• Journal of Microbiology and Biotechnology
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• v.34 no.8
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• pp.1671-1679
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• 2024

Aspergillus flavus, the primary mold that causes food spoilage, poses significant health and economic problems worldwide. Eliminating A. flavus growth is essential to ensure the safety of agricultural products, and extracellular compounds (ECCs) produced by Bacillus spp. have been demonstrated to inhibit the growth of this pathogen. In this study, we aimed to identify microorganisms efficient at inhibiting A. flavus growth and degrading aflatoxin B₁. We isolated microorganisms from soil samples using a culture medium containing coumarin (CM medium) as the sole carbon source. Of the 498 isolates grown on CM medium, only 132 bacterial strains were capable of inhibiting A. flavus growth. Isolate 3BS12-4, identified as Bacillus siamensis, exhibited the highest antifungal activity with an inhibition ratio of 43.10%, and was therefore selected for further studies. The inhibition of A. flavus by isolate 3BS12-4 was predominantly attributed to ECCs, with a minimum inhibitory concentration and minimum fungicidal concentration of 0.512 g/ml. SEM analysis revealed that the ECCs disrupted the mycelium of A. flavus. The hydrolytic enzyme activity of the ECCs was assessed by protease, β-1,3-glucanase, and chitinase activity. Our results demonstrate a remarkable 96.11% aflatoxin B1 degradation mediated by ECCs produced by isolate 3BS12-4. Furthermore, treatment with these compounds resulted in a significant 97.93% inhibition of A. flavus growth on peanut seeds. These findings collectively present B. siamensis 3BS12-4 as a promising tool for developing environmentally friendly products to manage aflatoxin-producing fungi and contribute to the enhancement of agricultural product safety and food security.

• Journal of the Korean Society of Groundwater Environment
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• v.5 no.4
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• pp.215-222
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• 1998

The objective of this study was to investigate the effects of physico-chemical environmental factors on the changes of bacterial population from two sites used for drinking water and eight sites polluted with various pollutant in Seoul city. In all the stations except for two sites used for drinking water, the concentrations of nitrate- nitrogen and ammonia were in excess of the criteria of groundwater quality by the result of analysis of 40 variations including physicochemical environmental factors, heavy metals, and bacterial populations. The numbers of total bacteria, heterotrophic bacteria and functional groups of bacteria were ranged from 5.1 to 41.4${\times}$10$\^$5/cells/ml and from 0.01 to 29.6${\times}$10$^4$cfu/ml, respectively. The activities of extracellular enzymes showed the ranges of 0.005∼11.3${\mu}$M/l/hr and its order to lipase, phophatase, ${\beta}$-glucosidase, cellulase, chitinase, amylase. The results of correspondence and multidimensional scaling analysis between bacterial populations and its physico-chemical environmental factors were explained the effects of physico-chemical environmental factors according to site characters and separated four group, which is accord with potential pollutants at wells.

• The Plant Pathology Journal
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• v.20 no.1
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• pp.75-80
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• 2004

In an attempt to develop effective biocontrol system for management of stem rot disease in groundnut, 57 bacterial isolates and 13 isolates of Trichoderma spp. were evaluated for their antagonistic activity against Sclerotium rolfsii. The antagonists were selected based on their ability to inhibit the external growth of S. rolfsii from infected groundnut seeds. Four isolates of Pseudomonas fluorescens, GB 4, GB 8, GB 10 and GB 27, and T. viride pq 1 were identified as potent antagonists of S. rolfsii. T. viride pq 1 produced extracellular chitinase and parasitized the mycelium of S. rolfsii. Under controlled environment conditions, P. fluorescens GB 10, GB 27, T. viride pq 1 and the systemic fungicide Thiram(equation omitted) reduced the mortality of S. rolfsii inoculated to groundnut seedlings by 58.0％, 55.9％, 70.0％ and 25.9％, respectively compared to control. In vitro growth of P. fluorescens GB 10 and GB 27 was compatible with T. viride pq 1 and Thiram(equation omitted). Integrated use of these two bacterial isolates with T. viride pq 1 or Thiram(equation omitted) improved their biocontrol efficacy. Combined application of either GB 10 or GB 27 with T. viride pq 1 was significantly effective than that with Thiram(equation omitted) in protecting groundnut seedlings from stem rot infection.

• Mycobiology
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• v.50 no.4
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• pp.244-253
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• 2022

Trichoderma fungi have been intensively studied for mycoparasitism, and the latter is closely related to their cell-wall degrading enzymes including chitinase. Here, we studied marine-derived Trichoderma spp., isolated from distinct sources and locations, for chitinolytic and antifungal activity. Based on morphological and phylogenetic analyses, two strains designated GJ-Sp1 and TOP-Co8 (isolated from a marine sponge and a marine alga, respectively) were identified as Trichoderma bissettii. This species has recently been identified as a closely related species to Trichoderma longibrachiatum. The extracellular crude enzymes of GJ-Sp1 and TOP-Co8 showed activities of chitobiosidase and b-N-acetylglucosaminidase (exochitinase) and chitotriosidase (endochitinase). The optimum chitinolytic activity of the crude enzymes was observed at 50 ℃, pH 5.0, 0-0.5% NaCl concentrations, and the activities were stable at temperatures ranging from 10 to 40 ℃ for 2 h. Moreover, the crude enzymes showed inhibitory activity against hyphal growth of two filamentous fungi Aspergillus flavus and Aspergillus niger. To the best of our knowledge, this is the first report of the chitinolytic and antifungal activity of T. bissettii.

• Journal of Life Science
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• v.23 no.8
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• pp.1010-1018
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• 2013

The greenhouse whitefly, Bemisia tabaci, is considered one of the most destructive pests of crops. In this study, we aimed to determine the optimal liquid culture conditions in shake flasks for maximal sporulation of Beauveria bassiana M130 using rice bran. The optimal initial pH for the spore production of B. bassiana using extracted rice bran medium was 5.2 and $28^{\circ}C$. The screening in shake flasks of carbon and nitrogen sources resulted in the identification of an optimal medium based on 0.5% $(NH_4)_2SO_4$, with extracted rice bran 8:1. Using this medium, a production level of $2.15{\times}10^9$ spores per ml was obtained after six days from culture inoculation at $28^{\circ}C$ in a rotary shaking incubator at 130 rpm. In addition, the specific activities of extracellular enzymes of chitinase and protease were $4,296{\mu}mol$ and $375{\mu}mol$, respectively. These results suggest that Beauveria bassiana M130 could be a bio-controller for the greenhouse whitefly.

• Journal of Microbiology and Biotechnology
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• v.20 no.11
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• pp.1577-1584
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• 2010

In total, 140 halotolerant bacterial strains were isolated from both the soil of barren fields and the rhizosphere of six naturally growing halophytic plants in the vicinity of the Yellow Sea, near the city of Incheon in the Republic of Korea. All of these strains were characterized for multiple plant growth promoting traits, such as the production of indole acetic acid (IAA), nitrogen fixation, phosphorus (P) and zinc (Zn) solubilization, thiosulfate ($S_2O_3$) oxidation, the production of ammonia ($NH_3$), and the production of extracellular hydrolytic enzymes such as protease, chitinase, pectinase, cellulase, and lipase under in vitro conditions. From the original 140 strains tested, on the basis of the latter tests for plant growth promotional activity, 36 were selected for further examination. These 36 halotolerant bacterial strains were then tested for 1-aminocyclopropane-1-carboxylic acid (ACC) deaminase activity. Twenty-five of these were found to be positive, and to be exhibiting significantly varying levels of activity. 16S rRNA gene sequencing analyses of the 36 halotolerant strains showed that they belong to 10 different bacterial genera: Bacillus, Brevibacterium, Planococcus, Zhihengliuella, Halomonas, Exiguobacterium, Oceanimonas, Corynebacterium, Arthrobacter, and Micrococcus. Inoculation of the 14 halotolerant bacterial strains to ameliorate salt stress (150 mM NaCl) in canola plants produced an increase in root length of between 5.2% and 47.8%, and dry weight of between 16.2% and 43%, in comparison with the uninoculated positive controls. In particular, three of the bacteria, Brevibacterium epidermidis RS15, Micrococcus yunnanensis RS222, and Bacillus aryabhattai RS341, all showed more than 40% increase in root elongation and dry weight when compared with uninoculated salt-stressed canola seedlings. These results indicate that certain halotolerant bacteria, isolated from coastal soils, have a real potential to enhance plant growth under saline stress, through the reduction of ethylene production via ACC deaminase activity.

• The Plant Pathology Journal
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• v.31 no.1
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• pp.50-60
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• 2015

The use of novel isolates of Trichoderma with efficient antagonistic capacity against Fusarium oxysporum f. sp. lycopersici (FOL) is a promising alternative strategy to pesticides for tomato wilt management. We evaluated the antagonistic activity of 30 isolates of T. asperellum against 4 different isolates of FOL. The production of extracellular cell wall degrading enzymes of the antagonistic isolates was also measured. The random amplified polymorphic DNA (RAPD) method was applied to assess the genetic variability among the T. asperellum isolates. All of the T. asperellum isolates significantly reduced the mycelial growth of FOL isolates but the amount of growth reduction varied significantly as well. There was a correlation between the antagonistic capacity of T. asperellum isolates towards FOL and their lytic enzyme production. Isolates showing high levels of chitinase and ${\beta}$-1,3-glucanase activities strongly inhibited the growth of FOL isolates. RAPD analysis showed a high level of genetic variation among T. asperellum isolates. The UPGMA dendrogram revealed that T. asperellum isolates could not be grouped by their antagonistic behavior or lytic enzymes production. Six isolates of T. asperellum were highly antagonistic towards FOL and potentially could be used in commercial agriculture to control tomato wilt. Our results are consistent with the conclusion that understanding the genetic variation within Trichoderma isolates and their biochemical capabilities are required for the selection of effective indigenous fungal strains for the use as biocontrol agents.

• KOREAN JOURNAL OF CROP SCIENCE
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• v.43 no.3
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• pp.172-178
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• 1998

Freezing-resistant plants can survive subzero temperatures by withstanding extracellular ice formation. During cold acclimation, their leaves accumulate antifreeze proteins (AFPs) that are secreted into the apoplast and have the ability to modify the normal growth of ice crystals. Three barley, two wheat and two rye cultivars were grown under two different temperature regimes (20/16$^{\circ}C$ and 5/2$^{\circ}C$, day/night). Apoplastic proteins from winter cereals were separated by SDS-PAGE and detected with antisera to AFPs from winter rye. Apoplastic proteins accumulated to much higher levels in cold-acclimated (CA) leaves compared with nonacclimated (NA) ones in winter cereals. After cold acclimation, the protein concentration of apoplastic extracts increased significantly from 0.088 $mgmL^{-1}$ to 0.448 $mgmL^{-1}$, with about 5-fold increment. Also, the apoplastic protein content per gram leaf fresh weight in CA leaves ranged from 31 $\mu\textrm{g}$ $(gFW)^{-1}$ to 120 $\mu\textrm{g}$ $(gFW)^{-1}$ with an averaged value of 77 $\mu\textrm{g}$ $(gFW)^{-1}$, and coefficients of variation of 54.9%. The CA leaves in Musketeer (a Canadian winter rye cultivar) showed the greatest AFPs and antifreeze activity followed by 'Geurumil' (a Korean winter wheat cultivar), and 'Dongbori l' (Korean facultative barley cultivar). The proteins secreted into the wheat leaf apoplast at CA condition were more numerous than those observed in winter rye, where two $\beta$-1,3-glucanase-like proteins (GLPs), two chitinase-like proteins (CLPs) and two thaumatin-like proteins (TLPs) accumulated during cold acclimation. The proteins in barley leaf apoplast at CA conditions were a little different from those in wheat leaves. The AFPs were various among and within species. More freezing-resistant cultivars had more clear and numerous bands than less freezing-resistant ones. The high determination coefficient ($R^2$ =91 %) between freezing resistance and AFPs per gram leaf fresh weight indicated that the amount of AFPs was highly related to freezing resistance in winter cereal crops.

• Korean Journal of Soil Science and Fertilizer
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• v.45 no.6
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• pp.1126-1135
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• 2012

In order to produce high-quality fermenting composts, bacteria strains with high activities of extracellular enzymes (cellulase, chitinase, amylase, protease and lipase) were isolated from the soils in 6 provinces of Korea, and characterized by 16S rRNA gene sequence analysis and properties. The selected 7 stains inoculated to livestock manure for 2' fermenting time, and experimental treatment divided into 3 groups, B1, B2 and B3, according to microbial activity and enzyme type. Our results showed that microbe applications (B1, B2 and B3) can increase (p<0.05) both rhizomes (17-38%) and enzyme activities (50-81%) in compost after fermenting time, respectively, compared to non-microbe treatment (control). The microbe application also decreased significantly (p<0.05) the $NH_3$ and $H_2S$ gas contents 13.4 and 27.3% compared with control, and the Propionic acid and Butyric acid gas contents 14.5 and 19.6%, respectively, as compared to the control. The microbial degradation rate (%) of pesticides and heavy metals increased significantly (p<0.05) after fermenting time, respectively, as compared to the control. Especially, microbe applications were more effective in total rhizomes yields and bioactivities than non-microbe treatment. Thus the results of this study could help in development of potential bioinoculants and composting techniques that maybe suitable for crop production, and protectable for earth environment under various conditions.