【目的】明确草菇培养料二次发酵过程中真菌群落变化情况,确定发酵不同阶段的优势菌群,为能够在分子水平上准确高效监测发酵过程,解析发酵机制奠定基础。【方法】采用变性梯度凝胶电泳(denaturing gradient gel electrophoresis,DGGE)及克隆菌株18S r DNA序列分析技术对草菇培养料二次发酵不同阶段真菌群落结构进行分析。【结果】DGGE图谱显示,不同处理真菌群落多样性存在差异,发酵高温阶段条带多样性较高,而且优势条带及相对含量也在发生动态变化。回收克隆不同发酵阶段的20个优势菌株中,9个菌株为非培养未知真核生物或真菌,其余克隆菌株为非培养散子囊菌目(Eurotiales)、烟曲霉(Aspergillus fumigatus)、曲霉菌属(Aspergillus sp.)、米曲霉(Aspergillus oryzae)、Melanocarpus albomyces、炭疽菌属(Colletotrichum sp.)、根毛霉菌属(Rhizomucor sp.)、轮枝菌属(Verticillium sp.)、普通青霉(Penicillium commune)、三角孢小囊菌(Microascus trigonosporus)和Trichosporon lactis真菌,其中14株(70%)克隆菌株为耐热真菌。【结论】草菇培养料二次发酵过程中真菌群落结构及优势菌群在发生着动态的变化。
Abstract:[Objective] To understand the fungal community succession during the phase Ⅱ of Volvariella volvacea compost and clarify the predominant fungi in different fermentation stages,to monitor the dynamic compost at the molecular level accurately and quickly,and reveal the mechanism.[Methods] The 18S rDNA-denaturing gradient gel electrophoresis (DGGE) and sequencing methods were used to analyze the fungal community structure during the course of compost.[Results]The DGGE profile shows that there were differences in the diversity of fungal community with the fermentation progress.The diversity was higher in the stages of high temperature.And the dynamic changes of predominant community and relative intensity was observed. Among the 20 predominant clone strains,9 were unknown eukaryote and fungi, the others were Eurotiales,Aspergillus sp.,Melanocarpus albomyces,Colletotrichum sp.,Rhizomucor sp.,Verticillium sp.,Penicillium commune,Microascus trigonosporus and Trichosporon lactis.The 14 clone strains were detected in the stages of high and durative temperature.[Conclusion]The fungal community structure and predominant community have taken dynamic succession during the phase Ⅱ of Volvariella volvacea compost.