甘蓝与大白菜的原生质体融合

Protoplast Fusion of Brassica oleracea and Brassica pekinensis

[目的]研究两种近缘物种原生质体的最佳融合条件。[方法]利用培养好的甘蓝与白菜的幼苗在果胶酶与纤维素酶的作用下分离出原生质体,再利用PEG融合液将这2种原生质体融合。[结果]用6%甘露醇+8%纤维素酶+2%果胶酶处理材料,酶解4 h后,可以获得大量的游离原生质体,大部分原生质体呈圆形,散乱地游离在原生质体溶液各处。利用30%PEG融合液及0.1 mol/L CaC l2溶液(含9%甘露醇,pH值9.5)将这两种原生质体融合可以获得稳定的、可育的细胞杂种植株,可以直接作为育种的种质材料。[结论]该研究为远缘杂交和进一步研究原生质体的培养及再生奠定了基础。

[ Objective] The aim was to study the optimum fusion condition of protoplast from 2 relative species. [ Method ] The protoplasts were separated from the cultured seedlings of Brassica oleracea and Brassica pekinensis with the function of pectinase and cellulase and the 2 protoplasts were fused by PEG fusion liquid. [ Result] A great deal of free protoplasts could be obtained by using 6% mannitol ＋ 8% cellulose ＋ 2% pectinase to treat the material for 4 h. Most protoplasts were round and dissociated in the protoplast solution irregularly. Using 30% PEG fusion liquid and 0.1 moL/L CaCl2 solution （containing 9% mannitol, pH value 9.5 ） for protoplast fusion could obtain stable and fertile cell hybrid plants and the protoplast could be used as the germplasm materials for breeding. [ Conclusion] The research laid the foundation for distant hybridization and further study of the culture and regeneration of protoplast.