一株产香真菌的筛选鉴定及脂肪酶基因序列克隆分析  

Isolation,Identification and Analyses of Lipase Gene Sequence of a Aroma-producing Fungi Strain

作　　者：孙媛[1] 余玉莎[1] 李梦娇[1] 周斌[2] 张芬芬[3] 李玲[1] 王继英[1] 赵明富[1] 文国松[3] 

机构地区：[1]云南农业大学农业生物多样性与病虫害控制教育部重点实验室,昆明650201 [2]云南大学微生物研究所,昆明650091 [3]云南农业大学农学与生物技术学院,昆明650201

出　　处：《湖北农业科学》2015年第5期1227-1231,共5页Hubei Agricultural Sciences

基　　金：云南省烟草公司项目"微生物菌剂在烟草病害防治中的应用研究"(2011YN58)

摘　　要：从香料植物天竺葵上分离筛选到一株产香真菌,对该菌株进行鉴定,确定其分类地位,并克隆其产脂肪酶基因进行序列分析。结果表明,分离筛选得到的菌株TZK-1形态为菌落圆形,质地疏松,菌丝由白色变为灰褐色,菌丝无隔膜,有假根,孢囊梗基部膨大形成球形孢子囊,孢囊孢子椭圆形;该菌株18S r DNA序列与米根霉Rhizopus oryzae(KF717370)相应片段的核苷酸序列同源性为99.6%。利用PCR扩增得到脂肪酶基因,该基因编码区全长为1 108 bp,编码369个氨基酸,脂肪酶核苷酸序列与推导的氨基酸序列与Rhizopus oryzae(AF229435)和Rhizopus oryzae(JN689988)聚为一支,支持强度分别达到92%和95%。结合形态学初步确定,产生浓郁甜酒香味的菌株为米根霉,能产生脂肪酶,克隆其脂肪酶基因全长。A strain of aroma-producing fungi isolated from spices plant geranium and identified. Its classification status was determined. The lipase gene was cloned and sequenced. The results showed that colonies of isolated strain TZK-1 were round, loose texture, hyphae from white to gray brown. Microscopic examination of hyphae were observed without the diaphragm, but had a rhizoid, sporangiophore base expanded to form spherical sporangia, sporangiospores oval. 18S rDNA sequence of the strain and Rhizopus oryzae （KF717370） had homology of 99.6%. The lipase gene was amplified by PCR method. The full-length coding region of the gene was 1 108 bp, encoding 369 amino acids. Amino acid sequences of nu- eleotide sequence of the lipase and Rhizopus oryzae （AF229435） and Rhizopus oryzae （JN689988） were elusteried within a branch, with homology of 92% and 95%, respectively. Combined with a preliminary determination of morphology, the strains of Rhizopus oryzae had strong wine flavor, produced lipase. The full-length of lipase gene was cloned.

关 键 词：真菌 筛选 脂肪酶 PCR扩增 

分 类 号：S188[农业科学—农业基础科学]