• Title/Summary/Keyword: tissue cultures

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Somatic Embryogenesis and Plant Regeneration in Leaf Explant Cultures of Gentiana scabra var buergeri (용담(Gentina scabra var. buergeri)의 잎 절편 배양에서 체세포배발생에 의한 식물체 재분화)

  • 방재욱;이미경;정성현
    • Korean Journal of Plant Tissue Culture
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    • v.21 no.4
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    • pp.233-237
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    • 1994
  • Plant regeneration system via somatic embryogenesis in leaf explant cultures of Gentiana scabra var. buergeri has been established. Leaf segments formed calli when cultured on MS medium supplemented with 0.5 mg/L 2,4-D and 2 mg/L BAP After transferred to SH medium supplemented with 0.5 mg/L 2,4-D, 2 mg/L CPA and 0.5 mg/L kinetin, the callus became embryogenic. The embryogenic callus was subcultured every 3 to 4 weeks. Upon transfer onto SH basal medium the embryogenic callus gave rise to numerous somatic embryos, which subsequently developed into plantlets. The regenerated plants were potted in an artificial soil with mixture (peatmoss : pearlite : vermiculite : 2 : 1 : 1) and transplanted to the soil after kept under a high humidity for two weeks. A total of 78 plants out of 105 regenerated plants survived in the soil. Phenotypic variations in height, number of stems and the flowering time were observed in tile regenerated plants. Cytogenetical analyses showed no chromosomal variation.

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In Vitro Propagation of Narcissus pseudonarcissus by Scale Cultures Using Thidiazuron (인편의 Thidiazuron처리에 의한 나팔수선의 기내증식)

  • 이병기;김영숙;박병모
    • Korean Journal of Plant Tissue Culture
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    • v.22 no.1
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    • pp.53-57
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    • 1995
  • This study was carried out to investigate the effect of TDZ (thidiazuron) treatment on Propagation in vivo through scale cultures of Narcissus pseudonarcissus. Scales with disk (5 to 7 m in size) were cultured on MS medium containing NAA, BA and TDZ. Bulbs and shoots were directly formed when scales were cultured on medium containing 5 mg/L NAA and 1 mg/L BA. In addition, combination of 3 mg/L NAA and 0.02 mg/L TDZ promoted effectively the direct formation of bulbs and shoots. The shoots were rooted when cultured on medium containing 5 mg/L NAA and 0.02 mg/L TDZ. non scales obtained from regenerates were cultured on medium containing 1.0 mg/L NAA and 0.5 mg/L TDZ, they gave rise to numerous bulbs and shoots . The overall results suggest that TDZ is an effective plant growth regulator in vitro propagation of N. pseudonarcissus.

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The Relation Between Sox9, TGF-${\beta}1$, and Proteoglycan in Human Intervertebral Disc Cells

  • Lee, Yong-Jik;Kong, Min-Ho;Song, Kwan-Young;Lee, Kye-Heui;Heo, Su-Hak
    • Journal of Korean Neurosurgical Society
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    • v.43 no.3
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    • pp.149-154
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    • 2008
  • Objective: The aim of this study is to elucidate the effects of transforming growth factor-${\beta}$ (TGF-${\beta}$)1 and L-ascorbic acid on proteoglycan synthesis, and the relationship between Sox9, proteoglycan, and TGF-${\beta}1$ in intervertebral disc cells. Methods: Human intervertebral disc tissue was sequentially digested to 0.2% pronase and 0.025% collagenase in DMEM/F-12 media and extracted cells were cultured in $37^{\circ}C$, 5% $CO_2$ incubator. When intervertebral disc cells were cultured with TGF-${\beta}1$ or L-ascorbic acid, the production level of sulfated glycosaminoglycan (sGAG) was estimated by dimethyl methyleneblue (DMMB) assay. The changes of Sox9 mRNA and protein levels via TGF-${\beta}1$ were detected by RT-PCR and Western blot analysis in each. Results: The amount of sGAG was increased with the lapse of time during incubation, and sGAG content of pellet cultured cells was much larger than monolayer culture. When primary cultured intervertebral disc cells in monolayer and pellet cultures were treated by TGF-${\beta}1$ 20 ng, sGAG content of experimental group was increased significantly compared to control group in both cultures. L-Ascorbic acid of serial concentrations (50-300 ug/ml) increased sGAG content of mono layer cultured intervertebral disc cells significantly in statistics. The co-treatment of TGF-${\beta}1$ and L-ascorbic acid increased more sGAG production than respective treatment. After treating with TGF-${\beta}1$, Sox9 mRNA and protein expression rates were significantly increased in disc cells compared with the control group. Conclusion: This study suggests that TGF-${\beta}1$ would increase sulfated glycosaminoglycan (sGAG) and other proteoglycans such as versican by elevating Sox9 mRNA and protein expressions in order.

Production of Ubiquinone 10 from the Callus Culture of Tabacco(Nicotiana tabacum cv Xanthi) (연초(Nicotiana tabacum cv Xanthi) 세포배양에 의한 Ubiquinone 10의 생산)

  • 양덕춘;박지창;최광태
    • Korean Journal of Plant Tissue Culture
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    • v.21 no.6
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    • pp.341-345
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    • 1994
  • The effect of phytohormones on in vitro production of ubiquinone 10 from the callus cultures of Nicotiana tabacum cv Xanthi was investigated. The growth of callus cultures of Xanthi was in proved by addition of NAA and 2,4-D, especially NAA 0.5 mg/L alone, at the light condition. Ubiquinone 10 was detected by HPLC, and confirmed from Xanthi callus cultured on the all of uppermedia. The ubiquinone 10 content in Xanthi tobacco callus cultured on the medium with NAA 0.5 mg/L only was higher than that of other mixed medium with NAA and 2,4-D. However addition of IBA 1 mg/L and NAA 0.5 mg/L to the medium was more effective in promoting ubiquinone 10 formation than that of NAA 0.5 mg/L only As the callus growth of Xanthi was considerabley restrained at concentration of kinetin, Content and production of ubiquinone low as the highest at kinetin 0.5mg/L and 2,4-D 0.5mg/L in the light.

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Production of Herbicide-resistant Transgenic Plants from Embryogenic Suspension Cultures of Cucumber (오이의 배발생 현탁 배양세포로부터 제초제 저항성 형질전환 식물체 생산)

  • 우제욱;정원중;최관삼;박효근;백남긴;유장렬
    • Korean Journal of Plant Tissue Culture
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    • v.28 no.1
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    • pp.53-58
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    • 2001
  • To develop herbicide-resistant cucumber plants (Cucumis sativus L. cv Green Angle) embryogenic suspension cultures were co-cultured with Agrobacterium tumefaciens strain LBA4404 carrying a disarmed binary vector pGA-bar. The T-DNA region of this binary vector contains the nopalin synthase/neomycin phosphotransferase Ⅱ (npt Ⅱ) chimeric gene for kanamycin resistance and the cauliflower 35S/phosphinothricin acetyltransferase (bar) chimeric gene for phosphinothricin (PPT) resistance, After co-cultivation for 48 h, embryogenic calli were placed on maturation media containing 20 mg/L PPT. Approximately 200 putatively transgenic plantlets were obtained in hormone free media containing 40 mg/L PPT. Northern blot hybridization analysis confirmed the expression of the bar gene that was integrated into the genome of five transgenic plants. Transgenic cucumber plants were grown to maturity. Mature plants in soil showed tolerance to the commercial herbicide (Basta) of PPT at the manufacturer's suggested level (3 mL/L).

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Effects of Light on Activities of Antioxidative Enzymes in Hairy Root Cultures of phytolacca esculenta Houtte (자리공(Phytolacca esculenta van Houtte) 모상근배양에서 항산화효소의 활성에 미치는 광의 영향)

  • 양덕조;김용해;권진이;최철희;양덕춘
    • Korean Journal of Plant Tissue Culture
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    • v.22 no.2
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    • pp.71-76
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    • 1995
  • The effects of light on the activities of several antioxidative enzymes, catalase (CAT), superoxide dismutase(SOD), ascorbate oxidase(AO), and peroxidase(POD) were examined in the hairy root cultures of Phytolacca esculenta van Houtte induced by Agrobacterium tumefaciens $A_4$T. Activities of CAT, SOD, and AO were significantly decreased with incresing light intensity (500-2,000 lx). The activity of AO under high light condition (2,000 lx)was decreased by 92% compared to the dark condition. The activities of glutathoine peroxidase (GPO), ascorbate peroxidase (APO) and general POD were increased under lower light intensify below 500 lx. The activity of GPO under 2,000 lx was decreased by 85% compared to the dark condition. The activities of antioxidative enzymes were more decreased in blue light (400-500nm). The activities of antioxidative enzymes in blue light intensity were increased in lower light intensity below 30 lx, but decreased 21-70% under 200 lx. The activity of AO was decreased by 70% under 200 lx with increasing blue light intensity. Our results suggest that the activities of antioxidative enzymes in hairy roots might be inhibited by endogenous oxidants generated under the high blue light conditions.

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Effect of Medium Component on Plant Regeneration via Adventitious Bud Formation from Leaf Explant Cultures of Strawberry (Fragaria ananassa Duch.) (딸기 (Fragaria ananassa Duch.) 잎 절편체 배양으로부터 부정아 형성을 통한 식물체 재생)

  • 조덕이;소웅영;정원일
    • Korean Journal of Plant Tissue Culture
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    • v.28 no.3
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    • pp.171-178
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    • 2001
  • This study was investigated to establish a regeneration system of plant via adventitious bud formation from leaf explant cultures of strawberry (Fragaria ananassa Duch). Effects of plant growth regulators (2,4-D, BAP), agar sucrose and myo-inositol on adventitious bud formation were investigated. When the leaf explants were cultured on MS medium supplemented with 0.1 mg/L 2,4-D and 3 mg/L BAP, the adventitious bud formation was most promoted. The adventitious bud formation was not induced from leaf explants cultured on MS medium containing 2,4-D alone. Adventitious bud formation was enhanced to almost 3 times on medium with low level of agar concentration (0.4%) in comparison with those on the medium with high level of agar (1%), but almost of shoot was vitrificated on the medium. Therefore, the normal adventitious bud formation from leaf explants was most effective on the medium containing 0.05 mg/L 2,4-D,1 mg/L BAP, 0.8% agar, 30 g/L sucrose and 100 mg/L myo-inositol. Thus, the mass propagation of healthy strawberry could be established using leaf explants.

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Effects of Growth Regulators on Shoot Differentiation and Bulblet Formation in Shoot-Tip and Bulb-Scale Cultures of Lilium longiflorum (백합 경단 및 인편배양으로부터 유식물체 분화 및 자구형성에 미치는 생장조절제의 영향)

  • 이은모;정해준;민병훈;이영복
    • Korean Journal of Plant Tissue Culture
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    • v.22 no.2
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    • pp.83-87
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    • 1995
  • Regulation of organ differentiation by growth regulators was investigated through the shoot-tip and bulb-scale cultures of Lilium longiflorum (cv Georgia). When shoot tips were placed on MS medium supplemented with 0.1 mg/L NAA alone or 0.1 mg/L NAA and 0.1 mg/L BA, axillary shoots were proliferated. Root diffentiation and growth were stimulated on the basal medium. Although growth regulation did not seem to be necessary when bulb scales were used as explants for shoot differentiation, its differentiation was promoted vigorously by 0.2 mg/L NAA, but suppressed by BA. Bulblets were formed from bulb-scale-derived plantlets cultured on MS medium supplemented with 0.1 mg/L IBA. And more bulblets were formed from the plantlet in MS medium supplying 0.2 mg/L NAA with 6% than3% sucrose

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Effect of Salicylic Acid on Anthocyanin Synthesis in Cell Suspension Cultures of vitis vinifera L. (포도의 현탁세포배양에서 안토시아닌 생합성에 미치는 Salicylic Acid의 영향)

  • 신동호;유상렬;최관삼
    • Korean Journal of Plant Tissue Culture
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    • v.22 no.2
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    • pp.59-64
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    • 1995
  • Effects of salicylic acid (SA) on anthocyanin synthesis in cell suspension cultures of grapes (Vitis vinifera L.) were investigated. tow concentrations (0.1 to 1$\mu$M) of SA did not affect the cell growth and anthocyanin accumulation whereas high concentrations (5 to 10$\mu$M ) of SA inhibited cell growth with increasement of anthocyanin synthesis. Five micromoles of SA promoted anthocyanin accumulation 4 folds compared to control cells. When SA was treated on the different culture times (0 to 7day), the highest pigment accumulation was obtained at the cells of second day. A low productivity of anthocyanin under continuous dark incubation was also recovered by adding SA which mimicked light irradiation effect These results suggest that SA is one of essential agents in anthocyanin biosynthesis.

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Elicitation of Seedlings and Cultured Cells for the Production of Capsidiol in Capsicum annum L. (고추 (Capsicum annum L.)식물체 및 배양세포의 Capsidiol 생산 유도)

  • 권순태;정은아;박해영;손건호
    • Korean Journal of Plant Tissue Culture
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    • v.28 no.5
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    • pp.249-254
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    • 2001
  • Effects of ultraviolet stress and elicitors, cellulase and jasmonic acid (JA), for the production of capsidiol, sesquiterpenoid phytoalexin, in seedlings and suspension cultures of pepper (Capsicum annum L. cv, Soobicho) were examined. Extracellular capsidiol in the medium of suspension cultures was absent from control cells, but accumulated in the elicitor treated cells with 0.05 $\mu\textrm{g}$/mL of cellulase or 0.1 $\mu\textrm{g}$/mL JA. Elicited cells gradually decreased their viability and eventually died within 48 hours of elicitor treatment by the toxicity of capsidiol accumulated in the culture medium. Capsidiol production in the leaves of pepper seedlings was markedly increased by the treatment of ultraviolet stress and reached maximum level at 48 hours of irradiation. Infiltration of elicitors, 0.05 $\mu\textrm{g}$/mL cellulase or 1.0 $\mu\textrm{g}$/mL JA, to the surface of leaf or fruit, stimulated the elicitation of the cells which resulted in the production of capsidiol and expansion of pathogene-like lesion around the elicitor treated region.

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