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作 者 ：黄振1,2；李赟1,2；李媛1,2；王沛琦1,2；康向阳1,2*

期 刊 ：植物研究 2015年 35卷 5期 页码：696-703

关键词：杨树；多基因型；悬浮细胞系；球形愈伤；体胚发生；

Keywords:Populus, multiple genotypes, suspension cell line, round callus, somatic embryogenesis,

摘 要 ：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。

Abstract:We used the “solid-liquid-liquid-solid” culture method to establish suspension cell line and embryonic suspension cell line from seeds, cotyledons and hypocotyls of Populus hybrids［(Populus pseudo-simonii×P.nigra ‘Zhenyin3#’)×P.×beijingensis］. Seed callus induction rates of different genotypes were significantly different, while cotyledon callus and hypocotyl callus induction rates of different genotypes had no significantly different, and the packed cell volume among different genotypes had significantly different. Double filtration with cytoscreeners after standing and layering process could well enrich uncombined embryonic suspension cell, and the number of uncombined embryonic cells among different genotypes had no significantly different. Only in the media without NH⁺₄, we could obtain embryonic suspension cell lines which were originated from seeds and cotyledon, seed and cotyledon round calli regenerated from differentiation medium including CPPU, and 20 genotypes of the regenerated plants from seed round calli and other 20 genotypes of the regenerated plants from cotyledon round calli.

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