【摘要】 对白黄小脆柄菇和阴沟肠杆菌的原生质体跨界融合进行条件优化,并对筛选出的融合子进行扫描电子显微镜观察,测定融合子的细胞形态。首先进行原生质体融合条件优化,在单因素实验的基础上采用三因素三水平的响应面分析,根据回归分析确定较优的融合条件为:PEG 34.9%、时间10.4min、温度36.4℃。然后用含抗生素的培养基对融合子进行初筛,将筛选出的11株融合子进行降解木质素研究,其中融合子PE-9经2天的培养后,木质素去除率达到了27.3%,具有良好的降解木质素潜力。最后将复筛的高效融合子PE-9与亲本菌株进行扫描电镜SEM比较观察。更多还原

【Abstract】 In this study,protoplasts of Psathyrella candolleana and Enterobacter cloacae were prepared and fused.After screening,one of the fusants was identified by the morphology with scanning electron microscope.Experiment factors and levels were firstly selected by one-factor test,then Box-Bennken design and response surface analysis with three factors and three levels,and factors affecting the protoplast fusion condition were determined by regression analysis.The results showed that the optimum conditions were: PEG 34.9%,10.4 min,36.4℃.The fusants have been regenerated on nystatin and streptomycin selective medium preliminarily and 11 fusants were selected for further studies.In comparison with their parental strains and fusants,fusion recombinant PE-9 showed higher lignin-degrading ability,which can achieve 27.3% after 2 d incubation.Finally,the morphological characteristics of their parental strains and Fusant PE-9 were observed with scanning electron microscope.更多还原