摘要
[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.
出处
《安徽农业科学》
CAS
2016年第14期11-14,100,共5页
Journal of Anhui Agricultural Sciences
基金
安徽省农业科学院院长青年基金项目(13B0941)
安徽省烟草公司科研项目(20120551008)
公益性行业(农业)科研专项(201203091)
关键词
烟叶
烟花
原生质体
分离纯化
完整率
Tobacco leaf
N.tabacum petal
Protoplast
Isolation and purification
Integrity
