Isolation and Evaluation of Protective Effect against Fusarium Wilt of Sesame Plants of Antibiotic Substance from Bacillus polymyxa KB-8

An antibiotic compound was isolated from the culture of an antagonist against Fusarium oxysporum f. sp. sesami, Bacillus polymyxa strain KB-8, and tested for the control of Fusarium wilt of sesame in greenhouse conditions. Optimum conditions for culturing the antagonist to obtain the maximum antibiotic activity were determined using different culture media, initial medium acidity, and incubation periods for which yeast -malt extract agar with the initial acidity of pH 5 and over 13 days culture were best. Antibiotic substances extracted by methanol had 2 main fractions, KB-8A and KB-8B, in thin layer chromatography (OLC) with Rf values of 0.35 and 0.67 in a solvent system of chloroform : methanol = 7 : 3. The fraction KB-8A wa purified further by XAD-2, silica gel and Sephadex LH-20 column chromatography, and crystalization. Its minimum inhibitory concentrations (MICs) were $12.8\mu\textrm{g}$/ml for F. oxysporum and Alternaria mali, $6.4\mu\textrm{g}$/ml for Colletotrichum gloeosporioides and Rhizoctonia solani, and $3.2\mu\textrm{g}$/ml for Phytophthora capsici. Soil drenching of antibiotic KB-8A in the concentrations of $13.0\mu\textrm{g}$/ml and $26.0\mu\textrm{g}$/ml effectively inhibited the Fusarium wilt of sesame in a greenhouse test, which appeared to be comparable to the fungicide benlate of $6.5\mu\textrm{g}$ a. i./ml.