野生茄子原生质体制备及电融合条件研究

Studies on Protoplast Preparation and Electrofusion Conditions in Wild Eggplant

以野生茄子蒜芥茄(Solanum sisymbriifolium)和水茄(S.torvum)叶片为原生质体融合材料,对原生质体制备中酶解液浓度、酶解时间和电融合参数进行研究。结果显示:在0.3%(m/V)纤维素酶+0.1%(m/V)离析酶浓度条件下镜检观测到的单细胞数量最多;蒜芥茄酶解最适时间为14h,原生质体产量达最大值15.06×10^6个·g^-1,水茄酶解最适时间为13h,原生质体产量达最大值15.92×10^6个·g^-1;最适电融合参数为交流电场强度80V·cm^-1,作用时间10s,直流脉冲强度1250V·cm^-1,作用时间45μs,次数1次时,显微镜观测到融合效果最佳。

Taking the leaves of Salanum sisymbriifolium and S. torvum as protoplast fusion materials,this paper studied the enzymatic hydrolysate concentration,enzymatic hydrolysis time and electrofusion parameters. The results showed that the optimal concentration was 0.3%(m/V)cellulase R-10+0.1%(m/V)macerozyme R-10.Under this condition,the maximum number of single cells could be observed by microscope. The optimal enzymolysis time were 14 hours for S. sisymbriifolium and 13 hours for S. torvum,which the protoplasts yield was 14.06×10^6 protoplasts per gram and 15.92×10^6 protoplasts per gram,respectively.The optimum electric fusion parameters of field strength was 80 V·cm^-1,for 10 s action time.The D.C. square pulses was 1 250 V·cm^-1 for 45 μs.Just once the best fusion effect could be observed by microscope.