欢迎访问《中国农学通报》,

中国农学通报 ›› 2010, Vol. 26 ›› Issue (21): 49-53.

所属专题: 生物技术 小麦

• 生物技术科学 • 上一篇    下一篇

小麦愈伤组织诱导及原生质体的分离与纯化

张小红   

  • 收稿日期:2010-03-26 修回日期:2010-07-06 出版日期:2010-11-05 发布日期:2010-11-05

Wheat Callus Induction and Protoplasts of Separation and Purification

  • Received:2010-03-26 Revised:2010-07-06 Online:2010-11-05 Published:2010-11-05

摘要:

摘 要:以普通小麦的幼穗和幼胚为外植体进行愈伤组织诱导,研究了外植体及低温预处理等因素对小麦愈伤组织诱导的影响;并由小麦幼胚形成的愈伤组织进行了原生质体分离和纯化实验,研究了酶浓度及配比、酶解时间、蔗糖浓度等因素对原生质体分离和纯化效果的影响。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3d,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。

关键词: 转基因741杨, 转基因741杨, 杨扇舟蛾, 生长发育, 抑制规律

Abstract:

Abstract: The three wheat varieties for the material, its immature spikes 、immature embryos and mature embryos for in vitro culture. 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 of the enzyme, time of enzymolysis and sucrose concentration on separate and pure protoplasts were been 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 low-temperature, long time will affect the callus induction. In protoplast separation experiments 2.0% cellulose +0.5% pectinase + 0.6M mannitol significantly better than the 3.0% cellulose +1 %Pectinase. The best time of protoplast isolated was 4-6 hours with higher protoplast vitality.