A marine bacterial strain producing agar-degrading enzymes was isolated from a mud flat in Jeboo-do (Korea) using a selective artificial sea water (ASW) agar plate containing agar as the sole carbon source. The isolate, designated as SH-1, was gram-negative, aerobic, and motile with single polar flagellum. 16S rRNA gene sequence similarity analysis showed the isolate SH-1 had the highest homology (96.5%) to marine bacterium Neiella marina J221. Cells could grow at $28-37^{\circ}C$ but not at $42^{\circ}C$, and the agarase activity of the cell culture supernatant was higher when grown at $28^{\circ}C$ than when grown at $37^{\circ}C$. Cells could grow when concentrations of 1-5% (w/v) NaCl were added to the growth media with the best growth observed at 3% NaCl, and the agardegrading enzyme activity of the cell culture supernatant was best when grown at 3% NaCl-containing growth media under the conditions we examined. The crude enzyme prepared from 48-h culture broth of strain SH-1 exhibited an optimum pH and temperature for agar-degrading activity at 7.0 and $40^{\circ}C$, respectively. Zymogram analysis of the crude supernatant and cell extract showed that strain SH-1 produced at least 3 agar-degrading enzymes with molecular weights of 15, 35, and 52 KD. Thinlayer chromatography (TLC) analysis also suggested that HS-1 produces ${\beta}$-agarase to degrade agarose to neoagarooligosaccharides.
Kim, Wi-Sik;Kim, Soo-Jin;Kim, Jeong-Ho;Jung, Sung-Ju;Kim, Do-Hyung;Oh, Myung-Joo
Journal of fish pathology
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v.23
no.3
/
pp.335-342
/
2010
In 2008, mass mortality was observed in paradise fish Macropodus opercularis which was imported from Indonesia. PCR of these fish found positive for megalocytivirus and Mycobacterium sp., while an unidentified virus was culture-isolated using CHSE-214 cells. In the present study, we investigated characterization of the unidentified virus and its pathogenicity to determine whether the virus was the causative agent of the mass mortality of paradise fish. The unidentified virus induced cytopathic effect (CPE) with syncytia in CHSE-214 and other fish cells, BF-2, GF, SSN-1, FSP and FFN. The virus was resistant against treatments with IUdR, chloroform, acidity at pH 3, basicity at pH 11 and high temperature at $56^{\circ}C$ for 3h. By electron microscopy, the viral particles were spherical having a double capsid structure with approximately 65 nm in external diameter. Viral genome was composed of at least 10-segmented RNA with sizes ranging from 0.7 kb to 3.6 kb. Based on these characters, this virus can be classified into family Reoviridae. This reovirus did not cause any mortality in an artificial experiment conducted by injecting the virus to paradise fish. This indicates that the reovirus is not only responsible for the mass mortality of paradise fish in 2008.
Jung, Won Kwon;Kim, Young Soo;Choi, Jin Kook;Kim, Seung-Han;Jang, Myeong-Hwan;Kwon, Tae Lyong;Jeon, Yongho
Research in Plant Disease
/
v.24
no.4
/
pp.332-338
/
2018
In August of 2011, a wilting disease of ginseng was observed at Bongwha, Gyeongbuk province, Korea. Affected plants initially show withering symptoms on leaves of ginseng. As the disease progresses, withering leaves spread downward, eventually encompassing the whole plant. Leaves lose vigor but remain pale green. Symptoms of roots were brown, and soft rots characterized by moist and watery decay of the whole ginseng root, which initiated as small brown, water-soaked lesions of hairy roots and enlarged to the entire roots. The causal organism isolated from the infected roots was identified as Serratia plymuthica based on its physiological and biochemical characteristics, by cellular fatty acid composition (GC-FAME), the utilization of carbon sources (BioLog System), and 16S rRNA sequence of the isolated bacterium were 99% homologous to those of Serratia plymuthica strains. Artificial inoculation of the bacterium produced the same brown or soft rot symptoms on the ginseng roots, from which the same bacterium was isolated. This is the first report of bacterial root rot caused by the Serratia plymuthica in ginseng in Korea. Serratia plymuthica has been used as antagonistic microorganism for biological control on several crop plants. But it was proved pathogen of ginseng at humid condition in this study.
Metabolism is essential for survival and reproduction, and there is a metabolic pathways entry in the clusters of orthologous groups of proteins (COGs) database, updated in 2020. In this study, the metabolic pathways of 1309 prokaryotes were analyzed using COGs. There were 822 COGs associated with 63 metabolic pathways, and the mean for each taxon was between 200.50 (mollicutes) and 527.07 (cyanobacteria) COGs. The metabolic pathway composition ratio (MPCR) was defined as the number of COGs present in one genome in relation to the total number of COGs constituting each metabolic pathway, and the number of pathways with 100% MPCR ranged from 0 to 26 in each prokaryote. Among 1309 species, the 100% MPCR pathways included murein biosynthesis associated with cell wall synthesis (922 species); glycine cleavage (918); and ribosomal 30S subunit synthesis (903). The metabolic pathways with 0% MPCR were those involving photosystem I (1263 species); archaea/vacuolar-type ATP synthase (1028); and Na+-translocation NADH dehydrogenase (976). Depending on the prokaryote, three to 49 metabolic pathways could not be performed at all. The sequence of most highly conserved metabolic pathways was ribosome 30S subunit synthesis (96.1% of 1309 species); murein biosynthesis (86.8%); arginine biosynthesis (80.4%); serine biosynthesis (80.3%); and aminoacyl-tRNA synthesis (82.2%). Protein and cell wall synthesis have been shown to be important metabolic pathways in prokaryotes, and the results of this study of COGs related to such pathways can be utilized in, for example, the development of antibiotics and artificial cells.
Researches on soil microbial community are increasing to assess ecosystem responses to anthropogenic disturbances and to provide an indicator of ecosystem recovery. Microbial communities are able to respond more rapidly to environmental changes than plants and therefore they may provide an early indication of the ecosystem recovery trajectory. This study was conducted using 16S rRNA gene pyrosequencing of soil samples to compare soil bacterial community composition between artificially covered soils of the Baedudaegan ridgeline and their adjacent forest soils in two restoration project sites, Ihwaryeong and Yuksimnyeong, which were completed in 2012 and 2013, respectively. Richness of the Phylum level was 29.3 in Ihwaryeong and 32.3 in Yuksimnyeong. Significant difference in the richness between artificial restored soils and adjacent forest soils(p<0.01) was observed, however no significant difference was observed for site location and soil depth. Acidobacteria(37.3%) and Proteobacteria(31.1%) were more abundant than any other phylum in collected soil samples. Also, we found the significant difference in the relative abundance of the two abundant phyla between artificially restored soils and their adjacent forest soils (Proteobacteria, 38.1% in restored soils vs 24.2% in adjacent forest soils, p<0.01; Acidobacteria, 55.4% in restored soils vs 19.2% in adjacent forest soils, p<0.001). The results support the previous researches indicating that soil bacterial community composition is affected by nutritional status of soils and that Acidobacteria is also strongly influenced by pH, thus favoring soils with lower pH. This study could be utilized to monitor and evaluate restoration success of forest soil environment quantitatively.
This study was aimed to characterize osteogenic potential of rat bone marrow stromal cells (BMSC) isolated with standard flushing method and investigate the plasticity of transdifferentiation between osteoblastic and adipocytic lineage of cultured BMSC. Unlike aspiration method in human, rat bone marrow was extracted by means of irrigation with culture media that elevates the possibility of co-extraction of committed osteoprogenitor, or preosteoblast or other progenitor cells of several types present inside bone marrow. The cultured stromal cells showed high ALP activity which is representative marker of osteoblast without any treatment. Osteogenic inducers such as Dex and BMP-2 were examined for the evaluation of their effect on osteogenic and adipocytic differentiation of stromal cells, because they function as osteoinductive agent in stromal cells, but simultaneously induce adipogenic differentiation. Osteogenic differentiation was evaluated by measuring alkaline phosphatase activity or mRNA expression of osteoblast markers such as osteopontin, bone sialoprotein, collagen type I and CbfaI, and in vitro matrix mineralization by von Kossa staining. Oil red staining method was used to detect adipocyte and adipocytic marker, aP2 and $PPAR{\gamma}2$ expression was examined using RT-PCR. It can be supposed that irrigation procedure resulted in high portion of already differentiation-committed osteoprogenitor cell showing elevated ALP activity and strong mineralization only under the supplement of $100{\mu}M$ ascorbic 2-phosphate and 10mM ${\beta}$-glycerophosphate without any treatment of osteogenic inducers such as Dex and BMP-2. Dex and BMP-2 seemed to transdifferentiate osteoprogenitor cells having high ALP activity into adipocytes temporarily, but continuous treatment redifferentiated into osteoblast and developed in vitro matrix mineralization. This property must be considered either in tissue engineering for bone regeneration, or in research of characterization of osteogenic differentiation, with rat BMSC isolated by the standard irrigation method.
Kim, Yoonha;Waqas, Muhammad;Khan, Abdul Latif;Mun, Bong-Gyu;Yun, Byung-Wook;Lee, In-Jung
Proceedings of the Korean Society of Crop Science Conference
/
2017.06a
/
pp.203-203
/
2017
The Earth's climate is rapidly changing because of increasing carbon dioxide content in atmosphere so, climate prediction models anticipate that earth surface temperature will rise by 3 to $5^{\circ}C$ in next 50 to 100 years. Therefore, frequency of un-expected weather events such as drought, salinity, low or high temperature and flooding etc. will be increasing worldwide. Furthermore, increased atmosphere temperature can influence pests and pathogens spread as well. Therefore, to protect enormous grain loss from unexpected weather conditions, studies related with combine stress conditions like abiotic plus biotic stress condition are really required. Thus, our research focused on physiological responses under combined abiotic and biotic stress condition in rice plant. To induce uniform stress condition, we used NaCl (100 mM) and salicylic acid (0.5 and 1.0 mM SA) as each stress a stimulator. Each artificial abiotic and biotic stress inducer was applied to hydroponically grown rice seedlings alone or together for four day. The data were collected in a time-dependent manner [1, 2, 3 and 4 day(s) after treatment (DAT)] and were matched with our anticipation that shoot length and shoot fresh weight was decreased in solo and combined abiotic and biotic stress condition. The lipid peroxidation content was significantly increased ($1.5{\pm}0.2$ to $2.7{\pm}0.1mg$ mg of $MDA\;g^{-1}FW$) in the first two days in both stress exposed plants, and showed the opposite trend ($0.5{\pm}0.01$ to $0.1{\pm}0.001mg$ of $MDA\;g^{-1}FW$) in last two days under multi stress condition. Superoxide dismutase (SOD) activity did not showed difference in only biotic stress condition (alone 0.5 and 1.0 mM SA) as compared to control however, it was significantly increased in multi stress condition or solo abiotic stress condition whereas, catalase (CAT), and ascorbate peroxidase (APX) activities were significantly decreased in solo biotic and combined abiotic and biotic condition. In particular, both enzymes activities were more decreased in multi stress condition as compared to solo biotic stress condition. The results for relative mRNA expression level of CAT and APX enzymes were in agreement with results of spectrophotometric values. Correlation value between each stress condition and phenotypic data showed that biotic stress condition showed high correlation with activity of CAT and APX whilst, abiotic stress condition revealed significant correlation with SOD activity.
19 strains, which could be identified as Lactobacillus sp. were isolated. The Cucurbita maxima has been known as a traditional healthy food and variable positive effects on the human body were already reported. In this study we tried to develop a production process for a healthy fermented drink with Cucurbita maxima and strains originated from Kimchi. Many kinds of lacctobacci species existed in the fermented food cannot survive in the acidic conditions in the stomach. So we tried to search and select a strain, which can arrive to the small intestine. A species of a Lactobacillus named as C332 was identifed as Lactobacillus plantarum and selected for the fermentation process. With the treatment with artificial gastric juice and artificial bile the survival rate of the cells could be calculated. The physiological characteristics at the variable conditions have been tested. After fermentation process the sensoric tests on the product with panels were tried. The most of the cells could survive in the acidic conditions and falcultive anaerobe. Especially some antibacterial effects aganinst E.coli were also found. With all kinds of the results from our research the fermented Cucurbita maxima drink can be a successful item in the market.
Kim, Jo-Min;Kim, Ki-Hwan;Kim, Song-Yi;Park, Young-Seo;Seo, Min-Jae;Yoon, Sung-Sik
Food Science and Biotechnology
/
v.14
no.4
/
pp.503-508
/
2005
Screening for antilisterial activity was performed in about three thousand isolates of lactic acid bacteria (LAB) from Chinese cabbage kimchi, and finally based on the relatively stronger antilisterial activities eight bacterial strains were selected. The bacteria were further characterized in terms of their tolerance to artificial gastric juice, pH 2.5, bile salts (0.3% oxgall), and to the different NaCl concentrations. Of the isolates, YK005 was especially investigated for its physiological characteristics due to its inhibitory activity against gram-positive Listeria monocytogenes as well as gram-negative Escherichia coli O157:H7, as they have been constantly reported to be resistant against bacteriocins produced by a number of LAB strains. YK005 was found to be rod-shaped, $3.8\;{\mu}m$ long ${\times}\;0.5\;{\mu}m$ wide, non-sporeforming, non-motile, catalase-negative, and produced gas from glucose (heterolactic). Based on the biochemical data obtained by API 50 CHL medium, the isolate was tentatively identified as Lactobacillus brevis. To validate the result obtained by the biochemical identification, rRNA-based PCR experiments using a pair of species-specific primers for L. brevis were conducted and a single band of 1400 bp was observed, which strongly indicated that YK005 belongs to L. brevis. The LAB isolates are potentially exploited as human probiotic organisms and are employed to control some food-borne pathogens like L. monocytogenes.
Journal of the Korean Society of Food Science and Nutrition
/
v.45
no.10
/
pp.1481-1487
/
2016
Lactic acid bacteria showing alginate-degrading and cellulolytic activity were isolated and identified as a starter for seaweed fermentation. A total of 331 strains of lactic acid bacteria isolated from various Korean traditional foods, such as Kimchi, Jeotgal, and Makgeolli, were examined alginate-degrading and cellulolytic activity by the plate assay method. Six strains showed strong alginate-degrading and cellulolytic activity among the isolated 331 strains. Among these six strains, four strains (strain No. 162, 164, 192, and 196) showed probiotic properties (antimicrobial activity, tolerance to simulated gastric juice, artificial bile acid, and NaCl). No. 192 strain (Gram-positive cocci, catalase negative, and homofermentative) showed the best probiotic properties among selected strains and was identified as Enterococcus faecium by 16S rRNA sequencing. Strain No. 192 (E. faecium) showed the best growth and antioxidative activity during seaweed (sea mustard and sea tangle) fermentation for 72 h at $37^{\circ}C$ among the four selected strains.
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