[Identification and characterization of a Bacillus amyloliquefaciens with high antifungal activity]

Plant disease can cause serious crop losses, and chemical control of disease is costly both to the environment and to the farmer. Some microorganism can produce the substance which has the preventing and exterminating functions to plant pathogens. These substances are valid to plant pathogens with only lower concentration, in addition the substances do not remain in soil and crops without being decomposed. If composization is performed with the microorganism, or the microorganism is mixed into compost, the functional compost having preventing and exterminating action will be made out and that can be more useful to environmental preservation. In order to screen antifungal bacteria for use in biological control, 200 compost samples were taken from different regions in China, over 10 bacterium with clear antifungal activity were isolated from composts, among them, strain Q-12 exhibited the highest antifungal activity which was strongly inhibits the growth of many plant pathogenic fungi such as Fusarium oxysporum and Rhizoctonia solan. According to the characteristics of morphology, physiology and biochemistry tests (API 50 CHB/E system) and the comparison of 16S rDNA sequence, the strain Q-12 was similar to B. subtilis and B. amyloliquefaciens. Some specific genes yyaR, yyaO and tetB, which have previously been shown to be effective for resolving these closely related taxa of the B. subtilis group, were analysed to clarify further the classification of Q-12, and two pairs of primers YyaR _ F/TetB _ R and YyaO _ F/TetB _ R were designed. From the analysis of fingerprints obtained with the two primers, strain Q-12 and B. amyloliquefaciens showed identical genomic fingerprints with primers YyaR _ F/TetB R, indicating their closely genetic relationship, and was identified as B. amyloliquefaciens. In the investigation of the culture condition, growth was carried out in a basal medium and gradually supplemented with the various ingredients to be investigated. The major ingredients being investigated included carbon sources, nitrogen sources and inorganic salts. The optimum medium and culture conditions of the strain all were studied with single factor test. The strain is easy to cultivate, and the optimal antibiotic production condition is growth in a medium (5 g/L glucose, 1 g/L NH4Cl, 0.8 g/L beef extract, 5 g/L MgCl2, pH 6.0) at 33 degrees C for 40 h. A number of Bacillus strains have proven safe over many years as a nonpathogenic species and are consumed in ton quantities in several human food preparations. Strains of Bacillus have several advantages over other biocontrol bacteria in that they are often soil isolates capable of sporulation, easy to cultivate and store, and some strains have been shown to increase yields of various crops.