紫罗兰下胚轴原生质体分离条件的研究

Study on Hypocotyl Protoplast Isolation of Matthiola incana

以紫罗兰(Matthiolaincana)无菌苗下胚轴为材料,对光照条件、无菌苗日龄、酶液渗透压、酶液组合和酶解时间对其原生质体分离的影响进行了研究。结果表明,种子在20℃黑暗条件下萌发4d后,转移到25℃,2000lx,14h光照/日条件下约14日苗龄,用1.2%Celluase(OnozukaR-10)+0.8%Macerozyme(OnozukaR-10)+3mmol/LMES+CPW-10mol/L酶组合处理约12h,可以高效分离出有活力的原生质体。这为有效从紫罗兰下胚轴原生质体再生植株和进行体细胞杂交奠定了基础。

In the study, the hypocotyls of Mathiola incana seedlings were used as materials. The effects of some factors such as the condition of light, day-ageof M.incana seedlings, osmotic stabilizers,different combinations of enzyme mixture and digesting temperature and time, on protoplast isolation from the hypocotyls of M.incana were investigated. The results showed that the protoplasts with higher yield and quality were obtained by treating the hypocotyls with a mixture of 1.2% Celluase (Onozuka R-10)+0.8%Macerozyme(Onozuka R-10)+3mmol/L MES+CPW-10mol/L for 12h. The cotyleclons from tow-week-old seedling(at 25℃, 2000lx, 14h light/d) were appropriate for isolating protoplasts. This laid the foundation of efficient plant regeneration fron hypocotyls protoplasts of M.incana seedlings and cell fusion.