从土壤中分离出82株细菌,通过平板对峙法初筛和双层平板法复筛,筛选出对烟草赤星病菌有较强拮抗作用的细菌B11菌株,其无菌发酵液对烟草赤星病菌(Alternaria alternata)的抑菌带达到26.55 mm.对B11菌株进行生理生化和分子鉴定,结果表明B11菌株生理生化特征与Bacillus amyloliquefaciens最为接近;16S rRNA 基因序列分析结果表明该菌株与B. amyloliquefaciens(NR-116022)聚类呈一支,同源性为99%,因此把该菌株鉴定为解淀粉芽孢杆菌(B.amyloliquefaciens).进一步研究表明B11菌株对几种植物病原真菌的生长均有抑制作用.
82 bacterial strains were obtained from soil. By plate confrontation method and preliminary screening and double plate screening,strain B11 was screened to have the strongest inhibition against Alternaria alternata. The inhibitory zone of aseptic fermented liquid against A. alternata reached 26.55 mm. Physiological, biochemical and molecular identification of B11 strain were carried out. The physiological and biochemical characteristics of B11 strain were closest to that of Bacillus amyloliquefaciens. 16S rRNA sequence analysis of strain B11 showed that B. amyloliquefaciens(NR-116022)clustering and strain B11 became a branch(99% similarity). So, it was identified as B.amyloliquefaciens. Further studies showed that B11 strain had some inhibitory effects on several plant pathogenic fungi.
[1] 祖艳青. 河南省烟草赤星病菌及内生链格孢的鉴定和遗传多样性研究[D]. 郑州:河南农业大学,2013. ZU Y Q. Identification and genetic diversity study on brown spot pathogen and endogenous Alternaria of tobacco in henan province[D]. Zhengzhou: Henan Agricultural University,2013.
[2] 朱亚滨,方晓宇. 我国烟草病害及抗赤星病育种研究进展[J]. 现代化农业,2004(5):7-8. DOI:10.3969/j.issn.1001-0254. 2004.05.003. ZHU Y B, FANG X Y. Research progress of tobacco diseases and anti-Brown spot disease breeding in China[J]. Modernizing Agriculture, 2004(5):7-8. DOI:10.3969/j.issn.1001-0254. 2004.05.003.
[3] 谭仲夏,杨龙祥,李成杰. 烟草赤星病的生物防治研究进展[J]. 烟草科技,2005(7):41-45.DOI:10.3969/j.issn.1002-0861. 2005.07.013. TAN Z X, YANG L X, LI C J. Recent advance in biological control to Alternaria alternate[J]. Tobacco Science, 2005(7):41-45.DOI:10.3969/j.issn.1002-0861. 2005.07.013.
[4] 谭仲夏,杨龙祥. 烟草赤星病的生物防治研究现状及展望[J]. 中国烟草学报,2005,11(3):34-38. DOI:10.3321/j.issn:1004-5708. 2005.03.007. TAN Z X, YANG L X. Current situation and prospect of biological control of Tobacco Brown Spot disease[J]. Acta Tabacaria Sinica, 2005, 11(3):34-38. DOI:10.3321/j.issn:1004-5708. 2005.03.007.
[5] 张超,王祥红,徐涛,等. 一株解淀粉芽孢杆菌的分离鉴定及其抑菌条件研究[J]. 生物技术通报,2015(9):170-176. ZHANG C, WANG X H, XU T, et al. Isolation and identification of Bacillus amyloliquefaciens WTD and optimal conditions of its antimicrobial activity[J]. Biotechnology Bulletin, 2015(9):170-176.
[6] 祝学海,李玉权,陶小买,等. 贵州半夏块茎腐烂病拮抗细菌的筛选与鉴定[J]. 微生物学通报,2017(2):438-448. ZHU X H, LI Y Q, TAO X M, et al. Screening and identification of antagonistic bacteria against soft rot of Pinellia ternata in Guizhou Province[J]. Microbiology China,2017(2):438-448.
[7] 戈登R E,蔡妙英. 芽孢杆菌属[M]. 北京:农业出版社,1983.
[8] 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.
[9] 陈成,崔堂兵,于平儒. 一株抗真菌的解淀粉芽孢杆菌的鉴定及其抗菌性研究[J]. 现代食品科技,2011, 27(1):36-39. DOI:10.3969/j.issn.1673-9078. 2011.01.009. CHEN C, CUI T B, YU P R. Identification of an anti-fungal strain of Bacillus amyloliquefaciens and the properties of the antifungal substance[J]. Modern Food Science and Technology, 2011, 27(1):36-39. DOI:10.3969/j.issn.1673-9078. 2011.01.009.
[10] 陈雪,万科,张传萍,等. 烟草赤星病拮抗细菌的筛选、鉴定及机制初步研究[J]. 生物学通报,2013, 48(7):51-54. DOI:10.3969/j.issn.0006-3193. 2013.07.016. CHEN X,WANG K,ZHANG C P, et al. Screening, identification and mechanism of antagonistic bacteria against tobacco brown spot disease[J]. Bulletin of Biology,2013, 48(7):51-54. DOI:10.3969/j.issn.0006-3193. 2013.07.016.
[11] 徐滔明,罗志威,易子霆,等. 拮抗微生物在防治烟草病害中的研究与应用[J]. 江西农业学报,2016(1):41-44. DOI:10.3969/j.issn.1001-8581. 2016.01.009. XU T M, LUO Z W, YI Z T, et al. Research and application of antagonistic microorganisms in control of tobacco diseases[J]. Acta Agriculturae Jiangxi, 2016(1):41-44. DOI:10.3969/j.issn.1001-8581. 2016.01.009.
[12] 刘文波,秦春秀,熊燕红,等. 解淀粉芽孢杆菌HAB-7对18株植物病原真菌的抑制作用[J]. 热带农业科学,2017(2):52-57+66. DOI:10.12008/j.issn.1009-2196.2017.02.013. LIU W B, QIN C X, XIONG Y H, et al. Analysis of antifungal activity of Bacillus amyloliquefaciens HAB-7 against 18 phytopathogens[J]. Chinese Journal of Tropical Agriculture, 2017(2):52-57. DOI:10.12008/j.issn.1009-2196.2017.02.013.
[13] 卢钰升,顾文杰,蒋瑞萍,等. 一株生防细菌GB58的鉴定与抑菌能力测定[J]. 中国农学通报,2016, 32(6):198-204. DOI:10.11924/j.issn.1000-6850. Casb15100004. LU Y S, GU W J, JIANG R P, et al. Identification and antimicrobial broad-spectrum analysis of a biocontrol bacteria strain GB58[J]. Chinese Agricultural Science Bulletin, 2016, 32(6):198-204. DOI:10.11924/j.issn.1000-6850. Casb15100004.
