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烟草疫霉拮抗菌枯草芽孢杆菌21b菌株的分离鉴定及其生物学特性研究
引用本文:王进,黄艳飞,汪汉成,王茂胜,陆宁,余知和.烟草疫霉拮抗菌枯草芽孢杆菌21b菌株的分离鉴定及其生物学特性研究[J].微生物学通报,2014,41(12):2481-2487.
作者姓名:王进  黄艳飞  汪汉成  王茂胜  陆宁  余知和
作者单位:1. 长江大学 生命科学学院 湖北 荆州 434025;2. 长江大学 农学院 湖北 荆州 434025;3. 贵州省烟草科学研究院 贵州 贵阳 550081;3. 贵州省烟草科学研究院 贵州 贵阳 550081;3. 贵州省烟草科学研究院 贵州 贵阳 550081;1. 长江大学 生命科学学院 湖北 荆州 434025
基金项目:国家自然科学基金项目(No. 1360448);贵州省科学技术基金项目(No. 黔科合J字[2011]2337号);贵州省烟草公司资助项目(No. 201305,201336)
摘    要:【目的】为了控制烟草疫霉(Phytophthora nicotianae)引起的烟草黑胫病对烟草生产造成的危害。【方法】采用稀释平板法从贵州省毕节地区烟草根际土壤中分离筛选拮抗烟草疫霉的细菌菌株,然后经形态观察、Biolog鉴定及16S rRNA基因序列分析,对分离菌株进行鉴定,同时测定抗菌谱,单因子变量分析、优化生长条件。【结果】共分离得到44株拮抗烟草疫霉的细菌菌株,其中菌株21b对烟草黑胫病菌菌丝生长的抑制率达78.33%,鉴定为枯草芽孢杆菌(Bacillus subtilis)。该菌株对烟草青枯病菌(Ralstonia solanacearum)、烟草灰霉病菌(Botrytis cinerea)、烟草赤星病菌(Alternaria alternate)和烟草炭疽病菌(Colletotrichum destructivum)均具有拮抗作用,抑菌圈大小分别为19.5、18.2、14.6和13.4 mm,最佳的发酵条件为:温度30°C、p H 7.0–8.0、装液量12%、盐浓度0.5%。【结论】分离筛选到一株对烟草寄生疫霉有较强拮抗活性的细菌菌株,为进一步开发烟草黑胫病的生防菌剂提供了菌种资源。

关 键 词:烟草黑胫病  枯草芽孢杆菌b  拮抗活性  菌株鉴定  液体发酵

Isolation, identification and biological characteristics of a bacterial strain Bacillus subtilis 21b against Phytophthora nicotianae
WANG Jin,HUANG Yan-Fei,WANG Han-Cheng,WANG Mao-Sheng,LU Ning and YU Zhi-He.Isolation, identification and biological characteristics of a bacterial strain Bacillus subtilis 21b against Phytophthora nicotianae[J].Microbiology,2014,41(12):2481-2487.
Authors:WANG Jin  HUANG Yan-Fei  WANG Han-Cheng  WANG Mao-Sheng  LU Ning and YU Zhi-He
Institution:1. College of Life Sciences, Yangtze University, Jingzhou, Hubei 434025, China;2. College of Agriculture, Yangtze University, Jingzhou, Hubei 434025, China;3. Guizhou Academy of Tobacco Sciences, Guiyang, Guizhou 550081, China;3. Guizhou Academy of Tobacco Sciences, Guiyang, Guizhou 550081, China;3. Guizhou Academy of Tobacco Sciences, Guiyang, Guizhou 550081, China;1. College of Life Sciences, Yangtze University, Jingzhou, Hubei 434025, China
Abstract:Objective] Control of the spread of tobacco black shank caused by Phytophthora nicotianae in tobacco production. Methods] With Phytophthora nicotianae used as the assay fungus, bacterial strains antagonistic to P. nicotianae were isolated from tobacco rhizosphere soil obtained from Bijie District, Guizhou Province using dilution-plate methods and identified based on morphology, Biolog identification and 16S rRNA gene sequence analysis. The antimicrobial spectrum of these strains was further tested against Ralstonia solanacearum, Botrytis cinerea, Alternaria alternate and Colletotrichum destructivum in vitro. Optimization of growth conditions was also investigated through single factor variable analysis. Results] Of 44 antagonistic strains obtained, strain 21b showed an inhibition rate of 78.33% to P. nicotianae and was identified as Bacillus subtilis. The size of inhibition zone of this strain against Ralstonia solanacearum, Botrytis cinerea, Alternaria alternate, and Colletotrichum destructivum were 19.5, 18.2, 14.6 and 13.4 mm, respectively. The optimal fermentation conditions for 21b was a temperature of 30 °C, pH 7.0?8.0, media/flask volume 12%, and 0.5% salt. Conclusion] One bacterial strain with antagonistic activity against P. nicotianae isolated in this study could provide microbiological resources for the development of biocontrol of tobacco black shank in the future.
Keywords:Black shank of tobacco  Bacillus subtilis 21b  Antagonistic activity  Bacterial identification  Liquid fermentation
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