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2025年4月5日 星期六
  2016, Vol. 24 Issue (2): 261-269    
  研究论文与报告 本期目录 | 过刊浏览 | 高级检索 |
芽胞杆菌FJAT-28592抗真菌脂肽的研究
陈倩倩1,刘波2,王阶平3,车建美 1,刘国红3,龚海艳3,关雄4
1. 福建省农业科学院农业生物资源研究所
2. 福建省农业科学院生物技术研究所
3. 福建省农业科学院 农业生物资源研究所
4. 福建农林大学生命科学院
Anti-fungal Lipopetides Produced by Bacillus siamensis FJAT-28592
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摘要 为研究产表面活性剂芽胞杆菌(Bacillus)的生防作用,本研究筛选了一株能够抑制尖孢镰刀菌(Fusarium oxysporum)的芽胞杆菌 FJAT-28592,经16S rRNA鉴定为暹罗芽胞杆菌(Bacillus siamensis)。菌株FJAT-28592的生长温度范围为25~50 ℃,最佳生长温度为35 ℃;pH 5.0~9.0,最佳pH 6.0~8.0;可在NaCl浓度为0~6%的条件下生长,最佳生长盐浓度为2%。透射电镜显示,菌株呈杆状,具有鞭毛,宽度为0.76~0.94 μm,长度为1.76 ~2.94 μm。暹罗芽胞杆菌FJAT-28592发酵液和提取的发酵液粗产物对尖孢镰刀菌有很好的抑菌效果。通过血平板和排油圈实验,初步认定暹罗芽胞杆菌FJAT-28592能分泌表面活性剂至发酵液。通过酸沉淀处理,甲醇和丙酮抽提,分离和收集发酵液中表面活性剂。高效液相色谱(high performance liquid chromatography, HPLC)对初步纯化的粗品与iturin A的标准品进行分析,粗品与标准品iturin A的主峰保留时间均为10.687 min,初步判断粗品含有iturin A。克隆iturinA类脂肽合成的相关基因,获得丙二酰辅酶a转酰酶(malonyl-CoA-transacylase, ituD)和 4'-磷酸泛酰巯基乙胺基转移酶(4 '- phosphate generic acyl cysteamine transferase, lpa-14)基因序列,得出此表面活性剂为iturin A类的脂肽。本研究为芽胞杆菌生防菌的开发,分析获得抑菌物质提供基础资料。
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陈倩倩
刘波
王阶平
车建美
刘国红
龚海艳
关雄
关键词 暹罗芽胞杆菌生物表面活性剂高效液相色谱(HPLC)透射电镜    
Abstract:Recently, the use of biological control agents has received great attention because of their ability of suppressing different plant diseases. Some species of the genus Bacillus are the most promising candidates for bacterial biocontrol agents, as the species are well known as safe micro-organisms in the environment, versatility to protect plants effectively against plant pathogens. Bacillus species also have an outstanding ability to sporulation, which assures their prevalence in the environment. Bacillus strains often produce a range of antimicrobial cyclic lipopeptides, including iturins, fengycins and surfactines antibiotics in the family iturin, especially iturin A, is most famous for biocontrol activity. Iturin A is heptapeptide with a β-amino fatty acid that exhibits strong antifungal activity, and shows a strong antibiotic activity with a broad antifungal spectrum. A Bacillus strain FJAT-28592 kept in Microbiological Culture Collection Center of Fujian Academy of Agriculture Sciences was selected to search for naturally occurring biological control agents. The Bacillus strain FJAT-28592 identified as Bacillus siamensis by 16S rRNA analysis had antifungal activity against fungal pathogens Fusarium oxysporum. FJAT-28592 growed at 25~50 ℃, pH 5.0~9.0 and in the presence of 0~6% (W/V) NaCl. The optimal culture conditions was 35 ℃, pH 6.0~8.0 and salt concentration of 2%. Colonies grown on LB agar for 2 d were white, dry and flat. Cell morphology was observed with transmission electron microscopy, B. siamensis FJAT-28592 was motile rods, 0.76~0.94 μm in diameter and 1.76~2.94 μm in length after 2 d culture on LB agar. By blood agar plate and oil spreading test,the strain was confirmed to produce biosurfactant. The crude extract was gained by acid precipitation. Menthol extraction was compared with standard iturin A by high performance liquid chromatography (HPLC). Iturin A synthesis related malonyl-CoA-transacylase gene (ituD) and 4 '- phosphate generic acyl cysteamine transferase gene (lpa-14) were cloned by PCR. The chemical and molecular genetic evidence thus indicated that the biosurfactant from FJAT-28592 was a kind of iturin A. The fermentation crude extract containing iturin A possessed significantly inhibitory activities against F. oxysporum. B. siamensis FJAT-28592 was an ideal potential biological control agent with the aim of reducing the use of chemical pesticides in agriculture. This study provides basic data on the developing biocontrol Bacillus and exploring bacterial biocontrol substances.
Key wordsBacillus siamensis    Biosurfactant    High performance liquid chromatography (HPLC)    Transmission electron microscopes
收稿日期: 2015-08-13      出版日期: 2015-12-29
基金资助:福建省自然科学基金
通讯作者: 关雄     E-mail: guanxfafu@126.com
引用本文:   
陈倩倩 刘波 王阶平 车建美 刘国红 龚海艳 关雄. 芽胞杆菌FJAT-28592抗真菌脂肽的研究 [J]. , 2016, 24(2): 261-269.
链接本文:  
http://journal05.magtech.org.cn/Jwk_ny/CN/     或     http://journal05.magtech.org.cn/Jwk_ny/CN/Y2016/V24/I2/261
 
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