玫瑰黄链霉菌Men-myco-93-63原生质体形成和再生条件的初探  

The Formation and Regeneration of Protoplast of Stremptomyces roseoflavus Men-myco-93-63

作　　者：刘金艳[1] 张立荣[1] 李亚宁[1] 张汀[1] 刘大群[1] 

机构地区：[1]河北农业大学植物病理系河北省农作物病虫害生物防治工程技术研究中心,保定071001

出　　处：《中国农学通报》2007年第6期473-476,共4页Chinese Agricultural Science Bulletin

基　　金：国家863项目"农作物重大病害多功能广谱生防菌剂研究和创制"(2006AA10A211);河北省科学技术研究与发展计划"防治草莓白粉病和蔬菜白粉病的生物农药的研制"(05547005D-2)

摘　　要：玫瑰黄链霉菌Men-myco-93-63在温室及大田试验中都对棉花黄萎病菌表现出很强的拮抗作用,是一株很有应用潜力的生防菌株。不论是菌丝体本身还是其代谢产物都对多种植物病原菌有很强的抑制作用。对该生防菌株进行深入研究,尤其是分子水平的研究尤为重要。笔者通过研究影响链霉菌原生质体形成的多种条件和因素,包括培养基的选择、培养时间、甘氨酸浓度、溶菌酶浓度、酶解时间等,最终成功的制备出了生防菌Men-myco-93-63的原生质体,初步确定了其形成条件:TSB培养基一级培养24h,用SGGP作为二级培养基培养菌丝体40h(补加甘氨酸2%),将菌丝体加入2mg/ml溶菌酶37℃水浴60min后即可得到原生质体,涂在再生培养基R5上再生培养,再生数可达4.8×106个/ml。为该生防菌原生质体的转化、基因表达等研究奠定了重要的基础。Both of the Stremptomyces roseoflavus strain Men-myco-93-63 itself and its antibiotics produced show very strong antibiosis activities against Verticillium dahliae in the lab and field. The formation and regeneration of protoplasts of Men-myco-93-63 were developed in this study. Different effects of protoplast formation of Men-myco-93-63 were tested, including the sub-culture medium, the sub-culture time, the concentration of Lysozyme or Gly, et al. Methods for the formation and regeneration of Stremptomyces roseoflavus Men-myco-93-63 protoplasts are described： Spores shake cultured in TSB for 24h,Add ω=2%Gly to SGGP （culture medium）, continue shake cultured for 40h, Collect mycelium, Protoplasts of Stremptomyces roseoflavus Men-myco-93-63 strains were obtained by treatment with lysozyme whose concentration is 2mg/ml in a protoplast formation buffer. The protoplast of Men-myco-93- 63 has been prepared successfully and Cells were regenerated on a complex medium, regenerated with 4.8×10^6/ml on Rs.The methods were applicable to other species of Stremptomyces roseoflavus as well. It laid a foundation for protoplast transform and gene expression.

关 键 词：玫瑰黄链霉菌MEN-MYCO-93-63 原生质体 制备 再生 

分 类 号：S188[农业科学—农业基础科学]