三生烟叶片与烟花的原生质体制备研究

Preparation of Protoplast of Leaf and Petal of Nicotiana tabacum L.,cv.Samsun NN

[目的]探讨三生烟叶片及烟花的高质量原生质体制备方法。[方法]通过优化纤维素酶和离析酶浓度配比、酶解时间、渗透调节剂甘露醇浓度等参数,研究三生烟叶片与烟花原生质体的最优制备方法。[结果]三生烟烟叶材料最佳制备条件为1%纤维素酶和0.5%离析酶、甘露醇浓度0.6 mol/L、酶解时间4 h,完整率达88.22%。烟花材料最佳制备条件为0.8%纤维素酶和0.4%离析酶、甘露醇浓度0.7 mol/L、酶解时间2 h,完整率达83.59%。[结论]三生烟叶片与烟花原生质体由于材料来源不同,酶解条件存在差异,制备时需要针对不同组织材料予以优化。

Objective] To discuss the preparation method of protoplast of leaf and petal of Nicotiana tabacum L., cv.Samsun NN.[Method] Several parameters were optimized, such as cellulase concentration, macerozyme concentration, enzymatic hydrolysis time and mannitol concen-tration.The optimal preparation method of N.tabacum leaf and petal was researched.[Result] The optimal preparation condition for N.tabacum leaf was 1% cellulose, 0.5% macerozyme, 0.6 mol/L mannitol concentration, and 4 h enzymatic hydrolysis time.Under this condition, the in-tegrity rate was 88.22%.The optimal preparation condition for N.tabacum petal was 0.8% cellulose, 0.4% macerozyme, 0.7 mol /L mannitol concentration, and 2 h enzymatic hydrolysis time.Under this condition, the integrity rate was 83.59%.[Conclusion] Due to the different sources of N.tabacum leaf and petal, there are certain differences in enzymatic hydrolysis condition.Enzymatic hydrolysis condition should be optimized according to the tissue materials.