• Microbiology and Biotechnology Letters
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• v.30 no.2
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• pp.135-141
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• 2002

This study was carried out to isolate antagonistic bacterium against Sclerotium cepivorum causing Allium fistulosum white rot. Total of 146 strains were isolated from A. fistulosum roots. The isolates were screened for antagonism to S. cepivorum and the isolated strain No. AL-1 was selected among these bacteria. It was identified as Serratia plymuthica based on morphological and physiological characteristics according to the Bergey's mannual of systematic bacteriology and 16S rDNA sequences methods. Serratia plymuthica AL-1 showed broad spectrum of antifungal activities against plant pathogenic fungi Alternaria altrata, Colletotrichum gleosporioids, Phoma sp., Rhizoctonia solani, Sclerotinia sclerotiorum, Stemphylium solani, Fusarium oxysporium niveum but not inhibited Didymella bryoniae. When S. plymuthica AL-1 cultivated in the TSB medium containing 1％ colloidal chitin, the high molecular fraction (>10 kDa) have chitinase activity (3.2 units/ml) and the low molecular fraction (<10 kDa) have not chitinase activity. Oppositely, after heat treatment (80℃ for 30 min) of the cultivation supernatant, the high molecular fractions have not antifungal activity but the low molecular fractions have antifungal activity.

• Journal of Life Science
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• v.19 no.11
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• pp.1672-1678
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• 2009

The purified antifungal substances produced by Bacillus amyloliquefaciens IUB158-03 was positive to ninhydrin but negative to aniline, suggesting that the antifungal substance could be a peptide. FAB-MS, UV adsorption spectrum, and amino acid composition analysis revealed that the molecular weight of the antifungal substance was 1042 and that maximal adsorption was at 220 nm and 277 nm. The antifungal substance was composed of $Asn_3$, $Gln_2$, $Ser_1$, $Gly_1$, and $Tyr_1$. The composition and structural characteristics of antifungal substance were analysed by $^1H$-NMR spectrum, $^1H$-COSY, HMQC, which revealed that the compound belongs to the iturin A family. Temperature and pH had little effect on the stability of the antifungal substance in the ranges of $-70{\sim}121^{\circ}C$ and pH 6.0~10.0, respectively. It showed strong antibiotic activity against fungi. An in vitro cytotoxicity test using NIH3T3 cell showed that the antifungal substance does not have cytotoxicity. The number of circulating leukocytes and the hematobiological analysis of the mice administered with the antifungal substances was similar to those of the control group, indicating no cytotoxicity in vivo. Therefore, the antifungal substances extracted from culture broth of Bacillus amyloliquefaciens IUB158-03 have future potential as biocontrol agents against plant diseases caused by fungi.

• Research in Plant Disease
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• v.29 no.4
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• pp.390-398
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• 2023

Apple white rot, caused by Botryosphaeria dothidea, is one of the important diseases in Korea. B. dothidea can cause pre- and postharvest decay on apple fruit as well as canker and dieback of apple trees. In this study, we isolated bacteria from the trunk of apple trees and tested their antagonistic activity against B. dothidea. Five bacterial isolates (23-168, 23-169, 23-170, 23-172, and 23-173) were selected that were most effective at inhibiting the mycelial growth of the pathogens. The isolate 23-172 was identified as Bacillus amyloliquefaciens and four isolates 23-168, 23-169, 23-170, and 23-173 were identified as Bacillus velezensis by RNA polymerase beta subunit (rpoB) and DNA gyraseA subunit (gyrA) gene sequencing. All isolates showed strong antagonistic activity against B. dothidiea as well as Colletotrichum fructicola and Diaporthe eres. All isolates exhibited cellulolytic, proteolytic and phosphate solubilizing activities. In particular, two isolates 23-168, 23-169 were shown to significantly reduce the size of white rot lesions in pretreated apple fruits. These results will provide the basis for the development of a fungicide alternative for the control of white rot of apple.

• Research in Plant Disease
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• v.18 no.3
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• pp.201-209
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• 2012

The multi-function of 18 Bacillus subtilis isolates collected from agricultural extension centers of local government and National Academy of Agricultural Science was investigated by measuring their antifungal activities against five plant pathogens, such as Rhizoctonia solani, Colletotrichum acutatum, Fusarium oxysporum, Magnaporthe oryzae and Phytophthora capsici, phosphorus solubilization ability, production of indole acetic acid (IAA) and siderophore, and nitrogen fixation. The B. subtilis isolates showed antifungal activity against several plant pathogens and nitrogen fixation activity, and produced siderophore and IAA. They could control pepper powdery mildew (Leveillula taurica), but there was no difference in control efficacy among the B. subtilis isolates. In fields, the control efficacy of B. subtilis R2-1 ($10^8$ cells/ml) was compared with two microbial fungicides, Q-pect and Topsid. In 2009, the control efficacy of B. subtilis R2-1 (37.7%) was lower than that of Topsid (47.6%), but higher than that of Q-pect (25.7%). In 2010, the control efficacy of B. subtilis R2-1 (83.3%) was higher than that of Topsid (67.9%). In order to elucidate mode of action of B. subtilis R2-1 for controlling pepper powdery mildew, spore germination rates of pepper powdery mildew pathogen collected on treated leaves was investigated when suspensions of B. subtilis R2-1 and two microbial fungicides (Q-pect and Topsid) were foliar-sprayed. They highly suppressed spore germination of the pathogen with inhibition values of 84.2% for B. subtilis R2-1, 97.9% for Q-pect and 94.7% for Topsid. Further study on the mass-culturing method and formulation is needed for development of a microbial fungicide.

• Korean Journal of Food Science and Technology
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• v.45 no.3
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• pp.285-292
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• 2013

This study was carried out to validate the safety of ametoctradin residues in agricultural commodities by developing an official analysis method. An analytical method was developed and validated using HPLC-PDA detectors. The samples were extracted with methanol, subsequently partitioned with dichloromethane and purified with florisil column chromatograph using acetone/hexane (30/70, v/v) as solvent. The method was validated by using grape, hulled rice, mandarin, and potato spiked with ametoctradin at 0.05 and 5.0 mg/kg, and pepper at 0.05 and 2.0 mg/kg. Average recoveries were 76-114.8% with relative standard deviation less than 10%, and the limit of detection and limit of quantification were 0.0125 and 0.05 mg/kg, respectively. The result of recoveries and overall coefficient of variation of the laboratory results from Gwangju regional Food and Drug Administration (FDA) and Daejeon regional FDA was accorded with Codex Alimentarius Commission Guideline (CAC/GL 40). Based on these results, this method was found to be appropriate for ametoctradin residue determination and can be used as the official method of analysis.