• Title/Summary/Keyword: Liquid inoculation

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Selection of KYC 3270, a Cellulolytic Myxobacteria of Sorangium cellulosum, against Several Phytopathogens and a Potential Biocontrol Agent against Gray Mold in Stored Fruit

  • Kim, Sung-Taek;Yun, Sung-Chul
    • The Plant Pathology Journal
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    • v.27 no.3
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    • pp.257-265
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    • 2011
  • During 2002-2008 in Korea, 455 extracts from myxobacteria consisting of 318 cellulolytic and 137 bacteriolytic myxobacteria were isolated, which were then screened for antifungal activity against the phytopathogens Botrytis cinerea, Colletotrichum acutatum, Penicillium sp., Pyricularia grisea, and Phytophthora capsici. 204 isolates had antifungal activity, causing both a clear zone due to blocked spore germination and inhibition of mycelial growth; most (199) were from cellulolytic (Sorangium cellulosum) and only five were from bacteriolytic myxobacteria. B. cinerea, the best controlled among the five tested pathogens, had a unique group of antifungal isolates of myxobacterial extracts compared to the other pathogens' groups. Among seventy-nine bioactive myxobacteria, four isolates, KYC 3130, KYC 3247, KYC 3248 and KYC 3270, were selected and all were cellulolytic. Liquid culture filtrates of these four myxobacteria were applied to tomato, cherry tomato, strawberry, and kiwi fruits 5 h before inoculation with gray mold conidia; then the treated fruits were placed in an airtight container and the experiment was repeated six to eight times. Incidence (%) of gray mold on fruit of the infected control treatment was 84-98%, whereas it was only 5-21% after the KYC 3270 treatment. After KYC 3270 treatment of the four fruits, mold control was 79-95%, which was highest among the filtrates and statistically the same as treatment with fludioxonil, a registered chemical against gray mold of stored fruits.

Isolation and Characterization of a Naphthalene-Degrading Strain,Alcaligenes sp,A111 (Naphthalene 분해균주 Alcaligenes sp. A111의 분리 및 특성)

  • Oh, Hee-Mock;Kang, Jung-Hyun;Lee, Chang-Ho;Park, Chan-Sun;Ahn, Sung-Ku;Yoon, Byung-Dae;Kho, Yung-Hee
    • Microbiology and Biotechnology Letters
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    • v.22 no.4
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    • pp.423-429
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    • 1994
  • A bacterial strain which formed a distinct colony on agar plate containing naphthalene as a vapor phase and grew well ina liquid minimal medium was isolated and identified as Alcaligenes sp. A111. Optimum temperature and pH for the cultivation of Alcaligenes sp. A111 were 30$\cir$C and 7.0, respectively. Cell growth increased dramatically from 12 hours after inoculation and revealed a stationary phase at about 48 hours. Relative growth rate ($\mu$')increased hyperbolically depending on the conceration of naphthalene up to 500 ppm and reached to the maximum value pf 2.8/day, but $\mu$' didn't change within a range of 500~4000 ppm naphthalene. NH$_{4}$Cl or NH$_{4}$NO$_{3}$ was preferrd as a nitrogen source and a P : N ratio by weight og 6 : 1 was favorable to cell growth. Alcaligenes sp. A111 utilized the intermediates of degradation of naphthalene and showed tolerance to benzene, toluene, and octane. therefore, it is suggested that Alcaligenes sp. A111 could be effectively used for the biological treatment of wastewater containing naphthalene in the presence of some aromatic compounds.

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Optimized Production of Biosurfactant by the Indigenous Bacterium, Pseudoalteromonas sp. HK-3 Originating from Oil-Spilled Areas (유류누출 지역에서 유래한 토착세균, Pseudoalteromonas sp. HK-3 배양에서 생물계면활성제의 최적 생산)

  • Cho, Su-Hee;Ma, Chae-Woo;Oh, Kye-Heon
    • KSBB Journal
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    • v.26 no.1
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    • pp.57-61
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    • 2011
  • The principal objective of this study was to determine the optimal conditions for the production of biosurfactant by the indigenous bacterium, Pseudoalteromonas sp. HK-3, originating from oil-spilled areas. The relationship between total biosurfactant production and the factors affecting biosurfactant production were evaluated by statistical analysis using SPSS software. The effects of various supplemental carbon sources (e.g., glucose, dextrose, mannitol, citrate, acetate) on the maximal production of biosurfactant by the test culture of Pseudoalteromonas sp. HK-3 was then evaluated. As a result, mannitol was found in this study to be the best supplemental carbon source for the production of biosurfactant. A spot inoculation of crude cultural liquid containing the HK-3 cells generated the largest clear zone, whereas only small clear zones appeared around the spots inoculated with either supernatant only or cell pellets following centrifugation. Our results demonstrated that the HK-3 test culture supplemented with 2% mannitol at an initial pH of 6 generated the maximal amount of biosurfactant within 72 h of incubation.

Production of Thaxtomin A by Korean Isolates of Streptomyces turgidiscabies and Their Involvement in Pathogenicity

  • Kim, Young-Sook;Cho, Jun-Mo;Park, Duck-Hwan;Lee, Heung-Goo;Kim, Jeom-Soon;Seo, Sang-Tae;Shin, Kwan-Yong;Hur, Jang-Hyun;Lim, Chun-Keun
    • The Plant Pathology Journal
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    • v.15 no.3
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    • pp.168-171
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    • 1999
  • Crude extracts of pathogenic Streptomyces turgidiscabies isolates produced necrotic reaction on potato slices. Necrosis was first visible 24 hr after application and increased in severity over several days. However, necrosis was not observed when crude extracts of nonpathogenic strains were applied onto tuber slices. Presence of thaxtomin A from crude extracts of pathogenic S. turgidiscabies was identified by thin layer chromatography and high performance liquid chromatography. Nonpathgenic strains did not produce thaxtomin A in oatmeal broth. Inoculation of potato tuber slices with thaxtomin A partially purified from crude extract of a pathogenic S. turgidiscabies ST5 reproduce necrotic reaction suggesting that thaxtomin A is a pathogenicity determinant in S. turgidiscabies.

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Regeneration Studies in Grateloupia filicina (J.V. Lamouroux) C. Agardh - An Important Carrageenophyte and Edible Seaweed

  • Baweja, Pooja;Sahoo, Dinabandhu
    • ALGAE
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    • v.24 no.3
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    • pp.163-168
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    • 2009
  • Grateloupia filicina (J.V. Lamouroux) C. Agardh (Halymeniaceae, Cryptonemiales, Rhodophyta) is an edible seaweed as well as an important source of carrageenan. In the present study, attempt has been made to develop a suitable protocol for effective regeneration of the seaweed and the rapid multiplication of the desired varieties. The young upright thallus of G. filicina was grown in axenic culture using both solid and liquid media. The various media tested were f/2, Provasoli’s Enriched Seawater (PES) and Enriched Seawater (ESW). The effect of glycerol (as a carbon source) and various plant growth regulators i.e. auxin (NAA) and cytokinins (Kinetin and BA) were tested. Although, regeneration of young thalli was observed from the cut ends in all the media, better growth was found in f/2, PES, f/2 (0.5% Glycerol), f/2 (NAA ${10^{-5}}_M)\;and\;f/2\;(BA\;{10^{-6}}_M$). On the other hand callusing was observed only in solid media supplemented with low concentration of Glycerol (0.5%) in f/2, NAA ${10^{-5}}_M\;in\;f/2,\;PES\;and\;BA\;{10^{-5}}_M$ in f/2. Young thalli were developed from the callus sub culture after 40 days of inoculation.

Effect of Microporosity on Tensile Properties of As-Cast AZ91D Magnesium Alloy

  • 이충도
    • Transactions of Materials Processing
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    • v.8 no.3
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    • pp.283-283
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    • 1999
  • In the present study, the effect of microporosity on the tensile properties of as-cast AZ91D magnesium alloy was investigated through experimental observation and numerical prediction. The test specimens were fabricated by die-casting and gravity-casting. For gravity-casting, the inoculation and use of various metallic moulds were applied to obtain a wide range of microporosity. The deficiency of the interdendritic feeding of the liquid phase acted as d dominant mechanism on the formation of the micropores in the Mg-Al-alloys, rather than the evolution of hydrogen gas. Although tensile strength and elongation has a nonlinear and very intensive dependence upon microporosity, the yield strength appeared to have a linear relationship with microporosity. However, it was possible to quantitatively estimate the linear contribution of microporosity on the individual tensile property far a range of microporosity, which was below about B %. The numerical prediction suggests that the effect of microporosity on fractured strength and elongation decreased as the strain hardening exponent increased. Furthermore. the shape and distribution of micropores may play a more dominant role than local plastic deformation on the tensile behavior of AZ9lD alloy.

Optimization of Indole-3-acetic Acid (IAA) Production by Bacillus megaterium BM5

  • Lee, Jae-Chan;Whang, Kyung-Sook
    • Korean Journal of Soil Science and Fertilizer
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    • v.49 no.5
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    • pp.461-468
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    • 2016
  • One of the important phytohormones produced by plant growth promoting bacteria is the auxin; indole-3-acetic acid (IAA), with L-tryptophan as the precursor. In this study, we focused on the investigation of optimal conditions for the production of IAA by Bacillus megaterium BM5. We investigated culturing conditions, such as incubation temperature, pH of the culture medium and incubation period, with varying media components such as inoculation volume, tryptophan concentration and carbon and nitrogen source. Besides, optimization study intended for high IAA production was carried out with fermentation parameters such as rpm and aeration. The initial yield of $42{\mu}g\;IAA\;ml^{-1}$ after 24 hr increased to $85{\mu}g\;ml^{-1}$ when 5% (v/v) of L-tryptophan was used in the culture broth. The maximum yield of $320{\mu}g\;IAA\;ml^{-1}$ was observed in trypticase soy broth (TSB) supplemented with starch and soybean meal as C and N sources with a C/N ratio of 3:1 (v/v) at $30^{\circ}C$, pH 8.0 for 48 hrs with 1.0 vvm and 250 rpm in 5 L working volume using 10 L scale fermenter. The bacterial auxin extracted from the culture broth was confirmed by thin layer chromatography and high-performance liquid chromatography and effect on plant growth was confirmed by root elongation test.

Coproduction of Enzymes and Beta-Glucan by Aspergillus oryzae Using Solid-State Fermentation of Brown Rice

  • Ji, Su Bin;Ra, Chae Hun
    • Journal of Microbiology and Biotechnology
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    • v.31 no.7
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    • pp.1028-1034
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    • 2021
  • The effect of medium composition on enzyme and β-glucan production by Aspergillus oryzae KCCM 12698 was investigated. Brown rice, rice bran, nitrogen, and ascorbic acid are key components of the synthetic medium used in liquid-state fermentation. To determine the optimal concentrations of these components for enzyme and β-glucan production, we conducted one factor at a time experiments, which showed that the optimal concentrations were 30 g/l brown rice, 30 g/l rice bran, 10 g/l soytone, and 3 g/l ascorbic acid. Pretreatment of brown rice for 60 min prior to inoculation enhanced fungal biomass, while increasing the production of enzymes and β-glucan using solid-state fermentation. Maximum fungal biomass of 0.76 mg/g, amylase (26,551.03 U/g), protease (1,340.50 U/g), and β-glucan at 9.34% (w/w) were obtained during fermentation. Therefore, solid-state fermentation of brown rice is a process that could enhance yield and overall production of enzymes and β-glucan for use in various applications.

Evidence of Greater Competitive Fitness of Erwinia amylovora over E. pyrifoliae in Korean Isolates

  • Choi, Jeong Ho;Kim, Jong-Yea;Park, Duck Hwan
    • The Plant Pathology Journal
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    • v.38 no.4
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    • pp.355-365
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    • 2022
  • Erwinia amylovora and E. pyrifoliae are the causative agents of destructive diseases in both apple and pear trees viz. fire blight and black shoot blight, respectively. Since the introduction of fire blight in Korea in 2015, the occurrence of both pathogens has been independently reported. The co-incidence of these diseases is highly probable given the co-existence of their pathogenic bacteria in the same trees or orchards in a city/district. Hence, this study evaluated whether both diseases occurred in neighboring orchards and whether they occurred together in a single orchard. The competition and virulence of the two pathogens was compared using growth rates in vitro and in planta. Importantly, E amylovora showed significantly higher colony numbers than E. pyrifoliae when they were co-cultured in liquid media and co-inoculated into immature apple fruits and seedlings. In a comparison of the usage of major carbon sources, which are abundant in immature apple fruits and seedlings, E. amylovora also showed better growth rates than E. pyrifoliae. In virulence assays, including motility and a hypersensitive response (HR), E. amylovora demonstrated a larger diameter of travel from the inoculation site than E. pyrifoliae in both swarming and swimming motilities. E. amylovora elicited a HR in tobacco leaves when diluted from 1:1 to 1:16 but E. pyrifoliae does not elicit a HR when diluted at 1:16. Therefore, E. amylovora was concluded to have a greater competitive fitness than E. pyrifoliae.

Proteomic and Phenotypic Analyses of a Putative YggS Family Pyridoxal Phosphate-Dependent Enzyme in Acidovorax citrulli

  • Lynn Heo;Yongmin Cho;Junhyeok Choi;Jeongwook Lee;Yoobin Han;Sang-Wook Han
    • The Plant Pathology Journal
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    • v.39 no.3
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    • pp.235-244
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    • 2023
  • Acidovorax citrulli (Ac) is a phytopathogenic bacterium that causes bacterial fruit blotch (BFB) in cucurbit crops, including watermelon. However, there are no effective methods to control this disease. YggS family pyridoxal phosphate-dependent enzyme acts as a coenzyme in all transamination reactions, but its function in Ac is poorly understood. Therefore, this study uses proteomic and phenotypic analyses to characterize the functions. The Ac strain lacking the YggS family pyridoxal phosphate-dependent enzyme, AcΔyppAc(EV), virulence was wholly eradicated in geminated seed inoculation and leaf infiltration. AcΔyppAc(EV) propagation was inhibited when exposed to L-homoserine but not pyridoxine. Wild-type and mutant growth were comparable in the liquid media but not in the solid media in the minimal condition. The comparative proteomic analysis revealed that YppAc is primarily involved in cell motility and wall/membrane/envelop biogenesis. In addition, AcΔyppAc(EV) reduced biofilm formation and twitching halo production, indicating that YppAc is involved in various cellular mechanisms and possesses pleiotropic effects. Therefore, this identified protein is a potential target for developing an efficient anti-virulence reagent to control BFB.