To evaluate the correlations of microbial populations with soil healthiness and crop production and establish the criteria for microbial population of soil types. We analyzed the microbial community structure of 13 soils which were different in physical and chemical properties and cultivation methods. According to the analysis of microbial population suing the dilution plate method, the large differences of the microbial population structures among soil types were shown: aerobic bacteria $2-27{\times}10^6$, fluorescent Pseudomonas $1-1,364{\times}10^5$, Gram negative bacteria $1-126{\times}10^4$, and mesophilic Bacillus $1-110{\times}10^5$. The density of Gram negative bacteria was highest on red pepper cultivating soils (sample no. 4 and 6) of Umsung and Gesan, Chungbuk, and the density of the fluorescent Pseudomonas was highest on greenhouse soil (sample no. 7) of Jinju, Kyungnam. The crop productivity of three soils was high as compared with those of other soils. It was supposed that the density of fluorescent Pseudomonas and mesophilic Bacillus were correlated with the incresed crop production. By MIDI analysis, 579 strains isolated from 13 soils composed of a variety of microbes including 102 isolates of Agrobacterium, 112 isolates of Bacillus, 32 isolates of Pseudomonas, 44 isolates of Kocuria, and 34 isolates of Pseudomonas. Among the 624 isolates of Gram negative bacteria, Pseudomonas including P. putida and p. fluorescens occupied the highest density (51%), and Stenotrophomonas maltophilia and Burkholderia cepacia also appeared at high density. From RAPD analysis, the fluorescent Pseudomonas strains isolated from 13 soil types showed a high level of strain diversities and were grouped into 2 - 14 patterns according to soil types. Many of unknown bacteria were recovered from the paddy soil, and needed to be further characterized on the molecular basis.
Mucin2 (MUC2), an important regulatory factor in the immune system, plays an important role in the host defense system against bacterial translocation. Probiotics known to regulate MUC2 gene expression have been widely studied, but the interactions among probiotic, pathogens, and mucin gene are still not fully understood. The aim of this study was to investigate the role of MUC2 in blocking effects of probiotics on meningitic E. coli-induced pathogenicities. In this study, live combined probiotic tablets containing living Bifidobacterium, Lactobacillus bulgaricus, and Streptococcus thermophilus were used. MUC2 expression was knocked down in Caco-2 cells by RNA interference. 5-Aza-2'-deoxycytidine (5-Aza-CdR), which enhances mucin-promoted probiotic effects through inducing production of Sadenosyl-L-methionine (SAMe), was used to up-regulate MUC2 expression in Caco-2 cells. The adhesion to and invasion of meningitic E. coli were detected by competition assays. Our studies showed that probiotic agents could block E. coli-caused intestinal colonization, bacteremia, and meningitis in a neonatal sepsis and meningitis rat model. MUC2 gene expression in the neonatal rats given probiotic agents was obviously higher than that of the infected and uninfected control groups without probiotic treatment. The prohibitive effects of probiotic agents on MUC2-knockdown Caco-2 cells infected with E44 were significantly reduced compared with nontransfected Caco-2 cells. Moreover, the results also showed that 5-Aza-CdR, a drug enhancing the production of SAMe that is a protective agent of probiotics, was able to significantly suppress adhesion and invasion of E44 to Caco-2 cells by upregulation of MUC2 expression. Taken together, our data suggest that probiotic agents can efficiently block meningitic E. coli-induced pathogenicities in a manner dependent on MUC2.
The Sea:JOURNAL OF THE KOREAN SOCIETY OF OCEANOGRAPHY
/
v.10
no.1
/
pp.19-30
/
2005
To investigate the seasonal distribution and grazing impacts of benthic protozoa in mud flat, their abundance, biomass and grazing rates of benthic protozoa were evaluated at interval of two or three month in Gangwha Island from April, 2002 to April, 2004. Heterotrophic flagellates and ciliates accounted for an average 98% of benthic protozoa biomass. Abundance and carbon biomass of heterotrophic flagellates ranged from $0.2{\times}10^5$ to $5.9{\times}10^5\;cells\;cm{-3}$ and from 0.02 to $9.2\;{\mu}gC\;cm^{-3}$, respectively. Biomass of heterotrophic flagellates was high in spring and fall, and showed no differences among stations. Abundance and biomass of heterotrophic flagellates decreased with the depth and were high within the surface 2.5 m sediment layer. The majority of heterotrophic flagellates were less than $10\;{\mu}m$ in length, and few euglenoid flagellates were larger than $20\;{\mu}m$. Abundance and carbon biomass of ciliates ranged from $0.1{\times}10^3$ to $17.8{\times}10^3\;cells\;cm^{-3}$ and from 0.02 to $9.1\;{\mu}gC\;cm^{-3}$, respectively, and those of ciliates were high in spring and fall. Biomass of ciliates was high within the surface 2.5 mm sediment layer and was higher at st. J2 and st. J3 than st. J1. Among the revealed benthic ciliates, the hypotrichs were the most important group in terms of abundance and biomass. During the sampling periods, an average 66% of benthic protozoa biomass was covered by ciliates. The seasonal distribution of benthic protozoa showed an almost similar fluctuation pattern to that of chlorophyll-a. The results suggest that the biomass of benthic protozoa were mainly controlled by prey abundance, for example, diatoms. Based on ingestion rates, benthic protozoa removed from 13.4 to 40.7% of bacterial production and from 20.1 to 36.4% of primary production. Ingestion rates of benthic protozoa on bacteria and microphytobenthos were high in April. Benthic protozoa in this study area may play a pivotal role in the carbon flow of the benthic microbial food web during spring.
In this study, to retain a stable bacterial inoculant, Bacillus strains showing antifungal activity were screened. The improved production, antifungal mechanism, and stability of the antifungal metabolite by a selected strain, AF4, a potent antagonist against phytopathogenic Botrytis cinerea, were also investigated. The AF4 strain was isolated from rhizospheric soil of hot pepper and identified as Bacillus subtilis by phenotypic characters and 16S rRNA gene analysis. Strain AF4 did not produce antifungal activity in the absence of a nitrogen source and produced antifungal activity at a broad range of temperatures (25-40℃) and pH (7-10). Optimal carbon and nitrogen sources for the production of antifungal activity were glycerol and casein, respectively. Under improved conditions, the maximum antifungal activity was 140±3 AU/ml, which was higher than in the basal medium. Photomicrographs of strain AF4-treated B. cinerea showed morphological abnormalities of fungal mycelia, demonstrating the role of the antifungal metabolite. The B. subtilis AF4 culture exhibited broad antifungal activity against several phytopathogenic fungi. The antifungal activity was heat-, pH-, solvent-, and protease-stable, indicating its nonproteinous nature. These results suggest that B. subtilis AF4 is a potential candidate for the control of phytopathogenic fungi-derived plant diseases.
A bacterial strain YC4963 with antifungal activity against Colletotrichum orbiculare, a causal organism of cucumber anthracnose was isolated from the rhizosphere soil of Siegesbeckia pubescens Makino in Korea. Based on physiological and biochemical characteristics and 16S ribosomal DNA sequence analysis, the bacterial strain was identified as Pseudomonas aurantiaca. The bacteria also inhibited mycelial growth of several plant fungal pathogens such as Botrytis cinerea, Fusarium oxysporum and Rhizoctonia solani on PDA and 0.1 TSA media. The antifungal activity was found from the culture filtrate of this isolate and the active compound was quantitatively bound to XAD adsorption resin. The antibiotic compound was purified and identified as phenazine-l-carboxylic acid on the basis of combined spectral and chemical analyses data. This is the first report on the production of phenazine-l-carboxylic acid by Pseudomonas aurantiaca.
Recently, Psychrobacter sp. ArcL13 strain showing the extracellular lipase activity was isolated from the Chuckchi Sea of the Arctic Ocean. However, due to the low expression levels of the enzyme in the natural strain, the production of recombinant lipase is crucial for various applications. Identification of the gene for the enzyme is prerequisite for the production of the recombinant protein. Therefore, in the present study, a novel lipase gene (ArcL13-Lip) was isolated from Psychrobacter sp. ArcL13 strain by gene prospecting using PCR, and its complete nucleotide sequence was determined. Sequence analysis showed that ArcL13-Lip has high amino acid sequence similarity to lipases from bacteria of some Psychrobacter genus (84-90%) despite low nucleotide sequence similarity. The lipase gene was cloned into the bacterial expression plasmid and expressed in E. coli. SDS-PAGE analysis of the cells showed that ArcL13-Lip was expressed as inclusion bodies with a molecular mass of about 35 kDa. Refolding was achieved by diluting the unfolded protein into refolding buffers containing various additives, and the highest refolding efficiency was seen in the glucose-containing buffer. Refolded ArcL13-Lip showed high hydrolytic activity toward p-nitrophenyl caprylate and p-nitrophenyl decanoate among different p-nitrophenyl esters. Recombinant ArcL13-Lip displayed maximal activity at $40^{\circ}C$ and pH 8.0 with p-nitrophenyl caprylate as a substrate. Activity assays performed at various temperatures showed that ArcL13-Lip is a cold-active lipase with about 40% and 73% of enzymatic activity at $10^{\circ}C$ and $20^{\circ}C$, respectively, compared to its maximal activity at $40^{\circ}C$.
Ahn Sun Hee;Lee Eun Mi;Kim Dong Gyun;Hong Gyoung Eun;Park Eun Mi;Kong In Soo
Journal of Life Science
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v.16
no.1
/
pp.131-135
/
2006
Oligopeptide is known to be an essential nitrogen nutrient for bacterial growth. Oligopeptide can be transported into cytoplasm by a specific transport system, Opp system. Opp system is composed of five proteins, which are transcribed by an operon. These are responsible for oligopeptide binding protein (OppA), permease (OppB and OppC) and energy generation system (OppD and OppF), respectively. Previously, we isolated the opp operon from Vibrio fluvialis and constructed the oppA mutant by allelic exchange method. In this study, we investigated the growth pattern and biofilm production under the different growth condition. When the cells were cultivated using brain heart infusion(BHI) medium, the wild type was faster than the mutant in growth during the exponential phase. However, it showed that the growth pattern of two strains in M9 medium is very similar. The growth of wild type showed better than that of the mutant grown at pH 8. At pH 7, there was no an obvious difference in growth. After 5 mM $H_2O_2$ was treated to the cells $(OD_{600}=1.2)$, the cell survival was examined. The oppA mutation did not affect in survivability. In the presence of $10{\mu}g/ml$ polymyxin B, the biofilm production of the oppA mutant was higher than that of the wild type.
It is urgently required to construct safety data on agricultural by-products imported for use as medium materials for domestic mushroom production. However, research on microorganisms is insufficient. This study was conducted to investigate the presence of bacteria that have the possibility of harmful effects on human, plants and mushroom in wheat straw, peatmoss, cottonseed hull, cottonseed meal, and beet pulp imported from Australia, Canada, China, Egypt, Germany. Bacteria were found in the range of $1.35{\times}10^2$ to $8.34{\times}10^6CFU/g$. As a result of 16S rDNA sequence analysis, total of 19 genera and 45 species of bacteria were identified. Bacillus genus was dominant, followed by Paenibacillus genus. At the species level, diverse species was in the order of Firmicute, Proteobacteria and Actinobacteria. Regarding the agricultural by-products, straw and peat moss had more diverse bacteria than other agricultural by-products. Among the indentified bacteria, 6 species of 5 genera (Enterobacter asburiae, Enterobacter ludwigii, Stenotrophomonas maltophilia, Pseudomonas monteilii, Bacillus anthracis, and Cellulosimicrobium funkei) were present as potent harmful bacteria to human. Surprisingly, both the human and plant pathogenic Klebsiella pneumoniae subsp. pneumonia was present. Bacillus altitudinis was present as a plant pathogen. Lysinibacillus sphaericus, an insect pathogen, and Ochrobactrum pseudogrignonense, a mushroom pathogen, were also present. The results of this study confirmed that several kinds of pathogenic bacteria were present in the agricultural by-products for the mushroom cultivation medium imported into Korea. Our work suggests that hygiene inspection and management is urgently needed for imported agricultural by-products to be safely used for mushroom production.
Kim, Hyun Tae;Ko, Jong Min;Baek, In Youl;Han, Won Young;Yun, Hong Tai;Lee, Byoung Won;Shin, Sang Ouk;Seo, Jeong Hyun;Kim, Hong Sik;Kwak, Do Yeon
Korean Journal of Breeding Science
/
v.51
no.4
/
pp.475-481
/
2019
The soybean cultivar 'Neulchan' was developed for production of soy-paste and tofu. SS91501-9-1-1 and SS96205 (F2) were crossed in 1998, and F3 to F7 were selected by the pedigree method. A preliminary yield trial (PYT) and an advanced yield trial (AYT) were conducted from 2006 to 2008, and a regional yield trial (RYT) in nine regions was conducted from 2009 to 2011. In the RYT, 'Neulchan' was stable in variable environments and generated high yield. 'Neulchan' was determinate with white flower, light brown pod color, yellow spherical seed, and yellow hilum. Its flowering date and maturity date were Jul. 30 and Oct. 9, respectively. The plant height was shorter than that of 'Daewonkong' (a standard cultivar). 'Neulchan' had the same node number (14), higher first-pod height (12 cm), and lighter seed weight (21.7 g/100-seed weight) than those of 'Daewonkong' (14, 11, and 24.2 g/100-seed weight, respectively). 'Neulchan' had high resistance to bacterial pustule, and its resistance to soybean mosaic virus was similar to that of 'Daewonkong'. The yield and color of 'Neulchan' tofu were similar to those of 'Daewonkong' tofu, but the hardness was lower than that of 'Daewonkong' tofu. The soybean malt scent, fermented soybean yield, and γ-polyglutamic acid (γ-PGA) of 'Neulchan' were 3, 215%, and 24.6 mg/g, respectively. Its yield in adaptable regions was 307 kg/10a, higher than that of 'Daewonkong'. 'Neulchan' was expected to be cultivated and used widely for soy-paste and tofu production. (Registration No. 4904).
Chan-Jung Lee;Hye-Sung Park;Seong-Yeon Jo;Gi-Hong An;Ja-Yun Kim;Kang-Hyo Lee
Journal of Mushroom
/
v.22
no.2
/
pp.60-66
/
2024
This study was conducted to selection and investigate appropriate conditions for mass production of antagonistic microbes to control cobweb disease caused by Cladobotryum mycophilum. A grampositive bacterium was isolated from spent substrate of Agaricus bisporus and showed significant antagonistic activity against Cladobotryum mycophilum. The bacterium was identified as Bacillus altitudinis. based on the cultural, biochemical and physiological characteristics, and 16S rRNA sequence. The isolate is saprophytic, but not parasitic nor pathogenic to cultivated mushroom whereas it showed strong inhibitory effects against C. mycophilum cells in vitro. The control efficacy of B. altitudinis HC7 against cobweb disease of C. mycophilum was up to 78.2% on Agaricus bisporus. The suppressive bacterium may be useful for the development of biocontrol system. To define the appropriate conditions for the mass production of the Bacillus altitudinis HC7, we have investigated appropriate culture conditions and effects of various nutrient source on the bacterial growth. The appropriate initial pH and temperature were determined as pH 6.0 and 30℃, respectively. The appropriate concentration of medium elements for the growth of pathogen inhibitor bacterium(Bacillus altitudinis HC7) was determined as follows: 3.0% soluble startch, 10% soytone, 1.0% (NH4)2HPO4, 1.0 mmol KCl, and 0.5% L-asparagine.
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