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利用胚状体再生植株,可以获得遗传稳定性好且数量多的再生植株,这对于建立高频稳定的遗传转化体系具有重要意义。该试验以苹果新品种“鲁加5号”为试材,采用叶片再生方法,研究了其胚状体再生体系,为建立遗传转化体系奠定了基础。结果表明:MS+6-BA1.0mg·L~(-1)+NAA 0.20mg·L~(-1)为“鲁加5号”最适宜的继代增殖培养基。叶片的最佳放置方式为近轴面接触培养基。胚胎发育阶段最佳暗培养时间为10d。最佳体细胞胚诱导培养基为MS+6-BA 1.0mg·L~(-1)+NAA 5.0mg·L~(-1)+TDZ 0.5mg·L~(-1)。胚胎发育阶段,通过胚状体不同时期形态学组织学观察,显示胚状体发育时期经历原胚、球形胚、心形胚、鱼雷形胚、子叶胚和成熟胚阶段。
Using embryoid body regenerated plants, regenerated plants with good genetic stability and large numbers can be obtained, which is of great significance for establishing a high-frequency stable genetic transformation system. In this experiment, a new apple cultivar “Lu Jia 5” was used as experimental material, and the embryo regeneration system was studied by blade regeneration method, which lays the foundation for the establishment of genetic transformation system. The results showed that MS + 6-BA 1.0 mg · L -1 and NAA 0.20 mg · L -1 were the most suitable subculture medium for “Luga No.5”. The best way to place leaves is paraxial contact medium. Optimal dark incubation time during embryonic development was 10 days. The best somatic embryo induction medium was MS + 6-BA 1.0 mg · L -1 NAA 5.0 mg · L -1 TDZ 0.5 mg · L -1. Embryonic development stage, through the embryoid body at different stages of morphological and histological observation showed embryoid body during embryonic development experience through the embryo, spherical embryo, heart-shaped embryo, torpedo-shaped embryo, cotyledon and mature embryo stage.