• Title/Summary/Keyword: tissue cultures

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Somatic Embryogenesis and Plant Regeneration in Immature Flower Bud Cultures of Carnation (카네이션의 미숙화뢰 배양을 통한 체세포배 발생 및 식물체 재분화)

  • 안병준
    • Korean Journal of Plant Tissue Culture
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    • v.24 no.6
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    • pp.369-374
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    • 1997
  • Immature flower buds of 'Desio' carnation were cultured on MS agar medium supplemented with 1 ㎎/L 2,L-D. Embryogenic calli were formed from 5-10% of the buds less than 20 ㎜ in length, but only non-embryogenic calli were produced from explants of shoot apex leaf, internode, and flowere buds larger than 20 ㎜. The same method was applied to 16 cultivars of cut Sower carnation and embryogenic calli were obtained in 7 cultivars. Several embryogenic callus lines were selected and maintained through subcultures over 120 weeks without loss of embryogenic competence. The embryogenic cultures were also proliferated rapidly in liquid agitation cultures using MS medium supplemented with 1mg/L 2,4-D. Numerous embryos were formed on the periphery of the cell aggregates upon transfer to auxin-free MS agar medium. Plantlets were transplanted in potting soil and grown to bloom in six months.

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Saponin Production in Tissue Culture of Ginseng (Panax ginseng C.A. Meyer) (고려인삼의 조직배양에 의한 사포닌 생산)

  • Choi, Kwang-Tae;Park, Ji-Chang;Ahn, In-Ok
    • Journal of Ginseng Research
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    • v.14 no.2
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    • pp.107-111
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    • 1990
  • Ginseng root explants and calli were cultured on modified Murashine and Skoog's media supplemented with different concentrations of organic or inorganic compounds and plant growth requlators to clarify the effects of chemical compositon and plant growth regulators in the medium on the growth of ginseng calli and the production of ginseng saponin. For optimum growth of ginseng calli, the concentrations of 2, 4-D and sucrose were in the range of 1 to 5 mg/l and 1 to 3%, respectively. And it was clarified that sucrose, nitrogen, phosphate, calcium, magnesium, plant growth regulators and their concentrations influcenced the relative biosynthesis of saponin in tissue cultures of Panax ginseng.

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Effect of Polyamines on Formation of Adventitious Roots, Trichomes and Calli by NAA in Leaf Segment Cultures of Arabidopsis thaliana (애기장대 잎 절편 배양시 NAA 농도에 따른 부정근, 모용 및 캘러스 형성에 미치는 Polyamine의 영향)

  • 한태진;홍종필;김준철;임창진;진창덕
    • Korean Journal of Plant Tissue Culture
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    • v.27 no.2
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    • pp.117-123
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    • 2000
  • In order to study the role of polyamines on the formation of adventitious roots, trichomes and calli, the effects of putrescine, spermidine, spermine, cyclohexylamine (CHA) and methylglyoxal-bis(guanylhydrazone) (MGBG) were investigated in the leaf segment cultures from ecotype Columbia of Arabidopsis thaliana. When the leaf segments were cultured on the media for forming adventitious roots (0.1 mg/L NAA), trichomes (2.0 mg/L NAA) and calli (10.0 mg/L NAA), and then each cultures was treated with 1-100 mg/L of putrescine, spermidine and spermine, respectively. On the adventitious root-forming medium treated with polyamines the trichomes were induced with adventitious roots. And on the trichome-forming medium with polyamines calli were induced with trichomes. In orther hand each cultures was treated with 1-100 mg/L of CHA and MGBG, respectively. CHA promoted adventitious roots on the medium for adventitious roots, was not effected on media for trichomes and calli. MGBG inhibited adventitious roots, trichomes and calli in all cultures, and induced adventitious roots on medium for trichomes in high concentration.

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High Frequency Plant Regeneration in Embryogenic Cell Suspension Cultures of Cucumber (오이 배발생세포의 현탁배양을 통한 고빈도 식물체 재분화)

  • 정원중;우제욱;박효근;최관삼;유장렬
    • Korean Journal of Plant Tissue Culture
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    • v.26 no.4
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    • pp.289-291
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    • 1999
  • Hypocotyl explants from 7 days old seedlings of one $F_1$ hybrid cultivar and two pure lines of cucumber formed embryogenic calli at frequencies of up to 8% when cultured on Murashige and Skoog medium (MS) supplemented with 1 mg/L 2,4-D for 3 weeks. Embryogenic calli gave rise to somatic embryos. When slices of somatic embryos were cultured on the same medium for 4 weeks, they formed embryogenic calli. Embryogenic cell suspension cultures were established with embryogenic calli in MS liquid medium with 1 mg/L 2,4-D. Embryogenic potential of cell suspension cultures was maintained by subculturing every seven days. When the level of 2,4-D in the medium was lowered to 0.2 mg/L by diluting with liquid MS basal medium, embryogenic cell suspension cultures underwent development into numerous somatic embryos. When plated onto MS basal medium, over 95% of somatic embryos developed into plantlets. Plantlets were transplanted to potting soil and grown to maturity.

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Inhibitory Effects of Pepper Mild Mottle Virus Infection by Supernatants of Five Bacterial Cultures in Capsicum annuum L.

  • Venkata Subba Reddy, Gangireddygari;In-Sook, Cho;Sena, Choi;Ju-Yeon, Yoon
    • The Plant Pathology Journal
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    • v.38 no.6
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    • pp.646-655
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    • 2022
  • Pepper mild mottle virus (PMMoV), one of the most prevalent viruses in chili pepper (Capsicum annuum L.) is a non-enveloped, rod-shaped, single-stranded positive-sense RNA virus classified in the genus Tobamovirus. The supernatants of five bacterial cultures (Pseudomonas putida [PP], Bacillus licheniformis [BLI], P. fluorescens [PF], Serratia marcescens [SER], and B. amyloliquifaciens [BA]) were analyzed to find novel antiviral agents to PMMoV in chili pepper. Foliar spraying with supernatants (1:1, v/v) obtained from Luria-Bertani broth cultures of PP, BLI, PF, SER, and BA inhibited PMMoV infection of chili pepper if applied before the PMMoV inoculation. Double-antibody sandwich enzyme-linked immunosorbent assay showed that treatments of five supernatants resulted in 51-66% reductions in PMMoV accumulation in the treated chili pepper. To identify key compounds in supernatants of PP, BLI, PF, SER, and BA, the supernatants were subjected to gas chromatography-mass spectrometry. The 24 different types of compounds were identified from the supernatants of PP, BLI, PF, SER, and BA. The compounds vary from supernatants of one bacterial culture to another which includes simple compounds-alkanes, ketones, alcohols, and an aromatic ring containing compounds. The compounds triggered the inhibitory effect on PMMoV propagation in chili pepper plants. In conclusion, the cultures could be used to further conduct tissue culture and field trial experiments as potential bio-control agents.

TGF-β1 upregulates the expression of hyaluronan synthase 2 and hyaluronan synthesis in culture models of equine articular chondrocytes

  • Ongchai, Siriwan;Somnoo, Oraphan;Kongdang, Patiwat;Peansukmanee, Siriporn;Tangyuenyong, Siriwan
    • Journal of Veterinary Science
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    • v.19 no.6
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    • pp.735-743
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    • 2018
  • We investigated the effect of transforming growth factor beta 1 ($TGF-{\beta}1$) on equine hyaluronan synthase 2 (HAS2) gene expression and hyaluronan (HA) synthesis in culture models of articular chondrocytes. Equine chondrocytes were treated with $TGF-{\beta}1$ at different concentrations and times in monolayer cultures. In three-dimensional cultures, chondrocyte-seeded gelatin scaffolds were cultured in chondrogenic media containing 10 ng/mL of $TGF-{\beta}1$. The amounts of HA in conditioned media and in scaffolds were determined by enzyme-linked immunosorbent assays. HAS2 mRNA expression was analyzed by semi-quantitative reverse transcription polymerase chain reaction. The uronic acid content and DNA content of the scaffolds were measured by using colorimetric and Hoechst 33258 assays, respectively. Cell proliferation was evaluated by using the alamarBlue assay. Scanning electron microscopy (SEM), histology, and immunohistochemistry were used for microscopic analysis of the samples. The upregulation of HAS2 mRNA levels by $TGF-{\beta}1$ stimulation was dose and time dependent. $TGF-{\beta}1$ was shown to enhance HA and uronic acid content in the scaffolds. Cell proliferation and DNA content were significantly lower in $TGF-{\beta}1$ treatments. SEM and histological results revealed the formation of a cartilaginous-like extracellular matrix in the $TGF-{\beta}1$-treated scaffolds. Together, our results suggest that $TGF-{\beta}1$ has a stimulatory effect on equine chondrocytes, enhancing HA synthesis and promoting cartilage matrix generation.

Difference of Microbiology according to Tissue Sampling in Bone Involved Diabetic Ulcers (골조직 병변이 있는 당뇨발 궤양에서 표재조직 및 골조직 세균배양 검사의 비교)

  • Rhee, Sung-Mi;Han, Seung-Kyu;Kim, Woo-Kyung
    • Archives of Plastic Surgery
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    • v.37 no.4
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    • pp.335-339
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    • 2010
  • Purpose: Diabetic foot ulcer with osteomyelitis is notorious with its complexity and healing difficulties. Bone biopsy is considered to be the gold standard method of guidance for antibiotic therapy. However, it is often replaced by cultures of ulcer swabs or by superficial samples because of the technical difficulties and possible adverse events. In this study, we compared microbiologic results of bone biopsy with those of superficial tissue biopsy or swab culture to investigate concordance and diagnostic value in bone involved diabetic foot ulcers. Methods: This study involved 106 patients with diabetic foot ulcers who showed positive results in bone probing test. Tissue samples for microbiologic tests were collected from all the patients by using superficial cotton swab, superficial tissue biopsy, and bone biopsy. The microbiologic results of bone biopsy were compared with swab culture and superficial tissue biopsy statistically. Results: The positive predictive value of bone probing test for underlying osteomyelitis was 82.1%. Microbiology of the bone biopsy showed same results with those of the swab culture and superficial tissue in 64% and 63%, respectively. Statistical analysis demonstrated that the microbiology of the swab culture or superficial tissue did not coincide with that of the bone biopsy. Conclusion: These results suggest that the microbiologic results of superficial tissue or swab culture do not coincide with those of bony tissue. To select appropriate antibiotic regimen for diabetic ulcer with bone involvement, the specimen for the microbiologic test should be obtained from involved bone.

Simvastatin as a Modulator of Tissue Remodeling through Inhibition of Matrix Metalloproteinase (MMP) Release from Human Lung Fibroblasts

  • Ra, Ji-Eun;Lee, Ji-Kyoung;Kim, Hui-Jung
    • Tuberculosis and Respiratory Diseases
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    • v.71 no.3
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    • pp.172-179
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    • 2011
  • Background: Statins can regulate the production of pro-inflammatory cytokines and inhibit MMP production or activation in a variety of types of cells. This study evaluated whether statins would inhibit MMP release from human lung fibroblasts, which play a major role in remodeling processes. Methods: This study, using an in-vitro model (three-dimensional collagen gel contraction system), evaluated the effect of cytokines (tumor necrosis factor-${\alpha}$, TNF-a and interleukin-$1{\beta}$, IL-1b) on the MMP release and MMP activation from human lung fibroblasts. Collagen degradation induced by cytokines and neutrophil elastase (NE) was evaluated by quantifying hydroxyproline. Results: In three-dimensional collagen gel cultures (3D cultures) where cytokines (TNF-a and IL-1b) can induce the production of MMPs by fibroblasts, it was found that simvastatin inhibited MMP release. In 3D cultures, cytokines together with NE induced collagen degradation and can lead to activation of the MMP, which was inhibited by simvastatin. Conclusion: Simvastatin may play a role in regulating human lung fibroblast functions in repair and remodeling processes by inhibiting MMP release and the conversion from the latent to the active form of MMP.

Rapid Propagation through Tissue Culture of Cudrania tricuspidata, Medicinal Plant

  • Lee, Cheul-Ho;Min, Ji-Yun;Jung, Ha-Na;Kim, Kyu-Sick;Choi, Myung-Suk
    • Korean Journal of Medicinal Crop Science
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    • v.15 no.5
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    • pp.315-318
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    • 2007
  • An effective rapid propagation method was established through in vitro cultures of the medicinal plant, Cudrania tricuspidata. In vitro plantlets were obtained from in vitro germinated seeds. The various levels of cytokinins (BAP, Kinetin and TDZ) were tested on multiple shoot formation from plantlets. BAP (1.0 mg/l) treatment induced highest number of multiple shoots. Single shoot cultures gave higher initial shoot numbers than 5 shoots per culture. Among the various culture media, the shoot elongation was optimal on 2 MS basal medium without growth regulators. The IAA (2.0 mg/l) treatment induced highest number of roots. IBA (2.0 mg/l) treatment more promoted in vitro root growth than other concentrations. Rooted shoots were transferred directly to small pots with an artificial soil and successfully acclimatized.

Effect of Sugar Starvation on the Sugar Transport System in Suspension Cultures of Streptanthus trotus (Streptanthus tortus 배양 세포에서 당류고갈이 당류 수송계에 미치는 영향)

  • 조봉희
    • Korean Journal of Plant Tissue Culture
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    • v.27 no.1
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    • pp.47-50
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    • 2000
  • In suspension cultures of Streptanthus. the uptake rate of sugar was increased during the ceil starvation of sugar in the medium. The maximal uptake rate obtained with 3 days of cell starvation. Sugar transport system induced by the sugar starvation was completely inhibited by 10 $\mu$M cycloheximide. Plant cells are known to possess only one sugar transport system, but the uptake rate of glucose obtained a saturated kinetic while the one of sucrose had two different kinetics after the sugar starvation. Induced sugar transport systems had different kinetics compared to plant cell. These results showed that higher plants have adaptable ability to induce new sugar transport systems when the environment changed unsuitable.

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