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采用“固—液—液—固”培养方法,分别以[‘哲引3号杨’(Populus pseudo-simonii×P.nigra‘Zhenyin3#’)ב北京杨’(P.×beijingensis)]杂交子代的种子、子叶和下胚轴为材料,开展了多个杂交子代悬浮细胞系建立和培养研究,结果显示:(1)不同基因型的种子愈伤诱导率差异显著,不同基因型的子叶、下胚轴愈伤诱导率差异不显著;不同基因型的初始悬浮细胞系密实体积差异均显著。(2)采用静置分层和细胞筛双层过滤结合的方法能把游离胚性细胞从初始悬浮细胞系中分离出来,不同基因型来源的游离胚性细胞数量差异不显著。(3)种子和子叶来源的胚性悬浮细胞系能在不含NH+4的液体培养基中诱导出球形愈伤,这些愈伤薄片能在含有CPPU的分化培养基上实现植株再生,最后共获得了20个基因型的来自种子球形愈伤的再生植株和另外20个基因型的来自子叶球形愈伤的再生植株。
In this study, Populus pseudo-simonii × P. nigra’Zhenyin3 # ’×’ P. beijingensis’ ] Hybrid progeny seed, cotyledon and hypocotyls as materials, carried out a number of hybrids on behalf of the suspension cell line establishment and culture research results showed: (1) different genotypes of callus induction rate significantly different, different genes Type cotyledon, hypocotyl callus induction rate difference was not significant; different genotypes of initial suspension cell line density volume were significant differences. (2) The separation of free embryogenic cells from the initial suspension cell line by static stratification and double-layer filtration combined with cell sieve method showed that there was no significant difference in the number of free embryogenic cells from different genotypes. (3) Seed and cotyledon derived embryogenic suspension cell lines capable of inducing bulb-shaped callus in NH4-free liquid medium are capable of regenerating plants on a differentiation medium containing CPPU and finally 20 genotypes of regenerated plants from seed bulb callus and another 20 genotypes of regenerated plants from cotyledon bulbous calluses were obtained.