陆地棉体细胞胚胎的发生和遗传转化条件初探

以陆地棉（冀棉20）下胚轴为外植体，在含KT0．3mg／L和2，4－D0．08mg／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20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基域瞬间进行检测加以证实，得出农杆菌的浓度为O．D600＝0．8时，浸染时间为5min，共培养时间为3d，筛选培养基中卡那霉素浓度为75mg／L，对冀棉20胚状体及胚性愈伤组织转化是较为适合的。

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

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.