Aims:To isolate an antagonist for use in the biological control of phytopathogenic fungi including Colletotrichum gloeosporioides, then to purify and characterize the biocontrol agent produced by the antagonist. Methods and Results: Bacteria that exhibited antifungal activity against the causative agent pepper anthracnose were isolated from soil, with Bacillus thuringiensis CMB26 showing the strongest activity. A lipopeptide produced by B. thuringiensis CMB26 was precipitated by adjusting the pH 2 with 3 N N HCl and extracted using chloroform/ methanol (2 : 1, v/v) and reversed-phase HPLC. The molecular weight was estimated as 1447 Da by MALDI-TOF mass spectrometry. Scanning electron and optical microscopies showed that the lipopeptide has activity against Escherichia coli O157:ac88, larvae of the cabbage white butterfly (Pieris rapae crucivora) and phytopathogenic fungi. The lipopeptide had cyclic structure and the amino acid composition was LL-Pro, and L L-Ile in a molar ratio of 3 : 1 : 2 : 1 : 1 : 2 : 1 : 1. The purified lipopeptide showed the same amino acid composition as fengycin, but differed slightly in fatty acid composition, in which the double bond was at carbons 13-14 (m/z 303, 316) and there was no methyl group. Conclusion: A lipopeptide was purified and characterized from B. thuringiensis CMB26 and found to be similar to the lipopeptide fengycin. This lipopeptide can function as a biocontrol agent, and exhibits fungicidal, bactericidal, and insecticidal activity. Significance and Impact of the Study: Compared with surfactin and iturin, the lipopeptide from B. thuringiensis CMB26 showed stronger antifungal activity against phytopathogenic fungi. This lipopeptide is a candidate for the biocontrol of pathogens in agriculture.
Environmentally friendly control measures are needed for suppression of soilborne pathogens of vegetable crops in the Republic of Korea. In vitro challenge assays were used to screen approximately 500 bacterial isolates from 20 Korean greenhouse soils for inhibition of diverse plant pathogens. One isolate, Bacillus subtilis ME488, suppressed the growth of 39 of 42 plant pathogens tested. Isolate ME488 also suppressed the disease caused by Fusarium oxysporum f. sp. cucumerinum on cucumber and Phytophthora capsici on pepper in pot assays. Polymerase chain reaction was used to screen isolate ME488 for genes involved in biosynthesis of 11 antibiotics produced by various isolates of B. subtilis. Amplicons of the expected sizes were detected for bacD and bacAB, ituC and ituD, and mrsA and mrsM involved in the biosynthesis of bacilysin, iturin, and mersacidin, respectively. The identity of these genes was confirmed by DNA sequence analysis of the amplicons. Bacilysin and iturin were detected in culture filtrates from isolate ME488 by gas chromatography coupled with mass spectroscopy and by thin layer chromatography, respectively. Detection of mersacidin in ME488 culture filtrates was not attempted. Experiments reported here indicate that B. subtilis ME488 has potential for biological control of pathogens of cucumber and pepper possibly due to the production of antibiotics.
Lewis, Jack A.; and Chung, Soohee, "Biocontrol agents applied individually and in combination for suppression of soilborne diseases of cucumber" (2005 AbstractThe soilborne pathogens Rhizoctonia solani, Pythium ultimum, and Meloidogyne incognita can cause severe economic losses to field-and greenhouse-grown cucumber. A collection of bacterial isolates and isolates GL3 and GL21 of Trichoderma virens were screened for suppression of diseases caused by these pathogens. T. virens isolates GL3 and GL21 provided the most effective suppression of damping-off caused by R. solani in greenhouse bioassays. Burkholderia ambifaria BC-F, B. cepacia BC-1, and Serratia marcescens N1-14 also provided significant suppression of R. solani relative to the pathogen check in some experiments. T. virens isolates GL3 and GL21 and S. marcescens isolates N1-6, N1-14, and N2-4 provided the most consistent and effective suppression of damping-off of cucumber caused by P. ultimum in growth chamber experiments. No microbial treatment containing individual or combined microbes significantly suppressed populations of M. incognita on cucumber or improved plant vigor in greenhouse bioassays. T. virens GL21 applied as a granular formulation, in combination with B. cepacia BC-1 or B. ambifaria BC-F applied as a seed treatment, significantly improved suppression of damping-off caused by R. solani over individual applications of these microbes in at least one experiment. Treatments combining B. cepacia BC-1, B. ambifaria BC-F, or S. marcescens isolates N1-14 or N2-4 with T. virens GL21 in R. solani biocontrol assays always resulted in plant stands that were similar or greater than treatments containing individual applications of these microbes. B. ambifaria BC-F combined with T. virens GL21 in seed treatments resulted in significantly improved suppression of damping-off caused by P. ultimum in two of three experiments. Populations of T. virens GL3 and GL21 were both substantially reduced after coincubation with B. cepacia BC-1, or S. marcescens isolates N1-14 or N2-4 for 10 to 12 d in cucumber rhizospheres. Populations of T. virens GL21 were slightly reduced after coincubation with B. ambifaria BC-F. Results presented here substantiate other studies reporting enhanced biocontrol performance This article is a U.S. government work, and is not subject to copyright in the United States.with certain combinations of biocontrol agents. These results also indicate that antagonism among combinations of biocontrol agents can vary with the assay system employed. r
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