解淀粉芽胞杆菌SSY2分离鉴定及其拮抗霉菌特性研究
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摘要
为了分离出可拮抗玉米常见霉菌的芽胞杆菌,通过分离、培养和形态学特征方法,从黑龙江齐齐哈尔地区玉米种植地土壤样品分离芽胞杆菌,通过平板对峙试验检测分离菌对黄曲霉菌、黑曲霉菌、卷枝毛霉菌、链格孢菌等指示菌的拮抗作用,通过生理生化鉴定和分子生物学方法鉴定分离菌,测定其安全性和药物敏感性。结果表明,筛选到革兰阳性芽胞杆菌1株,鉴定为解淀粉芽胞杆菌,编号为SSY2。该菌对黄曲霉菌、黑曲霉菌、卷枝毛霉菌、链格孢菌均具有较强的抑制作用,在测定的16种临床常用抗生素中仅对氨曲南和林可霉素表现为耐药,对其他抗生素均表现为敏感。急性毒性试验和慢性毒性试验结果表明该菌株安全性良好。分离的解淀粉芽胞杆菌具有安全、抑霉菌作用强、抑菌谱广等优点,在生物脱霉及生物饲料添加剂领域具有潜在的开发价值。
The goal of this work was to isolate Bacillus,which inhibits common mold in corn.In this study,Bacillus was isolated from soil samples from maize field in Qiqihar,Heilongjiang province by means of isolation,culture and morphological characteristics.The inhibitory effects of the isolate on the indicator fungi,such as Aspergillus flavus,Aspergillus niger,Mucor circinelloides,Alternaria alternate were detected by the agar disk diffusion method.The isolate was identified according to its physiological and biochemical characteristics and molecular biology,and the safety and drug susceptibility of the isolate were detected.One isolate was identified as Gram-positive,Bacillus amyloliquefaciens numbering SSY2 according to its physiological and biochemical characteristics and molecular biology identification.The isolate was safe and had strong inhibitory effects on Aspergillus flavus,Aspergillus niger,Mucor circinelloides and Alternaria alternata.Drug susceptibility test results showed it was only resistant to aztreonam and lincomycin,and sensitive to other 16 commonly clinically used antimicrobials tested.The Bacillus amyloliquefaciens isolated in this study had the advantages of safety,strong inhibition to mold and wide range of antimicrobial spectrum.It had potential value in the field of biological demoldy and bio-feed additives.
The goal of this work was to isolate Bacillus,which inhibits common mold in corn.In this study,Bacillus was isolated from soil samples from maize field in Qiqihar,Heilongjiang province by means of isolation,culture and morphological characteristics.The inhibitory effects of the isolate on the indicator fungi,such as Aspergillus flavus,Aspergillus niger,Mucor circinelloides,Alternaria alternate were detected by the agar disk diffusion method.The isolate was identified according to its physiological and biochemical characteristics and molecular biology,and the safety and drug susceptibility of the isolate were detected.One isolate was identified as Gram-positive,Bacillus amyloliquefaciens numbering SSY2 according to its physiological and biochemical characteristics and molecular biology identification.The isolate was safe and had strong inhibitory effects on Aspergillus flavus,Aspergillus niger,Mucor circinelloides and Alternaria alternata.Drug susceptibility test results showed it was only resistant to aztreonam and lincomycin,and sensitive to other 16 commonly clinically used antimicrobials tested.The Bacillus amyloliquefaciens isolated in this study had the advantages of safety,strong inhibition to mold and wide range of antimicrobial spectrum.It had potential value in the field of biological demoldy and bio-feed additives.
引文
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[9]曹广添,占秀安,张玲玲,等.解淀粉芽胞杆菌对肉鸡生长性能、小肠发育和血清生化指标及免疫功能的影响[J].中国畜牧杂志,2016,52(3):31-35.
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[11]王岩,陈楠楠,侯美如,等.解淀粉芽胞杆菌SSY1株的安全性试验[J].中国畜牧兽医,2017,44(3):928-934.
[12]白雪莲,吴慧玲,张虹,等.拮抗菌246-1活性物质合成条件优化及其抑菌作用[J].科学技术与工程,2017,33(17):69-76.
[13]汪敬鑫.棉花黄萎病菌拮抗菌的筛选及XSZ5菌株抑菌机制的初步研究[D].安徽合肥:安徽农业大学,2009.
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[15]东秀珠,蔡妙英.常见细菌系统鉴定手册[M].北京:科学出版社,2001:43-65.
[16]曹凤明,沈德龙,李俊,等.应用多重PCR鉴定微生物肥料常用芽胞杆菌[J].微生物学报,2008,48(5):651-656.
[17]陈楠楠,侯美如,王岩,等.解淀粉芽胞杆菌SSY1菌株的稳定性研究[J].中国微生态学杂志,2017,29(1):1-5.
[18]张雪花,李琳玲,程华,等.解淀粉芽胞杆菌CMN1308抗真菌特性研究[J].北方园艺,2016(2):117-121.
[19]冯志珍.解淀粉芽胞杆菌FB-16的筛选及其对作物疫霉菌的抑制作用[D].陕西杨凌:西北农林科技大学,2012.
[20]韩玉竹,邓钊,张宝,等.解淀粉芽胞杆菌H15产抗菌肽的发酵条件优化和提取方法比较研究[J].食品科学,2015,36(15):135-141.
[21]Etcheverry M G,Scandolara A,Nesci A,et al.Biological interactions to select biocontrol agents against toxigenic strains of Aspergillus flavus,and Fusarium verticillioides,from maize[J].Mycopathologia,2009,167(5):287-295.
[22]张宏梅,黄绍松,李发俊,等.发酵食品中益生菌对抗生素耐药性的研究进展[J].食品与发酵工业,2009(7):119-122.
[23]安健,曹海鹏,陈百尧,等.解淀粉芽胞杆菌的安全性分析[J].动物医学进展,2013,34(1):16-19.
[24]张超,王祥红,徐涛,等.一株解淀粉芽胞杆菌的分离鉴定及其抑菌条件研究[J].生物技术通报,2015,31(9):70-176.
[2]毕重朋,侯晓亮,张广宁,等.发酵饲料中霉菌毒素的危害及其防控措施[J].中国饲料,2017(15):5-7.
[3]胡淑荣,应光耀,胡玉莉,等.赭曲霉毒素A快速检测方法研究进展及其在中药中的应用前景[J].中国中药杂志,2017,42(11):2032-2037.
[4]刘瑜,郭雪霞,冉国伟,等.食物霉变优势菌筛选及霉变防治技术研究进展[J].食品工业,2017,38(12):219-222.
[5]Chowdhury S P,Uhl J,Grosch R,et al.Cyclic lipopeptides of Bacillus amyloliquefaciens subsp.plantarum colonizing the lettuce rhizosphere enhance plant defense responses toward the bottom rot pathogen Rhizoctonia solani[J].MPMI,2015,28(9):984.
[6]Kang S M,Radhakrishnan R,Lee I J.Bacillus amyloliquefaciens subsp.plantarum GR53,apotent biocontrol agent resists Rhizoctonia disease on Chinese cabbage through hormonal and antioxidants regulation.[J].World J Microbiol Biotechnol,2015,31(10):1517-1527.
[7]Kadaikunnan S,Rejiniemon T S,Khaled J M,et al.In-vitro antibacterial,antifungal,antioxidant and functional properties of Bacillus amyloliquefaciens[J].Annals Clin Microbiol Antimicrob,2015,14(1):9.
[8]蔡国林,徐铭乾,李秋,等.解淀粉芽胞杆菌黄曲霉毒素B1裂解酶的分离纯化及鉴定[J].食品与发酵工业,2015,41(12):26-30.
[9]曹广添,占秀安,张玲玲,等.解淀粉芽胞杆菌对肉鸡生长性能、小肠发育和血清生化指标及免疫功能的影响[J].中国畜牧杂志,2016,52(3):31-35.
[10]朱博.解淀粉芽胞杆菌对仔猪生长性能、抗氧化能力、免疫功能和肠道菌群的影响[D].江苏南京:南京农业大学,2016.
[11]王岩,陈楠楠,侯美如,等.解淀粉芽胞杆菌SSY1株的安全性试验[J].中国畜牧兽医,2017,44(3):928-934.
[12]白雪莲,吴慧玲,张虹,等.拮抗菌246-1活性物质合成条件优化及其抑菌作用[J].科学技术与工程,2017,33(17):69-76.
[13]汪敬鑫.棉花黄萎病菌拮抗菌的筛选及XSZ5菌株抑菌机制的初步研究[D].安徽合肥:安徽农业大学,2009.
[14]布坎南R E,吉本斯N E.伯杰细菌鉴定手册[M].8版.北京:科学出版社,1984:729-797.
[15]东秀珠,蔡妙英.常见细菌系统鉴定手册[M].北京:科学出版社,2001:43-65.
[16]曹凤明,沈德龙,李俊,等.应用多重PCR鉴定微生物肥料常用芽胞杆菌[J].微生物学报,2008,48(5):651-656.
[17]陈楠楠,侯美如,王岩,等.解淀粉芽胞杆菌SSY1菌株的稳定性研究[J].中国微生态学杂志,2017,29(1):1-5.
[18]张雪花,李琳玲,程华,等.解淀粉芽胞杆菌CMN1308抗真菌特性研究[J].北方园艺,2016(2):117-121.
[19]冯志珍.解淀粉芽胞杆菌FB-16的筛选及其对作物疫霉菌的抑制作用[D].陕西杨凌:西北农林科技大学,2012.
[20]韩玉竹,邓钊,张宝,等.解淀粉芽胞杆菌H15产抗菌肽的发酵条件优化和提取方法比较研究[J].食品科学,2015,36(15):135-141.
[21]Etcheverry M G,Scandolara A,Nesci A,et al.Biological interactions to select biocontrol agents against toxigenic strains of Aspergillus flavus,and Fusarium verticillioides,from maize[J].Mycopathologia,2009,167(5):287-295.
[22]张宏梅,黄绍松,李发俊,等.发酵食品中益生菌对抗生素耐药性的研究进展[J].食品与发酵工业,2009(7):119-122.
[23]安健,曹海鹏,陈百尧,等.解淀粉芽胞杆菌的安全性分析[J].动物医学进展,2013,34(1):16-19.
[24]张超,王祥红,徐涛,等.一株解淀粉芽胞杆菌的分离鉴定及其抑菌条件研究[J].生物技术通报,2015,31(9):70-176.