| 李晨楚,谭志琼,康迅,靳鹏飞,张荣意.枯草芽孢杆菌B43对香蕉枯萎病菌抑菌活性及其活性成分分析[J].中国南方果树,2017,46(3): |
| 枯草芽孢杆菌B43对香蕉枯萎病菌抑菌活性及其活性成分分析 |
| Characterization of the active substances of Bacillus subtilis B43 against Banana Fusarium Wilt |
| 投稿时间：2016-12-16 修订日期：2017-02-24 |
| DOI： |
| 中文关键词: 枯草芽孢杆菌B43 酸沉淀法 抑菌试验 活性成分 |
| 英文关键词:Bacillus subtilis B43 hydrochloric acid precipitation antifungal test active substances |
| 基金项目:海南大学博士启动基金（kyqd 1330）、海南省自然科学基金项目（314061）、海南省耕地改良关键技术研究与示范专项“水稻-瓜菜-绿肥等轮作模式研究与推广”（HNG Dg1201504）、海南大学2016年研究生实践创新项目、2016年海南省研究生创新基金项目（HyS-2016-20） |
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| 中文摘要: |
| 为明确拮抗菌枯草芽孢杆菌B43菌株拮抗香蕉枯萎病菌4号小种(Fusarium oxysporum f.sp. cubense race 4，FOC-4)的能力及其抑菌活性物质的性质，本研究采用平板对峙法进行抑菌谱试验；酸沉淀、有机溶剂萃取法提取B43菌株中的抑菌活性物质并作稳定性测定；根据脂肽类抗生素合成相关基因序列对B43基因组进行PCR扩增检测，运用LC-MS对粗提物作成分分析。结果表明，B43菌株对21种植物病原真菌均有较好的抑菌活性；酸沉淀法粗提物具有较好的耐热及耐紫外线特性，但不耐强酸强碱；通过特异性引物PCR检测，B43菌株基因组中含有srfAB、ituA、ituB、ituD、fenD、yndJ、sfp、mycB、lpa-14或fenB等脂肽类合成相关基因；LC-MS分析发现粗提物中含有Surfactin、Iturin、Mycosubtilin、Bacillomycin和Fengycin 5类脂肽类抗生素的同系物24种。枯草芽孢杆菌B43菌株抑菌谱广，其发酵液酸沉淀粗提物对FOC-4具有良好的抑菌活性以及较好的稳定性，本研究不仅检测推断出粗提物中含有多种脂肽类抗生素及其同系物，且于菌株基因组中扩增出多种脂肽抗生素基因，说明该菌株可能能够产生多种脂肽类抗生素。B43菌株与已报道的多数拮抗FOC-4的芽孢杆菌菌株相比，效果好、活性物质种类更多，因此具有良好的生物农药开发应用前景。 |
| 英文摘要: |
| In order to determine the antifungal substances of Bacillus subtilis strain B43 against Fusarium oxysporum f.sp. cubense race 4 and analyze their properties, the antifungal property of strain B43 against 21 plant pathogens was measured by using the plate dual-culture antagonistic method. The hydrochloric acid precipitation and organic solvent were used to extract the antifungal substances from strain B43, then screened out an effective way and their stability was analyzed. In addition, seventeen primers designed based on the known lipopeptide genes were used to amplify the corresponding genes from the genome of B43 strain. Lipopeptides were identified by LC-MS techniques. The results showed that strain B43 exhibited an effective inhibitory activity against 21 plant pathogens, with the inhibition rates ranging from 38.03% to 85.51%. Among the different extract methods, the hydrochloric acid precipitation exhibited potent antifungal activity. To a certain degree, antifungal substances exhibited pH stability among 4 to 10, and strong stability with heat and UV. The PCR products with seventeen primer pairs were cloned and sequenced. The BLAST analysis showed that the srfAB、ituA、ituB、ituD、fenD、yndJ、sfp、mycB、lpa-14 or fenB genes exist in the genome of strain B43. Twenty-four kinds of lipopeptides surfactin、Iturin、Mycosubtilin、bacillomycin and fengycin were identified in the antimicrobial extract by the LC-MS analysis. B. subtilis strain B43 demonstrated a broad spectrum of antifungal activity. The hydrochloric acid precipitation extract from strain B43 exhibited strong antifungal activity against FOC-4 and favorable stability. Strain B43 could produce many kinds of lipopeptides, exhibiting promising prospects for its further study and application. |
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