• Korean journal of applied entomology
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• v.34 no.4
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• pp.344-349
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• 1995

To isolate naturally occurring novel Bacillus thuringiensis strains for useful pest con시 agents, A total of 64 isolates of B. thuringiensis producing spore and crystal were obtained from 583 soil samples in Korea. Toxicity of isolates was assayed against Lepidoptera (Bombyx ori), Diptera (Culex pipens) and Coleoptera (Sitophilus oryzae), respectively. The results showed that B. thuringiensis isolates toxic Lepidoptera are 42.2% , both Lepidoptera and Diptera are 31.3% and Diptera are 20.3%, demonstrating that spectrum of insecticidal activity of B. thuringiensis isolates is various. But no B. thuringiensis isolates have toxicity against Coleoptera.

• The Plant Pathology Journal
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• v.37 no.3
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• pp.243-257
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• 2021

Bacillus pumilus is the causal agent of trunk bulges disease affecting rubber and rubberwood quality and yield production. In this study, B. pumilus and other closely related species were included in B. pumilus group, as they shared over 99.5% similarity from 16S rRNA analysis. Multilocus sequence analysis (MLSA) of five housekeeping genes and repetitive elements-based polymerase chain reaction (rep-PCR) using REP, ERIC, and BOX primers conducted to analyze the diversity and systematic relationships of 20 isolates of B. pumilus group from four rubber tree plantations in Peninsular Malaysia (Serdang, Tanah Merah, Baling, and Rawang). Multi rep-PCR results revealed the genetic profiling among the B. pumilus group isolates, while MLSA results showed 98-100% similarity across the 20 isolates of B. pumilus group species. These 20 isolates, formerly established as B. pumilus, were found not to be grouped with B. pumilus. However, being distributed within distinctive groups of the B. pumilus group comprising of two clusters, A and B. Cluster A contained of 17 isolates close to B. altitudinis, whereas Cluster B consisted of three isolates attributed to B. safensis. This is the first MLSA and rep-PCR study on B. pumilus group, which provides an in-depth understanding of the diversity of these rubber-pathogenic isolates in Malaysia.

• The Korean Journal of Mycology
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• v.44 no.1
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• pp.36-47
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• 2016

This study was conducted to evaluate five different strains of rhizobacterial isolates viz. PA1, PA2, PA4, PA5 and PA12 for biological control against Colletotrichum acutatum, C. coccodes, C. gloeosporioides, C. dematium, Botrytis cinerea, Rhizoctonia solani, Sclerotinia minor and Fusarium sp. In vitro inhibition assay was performed on three different growth mediums, potato dextrose agar (PDA), tryptic soy agar (TSA), and PDA-TSA (1:1 v/v) for the selection of potential antagonistic isolates. According to the result, isolate PA2 showed the highest inhibitory effect with 65.5% against C. coccodes on PDA and with 96.5% against S. minor on TSA. However, the same isolate showed the highest inhibition with 58.5% against C. acutatum on PDA-TSA. In addition, an in vivo experiment was performed to evaluate these bacterial isolates for biological control against fungal pathogens. Plants treated with bacteria were analyzed with phytopathogens and plants inoculated with phytopathogens were treated with isolates to determine the biological control effect against fungi. According to the result, all five isolates tested showed inhibitory effects against phytopathogens at various levels. Mode of action of these rhizobacterial isolates was evaluated with siderophore production, protease assay, chitinase assay and phosphate solubilizing assay. Bacterial isolates were identified by 16S rDNA sequencing, which showed that isolates PA1 and PA2 belong to Bacillus subtilis, whereas, PA4, PA5, and PA12 were identified as Bacilus altitudinis, Paenibacillus polymyxa and Bacillus amyloliquefaciens, respectively. Results of the current study suggest that rhizobacterial isolates can be used for the plant growth promoting rhizobacteria (PGPR) effect as well as for biological control of various phytopathogens.

• Journal of Microbiology and Biotechnology
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• v.31 no.9
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• pp.1241-1255
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• 2021

This study was carried out to explore a non-chemical strategy for enhancing productivity by employing some antagonistic rhizobacteria. One hundred eighteen bacterial isolates were obtained from the rhizospheric zone of various crop fields of Gangwon-do, Korea, and screened for antifungal activity against Fusarium wilt (Fusarium oxysporum f. sp. lactucae) in lettuce crop under in vitro and in vivo conditions. In broth-based dual culture assay, fourteen bacterial isolates showed significant inhibition of mycelial growth of F. oxysporium f. sp. lactucae. All of the antagonistic isolates were further characterized for the antagonistic traits under in vitro conditions. The isolates were identified on the basis of biochemical characteristics and confirmed at their species level by 16S rRNA gene sequencing analysis. Arthrobacter sulfonivorans, Bacillus siamensis, Bacillus amyloliquefaciens, Pseudomonas proteolytica, four Paenibacillus peoriae strains, and Bacillus subtilis were identified from the biochemical characterization and 16S rRNA gene sequencing analysis. The isolates EN21 and EN23 showed significant decrease in disease severity on lettuce compared to infected control and other bacterial treatments under greenhouse conditions. Two bacterial isolates, EN4 and EN21, were evaluated to assess their disease reduction and growth promotion in lettuce in field conditions. The consortium of EN4 and EN21 showed significant enhancement of growth on lettuce by suppressing disease caused by F. oxysporum f. sp. lactucae respectively. This study clearly indicates that the promising isolates, EN4 (P. proteolytica) and EN21 (Bacillus siamensis), can be commercialized and used as biofertilizer and/or biopesticide for sustainable crop production.

• Microbiology and Biotechnology Letters
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• v.47 no.3
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• pp.359-363
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• 2019

Meju is a spontaneously fermented soybean product produced in Korea. We isolated the bacteria of the genera Bacillus, Enterococcus, and Staphylococcus from 12 meju samples collected from five regions of Korea using selective media, and we identified them based on the sequence analysis of 16S ribosomal RNA and gmk (guanylate kinase) genes. Bacillus and Enterococcus strains were identified from all samples and the numbers of Bacillus strains in the 11 samples were >15% of the total cell number. Therefore, the genus Bacillus was confirmed as the predominant bacterial group of meju. Staphylococcus strains were identified from six samples. The identified 151 Bacillus isolates predominated in the following order: B. velezensis, B. sonorensis, B. subtilis, and B. licheniformis. Among the 165 Enterococcus isolates, 163 strains were identified as E. faecium. Eighty-two Staphylococcus isolates were classified into six species of coagulase-negative Staphylococcus group and S. xylosus was the predominant species.

• Korean Journal of Food Science and Technology
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• v.42 no.3
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• pp.361-367
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• 2010

Bacterial contamination of cooked rice was analyzed to evaluate the microbial safety. Thirty raw rice samples were collected in Korea and cooked in an electric rice cooker. Mesophilic aerobe, food-poisoning Bacillus cereus group, and their toxin genes were determined on cooked rice. The percentage of total mesophilic aerobe based on 1-3 log CFU/g was 27% among the samples. Bacillus spp. in MYP selective medium was similar to the number of mesophilic aerobe, whileas Bacillus spp. was detected in most samples after enrichment. Thirty-seven isolates from 30 cooked rices were identified as B. thuringiensis, B. cereus, B. valismortis, B. pumilus, B. coagulans, B. licheniformis, Geobacillus stearothermophilus, and Brevibacillus laterosporus. Twenty isolates (54%), more than half of the isolates, were B. thuringiensis while nine (27%) were identified as B. cereus. All B. thuringiensis isolates possessed non-hemolytic toxin genes and interestingly, seven B. cereus among nine isolates possessed emetic toxin genes. More B. thuringiensis was present on the cooked rice than B. cereus and most B. cereus possessed emetic toxin genes rather than diarrheal toxin genes. Therefore, food-borne outbreak due to B.cereus on the cooked rice kept at room temperature might be examples of emetic food-poisoning.

• Journal of Microbiology and Biotechnology
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• v.22 no.6
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• pp.729-735
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• 2012

The PCR-RFLP method has been useful for detection of known genes and identification of novel genes. In the present study, degenerate primers were designed from five groups of cry1 genes for PCR-RFLP analysis. Bacillus thuringiensis (Bt) isolates from different regions were evaluated for PCR amplification of various cry1 genes using newly designed primers and cry2 genes using reported primers. PCR analysis showed an abundance of cry1A genes and especially cry1Ac genes in isolates from all regions. RFLP analysis revealed the presence of multiple cry1A genes in isolates from central and southern regions. Unique digestion patterns of cry1A genes were observed in isolates from each region. Few of the isolates represented a digestion pattern of cry1A genes that did match to any of the known cry1A genes. RFLP analysis suggested an abundance of cry2Ab along with a novel cry2 gene in Bt isolates from different regions of India. Sequence analysis of the novel cry2 gene revealed 95% sequence identity to cry2Ab and cry2Ah genes. Phylogenetic analysis revealed that the novel cry2 gene could have diverged earlier than the other cry2 genes. Our results encourage finding of more diverse cry2 genes in Bt isolates. Rarefaction analysis was used to compare cry1A gene diversity in isolates from different soil types. It showed a higher degree of cry1A gene diversity in isolates from central region. In the present study, we propose the use of novel degenerate primers for cry1 genes and the PCR-RFLP method using a single enzyme to distinguish multiple cry1A and cry2 genes as well as identify novel genes.

• Korean journal of applied entomology
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• v.36 no.3
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• pp.264-269
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• 1997

Four strains of Bacillus thuringiensis were isolated froin the dead larvae of mulberry longicorn beetle (Apriong germari) and dung beetle (Aphodius apicalis). One nf four B. thuringiensis isolates turned out to be subspecies darinstadiensis but the remains were not identified using 33 B. thuringiensts flgellar ( H ) antibodies. Furthermore. bioassays of spore-parasporal inclusion protein mixture conducted against third instar larvae of A. gerrntrri or A. apicalis, second instar larvae of Bombyx mori, and third instar larvae of Cu1ex pipiens pullens showed that the isolates were non-toxic. To further confirm, four isolates were characterized and analysed by SDS-PAGE and agarose gel electrophoresis. The results revealed that parasporal protein and plasmid DNA patterns of four isolates are different from those of darmstadiensis and 20 known non-toxic B. thuringiensis strains, suggesting that the four isolates are novel non-toxic B. thuringiensis.

• Korean Journal Plant Pathology
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• v.10 no.4
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• pp.297-304
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• 1994

This study was carried out to develop bioassay for detection of pesticide residues in agricultural products by using the soil microbial isolates sensitive to pesticides. One hundred bacterial isolates and eighty five fungal isolates were obtained from soil and their sensitivity to 10 ppm of several pesticides was examined in vitro. Five bacterial isolates and three fungal isolates were found sensitive to organochloride fungicide and two fungal isolates sensitive to organocopper fungicide. Among these isolates, B46, B93 and F67 were tested to find out the difference in sensitivity according to the methods of fungicide treatment. All of the isolates were found sensitive to 10 ppm of organochloride fungicides mixed directly in PDA. But they were found insensitive to the fungicide mixed in PDA after filtering through membrane filter. In case of organocopper fungicide, the isolates were found sensitive only when it was treated in PDA. And their sensitivity showed difference among various kinds of organochloride fungicides. B46 and B93 were employed to check the possibility as the agent for detection of the pesticidal residues in twenty eight agricultural products including rice. It was found that all samples had not residues because the samples did not inhibit the growth of isolates. When organochloride fungicides were applied to the above products, it was possible to detect the residues in fruits and vegetables at the concentration of 10 ppm, but not in starch-rich grains. B46 and B93 were identified as Bacillus sp. according to their bacterial characteristics in culture.

• Journal of Sericultural and Entomological Science
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• v.38 no.2
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• pp.154-159
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• 1996

To isolate Bacillus thuringiensis toxic to Spodoptera species, we collected soil samples in Korea. In these samples, we characterized 7 B. thuringiensis isolates toxic to spodoptera exigua or S. litura from soil, granary and sericultural farm samples. The 7 isolates were named B. thuringiensis STB-1, STB-2, STB-3, STB-4, STB-5, STB-6 and STB-7, respectively. The bioassay of these isolates against S. exigua and S. litura showed highly insecticidal activity. The serotypes of them were determined by agglutination tests using 33 antisera ; STB-1 an STB-2 are identical to B. thuringiensis subsp. kurastaki, and STB-3, STB-4 and STB-5 are identical to subsp. kenyae. STB-6 and STB-7 did not react with 33 antisera. STB-1 and STB-3 which have different gene types from B. thuringiensis subsp. kurastaki and subsp. kenyae are identified new isolates. STB-6 and STB-7 which show no agglutination in serological tests havd cryIA(a), cryIA(b), cryIC, and cryII genes are also identified new isolates. Molecular weights of parasporal inclusions of all isolates were determined approximately 130 kDa by SDS-polyacrylamide gel elctrophoresis.