激素对大豆悬浮细胞系愈伤组织诱导的影响

Effect of Hormones on the Induction of Soybean Callus Suitable for Establishment of Suspension Cell Lines

以大豆无菌苗的子叶和下胚轴为外植体,通过比较筛选2,4-二氯苯氧乙酸(2,4-D)和6-苄基腺嘌呤(6-BA)的浓度及培养条件,诱导出相应的愈伤组织,并成功建立大豆悬浮细胞培养体系.结果表明:单独使用2,4-D即可诱导出适于建立大豆悬浮细胞系的愈伤组织;加入6-BA导致子叶和下胚轴外植体剧烈膨大,出愈率低,形成的愈伤组织量明显减少且局部组织坚硬,难以建立悬浮细胞系.组织切片观察结果显示加入6-BA致使外植体和愈伤组织之间的部分组织细胞核消失、细胞壁显著增厚,是悬浮细胞体系难以建立的主要原因.

The effects of 2,4-Dichlorophenoxy acetic acid （2,4-D） and 6-Benzylaminopurine （6-BA） on induction of soybean callus were studied. The results indicated that 5 mg/L and 0.8 mg/L 2,4-D were suitable concentrations for the induction of soybean cotyledon callus and hypocotyl callus, respectively. Plenty of incompact and rapid growing suspension cells were obtained with these two kinds of calluses. Compared to the callus induced by culture medium only containing 2,4-D, Calluses induced by mediums supplemented with different concentrations of 6-BA were significantly fewer in quantity and harder in quality. Microscope observation of hypocotyl callus induced by medium containing 6-BA showed that part of callus cells＇ nucleus disappeared, and their cell wall markedly thickened. This might be the main reason for explaining why calluses induced by mediums supplemented with 6-BA were not suitable for establishment of suspension cell lines.