• Title/Summary/Keyword: shoot culture

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Cryopreservation of Capsicum annum var. grossum using encapsulation/dehydration of apices produced in vitro

  • Senarath, Wtpsk;Lee, Kui-Jae;Rehman, S.;Lee, Wang-Hyu
    • Proceedings of the Plant Resources Society of Korea Conference
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    • 2002.11b
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    • pp.53-53
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    • 2002
  • Shoot tips of in vitro propagated plantlets were cryopreserved using encapsulation/dehydration procedures. Shoot tips were excised under filter sterilized antioxidants solution (0.2M phosphate buffer, pH 5.7 supplemented with 5g/1 ascorbic acid and 15g/1 sodium borate). They were drawn up into a sterile 10 $\textrm{cm}^3$disposable pipette and were dropped into the culture medium with 2.5w/v Na-alginate, then into 100mM CaCl$_2$.2$H_2O$. Encapsulated shoot tips were transferred into 10㎤ of liquid culture medium with a range of sucrose concentrations (0.25-1.0M) and were incubated in dark for 24 hours in 18C at 40rpm. Beads were then dehydrated in silica gel for different time intervals (1-24 hours). Then they were freeze dried either rapidly (plunge directly into liquid N2 or in two stages (samples were kept at 20C for 10 minutes, then reduced to 35C at 1C per minute. Then, plunge into liquid $N_2$). The influence of sucrose and silica gel pre-treatment on pre- and post-freeze shoot growth was examined.(중략)

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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.

Establishment of Cell Suspension Cultures and Plant Regeneration in White Dandelion (Taraxacum coreanum NAKAI.)

  • Sun, Yan-Lin;Kim, Jae-Hak;Hong, Soon-Kwan
    • Korean Journal of Plant Resources
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    • v.24 no.3
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    • pp.280-285
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    • 2011
  • In this study, we established a novel somatic embryogenesis and plant regeneration system through cell suspension culture of white dandelion (Taraxacum coreanum NAKAI.). Embryogenic calli could be initiated from leaf and root explants of sterile seedlings on solid Murashige and Skoog (MS) medium supplemented with 1.0 mg/L 2,4-dichlorophenoxyacetic acid (2,4-D) after 3-week cultures. To proliferate embryogenic calli rapidly, cell suspension culture was performed with transferred to liquid MS medium with various combinations of plant growth regulators (PGRs) including 2,4-D, ${\alpha}$-naphthalene acetic acid (NAA), indole-3-acetic acid (IAA), $N^6$-benzylamino purine (BAP), thidiazuron (TDZ), and kinetin. During suspension cultures, embryogenic calli not only greatly proliferated, but shoot organogenesis also simultaneously occurred from the surface of somatic embryos. Among them, TDZ at lower concentration, 0.1 mg/L produced the highest efficiency of somatic embryo formation and shoot organogenesis. Rooting of embryogenic calli with adventitious shoots was done on solid MS medium containing 0.1 mg/L NAA and 0.3% activated carbon. Nearly 80% of embryogenic calli with shoot organogenesis could be rooted normal. Well-rooted plantlets were transferred into pots under a greenhouse condition, and plants derived from this system appeared phenotypically normal.

Plant Regeneration and Bulblet Formation of Allium wakegi Araki

  • Song, Won-seob;Yang, Deok-Chun;Yoon, Jae-Ho;Ryu, Sang-Hyun
    • Plant Resources
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    • v.7 no.1
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    • pp.7-14
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    • 2004
  • Allium wakegi was cultured shoot tip in the condition of light culture. The Allium wakegi added plant growth regulator was observed of plant regeneration and bulblet formation. Callus Induction and growing rate was the best of 78% when added alone 2,4-D 0.5mg/L. In the formation of shoot, its regeneration rate was 96% when added BA 0.5mg/L in the light culture condition. When BA 0.5mg/L and NAA 0.5mg/L mixed and BA 0.5 mg/L and NAA 1.0mg/L mixed, the rates were 99% and 97% respectively, and these conditions were suitable for forming shoot. In the formation of roots, when added NAA 2.0mg/L in the light culture condition, the regeneration rate was 90.6 % and the roots were abnormal. When added NAA 1.0mg/L, the rate was 82 % and the highest. In the formation of bulbs, when BA 05mg/L and NAA 1.0mg/L mixed, the root generantion and its size in the bulbs was the best compare to other treatment experiments.

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STUDIES ON THE TISSUE CULTURE OF PANAX GINSENG

  • Harn C
    • Proceedings of the Ginseng society Conference
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    • 1974.09a
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    • pp.9-22
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    • 1974
  • Unlike the tissue culture in animals and human being, in higher plants various parts of the plant are cultured for varied purposes, and they are named variously depending on which parts are used as explants or what purposes they are cultured for. Followings are some of the names of culture used frequently: organ culture, tissue culture, callus culture, single cell culture, meristem culture, mericlone culture, ovary culture, ovule culture, embryo culture, endosperm culture, anther culture, pollen culture, protoplast culture, etc.. As the names of the culture indicate, in some kinds of culture the explants used for culture are actually not tissues, but organs, single cells, or protoplasts. It seems, however, convenient to call all of the above-mentioned cultures grossly as tissue culture. Several kinds of tissue culture were attempted using Panax ginseng as material and some of the results were summarized below. 1. Callus culture After dormancy of the sed was broken, whole embryo or parts (hypocotyl, cotyledon and epicotyl) of partly grown embryo were cultured in the media supplemented with growth regulators. Rapid swelling occurred in a few weeks, but most of the swelling was observed only in the basal part of epicotyl, changes in the other parts of embryo appearing in much later stages. The swelling or increase in size, however, was resulted not from the divisions of cells, but from the mere expansion of cell. Real calli were formed about two months after inoculation of explants. Callus tissues developed from cortex, pith, and vascular bundle in the cases of hypo- and epicotyl, from mesophyl tissue in the case of cotyledon. Shoots developed more easily from cotyledons regardless of whether they are detached from or attached to the embryo proper. 2. Culture in the Knudson C medium When cotyledons, detached from or attached to the embryo proper, were cultured in the growth regulator-free Knudson C medium comprision only several kinds of mineral compounds and sucrose, shoot primordium or callus developed profusely and finally plantlets were produced directly from shoot primordium or indirectly through callus. In this medium epidermal cells as well as mesophyl cells of the cotyledon became meristematic and divided, changing into multinucleate cells or multicellular bodies, developing eventually into either shoot primordia or calli. 3. Anther culture Anthers were cultured in the media supplemented with various growth regulators applied singly or in combinations. Callus was formed mostly in the connective tissue of anther. Cells of anther wall layers changed in appearance, but no division occurred. Microspores of all stages in development were not changed, ruling out the possibility that microspore-originated callus might be formed. 4. Isolation of protoplast Protoplasts were isolated from young root, leaf, and epicotyl, using 0.7M D-mannitols as osmoticum and using macerozyme and cellulase respectively for maceration and digestion of the cell wall. Production in large number of naked intact protoplast was rather difficult as compared with other plant species. Fusion of protoplasts occurred infrequently mainly due to the fewer number of naked protoplasts in the solution.

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An Efficient Plant Regeneration System for Sorghum bicolor - a Valuable Major Cereal Crop

  • Baskaran P.;Jayabalan N.
    • Journal of Plant Biotechnology
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    • v.7 no.4
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    • pp.247-257
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    • 2005
  • An efficient, rapid and large-scale in vitro clonal propagation of agronomically important Indian cereal crop genotypes (NSH27 & K5) of Sorghum bicolor (L.) Moench. by enhanced shoot proliferation in shoot tip segments was designed. MS medium fortified with plant growth regulators and coconut water markedly influenced in vitro propagation of Sorghum bicolor. In vitro plantlet production system has been investigated on Murashige and Skoog (MS) medium with the synergistic combination of 6-benzyladenine ($22.2\;{\mu}M$), kinetin ($4.6\;{\mu}M$), adenine sulphate ($2.8\;{\mu}M$), 5% coconut water and 3% sucrose which promoted the maximum number of shoots as well as beneficial shoot length. Subculturing of shoot tip segments on a similar medium enabled continuous production of more than 100 healthy shoots with similar frequency. When the healthy shoot clumps were cultured on MS medium fortified with 6-benzyladenine ($22.2\;{\mu}M$), kinetin ($4.6\;{\mu}M$), adenine sulphate ($2.8\;{\mu}M$), ${\alpha}$-naphthaleneacetic acid ($2.7\;{\mu}M$), ascorbic acid ($30.0\;{\mu}M$) and 5% coconut water, a rapid production of axillary and adventitious buds was developed after 8 wk culture. More than 300 shoots were produced 10 wk after culture. Rooting was highest (100%) on half strength MS medium containing 22.8 mM IAA. Micropropagated plants established in garden soil, farmyard soil and sand (2:1:1) were uniform and identical to the donor plant with respect to growth characteristics. These plants grew normally without showing any traits.

Micropropagation from Corm Apical Meristems Culture of Freesia refrecta Hybrida (정단 분열 조직배양에 의한 후리지아의 미세번식)

  • 고정애;김명준;김현순;이진재;김영숙
    • Korean Journal of Plant Resources
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    • v.16 no.1
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    • pp.34-39
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    • 2003
  • Corm apical meristem cultures of thirteen glasshouse freesia cultivars were tested to investigate the possibility of micropropagation using MS basal medium supplemented with 2,4-D, NAA(0.1, 0.5, 1.0mg/L, respectively) and BA (0.5∼2.0mg/L). The majority of the tested cultivars could be induced callus and shoot buds in all culture condition. The combinations of NAA and BA appeared superior to that of 2,4-D and BA depending on cultivars for callus induction and shoot formation. Among the cultivars, 'Golden Yellow' showed the highest regeneration capacity on MS media with 0.5mg/L NAA and 1.0 mg/L BA. The highest percentage of regeneration and the greatest number of shoot from calli were obtained through successive subculture on MS medium supplimented with 0.5mg/L BA. In that condition, more than 60 % shoot regeneration and average of 25.1 shoots per explant was achieved. Transformed shoots on half-strength MS medium without plant growth regulators rooted easily.

Correlation between in vitro Plant Regeneration of Apple Cotyledon and Light Condition, Polarity of Explant (사과 자엽의 기내 식물체 재분화와 배양 광조건 및 절편체 극성과의 관계)

  • 안현주;예병우;임용표;신용억
    • Korean Journal of Plant Tissue Culture
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    • v.27 no.1
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    • pp.19-23
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    • 2000
  • The experiment was carried out to investigate the influence of light condition and polarity of explant on adventitious shoot formation from cotyledon of apple in vitro The treatment of light culture after 10days dark treatment showed effective result than other treatments on the rate of adventitious shoot formation on light intensity treatments and also the treatment of blue light treatment after 4days dark treatment showed more effective result than other treatments in light quality treatments. The polarity of explant influence on adventitious shoot formation. Adventitious multiple shoot formation occured at the proximal end of an excised cotyledon. In other words. shoot organogenesis occured at the proximal cut surfaces of both proximal and distal explants rather than at the distal cut surface of proximal explants.

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Cytohistological Study of Development of Callus and Adventitious Shoots from Cultured Stem of Vigna radiata (녹두 줄기 조직배양에서 캘러스와 부정아 형성에 관한 세포조직학적 연구)

  • Park, Jong-Bum
    • Journal of Life Science
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    • v.16 no.7 s.80
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    • pp.1141-1147
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    • 2006
  • This study was carried out to establish a reproducible culture system for callus formation and adventitious shoot development from young stem segments of Vigna radinta, and histological work for orgin of callus tissue and adventitious shoot. Induction of callus from young stem explants of Vigna radiata was very effective on MS inorganic salts supplemented with 0.5 mg/L 2,4-D and 1.0 mg/L kinetin. For the adventitious shoot regeneration from the callus tissues, the hormone combination of 0.75 mg/L NAA, 1.5 mg/L kinetin and MS salts resulted in about 21% efficiency. Histological examination showed that callus tissues originated from out-growths by callus cambium rings with do novo meristematic activities, which were localized at the outside of the vascular cambium. Adventitious shoots were developed from shoot apical meristem originated from the surface of callus masses. The shoot apical meristem produced leaf primordium, which then became leaf.

In vitro propagation of Spathiphyllum floribundum cv. Cupid (Spathiphyllum floribundum cv. Cupid의 기내번식)

  • 한봉희;예병우;유희주;신지수
    • Korean Journal of Plant Tissue Culture
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    • v.28 no.4
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    • pp.209-213
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    • 2001
  • Spathiphyllum floribundum cv. Cupid was micropropagated on LS medium containing various growth regulators. Shoot tips were cultured on media with different concentrations of thidiazuron (TDZ) or banzyladenine (BA). The medium containing 2.0 mg/L BA was most effective on the shoot multiplication and the formation of adventitious multi-shoot cluster from shoot tips among the different concentrations of TDZ and BA. The combined treatments of BA and paclobutrazol (PBZ) were stimulated the shoot multiplication and the formation of adventitious mu]ti-shoot clusters from shoot tips compared to the ones of BA alone. The adventitious multi-shoot clusters were culled with 5∼7 mm in size and cultured on media with BA or thidiazuron (TDZ). Shoot multiplication and shoot cluster proliferation from the shoot clusters were very favorable on media supplemented with 2.0∼3.0 mg/L BA. The shoots grew and rooted out vigorously on media with 2.0∼3.0 mg/L IBA. The rooted plantlets were acclimatized in the soil mixed with a rate of vermiculite 1 and perlite 1, and survived more than 95% in greenhouse.

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