湖北雪茄烟叶致霉菌分离鉴定及芽胞杆菌属拮抗菌株筛选

Isolation and identification of molds in Hubei cigar tobacco and screening of antagonistic strains of Bacillus spp.

为筛选可高效拮抗湖北雪茄烟叶中致霉菌的芽胞杆菌属(Bacillus)微生物,采用形态学和分子生物学相结合的菌种鉴定方法,从湖北省十堰市和宜昌市雪茄烟叶中分离、纯化得到5种霉菌,经鉴定为聚多曲霉(Aspergillus sydowii)、产黄青霉(Penicillium chrysogenum)、绳状篮状菌(Talaromyces funiculosus)、云南木霉(Trichoderma yunnanense)和黑曲霉(Aspergillus niger)。用5种霉菌对烟叶进行回接侵染,发现湖北省宜昌市发酵前茄衣烟叶更易被5种霉菌混合的孢子悬浮液侵染,且绳状篮状菌(Talaromyces funiculosus)与黑曲霉(Aspergillus niger)均对该烟叶有强致霉性。采用牛津杯抑菌法分析了不同芽胞杆菌属(Bacillus)拮抗菌株对5种霉菌的拮抗能力,其中解淀粉芽胞杆菌DY-9(Bacillus amyloliquefaciens DY-9)对各霉菌拮抗能力均为最强。浓度为109CFU/mL时菌株DY-9菌悬液对雪茄烟叶的防霉效果最佳。

In order to screen Bacillus spp.that can efficiently antagonize molds in Hubei cigar tobacco,using a combination of morphological and molecular biological strain identification methods,five molds were isolated and purified from cigar tobacco in Shiyan and Yichang,Hubei Province.They were identified as Aspergillus sydowii,Penicillium chrysogenum,Talaromyces funiculosus,Trichoderma Yunnanense,and Aspergillus niger.The five molds were used to back-infect the cigar tobacco,and it was found that the cigar tobacco before fermentation in Yichang was more susceptible to infection by the spore suspension mixed with the five kinds of molds,and Talaromyces funiculosus and Aspergillus niger had strong mildew-causing properties to the cigar tobacco.The Oxford cup inhibition method was used to analyze the antagonistic ability of different antagonistic strains of Bacillus spp.against the molds,among which Bacillus amyloliquefaciens DY-9 was the strongest antagonist against each of the molds.At a concentration of 109 CFU/mL,strain DY-9 suspension had the best anti-mold effect on the cigar tobacco.