• Korean Journal of Microbiology
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• v.50 no.3
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• pp.201-209
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• 2014

This study was carried out in order to develop a biological control of anthracnose of red pepper caused by fungal pathogens. In particular, this study focuses on the Colletotrichum species, which includes important fungal pathogens causing a great deal of damage to red pepper. Antagonistic bacteria were isolated from the soil of pepper fields, which were then tested for biocontrol activity against the Colletotrichum gloeosporioides anthracnose pathogen of pepper. Based on the 16S rRNA sequence analysis, the isolated bacterial strain CS-52 was identical to Bacillus sp. The culture broth of Bacillus sp. CS-52 had antifungal activity toward the hyphae and spores of C. gloeosporioides. Moreover, the substances with antifungal activity were optimized when Bacillus sp. CS-52 was grown aerobically in a medium composed of 0.5% glucose, 0.7% $K_2HPO_4$, 0.2% $KH_2PO_4$, 0.3% $NH_4NO_3$, 0.01% $MnSO_4{\cdot}7H_2O$, and 0.15% yeast extract at $30^{\circ}C$. The inhibition of spore formation resulting from cellulase, siderophores, and indole-3-acetic acid (IAA), were produced at 24 h, 48 h, and 72 h, respectively. Bacillus sp. CS-52 also exhibited its potent fungicidal activity against anthracnose in an in vivo test, at a level of 70% when compared to chemical fungicides. These results identified substances with antifungal activity produced by Bacillus sp. CS-52 for the biological control of major plant pathogens in red pepper. Further studies will investigate the synergistic effect promoting better growth and antifungal activity by the formulation of substances with antifungal activity.

• Horticultural Science & Technology
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• v.32 no.6
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• pp.872-878
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• 2014

This study aimed to enhance storage and freshness of strawberry fruits and foliage vegetables by spray treatment with Chlorella vulgaris as a bio-fertilizer. The tested strain, C. vulgaris CHK0008, was isolated from an organically cultivated rice paddy and identified as C. vulgaris by its morphology and 18S rDNA and 23S rDNA sequence homology. We successfully cultured C. vulgaris CHK0008 in BG11 modified medium (BG11MM) and adjusted $2.15{\times}10^6cell/mL$ C. vulgaris CHK0008 to one OD value by measuring the optical density at 680 nm using a UV-vis spectrophotometer. The soluble solid content of 'Seolhyang' and 'Yukbo' strawberry fruits treated by spray application with C. vulgaris CHK0008 was enhanced by 22.2% and 11.5% respectively, compared to untreated controls. Additionally, the decay rates of treated 'Seolhyang' and 'Yukbo' strawberry fruits decreased 63.8% and 74.4% respectively, compared to untreated control. Surface color changes and chlorosis of leaves in leaf vegetables such as lettuce, kale, red ornamental kale, white ornamental kale and beet were observed in samples treated with water spray for 10 days after cold storage. However, the decay rate of leafy vegetables treated with foliar application of 25% C. vulgaris CHK0008 liquid culture was significantly decreased compared to that of the untreated control during storage at $4^{\circ}C$.

• Journal of Life Science
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• v.18 no.12
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• pp.1625-1630
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• 2008

A microorganism capable of producing high level of poly-3-hydoxybutyrate (PHB) from xylose was isolated from soil. The isolated strain J-65 was identified as Bacillus megaterium based on the morphological, biochemical and molecular biological characteristics. The optimum temperature and pH for the growth of B. megaterium J-65 were $37^{\circ}C$ and 8.0, respectively. The optimum medium composition for the cell growth was 2% xylose, 0.25% $(NH_4)_2SO_4$, 0.3% $Na_2HPO_4{\cdot}12H_2O$, and 0.1% $KH_2PO_4$. The optimum condition for PHB accumulation was same to the optimum condition for cell growth. Copolymer of ${\beta}$-hydroxybutyric and ${\beta}$-hydroxyvaleric acid was produced when propionic acid was added to shake flasks containing 20 g/l of xylose. Fermenter culture was carried out to produce the high concentration of PHB. In batch culture, cell mass was 9.82 g/l and PHB content was 35% of dry cell weight. PHB produced by B. megaterium J-65 was identified as homopolymer of 3-hydoxybutyric acid by GC and NMR.

• Journal of Life Science
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• v.25 no.6
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• pp.680-685
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• 2015

Previously, cellulase and xylanase producing microorganism, Bacillus subtilis NC1, was isolated from soil. Based on the 16S rRNA gene sequence and API 50 CHL test the strain was identified as Bacillus subtilis, and named as B. subtilis NC1. We cloned and sequenced the genes for cellulase and xylanase. Plus, the deduced amino acid sequences from the genes of cellulase and xylanase were determined and were also identified as glycosyl hydrolases family (GH) 5 and 30, respectively. In this study to optimize the medium parameters for cellulase production by B. subtilis NC1 the RSM (response surface methodology) based on CCD (central composite design) model was performed. Three factors, tryptone, yeast extract, and NaCl, for N or C source were investigated. The cellulase activity was measured with a carboxylmethyl cellulose (CMC) plate and the 3,5-dinitrosalicylic acid (DNS) methods. The coefficient of determination (R²) for the model was 0.960, and the probability value (p=0.0001) of the regression model was highly significant. Based on the RSM, the optimum conditions for cellulase production by B. subtilis NC1 were predicted to be tryptone of 2.5%, yeast extract of 0.5%, and NaCl of 1.0%. Through the model verification, cellulase activity of Bacillus subtilis NC1 increased from 0.5 to 0.62 U/ml (24%) compared to the original medium.

• Research in Plant Disease
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• v.21 no.2
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• pp.50-57
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• 2015

Control of nematode has become difficult owing to the restricted use of effective soil fumigant, methyl bromide, and other non-fumigant nematicides. Therefore, it is urgently necessary to develop microbial nematicide to replace chemical nematicides. In this study, the 50% aqueous methanol extraction solution of fermentation broths of 2,700 actinomycete strains were tested for their nematicidal activity against second stage of juveniles (J2s) of Meloidogyne incognita. As the results, only the 50% aqueous methanol extraction solution of AN110065, at 20% equivalent to 10% fermentation broth, showed strong nematicidal activity with 78.9% of mortality 24 h after treatment and 94.1% of mortality at 72 h. The 16S rRNA gene sequencing showed that the strain sequence was 99.78% identical to Streptomyces netropsis. The extract of S. netropsis AN110065 fermentation broth was successively partitioned with ethyl acetate and butanol and then the ethyl acetate, butanol and water layers were investigated for their nematicidal activity against the M. incognita. At $1000{\mu}g/ml$, ethyl acetate layer showed the strongest activity of 83.5% of juvenile mortality 72 h after treatment. The pot experiment using the fermentation broth of AN110065 on tomato plant against M. incognita displayed that it evidently suppressed gall formation at a 10-fold diluent treatment. The tomato plants treated with the fermentation broth of S. netropsis AN110065 did not show any phytotoxicity. The results suggest that S. netropsis AN110065 has a potential to serve as microbial nematicide in organic agriculture.

• The Korean Journal of Mycology
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• v.49 no.3
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• pp.337-349
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• 2021

This study aimed to isolate wild yeasts from water and soil sample of the Nonsan stream and Sapgyoho (lake) in Chungcheonnam-do, Korea, and to further characterize previously unrecorded wild yeast strains. In total, 102 strains, representing 55 different species of wild yeast were isolated from 95 samples collected from the Jangseoncheon and Ipchoncheon of Nonsan stream in Jellabuk-do and Chungcheonnam-do. Among these, 33 strains were isolated from alkalophilic yeast extract-peptone-dextrose (YPD) medium (pH 9.0), and 9 strains were isolated concurrently on general YPD medium (pH 6.5) and alkalophilic medium. Seventeen strains of Cryptococcus laurentii were predominantly isolated. Additionally, 65 strains, representing 27 different species of wild yeast were isolated from 58 samples obtained from Sapgyoho (lake). Among the 82 isolated wild yeast strains, 8 strains, including Candida fructus JSC 72-1(JSL-GGU 015), had not previously been recorded. All 8 previously unrecorded yeasts were oval in shape except C. fructus JSC72-1(JSL-GGU-015), and only the Filobasidium chernovii JSC39-1(JSL-GGU-013) strain formed spores. All strains except Pseudosydowia eucalypti JSC23-6(JSL-GGU-012) grew well in yeast extract-peptone-dextrose (YPD) and yeast extract-malt extract media and grew in vitamin-free medium. Four strains, including P.eucalypti JSC23-6(JSL-GGU-012) grew well in 15% NaCl-containing YPD medium. F.chernovii JSC39-1(JSL-GGU-013) and Sirobasidium intermedium JSC7-3(JSL-GGU-014) assimilated lactose, and five strains, including F. chernovii JSC39-1(JSL-GGU-013) also assimilated starch. All strains were resistant to 800 ppm of Ca, Cu, Li, and Mg ions.

• Journal of Mushroom
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• v.18 no.4
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• pp.331-338
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• 2020

This study was conducted to cultivate new Lepista nuda varieties with shorter cultivation period and better fruiting body compared to that of wild strains, for mass production and commercial application. Eighteen genetic resources of L. nuda were collected and grown in boxes using rice straw-fermented growth medium. Four lines with fruiting bodies were formed and selected as cross-breeding lines. Although 657 combinations were crossed through monospore crossing, only 17 combinations were bred between the 'CBMLN-19' line and the 'CBMLN-30' line. Among them, 8 lines with fast mycelial growth and high density were selected. After inoculating the rice straw-fermented growth medium with 14 genetic resources and 8 cross-breeding lines, their incubation period was investigated. Six of the cross-breeding lines completed their incubation in 20 days, while 7 of the 14 genetic resources took more than 40 days to complete their incubation, reducing the incubation period by more than 20 days in most cross-breeding lines. After the incubations were completed, the clay loam soil was covered with for post-cultivation, and when the mycelial cultivation was complete, the formation of fruiting bodies was induced after scraping the mycelial bodies under these environmental conditions: 14℃, 95% relative humidity or higher, and 1,500 to 2,000 ppm CO2 concentration. The temperature was reduced to 6℃ at night, resulting in a low temperature shock. Thus, 4 lines of fruiting bodies occurred from two genetic resources 'CBMLN-31' and 'CBMLN-44' and two cross-bred lines 'CBMLN-96' and 'CBMLN-103'. After inoculation, the longest period for fruiting bodies to occur was 100 days for the control:, the genetic resource 'CBMLN-31', and the shortest period (45 days) was observed for the cross-breeding line 'CBMLN-103'. The result of the investigation of the fruiting body characteristics shows that the cross-bred line 'CBMLN-103' showed a small form with 1.9 g of individual weight and 123validstipes per box, which was the highest incidence among the four lines. Another cross-bred line, 'CBMLN-96', had an individual weight of 5.5 g, which is larger than that of 'CBMLN-103'; however, the number of valid stipes per box was 30 less than that of 'CBMLN-103'. Quantity analysis showed that the control, 'CBMLN-31', had the highest quantity of 783 g per box, followed by the cross-bred line, 'CBMLN-96' with 165 g per box, and then the 'CBMLN-103' with 232 g. The quantity of the two crossbred lines was lower than that of the control 'CBMLN-31'; however, the amount of fruiting bodies was higher, and the cultivation period was shortened by 32 to 33 days. Therefore, these two lines would be selected as superior lines.