嘧肽霉素生物合成阻断突变株原生质体制备和再生条件的研究  

Protoplasts Formation and Regeneration of A Mutant Blocked in Cytosinpeptidemycin Biosynthesis

作　　者：魏颖颖[1] 吴元华[1] 张晓雯[1] 赵秀香[1] 高芬[1] 

机构地区：[1]沈阳农业大学植物保护学院,沈阳110161

出　　处：《沈阳农业大学学报》2008年第2期174-177,共4页Journal of Shenyang Agricultural University

基　　金：高等学校博士点专项基金项目(20070157010);辽宁省科技厅攻关项目(2006215004)

摘　　要：针对嘧肽霉素的生物合成阻断突变株,研究了其最佳的菌体培养、原生质体制备和再生条件。采用改良的含15%蔗糖、0.5%甘氨酸的YEME液体培养基振荡培养36h,获得细腻丰富的菌丝体;对不同的酶浓度、酶解温度和酶解时间进行3因子4水平的正交试验,筛选最佳酶解条件为:溶菌酶浓度2mg·mL-1,28℃,作用90min,可获得最高的制备率为99%;再生培养基以R2YE为最佳,采取直接涂布方式,25.5℃培养可达48%的再生率。Targeted with a mutant of Streptomyces ahygroscopicus var. Liaoningensis blocked in cytosinpeptidemycin biosynthesis, its optimal condition for mycelium growth, protoplasts formation and regeneration were developed. Exquisite and abundant mycelium was typically grown in 40ml of adopted YEME medium containing 15% of glucose and 0.5% of glycine. The optimal lysozyme system, including lysozyme concentration, temperature and lysis time, was studied through orthogonal design analysis, and the result indicated that the highest formation ratio of 99% was achieved when the lysozyme solution was 2mg.mL^-1, incubated at 28℃ for 90min. The protoplasts were plated on dry plates of R2YE by spreading with a glass spreader, and the regen- eration ratio was over 48% when cultured at 25.5℃.

关 键 词：嘧肽霉素:不吸水链霉菌 生物合成阻断突变株 原生质体 制备和再生条件 

分 类 号：S646.190.3[农业科学—蔬菜学] TQ920.1[农业科学—园艺学]