一株产毒曲霉拮抗细菌的筛选、鉴定及抑菌活性研究

doi:10.13560/j.cnki.biotech.bull.1985.2019-0074

摘要/Abstract

摘要：

为获得同时抑制产毒赭曲霉（Aspergillus ochraceus）和黄曲霉（Aspergillus flavus）生长的拮抗菌株,充分开发利用有益微生物资源。从种植园土壤和赤豆（Vigna angularis）中分离筛选拮抗赭曲霉和黄曲霉的细菌,通过形态学、生理生化反应、16S rDNA序列同源性及特异PCR鉴定其种属;并研究菌株发酵上清液的抑菌稳定性等指标。结果显示,从种植园土壤中筛选出同时抑制赭曲霉和黄曲霉生长的拮抗细菌SC-B15,对赭曲霉和黄曲霉抑菌率分别达到47.30±13.17%和52.44±2.78%,鉴定结果为解淀粉芽孢杆菌（Bacillus amyloliquefaciens）。其上清液对赭曲霉和黄曲霉的抑菌圈直径分别为15.03±2.66 mm和13.95±2.62 mm。抑菌物质在20-40℃,pH 6-10条件下保持稳定,对胰蛋白酶和木瓜蛋白酶不敏感,显微检测赭曲霉和黄曲霉菌丝均出现异常。分离自土壤的B. amyloliquefaciens SC-B15菌株及其上清液可以有效抑制产毒赭曲霉和黄曲霉,对食品及饲料的防霉抑菌具有一定的开发应用价值。

关键词: 解淀粉芽孢杆菌, 抑菌, 黄曲霉, 赭曲霉

Abstract:

To obtain the antagonistic strains against Aspergillus ochraceus and Aspergillus flavus and to fully exploit the beneficial microbial resources,efficient antagonistic strains against Aspergillus ochraceus and Aspergillus flavus were isolated and purified from plantation soil and Vigna angularis. Then,the morphology,physiological and biochemical reactions,16S rDNA phylogenetic analysis and specific PCR analysis were applied for the identification of the antagonistic bacteria. Furthermore,the bacteriostatic stability of the fermentation supernatant,etc. were studied. Strain SC-B15,isolated from the plantation soil,had strong antagonistic effects on A. ochraceus and A. flavus growth,and the bacteriostatic rates against A. ochraceus and A. flavus reached 47.30±13.17% and 52.44±2.78%,respectively. SC-B15 was identified as Bacillus amyloliquefaciens. The inhibition zone diameters of the supernatant against A. ochraceus and A. flavus were 15.03±2.66 mm and 13.95±2.62 mm,respectively. Meanwhile,the bacteriostatic substances remained stable at 20-40℃ and pH=6-10,and were insensitive to trypsin and papain. The indicated strains appeared abnormal changes of cavitation,swelling and deformation with microscope. In sum,B. amyloliquefaciens SC-B15,isolated from plantation soil,and its supernatant,efficiently inhibit the A. ochraceus and A. flavus growth and demonstrate certain development application value as antifungal agent in the foodstuff and feedstuff.

Key words: Bacillus amyloliquefaciens, antifungal activity, Aspergillus flavus, Aspergillus ochraceus

刘端木, 吴怿, 刘沄, 梁志宏. 一株产毒曲霉拮抗细菌的筛选、鉴定及抑菌活性研究[J]. 生物技术通报, 2019, 35(8): 42-50.

LIU Duan-mu, WU Yi, LIU Yun, LIANG Zhi-hong. Screening,Identification and Antifungal Properties of a Bacterium with Antagonistic Activities Against Mycotoxin-producing Aspergillus spp.[J]. Biotechnology Bulletin, 2019, 35(8): 42-50.

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