• Title/Summary/Keyword: Explants

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In vitro shoot proliferation of Alnus japonica (Thunberg) Steudel

  • Kang, Ho-duck;Lee, Min-Soon
    • Plant Resources
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    • v.7 no.1
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    • pp.1-6
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    • 2004
  • In vitro proliferation system was achieved by using nodal segment excised from greenhouse grown juvenile stock plants of Alnus japonica. Stem explants were cultured on MS medium supplemented with different plant growth regulators of cytokinin and/or their combinations. The most effective cytokinin source was the combination of zeatin 2.0 mg/L and TDZ 0.05 mg/L producing the average number of shoots (16.8 $\pm$ 3.6). In addition, healthy roots were formed after small clumps of shoots were transferred to half strength of MS medium containing IBA 0.02 mg/L with optimal rooting capacity. Soil acclimatization was successfully conducted in cell tray containing artificially mixed soil with 92 % survival rate.

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Optimization of Shoot Induction, Histological Study and Genetic Stability of in vitro Cultured Pisum sativum cv. 'Sparkle'

  • Kantayos, Vipada;Bae, Chang-Hyu
    • Korean Journal of Plant Resources
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    • v.32 no.1
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    • pp.19-28
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    • 2019
  • An efficient shoot regeneration condition for pea cv. 'Sparkle' was developed by using optimum explant, plant growth regulator concentrations, and pretreatment of BA onto explant. The average shoot number per explant showed the highest on two kinds of shoot induction media (MSB5 media containing 2 mg/L BA and a combination of 2 mg/L BA and 1 mg/L TDZ) when cotyledonary node explants were cultured. Moreover, the pretreatment of explant in 200 mg/L BA solution was found to be more effective in shoot induction than that of non-pretreatment. By histological study, cell division and proto-meristem were formed near the surface of the sub-epidermal and epidermal cell layers of cotyledonary node in earlier than 3 days after culture. The analysis of genetic stability of regenerants by using thirteen ISSR markers showed that in vitro regenerated plants showed polymorphism with 8.3% compared with their mother plants.

In vitro Organogenesis and Propagation of Heloniopsis orientalis Thunb

  • Jong-Woo Nam;Yoon-Kyung Choi;Kyeong-Mi Cho;Young-Been Kim;Sung Hwan Yim;Kee Hwa Bae
    • Journal of Forest and Environmental Science
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    • v.39 no.3
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    • pp.150-154
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    • 2023
  • Heloniopsis orientalis (Liliaceae) is an important horticultural crop native to Korea. Under natural conditions, germination is poor and plant growth is delayed. Therefore, we have developed a vegetative propagation method to produce plants with vigorous growth characteristics via tissue culture. The regenerated shoots were then initiated directly from leaf explants on an MS medium containing either 0.5 to 2.0 mg/L 2,4-D or 1.0 to 3.0 mg/L BA. Healthy plantlets with adventitious roots were formed on the medium supplemented with 1.0 mg/L BA (81%). BA triggered callus initiation without caulogenesis or rhizogenesis, and callus formation was better on the half-strength MS medium than on the full-strength medium. This in vitro propagation protocol will be useful for conservation, as well as for mass propagation.

PRODUCTION OF GINSENOSIDES THROUGH IN VITRO CULTURE OF GINSENG(Panax ginseng C.A. MEYER)

  • Choi K.T.;Ahn I.O.;Park J.C.
    • Proceedings of the Ginseng society Conference
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    • 1993.09a
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    • pp.143-149
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    • 1993
  • Ginseng root explants and calli induced from selected cell lines were cultured on modified Murashige and Skoog's media supplemented with different concentrations of organic or inorganic compounds and plant growth requlators to clarify the effects of chemical composition and plant growth regulators in the medium on the growth of ginseng calli and the production of ginseng saponin. For optimum growth of calli, the concentrations of 2, 4-D and sucrose were the range of 1 to 3 mg/${\ell}$l and 1 to $3\%,$ respectively. And it was clarified that sucrose, nitrogen, phosphate, calcium, magmesian plant growth regulators and their concentrations influcenced the relative biosynthesis of saponin in tissue cultures of Panax ginseng. The patterns of ginsenosides, pharmacologically useful component, were different among the cell lines and contents of ginsenosides were much higher in selected cell lines than in original cell line.

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Adventitious Shoots Regeneration from Seed Explants of Xanthoceras sorbifolium

  • Hyunseok Lee
    • Proceedings of the Plant Resources Society of Korea Conference
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    • 2020.12a
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    • pp.58-58
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    • 2020
  • Xanthoceras sorbifolium Bunge (yellowhorn) is a woody tree in the soapberry family, Sapindaceae, native to northern China. This species has been identified as a major woody bioenergy plant for bio-diesel production because of high oil content in seed. But the flowers do not bear fruit well while the many flowers blooming. This study was performed to regenerate in vitro plantlet using adventitious shoot formation. To establish the protocol of plant regeneration, adventitious shoots formation rate in the culture of cotyledon of immature zygotic embryos was 68.6% in 1/2 MS medium with 0.5 mg l-1 BA and 3% sucrose (w/v). In the culture of cotyledons of mature zygotic embryos, induction of adventitious shoots was needed to contain high sucrose in pre-culture medium and the frequency of shoot induction was 64.4%. Multiple shoots were induced in 0.5 mg l-1 TDZ, and rooting of shoot was induced 4.0 mg l-1 IBA. Flow cytometry analysis revealed that all the regenerated plantlets were diploid.

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Development of a Protocol for Somatic Embryogenesis of Cnidium officinale M akino

  • Hui Yeong Jeong;Ji Ah Kim
    • Proceedings of the Plant Resources Society of Korea Conference
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    • 2021.04a
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    • pp.51-51
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    • 2021
  • This study was conducted to develop a somatic embryogenesis protocol for the Cnidium officinale Makino difficult to seed propagation. The immature flowers were used as explants. The concentration of a 2,4-D 1.0mg/L was found to be optimal concentration for induction of embryogenic callus and somatic embryos. Addition of 0.3mg/L, 0.5mg/L and 1.0mg/L to the embryo germination medium promoted somatic embryo germination. Among four concentrations, GA3 1.0mg/L were superior to others. Shoots were transferred to hormone-free MS medium after 2 months of culture in the dark. We obtained an optimized protocol for the regeneration of C. officinale.

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Autotrophic Growth of Dendranthema grandiflorum R. 'Bongwhang' Plantlets In Vitro as Affected by PPF, Air Exchange Rate and $\textrm{CO}_2$ Concentration (봉황국화의 자가영양배양시 광도, 환기횟수 및 $\textrm{CO}_2$농도가 기내생육에 미치는 영향)

  • 김영회;정병룡
    • Journal of Bio-Environment Control
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    • v.8 no.1
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    • pp.56-66
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    • 1999
  • Growth of Dendranthema grandiflorum R. ‘Bongwhang’plantlets, as affected by three levels of photosynthetic photon flux (PPF), 70, 150 and 220 $\mu$mol. $m^{-2}$ . $s^{-1}$ , three levels of C $O_{2}$ concentration, 400-500 (ambient), 1000 and 2000 $\mu$mol.mo $l^{-1}$ , and two levels of number of air exchanges per hour (NAEH), 0.1 $h^{-1}$ and 2.8 $h^{-l}$, was studied. Explants were obtained from photomixotrophically-micropropagated plantlets. Four explants were planted in each 3.7$\times$10$^{-4}$ $m^{3}$ polycarbonate box containing MS medium supplemented with 1.25 meq. $L^{-1}$ $H_{2}$P $O_{4}$$^{[-10]}$ and no added sugar. Explants were cultured under cool-white fluorescent lamps (16 h. $d^{-1}$ ), at 25$\pm$1$^{\circ}C$ temperature, and 70-80% relative humidity. In treatments of 2.8 $h^{-1}$ NAEH, a 10 mm round hole made on the vessel cap was sealed with a microporous filter For higher C $O_{2}$ concentrations in the culture room, C $O_{2}$ gas was provided from a tank of liquefied C $O_{2}$. Fresh and dry weights, height, length of the longest roots, number of leaves, and leaf area significantly increased with increasing PPF and especially, with increasing C $O_{2}$ concentration. Growth was enhanced with increased number of air exchanges per hour (2.8 $h^{-1}$ ). Overall, treatment of 220$\mu$mol. $m^{-2}$ . $s^{-1}$ PPF combined with 2000$\mu$mol.mo $l^{-1}$ C $O_{2}$ and 2.8 $h^{-1}$ NAEH gave the most vigorous growth of Dendranthema grandiflorum R. ‘Bongwhang’ plantlets in vitro.o.

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Introduction and Expression of PAP gene using Agrobacterium in Scrophularia buergeriana Miquel (Agrobacterium을 이용한 PAP 유전자의 현삼으로 도입 및 형질발현)

  • Yu, Chang-Yeon;Seong, Eun-Soo;Lim, Jung-Dae;Huang, Shan-Ai;Chae, Young-Am
    • Korean Journal of Medicinal Crop Science
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    • v.9 no.2
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    • pp.156-165
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    • 2001
  • Exogeneous application of pokeweed antiviral protein (PAP), a ribosomal-inacivating protein in the cell wall of Phytolacca americana (pokeweed) protects heterologous plants from viral and fungal infection. A cDNA clone of PAP introduced into Scrophularia buergeriana Miquel by thransformation with Agrobacterium tumefaciences. For plant transformation, explants were precultured on shoot induction medium without kanamycin for 2-5 day, and then they were cocultured with Agrobacterium for 10 minutes. The explants were placed on co culture medium in dark condition, $28^{\circ}C$ for 2days. After explants were washed in MS liquid medium, they were transferred into selection medium including kanamycin 50mg/L (MS salts+1mg/ l BAP+2mg/ l TDZ+0,2mg/ l NAA+MS vitamin+3% sucrose+0.8% agar, pH5.8). From PCR analysis, NPT II band was confirmed in transgenic plant genome and showed resistance against fungi in antifungal activity test. Micro assay to which protein extracted from transgenic line were added, revealed hyphae growth inhibition and no spore germination at high concentration. The characteristics of inhibited hyphae was represented transparent and thin. Expression of PAP in transgenic plants offers the possibility of developing resistance to viral and fungal infection.

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Effect of $CO_2$ Concentration, NAEH and Light Intensity on the Photoautotrophic Growth of Campanula punctata 'Rubriflora' Plantlets In Vitro (자주초롱꽃의 기내 자가영양배양시 $CO_2$농도, 환기횟수 및 광도가 생장에 미치는 영향)

  • Shim, Jae-Nam;Kim, Gyeong-Hee;Jeong, Byoung-Ryong
    • Journal of Bio-Environment Control
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    • v.14 no.4
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    • pp.233-238
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    • 2005
  • Growth of Campanula punctata 'Rubriflora' plantlets, as affected by three levels of photosynthetic photon flux (PPF), 70, 110, and $220{\mu}mol{\cdot}m^{-2}{\cdot}s^{-1}$, two levels of $CO_2$ concentration, 500 and $1,500{\mu}mol{\cdot}m^{-1}$, and two levels of number of air exchanges per hour (NAEH), 0.1 and $2.8 h^{-l}$, was studied. Explants were obtained from photomixotrophically-micropropagated plantlets. Four explants were planted in each $3.7{\times}10^{-4}m^3$ polycarbonate box containing MS basal medium and no added sucrose. Explants were cultured under cool-white fluorescent lamps for $16h{\cdot}d^{-1},\;at\;25\pm1^{\circ}C$ temperature, and $70\~80\%$ relative humidity In treatments of $2.8h^{-1}$ NAEH, a 10mm round hole made on the vessel cap was sealed with a microporous filter. For higher $CO_2$ concentrations in the culture room, $CO_2$ gas was provided from a tank of liquefied $CO_2$. Fresh and dry weights, length of the longest root, and number of leaves significantly increased with increasing PPF and especially $CO_2$ concentration. Length of the longest root, number of leaves, fresh and dry weights, and chlorophyll concentration were enhanced with increased NAEH. However, leaf area was the smallest in the $220{\mu}mol{\cdot}m^{-2}{\cdot}s^{-1}\;PPF\;2.8h^{-1}$ NAEH and especially, $1,500{\mu}mol{\cdot}mol^{-1}\;CO_2$ concentration treatment. Treatment effect became more produced with time. Overall, treatment with $220{\mu}mol{\cdot}m^{-2}{\cdot}s^{-1}\;PPF\;and\;1,500{\mu}mol{\cdot}mol^{-1}\;CO_2$ gave the most vigorous growth.

Callus Initiation and Organ Formation from in vitro Culture of Pelargonium spp. (In Vitro Culture에 의한 Pelargonium spp.의 Callus 유도 및 기관분화에 관하여)

  • Lee, Chun-Ha;Chung, Hae-Joon
    • The Journal of Natural Sciences
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    • v.4
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    • pp.143-159
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    • 1991
  • In order to investigate the micropropagation of Pelargonium, 2 cultivars of P. peltatum 'Pouletta' and P. zonale 'Pinto Red' were cultured in vitro on the MS basal medium supplemented with various concentrations of growth regulators. It attempted to study the induction of callus and the differentiation of organs from leaf disc, petiole segments, stem segments. hypocotyle segments and flower stalk segments. The results are summarized as follows; A. As for the initiation of callus, stem explant was proved to be the most suitable one among various explants of P. zonale 'Pinto Red'. The medium was supplemented with 1.0mg/1 BAP and 1.0mg/1 NAA. As NAA concentration increased, callus formation was enhanced, but higher concentration of NAA inhibited callus fromation. Leaf and hypocotyle explants showed less callus formation than stem and petiole explants. B. In P. zonale 'Pinto Red' petiole culture, the condition of cullus culture such as hormone concentration resulted in affecting shoots differentiation. The best result of shoots formation from the callus reculture were obtained from the combination of 0.5-1.0mg/1 BAP and 0.1-1.0mg/1 NAA when the callus was cultured in 1.0mg/1 BAP and 0.05mg/1 NAA. When the callus was cultured in medium without BAP, the shoot was not differentiated in subculture regardless to BAP and NAA concentration. and only callus was formed. C. Poly-phenol substance was observed in MS medium supplemented without PVP, in which callus was not formed from the leaf of P. peltatum 'Rouletta'. Polyphenol substance was not observed in MS medium supplemented with PVP, in which callus formation was increased. D. The callus formation of P. peltatum 'Rouletta' showed the stem explant being best result. The best result particularly in the stem explant among others. The optimal hormonal concentration was 0.1mg/1 NAA and 5.0mg/1 BAP. The shoot formation was observed at 0.05mg/1 NAA and 1.0mg/1 BAP, 0.1mg/1 NAA and 5.0mg/1 BAP. The shoot was malformed and the tissue recultured turned necrotic.

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