阮俊峰 李祥云 江胜滔 周银珠

摘要 利用组织分离法、研磨法、印迹法3种分离方法对双孢蘑菇内生菌进行分离，共分离出内生细菌7株、内生真菌1株;对分离出的菌株进行纯化，并进行抑菌试验。结果表明，内生细菌 AB-21菌株对青霉病原菌表现出强的抑制作用，进而对该菌株进行分类鉴定，形态学观察显示，该菌株为革兰氏阳性菌（G+），杆状，有芽孢;生理生化特性鉴定结果与病原库中芽孢杆菌属相符，同源率为95%;进一步对该菌株进行16S rDNA基因的扩增、测序，结果经美国国立生物技术信息中心（NCBI）数据库比对，与库中解淀粉芽孢杆菌（Bacillus amyloliquefaciens）相似度达100%。初步确定内生细菌AB-21为解淀粉芽孢杆菌（Bacillus amyloliquefaciens）。

關键词 双孢蘑菇;内生菌;分离鉴定

中图分类号 S-646.1+1  文献标识码 A

文章编号 0517-6611（2021）12-0043-04

doi：10.3969/j.issn.0517-6611.2021.12.013

开放科学（资源服务）标识码（OSID）：

Isolation and Identification of Endophytic Fungi from Agaricus bisporus

RUAN Jun feng1，2，LI Xiang yun1，JIANG Sheng tao1，2 et al （1.School of Life Sciences，Ningde Normal University，Ningde，Fujian 352100;2.Fujian University Engineering Research Center of Characteristic Biological Resources in Eastern Fujian，Ningde，Fujian 352100）

Abstract The endophytic bacteria of Agaricus bisporus were isolated by tissue separation method，grinding method and western blot method.A total of 7 strains of endophytic bacteria and 1 strain of endophytic fungus were isolated.The isolated strains were purified and tested against bacteria.The results showed that the endophytic bacteria AB 21 showed a strong inhibitory effect on the pathogenic bacteria of Penicillium.Then，the strain was classified and identified.Morphological observation showed that the strain was gram positive （G+），rod shaped，with spore.Physiological and biochemical characteristics were identified，and the results were consistent with the Bacillus genus in the pathogen database，with the homology rate of 95%.The 16S rDNA gene of the strain was amplified and sequenced.The results were compared with the database of National Center for Biotechnology Information （NCBI），and the similarity between the strain and Bacillus amyloliquefaciens in the library was 100%.The endophytic bacterium AB 21 was preliminarily identified as Bacillus amyloliquefaciens.

Key words Agaricus bisporus;Endogenous bacteria;Separation identification

双孢蘑菇（Agaricus bisporus），又称白蘑菇，欧美常称之为普通栽培蘑菇或纽扣蘑菇。双孢蘑菇子实体中等大，菌盖宽5～12 cm，初半球形，后平展，白色、光滑，略干渐变黄色，边缘初期内卷。菌肉白色、厚，伤后略变为淡红色，具蘑菇所特有的气味。菌褶初粉红色，后变褐色至黑褐色，密、窄，离生，不等长。菌柄长4.5～9.0 cm，粗1.5～3.5 cm，白色，光滑，近圆柱形，内部松软或中实。菌环白色单层，膜质，生菌柄中部，容易脱落，生长于公园、森林地、田野、草地、道路旁等处，分布极广泛，我国普遍栽培。

内生菌（Endophytic）是指在其生活史中某段时期生活在活的植物组织内，不引起植物组织产生明显病害症状的真菌和细菌[1-3]。内生菌与宿主间具有重要的生态关系，目前已研究出固氮作用、促进植物生长、能产生与宿主相同或相似的生物活性物质[4]。……