高寒草地土壤产漆酶真菌的筛选及分子鉴定

以高寒草地土壤分离出的9株真菌菌株为研究对象,采用rDNA-ITS方法对筛选出的产漆酶菌株进行分子鉴定,根据菌株在愈创木酚、蒽酮、邻苯二酚、邻联甲苯胺、邻甲苯胺、α-萘酚、联苯胺、没食子酸为底物的选择性培养基上的生长情况、菌落大小、漆酶催化氧化还原反应产生的变色圈直径大小及其颜色深浅程度,进行产漆酶真菌初筛,采用液体产酶发酵法选用ABTS为底物测定漆酶活力进行复筛。鉴定结果为菌株1.9为Uncultured fungus clone.,2.1a为Scytalidium,310b为Marasmius tricolor,2.1c为Saccharicola,2.3a为fungal sp,2.4d为Saccharicola.,10a为Marasmiusrotalis,3.7c为Alternaria.WB为Verticilliumlongisporum。筛选结果表明:除菌株3.7c以外,其余8株菌株在以愈创木酚、蒽酮、邻联甲苯胺、联苯胺为底物的培养基上基本都产生氧化带,在没食子酸底物培养基上均不产生氧化带;菌株1.9,2.1a,310b,2.4d,10a,2.1c,2.3a均具有产漆酶活力,其中菌株310b具有较强产漆酶活力。

Screening and Identification of Laccase Fungi in Alpine Meadow

Nine strains of fungi that isolated from alpine meadow were used as research objects. The laccase-producing strains were identified by rDNA-ITS method. According to the strains,guaiacol,anthrone,catechol and neighbors were identified. Growth on selective medium of toluidine,o-toluidine,α-naphthol,benzidine,gallic acid as substrate,colony size,size of the color circle produced by laccase-catalyzed redox reaction,and the degree of color depth,the laccase-producing fungus is firstly screened,and the laccase activity is determined by the liquid-producing enzyme fermentation method using ABTS as a substrate for re-screening. The results showed that strain 1.9 was Uncultured fungus clone. 2.1a was Scytalidium,310b was Marasmius tricolor,2.1c was Saccharicola,2.3a was fungal sp,2.4d was Saccharicola. 10a was Marasmiusrotalis,3.7c was Alternaria. WB was Verticilliumlongisporum. The screening results showed that except for strain 3.7c,the other 8 strains basically produced oxidative bands on the culture medium with guaiacol,anthrone,ortho-toluidine and benzidine as substrates,and cultured in gallic acid substrate. No oxidative bands were produced on the culture medium;strains 1.9,2.1a,310b,2.4d,10a,2.1c,and 2.3a all had laccase activity,and strain 310b had stronger laccase activity than the other strains.