The Plant Pathology Journal

The Korean Society of Plant Pathology (한국식물병리학회)
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Purification and Structural Analysis of Surfactin Produced by Endophytic Bacillus subtilis EBS05 and its Antagonistic Activity Against Rhizoctonia cerealis

  • Wen, Cai-Yi (Post-doctoral Work Station, Fujian Agriculture and Forestry University) ;
  • Yin, Zhi-Gang (College of Plant Protection, Henan Agricultural University) ;
  • Wang, Kai-Xuan (College of Plant Protection, Henan Agricultural University) ;
  • Chen, Jian-Guang (College of Plant Protection, Henan Agricultural University) ;
  • Shen, Shun-Shan (College of Plant Protection, Henan Agricultural University)
  • Received : 2011.03.09
  • Accepted : 2011.08.19
  • Published : 2011.12.01
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Abstract

Bacillus subtilis EBS05, an endophytic bacteria strain isolated from a medicinal plant Cinnamomum camphor, can produce antagonistic compounds that effectively inhibit plant pathogenic fungi. The greenhouse experiments showed that wheat sharp eyespot disease (WSED) was reduced by 91.2%, 88.2% and 43.0% after the treatment with fermentation broth, bacteria-free filter and a fungicide fludioxonil, respectively. The culture broth of strain EBS05 can more effectively control WSED than can fludioxonil. The fermentation broth and bacteria-free filter ability to suppress WSED was not significantly different, suggesting that an active secreted substance played a major role in controlling WSED. Separation and purification of the active compounds was carried out by serial processes, including hydrochloric acid (pH 2.0) treatment, methanol extraction and Sephadex LH-20 column chromatography, silica gel column chromatography and reverse-phase high-pressure liquid chromatography (HPLC), respectively. The purified compounds, one of active peaks in the HPLC spectrum, were obtained from the collection. Analysis of the chemical structures by time-of-flight mass spectrometry (TOF-MS) and electrospray ionization mass spectrometry/mass spectrometry (ESI-MS/MS) showed that the active substances produced by the endophytic bacteria EBS05 are mixture of the ${\beta}$-hydroxy-C12~C15-$Leu^7$ surfactin A isomers with 1035.65 Da, 1021.64 Da, 1007.63 Da and 993.65 Da molecular weights, respectively.

Keywords

References

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