潞酒酒窖壁及酒缸上可培养真菌的分离和鉴定

Isolation and identification of culturable fungi on the walls and winejar in Lujiu winery of Changzhi

采用察氏培养基对山西省长治市潞酒有限公司地下酒窖墙壁及酒缸外壁上采集的灰黑色霉状物进行分离纯化以获得真菌纯培养物,然后用扫描电镜和乳酸石炭酸棉蓝染色法在显微镜下观察纯培养物的菌丝体,再利用18S rDNA序列、β-tubulin微管蛋白基因(BenA)序列、Calmodulin钙调蛋白基因(CaM)序列分析技术分别对分离的纯培养物进行分子水平的鉴定并构建系统发育树。共分离到3株可培养真菌,分别是LJB-1、LJ3-2和LJ5-2。其中菌株LJB-1鉴定为地窖无孢霉Racodium cellare,该种在国内未见报道,属于国内新记录种;菌株LJ3-2和LJ5-2分别鉴定为小刺青霉Penicillium spinulosum和产黄青霉Penicillium chrysogenum,这两个种在酒窖菌中鲜见报道。

The gray-black moldy materials were collected on the wall of underground wine cellar and outer wall of the wine jar in Lujiu winery of Changzhi City of Shanxi Province, and the fungal pure cultures were obtained and purified in Czapek medium. The mycelium was observed by scanning electron microscope and stained by cotton blue with lactic acid carbonate. Based on 18 S rDNA, β-tubulin gene and calmodulin gene sequence analysis, the isolated fungal were identified on molecular level. Three cultivable fungal species were idenified. The strain LJB-1 was identified as Racodium cellare, which hasn’t been reported so far in China. The strains LJ3-2 and LJ5-2 were identified as Penicillum spinulosum and Penicillium chrysogenum, and were rarely reported in liquor cellars.