甘蓝型油菜与蔊菜的原生质体融合与植株再生

以甘蓝型油菜下胚轴和蔊菜叶片为外植体提取原生质体, 采用PEG-高pH、高Ca²⁺附加DMSO的原生质体融合方法, 用液体浅层静置培养融合体, 获得了10株融合杂种, 观察了杂种形态学和细胞学。结果表明：1%纤维素酶+0.2%离析酶+3 mmol/L MES 酶解14 h 可获得较高产率的油菜原生质体, 0.25% 纤维素酶+0.5%离析酶+5 mmol/L MES酶解12 h可获得较高产率的蔊菜原生质体; 30% PEG + 0.3 mol/L葡萄糖+50 mmol/L CaCl₂•2H₂O +15%DMSO的融合条件下, 获得了10.4%的融合率; 实验所获的原生质体融合材料可作为新种质。

Somatic hybrids between Rorippa indica (Linn.) Hiern and Brassica napus L. through protoplast-fusion system

High yield of protoplast isolation was achieved from hypocotyls of B. napus L. and leaves of Rorippa indica (Linn.) Hiern. The isolated protoplasts were used to establish an efficient protoplast-fusion system between the two cruciferous species by PEG-DMSO method and culturing with MS liquid medium. Ten somatic fusion hybrids between B. napus and R. indica were obtained. The enzyme combinations for isolating protoplast from B. napus L. and R. indica were 1% cellulase + 0.2% macerozyme + 3 mmol/L MES and 0.25% cellulase + 0.5% macerozyme + 5 mmol/L MES, respectively. Fusion percentage of 10.4% was obtained on the condition of 30% PEG + 0.3 mol/L glucose +50 mmol/L CaCl2.2H2O + 15% DMSO. Seeds plants obtained from protoplast fusion are new germplasm derived from R. indica.