• Korean Journal of Plant Tissue Culture
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• v.26 no.4
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• pp.251-257
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• 1999

Adventitious buds were produced from the cultures of mature zygotic embryos of Larix leptolepis with the highest frequency of 91.7% in SH medium containing 1.0 mg/L zeatin. The most effective cytokinin combination for inducing adventitious buds was 1.0 mg/L zeatin + 1.0 mg/L thidiazuron (40.3%). The highest mean number of adventitious buds induced in 1.0 mg/L zeatin + 1.0 mg/L 2iP or 1.0 mg/L zeatin + 1.0 mg/L kinetin combined treatments. Elongation of the adventitious buds occurred the best on half strength LM salts medium, on which the buds elongated upto 27 mm. Also, the supplement of activated charcoal in medium suppressed the elongation of the adventitious buds. The highest frequency (23.3%) of rooting from elongated shoots was obtained on the medium containing 5.0 mg/L IBA and 0.2 mg/L NAA. The highest number (3.5) of roots was induced on the medium containing 5.0 mg/L IBA alone.

• Journal of Plant Biotechnology
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• v.33 no.4
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• pp.297-302
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• 2006

In general, the proliferation of orchids via somatic embryos has been used for mass production of somatic clones because of high propagation efficiency. In spite of high propagation rate, this method often brings somaclonal variation, especially polyploid frequency. Therefor we here concentrated to investigate the relationship between endopolyploidization patterns of explants and the occurrence of tetraploid variant in clonally proliferated Doritaenopsis via somatic embryo regeneration system. In the fully developed somatic embryo, upper part contained 2C to 16C while middle and lower parts showed 2C to 32C DNA content. Two-week-old embryo contained 2C to 16C, whereas those regenerated after 4 to 10-week-old contained 2C to 64C nuclei. Results showed that endoreduplication was variable depending upon tissue types, ages, and parts in one species. lower part of somatic embryo having high endoreduplication degree increased the regeneration of tetraploid variants by about 3-fold comparing to upper part of somatic embryo culture. polyploid frequency occurrence might be closely related to the high levels of endoreduplication of somatic embryos used as explant. It suggested that the upper part of somatic embryo having comparatively low endoreduplication degree is suitable for the stable in vitro propagation system.

• Korean Journal of Plant Tissue Culture
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• v.28 no.5
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• pp.283-287
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• 2001

Differential display based on PCR was employed to identify genes expressed during tuber-developing stage of potato (Solanum tuberosum L. cv. Irish Cobbler). An eukaryotic initiation factor 5A (eIF-5A) clone isolated from a cDNA library constructed with developing micro-tuber using a probe of PCR fragment. We isolated three positive clones and ore of them contained open reading frame. This clone revealed high sequence similarity to tomato eIF 5A cDNA. At the DNA level, there is 94.8% identity with the tomato eIF-5A4, whereas at the protein level there is a high identity with 97.5%. The potato eIF 5A clone is 716 bp in length and contains a single open reading frame from 57 to 539 bp, a 56 bp 5'-untranslated region and a 177 bp 3'-untranslated region. The deduced protein composed of 160 amino acid residues, with a predicted molecular mass of 17.4 kD and an estimated pl of 5.5. The sequence of 12 (STSKTGKHGHAK) amino acids among eIF-5A proteins is perfectly conserved from yeast to human. That sequence in potato eIF-5A protein is also conserved at position 46 to 57 amino acid. This region embeds the post-translational modification site of the lysine residue (at the seventh K) to hypusine that is crucial to eIF-5A activity. The northern blot analysis of eIF5A has shown abundant expression, mainly in flower organs (stamen, ovary, petal, sepal), fruit and stolen.

• Journal of Plant Biotechnology
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• v.43 no.1
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• pp.12-20
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• 2016

Advances in DNA sequencing technologies have contributed to revolutionary understanding of many fundamental biological processes. With unprecedented cost-effective and high-throughput sequencing, a single laboratory can afford to de novo sequence the whole genome for species of interest. In addition, population genetic studies have been remarkably accelerated by numerous molecular markers identified from unbiased genome-wide sequences of population samples. As sequencing technologies have evolved very rapidly, acquiring appropriate individual plants or populations is a major bottleneck in plant research considering the complex nature of plant genome, such as heterozygosity, repetitiveness, and polyploidy. This challenge could be overcome by the old but effective method known as haploid induction. Haploid plants containing half of their sporophytic chromosomes can be rapidly generated mainly by culturing gametophytic cells such as ovules or pollens. Subsequent chromosome doubling in haploid plants can generate stable doubled haploid (DH) with perfect homozygosity. Here, classical methodology to generate and identify haploid plants or DH are summarized. In addition, haploid induction by epigenetic regulation of centromeric histone is explained. Furthermore, the utilization of haploid plant in the genomics era is discussed in the aspect of genome sequencing project and population genetic studies.

• Korean Journal of Veterinary Research
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• v.46 no.4
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• pp.395-398
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• 2006

A 2-month-old female Shih-tzu dog was referred because of lameness, exercise intolerance, depression, elbow and stifle joint swelling. Physical examination, complete blood counts, serum-chemistry, radiography, synovial fluid analysis, antinuclear antibody test, and rheumatoid factor measurement were initiated. On radiography, soft tissue swelling of elbow and stifle joints without erosiveness were founded. The results of synovial fluid analysis revealed severe neutrophilic pleocytosis (nondegenerative), decreased viscosity, increased turbidity, positive on mucin-clot test, and negative on bacterial culture. The results of rheumatoid factor measurement and antinuclear antibody test were negative and below 1 : 40, respectively. Based on all tests, we diagnosed this case as juvenile onset type I immune-mediated polyarthritis. Azathioprine (1 mg/kg body weight, per os q 24 h, for 4 weeks) was then administered and clinical signs improved gradually. Four weeks after azathioprine administration, clinical signs were disappeared. This report describes the clinical findings, imaging characteristics, synovial fluid findings, and other laboratory results of type I immune-mediated polyarthritis and successful management with azathioprine therapy.

• Journal of The Korean Society of Grassland and Forage Science
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• v.31 no.1
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• pp.1-8
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• 2011

Molecular techniques such as genetic transformation are powerful tools that can be used for the genetic modification of warm-season grasses. The P. meyerianum with high regeneration ability was used for establishing an Agrobacterium-mediated transformation system. We investigated various factors affecting Agrobacterium infection by examining GUS gene expression of pCAMBIA1304 vector. Among various concentration of acetosyringone and betaine tested for inoculation and co-cultivation, 10 mg/L acetosyringone and 60 mg/L betaine resulted in the highest transformation frequency in terms of GUS expression. The calli of 4 species of Panicum spp. with excellent tissue culture response were inoculated with Agrobacterium under the optimal infection conditions. The high activity of GUS gene was observed in all species and hygromycin-resistant calli expressing GFP were obtained in P. meyerianum, P. longijubatum, P. stapfianum and guineagrass Noh-PL1. Co-cultivated calli were transferred onto the selection medium containing hygromycin, and the hygromycin resistant calli were selected after 3 months. Hygromycin-resistant plantlets were then successfully regenerated from the calli and grown in a greenhouse. We confirmed stable insertion of hpt gene among the hygromycin-resistant plantlets of P. meyerianum by PCR analysis.

• Pediatric Infection and Vaccine
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• v.3 no.2
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• pp.154-162
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• 1996

Purpose : Adenoviruses(Ad) have been shown to play an important role in the etiology of severely acute respiratory diseases, particulary in infants and young children, and the occurrence of fatal outcome and chronic pulmonary sequelae in association with adenoviral infection has been a cause of great interest and concern. This report presents the resul of a retrospective analysis on 30 cases of lower respiratory infection from which adenovirus was isolated. Patients & Methods : The 30 patients in this study represent all detected cases of adenovial infection out of 240 children who were admitted to Sang Sung Medical Center between February to June 1996 showing signs and symptoms of lower respiratory tract infection. The diagnosis of adenovirus infection was based on microscopic visualization of typical cytopathic effect in HEp-2 tissue culture and used monoclonal Ab with nasopharyngeal aspiration. Results : The male/female ratio was 2:1 and the majority of age range was below 36months. Clinical diagnoses in all 30 patients were pneumonia(n=21), bronchitis and Bronchiolitis(n=5) and ARDS(n=4). We recieved the most of patients in the month of May. The chief complaints were fever(93.3%) and cough(80%) and extrapulmonary symptoms were diarrhea(n=5), seizure(n=4), abdominal pain(n=1). The mean duration of fever was $11.95{\pm}6.54$days. Physical examination on admission were crackles(73.3%), coarse breathing sounds(60%), hepatosplenomegaly(33.3%), decreased brething sounds(30%). In WBC counts, 8cases were below $4000/mm^3$ and 14 cases were above $10,000/mm^3$. In platelets counts, 4cases were below $150,000/mm^3$ and 10 cases were above $450,000/mm^3$. 21 cases were above 1 in CRP. GOT and GPT were abnormal in some cases. Chest X-ray revealed diffuse pulmonary infiltration(n=15), pleural effusion(n=6), consolidation(n=4) and hyperaeration(n=3). Seven patients were treated at the peditric intensive care unit with respiratory support and high dose of gammaglobulin. However, one patients died even through he was treated with NO ventilation and high frequency ventilation. Conclusion : Those with adenoviral pneumonia and respiratory infection having long fever duration and symptoms like bacterial pneumonia must be carefully differentiated in order to provide proper treatement and preventive measures due to possible fatal outcome.

• The Plant Pathology Journal
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• v.19 no.1
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• pp.13-18
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• 2003

From 1996 to 1999, potato-growing areas in Korea were surveyed for identification and distribution of potato scab pathogens. Potato scab was widely distributed in the mass cultivation areas, especially in Jriu island, southern areas of Chonnam and Gyounggi provinces, and the alpine area of Gangwon province. Jeju island was the most affected area by this disease. A total of 55 Streptomyces strains were isolated from potato scab lesions, among which 40 strains were pathogenic on progeny tubers. Among the pathogenic strain, 21 strains were identified as previously described S. scabies, 7 Strains as S. turgidiscabies, and 5 Strains as S. acidiscabies, while 7 strains were observed as having distinct phenotypic properties. These strains were classified into six distinct clusters based on phenotypic characteristics and selected representative strains for each cluster. S. scabies (S33) had grey spores in a spiral chain. Mean-while, S. turgidiscabies (S27) had grey spores, S. acidiscabies (S71) had white spores, S. luridiscabiei (S63) had yellow-white spores, S. puniciscabiei (S77) had purple-red spores, and S. niveiscabiei (S78) had thin and compact white spores, all in a rectiflexuous chain. Pathogenicity was determined by the production of thaxtomin A and homologs of necl and ORFtnp genes. In TLC, representative strains S27, S71, S63, S77, and S78 produced a yellow band that co-migrated with the authentic thaxtomin A. However, thaxtomin A was not detected in chloroform extracts from oatmeal broth culture and Slice tuber tissue of S. luridiscabiei (S63) and S. puniciscabiei (S77) by HPLC analysis. In addition, no homologs of necl and ORFtnp genes in S. acidiscabies (S71), S. luridiscabiei (S63), S. puniciscabiei (S77), and S. niveiscabiei (S78) were detected by PCR and Southern hybridization analysis.

• Korean Journal of Plant Tissue Culture
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• v.25 no.5
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• pp.289-293
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• 1998

We investigated the 1,1-diphenyl-2-picrylhydrazyl (DPPH) radical scavenging activity in methanol extracts of 64 cultured cell lines, which were derived from various plant species, and the ascorbate content in cell lines, which showed a high radical scavenging activity. Thirteen cell lines revealed the antioxidative activity ($IC_{50}$) by methanol extracts of less than 50 mg in cell fresh wt. Of them, six cell lines showed the same Rf value as ascorbate on the DPPH sprayed silica gel TLC. The ascorbate content in cell lines of Rosa multiflora, Scutellaria baicalensis, and Achyranthes japonica showed 48.5, 30.3, and $16.8\;\mu\textrm{g}$ per g cell fresh wt by HPLC analysis, respectively. In callus cultures of S. baicalensis, the concentration of ascorbate reached a maximun ($39{\pm}3.4\;\mu\textrm{g}/g$ cell fresh wt) on 30 days after subculture, which corresponded to the stationary growth phase, and subsequently decreased by successive culturing.

• Korean Journal of Plant Tissue Culture
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• v.26 no.3
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• pp.183-187
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• 1999

To establish an efficient screening system for new herbicides using plant cultured cells, responses of tobacco photomixotrophic cultured (PH) cells to various herbicides with different modes of action were surveyed by measuring the cell growth and ion conductivity in medium. The cells were cultured in Murashige and Skoog (MS) medium containing 0.7mg/L 2,4-D, 0.3mg/L kinetin and 30 g/L sucrose at $25^{\circ}C$ in the light (100 rpm). Chemicals were treated to suspension cultures of tobacco PH cells at the time of subculture. The cell growth and ion conductivity in the medium were investigated on 12 days after chemical treatment. The ion conductivity assay gave well correlated results to the cell growth inhibition data. The responses of tobacco PM cells were dependent on the modes of action of chemicals tested. Atrazine, an inhibitor of photosynthetic electron transport (PET), strongly inhibited both the cell membrane and cell growth ($IC_{50}$/, about 1 $\mu$M). Butachlor (an inhibitor of cell division), glufosinate (an inhibitor of amino acid biosynthesis), and fluridone (an inhibitor of carotenoid biosynthesis) showed a dose-dependent inhibition. However, Quinclorac, a herbicide with an auxin activity, did not affect the cell growth and ion leakage. These results suggested that tobacco PM cells is suitable materials for the simple screening of new herbicides such as PET, amino acid biosynthesis, ceil division inhibitors by measuring the cell growth and ion conductivity.