• Title/Summary/Keyword: regenerated shoots

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In vitro regeneration from cotyledon explants in figleaf gourd (Cucurbita ficifolia Bouch$\'{e}$), a rootstock for Cucurbitaceae

  • Kim, Kyung-Min;Kim, Chang-Kil;Han, Jeung-Sul
    • Plant Biotechnology Reports
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    • v.4 no.2
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    • pp.101-107
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    • 2010
  • An efficient plant regeneration system has been developed for figleaf gourd (Cucurbita ficifolia Bouch$\'{e}$), which is exclusively used as a rootstock for cucumber. The protocol is based on results obtained from a series of culture experiments involving different parts of the cotyledons and various media. The culture of cotyledon explants was critical for the enhancement of shoot regeneration frequency. The lower parts of the cotyledon excised at the plumule base were found to display a markedly enhanced production of adventitious shoots compared to other cotyledon regions. Culture in silver nitrate-supplemented Murashige and Skoog (MS) medium was not beneficial for shoot regeneration and suppressed root regeneration. Efficient shoot regeneration was obtained on MS medium containing 1.0 $mg\;l^{-1}$ zeatin and 0.1 $mg\;l^{-1}$ indole-3-acetic acid. Regenerated shoots successfully elongated and rooted in medium containing 0.1 $mg\;l^{-1}$ 1-naphthalene-acetic acid after 10-15 days of subculturing. The plantlets were satisfactorily acclimatized in a greenhouse and grew into normal plants without any morphological alterations.

Shoot Regeneration from Leaf Explants of Digitalis purpurea L. (디기탈리스의 잎 절편으로부터 신초의 재분화)

  • Hwang, Sung-Jin;Lee, Hye-Jung;Hwang, Baik
    • Korean Journal of Medicinal Crop Science
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    • v.11 no.5
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    • pp.397-401
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    • 2003
  • Digitalis purpurea L. is a medicinal herb and have been used to congestive heart failure, mycocardial infarction, edema, angina etc. A protocol has been developed for in vitro propagation of adventitious shoot buds directly from leaf segments of D. purpurea Leaf explants of D. purpurea directly formed shoot buds when cultured on a MS medium supplemented with $2\;mg/l$ BA and $0.1\;mg/l$ IAA for 5 weeks. Adventitious shoots were multiplied by subculturing on the $B_5$ medium and shoot elongation was developed by subculturing on the WPM medium. Root formation from the shoot regenerated was achieved on MS basal medium containing 1 mg/ IBA.

Microprogation And Environment Conditions Affecting On Growth Of In Vitro And Ex Vitro Of A. Formosanus Hay

  • Ket, Nguyen-Van;Paek, Kee-Yoeup
    • Proceedings of the Plant Resources Society of Korea Conference
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    • 2002.11b
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    • pp.6-7
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    • 2002
  • The goal of this research was to develop the effectiveness of in vitro culture method for A. formosanus and study the environment in vitro conditions affecting on growth. The first series of experiments were examined to investigate the response of three different basal media, MS (Murashige and Skoog, 1962), Knudson (KC; Knudson, 1946) and modified hyponex on growth and multiplication during in vitro culture. Multiple shoot proliferation was induced in shoot tip explants on Hyponex (H3) media supplemented with BA (1 mg1$^{-1}$) or TDZ (1-2 mg1$^{-1}$). Addition of activated charcoal (1%) to the TDZ containing medium promoted rapid shoot tip proliferation (11.1 shoots per explant) but the same medium had an opposite effect resulting in poor proliferation in the nodal explants. However, the regenerated shoots had slow growth rate and failed to elongate. This problem was overcome by transferring the shoot clumps to a hormone free H3 media supplemented with 2% sucrose and 0.5% activated charcoal. Using bioreactor culture for scaling up was also shown the best way for multiple shoot induction and growth of this plant.(중략)

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Plant Regeneration Derived from Leaf Disk Cultures in Purple Sweetpotato (자색고구마의 잎 조직배양을 통한 식물체 재생)

  • Park, Hyae-Jeong;Ahn, Young-Sup;Jeong, Byeong-Choon;Park, Hyeon-Yong
    • Journal of Plant Biotechnology
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    • v.30 no.3
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    • pp.245-249
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    • 2003
  • This study was carried out to establish a regeneration system from leaf explant of purple sweetpotato(Ipomoea batatas L.) The optimal concentrations of plant growth regulators for callus induction and shoot formation were determined. The optimal combination for callus formation was 1$\mu$M 2,4-D 5$\mu$M BM, and highest yield of embryogenic calli were observed on Murashige and Skoog basal medium containing 0.5$\mu$M 2,4-D under light condition after 4weeks of culture. Embryogenec callus was subcultured on medium supplemented with 5$\mu$M ABA for 4 days. Subsequently, regeneration of adventitious shoots occurred when these embryogenic calli were transferred onto medium with 3∼6$\mu$M gibberellic acid. Regenerated shoots were developed into normal plantlets.

Plant Regeneration from Callus and Adventitious Root Segments of Pulsatilla Koreana Nakai

  • Jung, Su-Jin;Jeong, Jae-Hun;Yoon, Eui-Soo;Choi, Yong-Eui
    • Journal of Plant Biotechnology
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    • v.34 no.2
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    • pp.153-159
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    • 2007
  • Plant regeneration of Pulsatilla koreana was achieved via adventitious shoot formation indirectly from callus and directly from adventitious root segments. For the callus induction from leaf or petiole explants, combination of 2,4- dichlorophenoxyaceticacid (2,4-D) with $2.22\;{\mu}M$ 6-benzyladenine (BA) was effective. Adventitious shoot induction from callus was enhanced by the combined treatment with $0.1\;{\mu}M$ polyvinylpyrrolidone (PVP) compared to cytokinin treatment alone. Adventitious roots were induced from the petiole segments on 1/2 MS medium with $4.93\;{\mu}M$ IBA. High frequency direct adventitious shoot formation from the segments of adventitious roots was achieved on medium with $4.92\;{\mu}M$ 2-isopentenyladenine (2-ip). Elongated shoots were rooted on half-strength MS medium containing $5.71\;{\mu}M$ indole acetic acid (IAA). Regenerated plantlets with well-developed shoots and roots were successfully transferred to soil. This in vitro propagation protocol might be useful for mass propagation as well as conservation of this plant.

MdMADS2 - transgenic chrysanthemum (Dendranthema grandiflorum (Ramat.) Kitamura) showing the reduction of the days to flowering

  • Han, Bong-Hee;Lee, Su-Young;Choi, Seong-Youl
    • Journal of Plant Biotechnology
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    • v.36 no.4
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    • pp.366-372
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    • 2009
  • This study was conducted to develop new lines expressing the characteristic of early flowering by introducing MdMADS2 gene in chrysanthemum (Dendranthema grandiflorum (Ramat.) Kitamura) ‘Zinba'. Transformation of chrysanthemum was conducted by Agrobacterium tumefaciens LBA4404 harboring the binary vector containing MdMADS2 controlled by double CaMV 35S promoters. Ninety three shoots were regenerated from 1,463 leaf segment explants cultured on the first selection medium (MS basal salts + 1.0 mg/L BA + 0.5 mg/L IAA + 10 mg/L kanamycin + 400 mg/L cefotaxime, pH 5.8) after co-cultivation, and 20 out of the 93 shoots rooted on the second selection medium containing 20 mg/L kanamycin and 400 mg/L cefotaxime. Many escapes (98.6%) were removed on the selection stage for rooting. Nineteen lines were confirmed as transgenic plant with transgene by PCR analysis. Six transgenic plants flowered 2-11 days earlier than non-transgenic plant without big change of phenotype, and especially, 3 (Mo-7, Mo-11, Mo-17) out of 6 transgenic lines showed a significant reduction in days to flowering compared to non-transgenic plant. Introduction and expression of MdMADS2 gene in them were confirmed by Southern and real-time PCR analyses, respectively.

Effect of Thidiazuron on Regeneration from Long-Term Cultured Callus of Solanum spp. (장기간 계대배양된 야생약초 까마중종 캘러스로부터 식물체 재분화)

  • Yu, Chang-Yeon;Chae, Young-Am;Ahn, Sang-Deuk;Cho, Dong-Ha
    • Korean Journal of Medicinal Crop Science
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    • v.2 no.1
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    • pp.38-43
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    • 1994
  • The effect of thidiazuron on callus growth and shoot regeneration of Solanumspecies was very positive. Increasing concentrations of thidiazuron stimulated callus growth of Solanum ptycanthum and Solanum nigrum. Shoot regeneration of S. ptycanthum and S. nigrum was greater on medium with thidiazuron than that with IAA and BA. Thidiazuron at $0.5{\mu}M$ or above increased the number of shoots regenerated from S. ptycanthum calli compared to the IAA with BA. High concentrations of thidiazuron $( >I{\mu}M)$ increased the number of shoots than BA or low levels of thidiazuron and IAA or BA The addition of IAA to thidiazuron media reduced S. ptycanthum shoot formation.

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Improved Micropropagation of Root Chicory, Cichorium intybus L. var. sativus.

  • Lim, Jung-Dae;Yang, Deok-Chun;Lee, Hyeon-Yong;Kim, Jong-Dai;Lee, Jin-Ha;Sung, Eun-Soo;Yu, Chang-Yeon
    • Plant Resources
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    • v.7 no.1
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    • pp.60-64
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    • 2004
  • The establishment of an efficient protocol for plant regeneration and micropropagation from leaf explant cultures of Chicory, Cichorium intybus L. var. sativus. is reported. Callus formation rate appeared 100% from explant in all growth regulators, but calli formed in the prensence of naphthaleneacetic acid (NAA) were appeared very compact and non-embryogenic state. The regenerated shoots were obtained from leaf explant cultures on solid MS medium containing different concentrations of cytokinins and auxin. The highest number of shoots (5.7) per explant and shoot growth (2.8cm) was obtained on MS medium containing 0.1 mg BAP L$^{-1}$ and 0.1 mg NAA L$^{-1}$ . Indole acetic acid was the most suitable auxin for root formation among three auxins tested. 2,4-D had no effect on shoot and root formation.

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Multiple Shoot Induction from Radicle-derived Callus and in Vitro Propagation of Silene Acaulis Subsp. Arctics (극지식물 Silene acaulis subsp. arctica의 유근 유래 캘러스로부터 다신초 유도와 기내 증식)

  • Seo, Hyo-Won;Yi, Jung-Yoon;Park, Young-Eun;Kang, Sung-Ho;Chung, Ho-Sung;Kim, Ji-Hee
    • Journal of Plant Biotechnology
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    • v.33 no.4
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    • pp.303-307
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    • 2006
  • We describe here an efficient in vitro propagation method of Silene acaulis subsp. arctica (Caryophyllaceae), one of the higher arctic angiosperms, through the multiple shoot regeneration after callus induction from the radicle. The seeds of S. acaulis subsp. arctica collected from Svalbard, the Norwegian Arctic, were germinated and calli were induced from the radicle on solid MS media supplemented with 0.25mg/L 2,4-D and 1mg/L $GA_3$ at both $10{\pm}1^{\circ}C\;and\;23{\pm}1^{\circ}C$ Two weeks after callus induction, the multiple shoots were efficiently regenerated on the MS media supplemented with 0.25 g/L BA and 0.05mg/L HPh. The total biomass increment of regenerated shoots increased most efficiently of S. acaulis subsp. afctica was showed the maximum efficiency in at $23{\pm}1^{\circ}C$ on 1/2 MS salt strength. The multiple regenerated plantlets of S. acaulis subsp. arctics were grown to normal plants on soil.

Perilla transformation using selection markers containing antibiotics and basta (항생제와 제초제 이중 선발 마커를 이용한 들깨 형질전환)

  • Kim, Kyung-Hwan;Lee, Jung-Eun;Ha, Sun-Hwa;Hahn, Bum-Soo;Park, Jong-Sug;Lee, Myung-Hee;Jung, Chan-Sik;Kim, Yong-Hwan
    • Journal of Plant Biotechnology
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    • v.35 no.4
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    • pp.299-306
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    • 2008
  • A modified method of Agrobacterium-mediated perilla transformation was developed using two selection markers of an antibiotics (either hpt or nptll) and an herbicidal (bar) gene. Perilla hypocotyl explants were cocultured with Agrobacterium tumefaciens EHA 105 strain harboring plasmid vector (either pMOG6-Bar or pCK-Bar) for three days, respectively. Primary shoots were selected with antibiotics of hygromycin (15 mg/L) or kanamycin (125 mg/L) and regenerated shoots were further selected with herbicide phosphinothricin (ppt,1.2 mg/L) to obtain authentic transformants. Roots were induced for the regenerated shoots on the MS medium without hormone and 80 putative transgenic plants were obtained. Transgene integration into perilla genome was confirmed by Southern blot and their expression was analyzed by Northern blot. T1 perilla seeds drived from To plants were tested 0.3% basta spray for identification of stable gene delivery to next generation.