Isolation, Screening and Identification of Native and New Bacillus subtilis with Strong Antifungal Compound against Fusarium oxysporum.

IF 0.9 4区 农林科学 Q4 BIOTECHNOLOGY & APPLIED MICROBIOLOGY Biocontrol science Pub Date : 2022-01-01 DOI:10.4265/bio.27.201
Fatemeh Rafiee, Mohammad Reza Fazeli, Abbas Akhavan Sepahi, Zahra Noormohammadi
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引用次数: 2

Abstract

The genus Fusarium causes a wide range of infections in human, animals and herbs. The purpose of this research was to investigate and identify the native strains of Bacillus subtilis playing an inhibitory role against Fusarium oxysporum by producing surfactin. B. subtilis was isolated from the soil of various parks in Tehran-Iran, and identified by biochemical tests. Growth inhibition zone, minimum inhibitory concentration (MIC) and minimum fungicidal concentration (MFC) of B. subtilis were determined. After purification of surfactin, quantitative and qualitative analysis of surfactin conducted using high performance liquid chromatography (HPLC) . Finally, two selected native strains with the highest production rate of surfactin identified using PCR for 16S rRNA and phylogenetic tree was drawn. Sixty strains of B. subtilis were isolated from soil, after identification through phenotypical and biochemical tests, the antagonistic activity of 27 different strains against F. oxysporum by Agar well diffusion assay determined and the highest inhibition zone was 13.66 mm. Six strains showing the best inhibitory effect, were isolated and their metabolite were purified by methanol. MIC and MFC values of different strains were in the range of 0.5-1.6 and 1.6-2.6 mg/mL. Using HPLC, the purified surfactin content in B. subtilis was about 56.7 - 131.9 μg/mL. Based on the curves of the chromatogram, the preferred strains with the highest production of surfactin, by molecular identification, displayed high similarity to B. subtilis. We got a maximum amount of yellow and transparent surfactin from native strains. Furthermore, the selected bacteria can be good candidates for biological control of fungal pathogens.

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具有抗尖孢镰刀菌强效化合物的原生与新型枯草芽孢杆菌的分离、筛选与鉴定。
镰刀菌属在人类、动物和草药中引起广泛的感染。本研究的目的是研究和鉴定通过生产表面素来抑制尖孢镰刀菌的天然枯草芽孢杆菌菌株。从伊朗德黑兰多个公园的土壤中分离到枯草芽孢杆菌,并进行了生化鉴定。测定了枯草芽孢杆菌的生长抑制带、最小抑菌浓度(MIC)和最小杀真菌浓度(MFC)。表面素纯化后,采用高效液相色谱(HPLC)对表面素进行定量和定性分析。最后,通过16S rRNA PCR鉴定了2株产surfactin率最高的本地菌株,并绘制了系统发育树。从土壤中分离到60株枯草芽孢杆菌,经表型和生化鉴定,通过琼脂孔扩散法测定了27株不同菌株对尖孢镰刀菌的拮抗活性,最高抑菌带为13.66 mm。分离得到6株抑菌效果最好的菌株,并用甲醇对其代谢产物进行纯化。不同菌株的MIC和MFC值分别为0.5 ~ 1.6和1.6 ~ 2.6 mg/mL。HPLC法测得枯草芽孢杆菌中纯化的表面素含量约为56.7 ~ 131.9 μg/mL。从色谱曲线上看,表面素产量最高的优选菌株与枯草芽孢杆菌具有较高的相似性。我们从本地菌株中获得了最大量的黄色和透明表面素。此外,所选择的细菌可以作为真菌病原体生物防治的良好候选者。
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来源期刊
Biocontrol science
Biocontrol science BIOTECHNOLOGY & APPLIED MICROBIOLOGY-
CiteScore
2.60
自引率
8.30%
发文量
21
审稿时长
>12 weeks
期刊介绍: The Biocontrol Science provides a medium for the publication of original articles, concise notes, and review articles on all aspects of science and technology of biocontrol.
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