2.4-D、6-BA对人参体细胞胚胎发生过程的影响研究

本实验以人参芽胞、二年生人参根、实生苗的茎、叶为外植体研究了体细胞胚的发生条件，并对其发生过程中可溶性蛋白、相关酶活性及内源激素的变化等进行了研究。结果表明，诱导愈伤组织的培养基为MS+2,4-D 4.0mg/L + BA 0.2mg/L；在MS+2,4-D 1.0mg/L + KT 0.2 mg/L培养基上继代培养，可获得胚性愈伤组织；在无2,4-D的培养基上可诱导出胚状体。将胚状体转入无任何激素的MS培养基上继续培养，之后转入1/2MS培养基上获得再生植株。组织细胞学观察表明人参胚状体的起源方式为单细胞起源。在体细胞胚胎发生过程中，多糖和淀粉含量在早期胚时较低，可溶性蛋白含量、POD及PPO活性在早期胚时最高；IAA在早期胚时期含量最高，在成熟胚时期ABA含量最高，而ABA/IAA比值在成熟胚时较高，利于体细胞胚的发育成熟。cDNA-AFLP 分析表明胚状体发育不同时期的人参培养物基因表达不同,从而导致了分化和发育。培养物HPLC分析表明胚胎发生试管苗总皂苷含量比子叶胚时期高4倍多。单体皂苷差异较大。

The influence of somatic embryogeny of Panax ginseng with 2.4-D and 6-BA in the process

In this article , rapid propagation of Panax ginseng in tissue culture was reported ; the process of morphogenesis through different path from shoots , stems , leaves and seeds as explants obtained regenerated plant . The callus was induced on MS medium supplemented with 4.0mg/L 2,4-D and 0.2mg/L BA .The callus was subcultured on MS medium supplemented with 2.0mg/L 2,4-D and 0.2mg/L KT. Embryogenic callus could be obtained and then lacking 2,4-D we can obtain embryoid. It can grow fast on MS medium with no growth regulators. Plantlets regenerated on 1/2MS. We assume that the origin of somatic embryo of Panax ginseng is from the single-cell. The origin of adventitious buds of Panax ginseng is from callus surface or inner. High sugar and starch involve in its embryoid callus and development. High activities of POD and PPO and protein level were necessary to earlier offer material and energy. The contents of IAA is higher in earlier embryo and ABA is higher in mature embryo. The ratios of ABA/IAA were the development of somatic embryogenesis.