• Korean Journal of Agricultural Science
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• v.47 no.3
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• pp.693-705
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• 2020

Grapes (Vitis spp. L.) are the third most produced fruit in the world. Crown gall disease caused by Agrobacterium vitis forms galls in the stems of the grapevines and reduces the vitality of the fruit trees, resulting in reduced yields. This pathogen has occurred in vineyards worldwide and caused serious economic losses. It is a soil-borne disease, so Agrobacterium vitis can survive for several years in vineyards and is difficult to control. Additionally, since there is no effective chemical control method, the most effective control method is the breeding of resistant varieties. To make the resistant variety, marker-assisted selection (MAS) enables fast breeding with low cost. In this study, we applied a genome-wide association study (GWAS), by combining phenotyping and genotyping-by-sequencing (GBS), for the development of a single nucleotide polymorphism (SNP) marker set related to crown gall disease using 350 grapevine varieties. As a result of the GBS based genotyping analysis, about 58,635 SNPs were obtained. In addition, the phenotypic analysis showed 35.2% resistance, 73% moderate susceptibility and 16.4% highly susceptibility. Moreover, after confirmation, two genes (VvARF4 and VvATL6-like) were shown to be related to crown gall disease based on the results of GWAS analysis, using the phenotypic data, and GBS. High-resolution melting analysis (HRMA) was performed using the Luna^® Universal Probe with real-time PCR to distinguish the melting peaks of the resistant and susceptible varieties. Our data show that these SNP markers are expected to be helpful in evaluating resistance against grapevine crown gall disease and in breeding.

• Archives of Plastic Surgery
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• v.34 no.1
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• pp.1-7
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• 2007

Purpose: Human adipose tissue-derived stromal cells(hADSCs) can be expanded in vitro and induced to differentiate into multiple mesenchymal cell types. In this study we have examined various neuronal phenotypes and gene expression profiles of the hADSCs in the neuronal induction. Methods: The hADSCs were isolated from human adipose tissue and they were characterized by the flow cytometry analysis using CD13, CD29, CD34, CD45, CD49d, CD90, CD105 and HLA-DR cell surface markers. We differentiated the hADSCs into the neuronal lineage by using chemical induction medium and observed the cells with contrast microscopy. The immunocytochemistry and western blotting were performed using the NSE, NeuN, Trk-A, Vimentin, N-CAM, S-100 and ${\beta}$-Tubulin III antibodies. Results: The hADSCs were positive for CD13($90.3{\pm}4%$), CD29($98.9{\pm}0.7%$), CD49d($13.6{\pm}6%$), CD90 ($99.4{\pm}0.1%$), CD105($96%{\pm}2.8%$) but negative for CD34, CD45 and HLA-DR. The untreated cultures of hADSCs predominately consisted of spindle shaped cells and a few large, flat cells. Three hours after the addition of induction medium, the hADSCs had changed morphology and adopted neuronal-like phenotypes. The result of immunocytochemistry and western blotting showed that NSE, NeuN, Trk-A, Vimentin, N-CAM, S-100 and ${\beta}$-Tubulin III were expressed. However, NSE, NeuN, Vimentin were weakly expressed in the control. Conclusion: Theses results indicate that hADSCs have the capabillity of differentiating into neuronal lineage in a specialized culture medium. hADSCs may be useful in the treatment of a wide variety of neurological disorders.

• Current Research on Agriculture and Life Sciences
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• v.32 no.3
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• pp.111-117
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• 2014

Phytophthora blight caused by Phytophthora capsici Leonian is a dangerous disease threatening pepper growers worldwide. The efficacy of chemical control is generally low as the pathogen is soil-borne and rapidly spread by zoospores during the rainy season. Thus, based on the demand for resistant varieties, various good resistant sources, such as CM334, AC2258, and PI201234, have been reported and their inheritance of resistance studied by many different authorities. However, the mode of inheritance remains unclear, as 1 or 2 independent dominant genes, 3 genes, or multiple genes have all been reported as responsible for resistance. Recently, QTL mappings of the gene factors for resistance have been reported, and molecular markers for resistance used in breeding programs. With the release of many resistant commercial hybrid cultivars, differentiation of pathotypes of the pathogen is attracting interest among breeders and plant pathologists. Various authorities have already classified the pathogen strains into different races according to the inter-action between resistant host plants, including the source of resistance, such as CM334 and PI201234, and resistant commercial varieties and P. capsici isolates. However, no standard differential host sets have yet been established, so the results are good only for the pathogen strains used in the experiments. Thus, for breeding varieties with durable resist-ance, it is important to introduce resistance from different sources and use diverse local pathogen strains collected in the target area for distribution in a breeding program.

• Polymer(Korea)
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• v.33 no.2
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• pp.118-123
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• 2009

Keratin contains regulatory molecules that can enhance neuronal cell activity. We fabricated keratin/ PLGA films using 0, 10, 20, and 50 wt% of keratin using solvent casting method. We measured the contact angle of each film and cell proliferation was assayed by counting the cells attached on the film. Adhered cell morphology was confirmed by scanning electron microscope. RT-PCR was conducted to evaluate the gene expression of NF, NSE, and S-100, the Schwann cell markers. The keratin content of 20 and 50 wt% provided higher wettability than PLGA. The 20 wt% keratin was better in cell adhesion and proliferation of SCs than other keratin/PLGA films. The phenotypic stability of SC was maintained with the keratin content of 10 and 20 wt%.

• Korean Journal of Breeding Science
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• v.41 no.4
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• pp.474-481
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• 2009

The purpose of this study was to identify the quantitative trait loci (QTL) associated with gel consistency (GC), and to evaluate the relationships between GC and physico-chemical properties related to eating quality in rice. A total of 120 doubled haploid (DH) lines derived from a cross between 'Samgang' and 'Nagdong' and 26 varieties were used for this study. The 120 DH lines were divided into four groups, soft (61~100 mm), medium (41~60 mm), hard (26~40 mm) and whole population, according to the gel length. A negative correlation between GC and amylose content was observed only in the soft group. The GC negatively correlated with lipid content in the medium, and the whole population, but positively correlated with lipid content in the soft group. The positive correlation between GC and protein content was observed in the soft group, while negative correlations were observed in the medium group and the whole population. Two significant QTLs regarding GC were detected on chromosomes 4 and 11. They collectively explained 23% of phenotypic variation with LOD score of 3.1 and 2.9, respectively. The DNA markers of S4026 and RM287 were tightly linked to GC in the DH population and 26 varieties, respectively.

• The Korean Journal of Physiology and Pharmacology
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• v.23 no.2
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• pp.151-159
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• 2019

Pruritus (itching) is classically defined as an unpleasant cutaneous sensation that leads to scratching behavior. Although the scientific criteria of classification for pruritic diseases are not clear, it can be divided as acute or chronic by duration of symptoms. In this study, we investigated whether skin injury caused by chemical (contact hypersensitivity, CHS) or physical (skin-scratching stimulation, SSS) stimuli causes initial pruritus and analyzed gene expression profiles systemically to determine how changes in skin gene expression in the affected area are related to itching. In both CHS and SSS, we ranked the Gene Ontology Biological Process terms that are generally associated with changes. The factors associated with upregulation were keratinization, inflammatory response and neutrophil chemotaxis. The Kyoto Encyclopedia of Genes and Genomes pathway shows the difference of immune system, cell growth and death, signaling molecules and interactions, and signal transduction pathways. Il1a, Il1b and Il22 were upregulated in the CHS, and Tnf, Tnfrsf1b, Il1b, Il1r1 and Il6 were upregulated in the SSS. Trpc1 channel genes were observed in representative itching-related candidate genes. By comparing and analyzing RNA-sequencing data obtained from the skin tissue of each animal model in these characteristic stages, it is possible to find useful diagnostic markers for the treatment of itching, to diagnose itching causes and to apply customized treatment.

• Journal of Korean Medicine for Obesity Research
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• v.20 no.2
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• pp.69-77
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• 2020

Objectives: In this study, we investigated the antiobesity and antidiabetic effects of polyherbal extract, DM2 consisting of Atractylodis Rhizoma, Anemarrhenae Rhizoma, Cinnamomi Cortex, and Moutan Radicles Cortex in high fat diet-induced obesity mice. Methods: DM2 extract was prepared with a hot water. Six-week-old male C57BL/6N mice were fed a high-fat diet (HFD) for 8 weeks and then administrated with DM2 extract (500 mg/kg, p.o.) for 4 weeks. The changes of physiological markers, body weight (BW), food and water intakes, and the levels of fasting blood glucose (FBG) were measured once a week for 4 weeks in mice. The the serum levels of glucose, insulin, aspartate aminotransferase (AST), alanine aminotransferase (ALT), total cholesterol (T-CHO), triglyceride, and low density lipoprotein cholesterol in sera were measured in mice using autometic chemical analyzer and enzyme linked immunosorbant assay. We also observed the histological changes of liver and pancreatic tissues with Hematoxylin & Eosin staining. Results: In physiological change, the increases of BW, calorie intake, and FBG in HFD-induced obese mice were significantly decreased after administration of DM2 extract for 4 weeks. The decrease of water intake was significantly increased in DM2 extract-administrated mice. In serological change, the administration of DM2 extract in obesity mice was significantly decreased the serum levels of glucose, insulin, T-CHO, AST, and ALT levels. We also found that DM2 extract inhibited the increase of lipid droplets in liver and the structural destruction of pancreatic tissues in obesity mice. Conclusion: Our study demonstrated that DM2 extract has antiobesity antidiabetic effects with body weight loss, decrease of glucose and insulin levels, and lipid accumulation on liver tissue.

• Proceedings of the Korean Society of Applied Pharmacology
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• 1997.04a
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• pp.107-107
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• 1997

Cytochrome P450 enzymes have been intensively investigated in hepatic tissues and several mammalian cell lines. Compared to most studies about cytochrome P450 isozymes in liver in vivo and hepatic, cell lines in vitro, the study of cytochrome P450IA1 in human breast cancer cells could be very important to understand the mechanism of the regulation of CYPIA1 gene expression and cell growth. MCF-7 human breast cancer cells are well characterized to study estrogen and antiestrogen action due to the fact that they contain high level of estrogen receptor and have biological markers characterized. And also MCF-7 cells express high level of arylhydrocarbon hydroxylase activity and human cytochrome P450IA1 cDNA was cloned from MCF-7 cells. Ah receptor was characterized in many breast cancer cell lines and polycyclic aromatic hydrocarbon such as 3-MC induced the expression of CYPIA1 gene and cytochrome P450- dependent monooxygenase activity. We undertook a study to examine the effect of estrogens and other chemicals on the regulation of human CYPIA1 gene expression in MCF-7 cells via RTPCR analysis, that might help us to understand the mechanism of the regulation of CYPIA1 gene expression and MCF-7 cell growth. Expression vector containing the functional 5'-regulatory region of human CYPIA1 fused to the CAT reporter gene was transfected into estrogen receptor positive MCF-T cells or estrogen receptor negative MDA-MB-231 cells. After these cells were treated with various chemicals, RTPCR was carried out to measure both CYPIA1 mRNA and CAT mRNA levels. 1nM 3-MC increased in both P450 and CAT mRNA levels over those of control by two folds in MCF-7 cells but does not in MDA-MB-231 cells. Estrogen or tamoxifen or retinoic acid or chrysin decreased in both P450 and CAT mRNA levels that were induced by 3-MC in MCF-7 when each chemical was administered with 3-MC concomitantly. These results suggested that the level of CYPIA1 gene expression is modulated with estrogen-related molecules and make it possible to speculate that ER is related to CYPIA1 gene expression and cell growth in breast cancer cells. [Supported by grants from the Korean Ministry of Education ]

• Journal of Korean Society of Forest Science
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• v.112 no.4
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• pp.554-560
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• 2023

To prevent illegal timber distribution, DNA markers have been used to identify the species and origin. However, extracting high-quality DNA from timber is difficult because of its physical and chemical properties. In this study, we investigated whether the age of timber tissue influences the yield of DNA extraction and the success rate of polymerase chain reaction (PCR) to understand the relationship between the establishment time of the wood annual ring and the extracted DNA concentration (ng/μl), purity (A260/A280), and PCR success rate (%) from pinewood, a major Korean domestic species. According to the results, it was observed that as the distance from the cambium increased, indicating that the tissue was older, the concentration and purity of the extracted DNA decreased significantly. For the trnM-trnV (285 bp) and rpoC1 (298 bp) regions, the PCR success rate was 100%. However, for the rbcL (1.3 kb) region, the PCR success rate was 66.67%. Moreover, PCR amplification of the rbcL region failed at all points older than 30 years. Thus, it is deduced that as time passes, along with the decay of timber cells, DNA is degraded, leading to a decrease in DNA concentration, purity, and PCR success rate. The results of this study are expected to be beneficial for future applications, such as the species identification of timber, providing valuable insights and potential utilization in this field.

• Proceedings of the Korean Society of Crop Science Conference
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• 2017.06a
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• pp.107-107
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• 2017

Sweet corns are enjoyed worldwide as processed products and fresh ears. Types of sweet corn are based on the gene(s) involved. The oldest sweet corn type has a gene called "sugary (su)". Sugary-based sweet corn was typically named "sweet corn". With its relatively short shelf life and the discovery of a complementary gene, "sugary enhanced (se)", the sweet corn (su only) was rapidly replaced with another type of sweet corns, sugary enhanced sweet corn, which has recessive homozygous su/su, se/se genotype. With the incorporation of se/se genotype into existing su/su genotype, sugary enhanced sweet corn has better shelf life and increased sweetness while maintaining its creamy texture due to high level of water soluble polysaccharide, phytoglycogen. Super sweet corn as the name implies has higher level of sweetness and better shelf life than sugary enhanced sweet corn due to "shrunken2 (sh2)" gene although there's no creamy texture of su-based sweet corns. Distinction between sh2/sh2 and su/su genotypes in seeds is phenotypically possible. The Involvement of se/se genotype under su/su genotype, however, is visually impossible. The genotype sh2/sh2 is also phenotypically epistatic to su/su genotype when both genotypes are present in an individual, meaning the seed shape for double recessive sh2/sh2 su/su genotype is much the same as sh2/sh2 +/+ genotype. Hence, identifying the double and triple recessive homozygous genotypes from su, se and sh2 genes involves a testcross to single recessive genotype, chemical analysis or DNA-based marker development. For these reasons, sweetcorn breeders were hastened to put them together into one cultivar. This, however, appears to be no longer the case. Sweet corn companies began to sell their sweet corn hybrids with different combinations of abovementioned three genes under a few different trademarks or genetic codes, i.g. Sweet $Breed^{TM}$, Sweet $Gene^{TM}$, Synergistic corn, Augmented Supersweet corn. A total of 49 commercial sweet corn F1 hybrids with B73 as a check were genotyped using DNA-based markers. The genotype of field corn inbred B73 was +/+ +/+ +/+ for su, se and sh2 as expected. All twelve sugary enhanced sweet corn hybrids had the genotype of su/su se/se +/+. Of sixteen synergistic hybrids, thirteen cultivars had su/su se/se sh2/+ genotype while the genotype of two hybrids and the remaining one hybrid was su/su se/+ sh2/+, and su/su +/+ sh2/+, respectively. The synergistic hybrids all were recessive homozygous for su gene and heterozygous for sh2 gene. Among the fifteen augmented supersweet hybrids, only one hybrid was triple recessive homozygous (su/su se/se sh2/sh2). All the other hybrids had su/su se/+ sh2/sh2 for one hybrid, su/su +/+ sh2/sh2 for three hybrids, su/+ se/se sh2/sh2 for three hybrids, su/+ se/+ sh2/sh2 for four hybrids, and su/+ +/+ sh2/sh2 for three hybrids, respectively. What was believed to be a classic super sweet corn hybrids also had various genotypic combination. There were only two hybrids that turned out to be single recessive sh2 homozygous (+/+ +/+ sh2/sh2) while all the other five hybrids could be classified as one of augmented supersweet genotypes. Implication of the results for extension service and sweet corn breeding will be discussed.