| 烟草黑胫病拮抗菌的筛选、鉴定及发酵条件优化 |
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| 引用本文： | 何明川,王志江,谢永辉,詹莜国,柯昌磊,李微杰,张忠,吴国星.烟草黑胫病拮抗菌的筛选、鉴定及发酵条件优化[J].中国生物防治学报,2022,38(2):428-439. |
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| 作者姓名： | 何明川 王志江 谢永辉 詹莜国 柯昌磊 李微杰 张忠 吴国星 |
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| 作者单位： | 1. 云南农业大学植物保护学院，昆明650201；2.云南省烟草公司昆明市公司，昆明650051；3. 山东第一医科大学生命科学学院，泰安 271016 |
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| 基金项目： | 云南省烟草公司科技计划项目(2020530000242026) |
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| 摘 要： | 为获得对烟草黑胫病（tobacco black shank）具有较好拮抗作用的菌株，从食腐昆虫美洲大蠊Periplaneta americana肠道分离纯化得到1株对烟草黑胫病具有较强抑制作用的D5-8菌株，拟进一步明确该菌株的分类地位，优化其发酵条件并测定其对烟草黑胫病的防治效果。利用形态特征、生理生化特性观察及16S rRNA基因序列分析对D5-8菌株进行鉴定；以细菌发酵液OD600值及对病原菌的抑制率为指标，对其发酵培养基和发酵条件进行单因素试验及正交试验；通过室内盆栽试验，明确菌株D5-8对烟草黑胫病的防治效果。结果表明，经平板对峙试验发现菌株D5-8对烟草寄生疫霉的抑制率达66.80%；通过形态特征和生理生化特征初步确定菌株D5-8为芽胞杆菌属Bacillus，结合16S rRNA基因序列分析鉴定菌株D5-8为特基拉芽胞杆菌Bacillus tequilensis；其最适培养基：牛肉浸膏8 g，酵母浸粉5 g，麦芽糖10 g，蒸馏水1000 mL；最佳的发酵条件为温度28℃、初始pH 6、转速210 r/min、装液量30 mL、接种量2%、培养时间60 h、光照时间4 h；盆栽试验结果表明，发酵条件优化后D5-8发酵液（108cfu/mL）对烟草黑胫病的保护防效和治疗防效可达66.89%和53.72%，均高于原始发酵液的61.84%和51.83%。研究结果为特基拉芽胞杆菌D5-8生防菌剂的开发及其对烟草黑胫病的防治应用提供了理论依据。
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| 关 键 词： | 美洲大蠊 烟草黑胫病 特基拉芽胞杆菌 鉴定 发酵条件优化 |
| 收稿时间： | 2021-04-26 |
Screening,Identification and Fermentation Conditions Optimization of Antagonistic Bacterium against Tobacco Black Shank |
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| HE Mingchuan,WANG Zhijiang,XIE Yonghui,ZHAN Youguo,KE Changlei,LI Weijie,ZHANG Zhong,WU Guoxing.Screening,Identification and Fermentation Conditions Optimization of Antagonistic Bacterium against Tobacco Black Shank[J].Chinese Journal of Biological Control,2022,38(2):428-439. |
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| Authors: | HE Mingchuan WANG Zhijiang XIE Yonghui ZHAN Youguo KE Changlei LI Weijie ZHANG Zhong WU Guoxing |
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| Affiliation: | 1. College of Plant Protection, Yunnan Agricultural University, Kunming 650201, China; 2. Yunnan Tobacco Company Kunming Company, Kunming 650201, China; 3. College of Life Sciences, Shandong First Medical University, Taian 271016, China |
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| Abstract: | A strain D5-8 with strong inhibitory effect on tobacco black shank was isolated and purified from the intestinal tract of Periplaneta americana. The strain D5-8 was identified as Bacillus tequilensis by morphological characteristics, physiological and biochemical characteristics and 16S rRNA gene sequence analysis. The fermentation conditions of strain D5-8 were optimized and its control effect on tobacco black shank disease was also determined. Single factor test and orthogonal test were carried out on the fermentation medium and fermentation conditions with the OD600 value of bacterial fermentation broth and the inhibition rate against pathogenic bacteria as indexes, and control effect of strain D5-8 on tobacco black shank was determined by pot experiment. The results showed that the inhibition rate of strain D5-8 against Phytophthora nicotiana reached 66.80%. The optimal medium was beef extract 8 g, yeast extract 5 g and maltose 10 g in distilled water 1000 mL. The optimal fermentation conditions were as follows: temperature 28 ℃, initial pH 6, rotational speed 210 r/min, liquid volume 30 mL, inoculation volume 2%, culture time 60 h and light time 4 h. Pot experiment showed that the control efficacy of D5-8 fermentation broth (108cfu/mL) on tobacco black shank disease was 66.89% and 53.72%, which were higher than that of original fermentation broth (61.84% and 51.83%). |
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| Keywords: | Periplaneta americana tobacco black shank Bacillus tequilensis identification optimization of fermentation conditions |
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