小麦愈伤组织诱导及原生质体的分离与纯化

以普通小麦的幼穗和幼胚为外植体进行了愈伤组织诱导,及由幼胚愈伤组织进行了原生质体分离和纯化试验,研究了幼穗和幼胚外植体及低温预处理等因素对愈伤组织诱导的影响,在原生质体分离中研究了酶浓度和配比、酶解时间及蔗糖浓度等因素对幼胚愈伤组织原生质体游离和纯化的影响。结果表明:小麦幼穗离体培养时,以1.0～1.5cm长度的幼穗有利于愈伤组织的诱导;小麦幼穗和幼胚外植体采后低温储藏时不易超过3天,时间太长会影响愈伤组织诱导;2.0%纤维素酶+0.5%果胶酶+0.6M甘露醇分离原生质体较好,最佳酶解时间为4～6h,原生质体产量和活力较高。

Wheat Callus Induction and Protoplasts of Separation and Purification

Immature spikes and immature embryos from the wheat variety Xiaoyan-22 were used for culture in vitro.The infection of explants and the low-temperature processing time before the vaccination in vitro culture characteristics were studied and discussed.The wheat protoplasts from callus of immature embryos were isolated and purified.The infection of concentration and proportioning of the enzyme,time of enzymolysis and sucrose concentration on separate and pure protoplasts were discussed.The results showed that：at 1.0-1.5 cm length of the immature spikes more conducive to the in vitro culture;when the immature spikes and immature embryos storied beyond 3 d at 4℃ low-temperature,long time would affect the callus induction.In protoplast separation experiments,2.0% cellulose ＋ 0.5% pectinase ＋ 0.6 M mannitol were significantly better than the 3.0% cellulose ＋ 1% pectinase.The best time of protoplast isolated was 4-6 hours with higher protoplast vitality.