小芽 .胚状体萌发过程与合子胚相似 ,但出现了一些畸形胚 .解剖学方法观察发现 ,这些畸形胚的维管束系统不明显或形态异常 .以农杆菌介导法对冀棉 2 0胚状体及胚性愈伤组织进行了雪花莲凝集素 (GNA)基因的遗传转化条件探索 ,并以GUS基域瞬间表达检测加以证实 ,得出农杆菌的浓度为 O.D60 0 =0 .8时 ,浸染时间为 5 min,共培养时间为 3d,筛选培养基中卡那霉素浓度为 75 mg/ L,对冀棉 2 0胚状体及胚性愈伤组织转化是较为合适的 .Hypocotyls of Gossypium hirsutum L.(Jimian20) cultured on the MS medium containing 0.08mg/L 2,4-D and 0.3mg/L KT produced embryogenetic callis. Embryogenetic callis differentiated embryoids and shoots without adding hormone in sudculture. The process of embryonic germination was similar to zygotic embryoid but presented some abnormal embryoids.The bundle system of these abnormal embryoids were unclear or abnormal shaped by dissect anatomy observation. It was studied that the embryogenetic callis and embryoids of Gossypium hirsutum L.(Jimian20) were transformed with Galanthus nivalis agglutinin agglutinin(GNA) gene by Agrobacterium tumefaciens. And GUS gene was transient expressed in the embryonic callis and embryoids. It was confirmed that the transformation conditions suitable for Gossypium birsutum L. (Jimian20) are O.D600=0.8 of Agrobacterium tumefeciens concentration, 5 minutes of infective time, 3 days cocultivation, 75mg/L Kanamycin in selective medium.

关 键 词：体细胞 陆地棉 胚状体 GNA基因 遗传转化 GUS基因 胚性愈合组织 胚胎发生 

分 类 号：Q943.2[生物学—植物学] S562[农业科学—作物学]