• Journal of Mushroom
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• v.7 no.3
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• pp.122-129
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• 2009

We determined the genetic relatedness among 81 commercial varieties of oyster mushrooms by either comparison of fruiting body characteristics or using PCR amplification with URP primers. Also, we assessed their temperature adaptation by using box cultivation. Based on our data, some varieties such as Wonhyung and variety 1, Suhan and variety 20, Wang-hukpyung and variety 4, Chunchu #2 and variety 6, Shinong #11 and variety 1, Samgu #1 and variety 2, showed the close genetic relatedness between each other. It is known that mushroom culture room is affected by outer temperature because mushroom need the adequate aeration for their growth. Therefore, in aspect of saving energy, the cultivation of season favorable mushroom species has been recommended. Eight different commercial varieties including Sa-chul, Yeoreum and Bunhong(Pink) grew well at high temperature ($16^{\circ}C$). Also, about 51 different oyster mushroom strains were good at relatively high temperature. However, other 64 different oyster mushroom varieties were grew well at low temperature ($11^{\circ}C$). Fourty-nine varieties were adapted well in most temperatures. The slow growth rate were observed in some varieties including Kun(P. eryngii ), Awi(P. nebrodensis ), and Cheunbok(P. abalonus ), which are not suitable for box cultivation. We believe that the culture characterization of each mushroom varieties should help many farmers in many aspects such as saving energy, quantity improvement and new variety development etc.

• Journal of Life Science
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• v.24 no.9
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• pp.967-972
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• 2014

In the present study, we prepared eighty-five different kinds of lees extracts and their solvent fractions and investigated their anti-proliferative activities against human colorectal cancer HCT116 cells. HCT116 cells were treated with eighty-five solvent fractions of lees extracts and then cell viability was measured using MTS assay. Among the treated solvent fractions, three solvent fractions (KSD-E1-3, KSD-E2-3, and KSD-E4-3) were selected based on cell viability assay. In addition, we performed an oligo DNA microarray analysis to analyze the gene expression changes by treatment of KSD-E1-3 in HCT116 cells. Among the upregulated genes, we selected 4 genes (NAG-1, ATF3, p21, and DDIT3) and performed RT-PCR using gene-specific primers. Among the treated solvent fractions, KSD-E1-3 dramatically induced the expressions of the four selected genes. In addition, we investigated whether the upregulations of those genes were dependent on the transcription factor p53's presence using p53 null HCT116 cells. The results indicate that the upregulations of NAG-1, ATF3, and DDIT3 are not dependent on the p53 presence, whereas p21 is dependent on the p53 presence. These findings may help to understand the molecular mechanisms of the anti-proliferative activity mediated by rice wine lees in human colorectal cancer cells.

• Journal of Plant Biotechnology
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• v.29 no.3
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• pp.151-159
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• 2002

Genetic map and molecular marker have a great importance in improving and facilitating crop breeding program as well as in genome analysis and map-based cloning of genes representing desirable characters. This study aimed at developing RAPD markers and constructing a genetic linkage map using 82 BC$_1$F$_1$individuals originated from the cross between '835' and B$_2$in radish (Raphanus sativus L.). One of the parents for genetic linkage map construction, '835'(P$_1$) of egg type is susceptible to Fusarium wilt and have medium resistance to virus infection and the other parent, B$_2$(P$_2$) of round type, is susceptible to Fusarium wilt and virus, Screening of 394 RAPD primers in BC$_1$F$_1$) population resulted in selecting 128 polymorphic markers which displayed 1:1 segregation pattern. Two markers failed to display 1:1 segregation and showed the segregation ratio skewed to maternal genotype. Selected markers were categorized into 14 linkage group based on LOD score represented by MAPMAKER/EXP program. Five groups composed of single marker among them were excluded from the linkage map, and consequently, the remaining groups are well matched with the number of radish chromosome (n＝9). The linkage map constructed with 128 markers covers 1,688.3 cM and the average distance between markers was 13.8 cM. For developing STS marker, we determined the partial nucleotide sequence of OPE10 marker at both ends and designed a oligonucleotide primer pair based on this sequence. STS PCR using the primer pair displayed a single, clear band of which segregation is perfectly matched with that of OPE10 marker. This implies that RAPD markers could readily convert into clear and reliable STS markers.

• Journal of Korean Society of Forest Science
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• v.90 no.1
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• pp.155-160
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• 2001

Changes in genetic diversity after seed-tree practice were simulated in test plantation of tulip tree at Sokdal-ri, Banwol-myon, Hwasung-gun, Kyungki Provence. I-SSR markers were analyzed in a total of 305 tulip trees. A total of 89 amplicon variants were observed by PCR with 9 I-SSR primers. Genetic diversity for 305 trees was relatively high (S. I. =0.4532). Individuals originated from the seed orchards in U.S.A. showed the highest level of genetic diversity (0.4530), those from Anyang showed the medium level (0.4152), and those from Cheonbuk showed the lowest (0.3929). Simulation of seed-tree practice accompanied by 2 consecutive thinnings was performed on the basis of morphological characteristics and planted distances of the individual trees, which left 37 trees as candidates for seed-trees. Decreasing rate of genetic diversity within seed sources was greatest for individuals from Cheonbuk (28.3%), moderate for those from Anyang (16.3%), and smallest for those from U.S.A. (8.0%). In spite of little difference in decreasing rate of individuals for the 3 seed sources (87.5~88.2%), large difference in decreasing rate of genetic diversity within seed sources might be due to difference in number of mother trees for the 3 seed sources. For example, whereas individuals originated from the seed orchard in U.S.A., which showed the smallest decreasing rate of genetic diversity, might be originated from relatively large number of mother trees, those from Anyang and Cheonbuk might be originated from relatively small number of mother trees. Although mean of 17.5% of the genetic diversity within seed sources was decreased through 2 consecutive thinnings, a decrease in genetic diversity for the whole individuals (37 trees) was only 6.1%. This observation suggests that the seed-tree practice on the basis of the criteria established in the present study may not result in great reduction in overall genetic diversity of the progenies.

• Journal of Chest Surgery
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• v.37 no.12
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• pp.967-977
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• 2004

Background: DNA methylation is one of the important gene expression mechanisms of the cell. When cytosine of CpG dinucleotide in promotor is hypomethylated, expression of some genes that is controlled by this promoter is altered. In this study, the author investigated the effect of DNA demethylating agent, 5-aza-2'-deoxycytidine (ADC), on the expressions of cancer antigen genes, MHC and B7 in 4 lung cancer cell lines, NCIH1703, NCIH522, MRC-5, and A549. Material and Method: After treatment of cell lines, NCIH1703, NCIH522, MRC-5 and A549 with ADC (1 uM) for 48 hours, RT-PCR was performed by using the primers of MAGE, GAGE, NY-ESO-1, PSMA, CEA, and SCC antigen gene. In order to find the optimal ADC treatment condition for induction of cancer antigen, we studied the effect of ADC treatment time and dose on the cancer antigen gene expression. To know the effect of ADC on the expression of MHC or B7 and cell growth, cells were treated with 1 uM of ADC for 72 hours for FACS analysis or cells were treated with 0.2, 1 or 5 uM of ADC for 96 hours for cell counting. Result: After treatment of ADC (1 uM) for 48 hours, the expressions of MAGE, GAGE, NY-ESO-1, and PSMA genes increased in some cell lines. Among 6 MAGE isotypes tested, and gene expression of MAGE-1, -2, -3, -4 and -6 could be induced by ADC treatment. However, CEA gene expression did not change and SCC gene expression was decreased by ADC treatment. Gene expression was generally induced 24 - 28 hours after ADC treatment and expression of MAGE, GAGE, and NY-ESO-1 was maintained at least 14 days after ADC ADC teatment, and expression of MAGE, GAGE, and NY-ESO-1 was maintained at least 14 days after ADC teatment in ADC-Free medium. Most gene expression could be induced at 0.2 uM of ADC, but gene expression increased dependently on ADC treatment dose. The expression of MHC and B7 was not increased by ADC treatment in all four cell lines, and the growth rate of 4 cell lines decreased significantly with the increase of ADC concentrations. Conclusion: Treatment of lung cancer cell lines with ADC increases the gene expression MAGE, GAGE and NY-ESO-1 that are capable of induction of cytotoxic T lymphocyte response. We suggest that treatment with 1 uM of ADC for 48 hours and then culturing in ADC-free medium is optimal condition for induction of cancer antigen. However, ADC has no effect on MHC and B7 induction, additional modification for increase of expression of MHC, B7 and cytokine will be needed for production of efficient cancer cell vaccine.

• Clinical and Experimental Reproductive Medicine
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• v.36 no.4
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• pp.265-274
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• 2009

Objective: MicroRNAs (miR) are known to repress target genes at post-transcriptional level and play important roles in development and maturation of cell. However, the expression profiles of miR during ovarian follicle maturation have not been fully elucidated. Here, we designed this study to investigate the expression profiles of miR in oocytes and granulose cells (G-cells) after in vitro culture according to gonadotropins and adding hCG. Methods: Ovaries from 12-day-old mice (C57BL6) were removed and preantral follicles were isolated and cultured in $20\;{\mu}L$-drop of culture media with supplementation of either rFSH, rLH, or rFSH+rLH. After their full maturation, follicles were incubated with rhCG and rEGF. RNA was isolated from oocytes and G-cells, and real-time PCR were performed with primers of miR known to be expressed in the mouse ovary (mmu-miR-16, -miR-27a, -miR-126, -miR-721). Results: FSH+LH group showed the highest ovulation and MII rates among gonadotropin groups. The profiles of miRs in oocytes and G-cells differed according to gonadotropin groups and adding hCG. The profiles of miRs showed divergent changes between oocytes and G-cells. Conclusion: miR expression profiles are altered by gonadotropins and supplementation of hCG during in vitro maturation of murine follicles. Target gene study must be necessary to validate these findings.

• Journal of Periodontal and Implant Science
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• v.28 no.4
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• pp.659-678
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• 1998

In the present study, oligonucleotide probes based on 16S rRNA were taken to investigate the diversity of oral spirochetes without culture method. This is the first study that revealed oral spirochetes of both presently cultivable and uncultured oral spirochetes in Korean adult periodontitis patients. Subgingival plaque samples were taken from diseased sites(probing depth ${\geq}6\;mm$, experimental group, n=116) and healthy sites(probing depth${\leq}3mm$, control 1 group, n=28) in 29 patients with adult periodontitis, and from 20 periodontally healthy subjects(probing depth${\leq}3mm$, control 2 group, n=100). Following being examined under phase-contrast microscope, all samples were submitted to dot-blot hybridization after polymerase chain reacton with eubacterial primers. 5 species-specific probes(TVIN, TDEN, TMAL, TSOC, and TPEC) and 7 group-specific probes(TRE I, TRE II, TRE III, TRE IV, TRE V, TRE VI, and TRE VII) were used one by one for the identification of both cultivable and so far uncultivable oral spirochetes. All probes were labeled with digoxigenin(DIG)-ddUTP and detected by chemilumininescence. The following results were obtained. 1. Under phase-contrast microscope, 91.37% and 14.28% of oral spirochetes were observed in the experimental and control 1 groups, respectively. None of oral spirochetes were observed in control 2 group. 2. With universal probe, 98.27%, 46.42%, and 22.0% of oral spirochetes were observed in experimental, control 1, and control 2 groups, respectively. 3. With specific probe, 95.68%, 35.71%, and 19.0% of oral spirochetes were observed in experimental, control 1, and control 2 groups, respectively. 4. With species-specific probes, T. socranskii were recovered in a high percentage of sites(81.89%) examined, followed by T. maltophilum(50.0%), T. vincentii(36.20%), T. denticola(13.79%), respectively. With group- specific probes, TRE IV was recovered in a high percentage of sites(85.34%) examined, followed by TRE II(77.58%), TRE I(56.89%), TRE III(25.86%), TRE VI(5.17%), and TRE V(2.58%), respectively. 5. T. vincentii were only observed in the diseased sites, not in the healthy sites. 6. Neither T. pectinovorum nor group VII oral spirochetes were observed in any sites. The findings warrant further investgations of the recovered spirochetes to elucidate the possible associations of oral spirochetal prevalence in race and types of periodontitis, pathogenesis of T. vincentii and the possible distributional change of oral spirochetes before and after treatments.

• The Korean Journal of Mycology
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• v.27 no.2 s.89
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• pp.124-131
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• 1999

This study was carried out to identify the phylogenetic relationship among Phellinus species by comparing the DNA sequences of the 5.8S ribosomal DNA (rDNA) and the internal transcribed spacers (ITSs), ITS1 and ITS2 regions. Two primers from the 3' end of 18S rDNA and the 5' end of 28S rDNA sequences were chosen to amplify the specific ITS regions of Phellinus spp. Phellinus strains used in the study were divided into four clusters by the phylogenetic tree based on the amplified regions of ITS and 5.8S rDNA sequences. The first cluster consist of Phellinus hartigii IMSNU 32041 and Phellinus robustus IMSNU 32068, and the second cluster consists of Phellinus linteus strains and Phellinus weirianus IMSNU 32021. Phellinus laevigatus KCTC 6229, KCTC 6230 and Phellinus igniarius KCTC 6227, KCTC 6228 belong to the third cluster. Finally, Phellinus chrysoloma KCTC 6225 and Phellinus chrysoloma KCTC 6226 are the fourth cluster. In the second cluster the differentiation between Phellinus linteus strains and Phellinus weirianus species were not possible by the comparison of the ITS sequences. These results revealed that Phellinus linteus and Phellinus weirianus cannot be established the concept of species level only by the ITS sequences. Therefore, both physiological and molecular biological methods as well as the sequences of type strains are necessary to classify the strains of these two species accurately. The comparison of the ITS sequences of four Phellinus species indicated that the sequences of the ITS1 generally are more divergent than those of the ITS2. Although the ITS sequences are varied in some species, the conserved regions in both ITS1 and ITS2 are useful tool to differentiate the species. Phellinus linteus and related species have their specific sequences in the ITS1 compared to the other species.

• Journal of fish pathology
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• v.30 no.2
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• pp.89-96
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• 2017

The majority of freshwater ornamental fish are imported and distributed domestically, causing high risk of exposure to exotic pathogens and drug resistant bacteria in Korea. Aeromonas hydrophila is known as a common species of fresh water bacteria and opportunistic fish pathogen, as well as a species causing zoonotic infection. In this study, we isolated motile aeromonads from various imported freshwater ornamental fish and studied the characters of the isolates. Imported freshwater ornamental fish were purchased on day 1 after the fish were deposited in the aquarium. Bacteria were isolated from the liver, kidney and spleen of fish using 0.5% NaCl containing tryptic soy agar medium. Bacteria were grouped on the basis of their morphological characteristics. The colonies with clear zone on starch-ampicillin agar (SA agar) were tentatively identified as Aeromonas spp. Two hundred and twenty-six strains, about 70% of total isolates were assumed to be Aeromonas spp. Nine isolates were further identified based on the result of the API 20E test and PCR using primers specific for A. hydrophila 16S rRNA gene. The isolates were identified as A. hydrophila and the API 20E test showed differences in trisodium citrate, D-sucrose, D-melibiose, amygdalin and L-arabinose availability between the nine isolates and standard A. hydrophila. The susceptibilities of the isolated bacteria to 10 antibacterial agents were confirmed by the disk diffusion method. Isolated strains were found to be resistant to amoxicillin and ampicillin and sensitive to florfenicol. However, 7 isolates showed multiple drug resistances to erythromycin, oxytetracycline, nalidixic acid etc. Pathogenicity of the isolates was determined by the artificial challenge test on goldfish (Carassius auratus). Three isolates caused 60 ~ 80% mortality in goldfish within 5 days after the initiation of challenge. These results indicate that multiple drug resistant, highly pathogenic and exotic A. hydrophila can spread to domestic aquarium and the preventive treatment of fish before sale is necessary.

• Journal of Mushroom
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• v.15 no.3
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• pp.129-133
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• 2017

Oyster mushroom is a type of mushroom that is commonly cultivated and consumed in Korea. P. ostreatus 'Suhan' is a preferred cultivar for many mushroom farmers because it has a dark pileus and thick stipe. However, as it is very sensitive to environmental conditions, farmers consistently demand an alternative cultivar. To develop a new cultivar, the parental strains KMCC01680 ('Suhan') and KMCC00478 ('Gosol') were selected from various collected P. ostreatus strains by cultivating genetic resources. P. ostreatus 'Heuksol' was developed by the method of Mon-Mon crossing between monokaryotic strains derived from 'Suhan' and 'Gosol'. Thirty strains of 174 crossed strains were initially selected by cultivation experiments. After bulk cultivation tests, 'Heuksol' was selected. The nuclear DNA profile of 'Heuksol' was similar to those of the parental strains, 'Suhan' and 'Gosol', when RAPD (random amplified polymorphic DNA) primers and UPF (Universal PCR Fingerprinting) 2, 3, and 4 were used. The optimum temperature for mycelial growth was $30^{\circ}C$ for 'Heuksol', but medium-high temperatures were also appropriate, especially $13-20^{\circ}C$. The fruiting body production per bottle (1,100 mL) was approximately 140.8 g. When compared to the control strain 'Suhan', the thickness of the stipe of 'Heuksol' was greater than that of 'Suhan' (13.5 mm vs 9.4 mm). The pileus diameter of 'Heuksol' was similar to that of 'Suhan' and the pileus thickness of 'Heuksol' and 'Suhan' was 19.7 mm and 21.8 mm, respectively. 'Heuksol' had more a productive stipe number than 'Suhan' and the pileus of 'Heuksol' was dark gray, even at high temperatures. Therefore, it was suggested that this new cultivar, 'Heuksol', could provide an alternative to 'Suhan' and contribute to the profit of oyster mushroom farms.