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journal1 ›› 2017, Vol. 33 ›› Issue (5): 667-674.DOI: 10.16409/j.cnki.2095-039x.2017.05.013

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Screening, Identification of Antagonistic Ocean Bacteria Bacillus methylotrophicus and Its Antimicrobial Substances to Botrytis cinerea pres.

WEI Xinyan1, HUANG Yuanyuan2, HUANG Yali2, DU Kejiu1   

  1. 1. College of Forestry, Hebei Agricultural University, Baoding 071001, China;
    2. Institute of Biology, Hebei Academy of Science, Shijiazhuang 050051, China
  • Received:2017-06-08 Online:2017-10-08 Published:2017-10-11

Abstract: 464 strains of ocean bacteria were isolated from 65 samples of sea water and sediment from Bohai using ZoBell's 2216E medium. Through plate confrontation method, 47 strains of marine bacteria with strong antagonism against Botrytis cinerea were obtained. Strain BH21 with high salt and methanol tolerance had the strongest antifungal effect and the inhibition zone diameter was 11.8 mm±0.3 mm. Strain BH21 was identified as Bacillus methylotrophicus by morphological, physiological, biochemical and molecular methods. Using the specific primers of lipopeptide gene, the related genes in strain BH21 were amplified and 7 lipopeptide antibiotics synthetic genes including ItuA, ituB, ituC, ituD, fend, srfAB and yndJ genes were identified in the strain. Crude extract of lipopeptide was obtained by the hydrochloric acid precipitation and methanol extraction. Oxford cup analysis demonstrated that the crude extract of lipopeptide could inhibit the mycelial growth of B. cinerea, with the inhibition rate of 73.9%.

Key words: Marine bacteria, Botrytis cinerea, Bacillus methylotrophicus, lipopeptide antifungal substance, antagonistic microbe

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