• Title/Summary/Keyword: Explant

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In vitro regeneration of Lycaste aromatica (Graham ex Hook) Lindl. (Orchidaceae) from pseudobulb sections

  • Mata-Rosas, Martin;Baltazar-Garcia, Rosario J.;Moon, Pamela;Hietz, Peter;Luna-Monterrojo, Victor E.
    • Plant Biotechnology Reports
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    • v.4 no.2
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    • pp.157-163
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    • 2010
  • A protocol for in vitro propagation from pseudobulb sections of Lycaste armomatica (Graham ex Hook) Lindl., an ornamental and fragrant orchid, was developed. The effect of four cytokinins: kinetin (K), metatopolin (mT), $N^6$-benzyladenine (BA), and thidiazuron (TDZ), in equimolar concentrations, was investigated. Shoot formation from apical and basal pseudobulb sections was obtained in all treatments. A few medial sections cultured in media supplemented with BA formed protocorm-like bodies. Shoot formation was greater from the basal section than the apical, and mainly occurred in explants cultured in media containing TDZ. The highest average numbers of shoots per explant were achieved from basal sections cultured in media supplemented with TDZ at 4.4, 8.87 and 2.2 ${\mu}M$, forming on average 9.9, 8.6 and 7.3 shoots per explant, respectively. Since the medial pseudobulb section was the worst explant for propagation of L. aromatica, we recommend that pseudobulbs be divided into two sections; the basal half should be cultured in MS medium supplemented with TDZ at 4.4 ${\mu}M$ and the apical half with TDZ at 2.2 ${\mu}M$. Subculturing individual shoots in MS medium without plant growth regulators allows further development and rooting. A survival rate of more than 90% under greenhouse conditions was achieved. This research represents a direct contribution to the conservation and sustainable use of this valuable natural resource.

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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Effect of Culture Method and Medium Composition on Shoot Regeneration from Sporophytes of Cyrtomium caryotideum var. coreanum Nakai. (참쇠고비(Cyrtomium caryotideum) 포자체로부터의 식물체 재생에 미치는 배양방법 및 배지구성물질의 영향)

  • Jeong Jin-A;Lee Cheol-Hee
    • Korean Journal of Plant Resources
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    • v.19 no.2
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    • pp.265-272
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    • 2006
  • This study was conducted to investigate the efficient propagation method of Cyrtomium caryoptideum var. coreanum by sporophyte culture. The influence of origin of explant sources (rhizome, blade, or stipe) and homogenization of culture materials on shoot regeneration were investigated. As a result, only rhizome explant exhibited the organogenic capacity and the shoot regeneration was promoted by homogenization of culture material. Vigorous and excellent growth of multiple shoots was induced on the half-strength of inorganic salts containing MS medium. It was appeared that optimum nitrogen content of shoot regeneration was half-strength of nitrogen containing MS medium (30mM) and optimum sucrose concentration was 1%. Addition of $NaH_2PO_4$ to culture medium generally enhanced shoot multiplication and promoted growth of the regenerants. The organogenic capacity of homogenized rhizomes was especially promoted on medium supplemented with $5{\mu}M$ kinetin plus $5{\mu}M$ IBA. The incorporation of $0.1\sim0.2%$ activated charcoal on medium supplemented with growth regulators prevented the formation of multiple bud primordia - nodule-like bud clusters and improved the normal morphogenesis of sporophytes.

In Vitro Plant Regeneration from Stolen Node Explant in Eremochloa Ophiuroides (Munro) Hack

  • Barampuram, Shyamkumar;Chung, Byung-Yeoup;Lee, Seung-Sik;An, Byung-Chull;Kim, Jae-Sung
    • Journal of Plant Biotechnology
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    • v.34 no.2
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    • pp.161-166
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    • 2007
  • In vitro shoot regeneration and multiple shoot induction has been obtained from the stolen node explants in Eremochloa ophiuroides (Munro) Hack. The highest number of shoots ($10.66{\pm}0.21$) was observed from initial explants after one month culture duration on Murashige and Skoog (MS) medium containing 6-benzyladenine (BA: 0.5 mg/l). First generation shoot was excised and sub-cultured on the same fresh media for further multiplication of shoots. An enhanced number of second round shoots ($15.33{\pm}0.21$) was obtained compared to the initial culture media containing BA (0.5 mg/l). The number of shoots/stolon node was higher among all the concentrations of BA than kinetin (KN). In vitro regenerated shoots were successfully rooted in the phytohormone free MS medium. Plantlets generated with roots were transferred to pots containing compound mixture of soil and kept in green house conditions. Acclimatized plants showed 100% survival rate with normal morphology in green house conditions. The present study demonstrates the effect of explant and different plant growth regulators towards in vitro response in E. ophiuroides. Moreover, the study reveals the effect of cytokinin on induction of shoot number per stolen node explant in E. ophiuroides.

Effects of Light, Temperature, and Sucrose on Plant Regeneration from the Flower Organ Explant in Iris ensata (꽃창포 화기조직 절편체 배양으로부터 식물체 분화에 미치는 광.온도.당의 영향)

  • Yoon, In-Kyung;Koh, Jae-Chul
    • Journal of Plant Biotechnology
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    • v.30 no.1
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    • pp.41-45
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    • 2003
  • A study was under taken to investigate the appropriate explant sources of flower organ and suitable cultural conditions such as light, temperature, and sucrose in plant regeneration of Iris ensata culture. Explants of perianth, ovary, pedicel, and peduncle of Iris ensata were cultured at different daylength (0, 8, 16, 24 hour), different temperatures (10, 15, 25, 3$0^{\circ}C$), and sucrose concentrations (1, 3, 6, 9%) on MS medium. Formation of adventitious roots from explants of Iris ensata was effective in the dark, while that of adventitous shoots was effective in the light. The optimum daylength for young plant regeneration was 16 hours. The optimum temperature for shoot formation of Iris ensata explants was $25^{\circ}C$ but the formation at 10 and 15$^{\circ}C$ was ineffective. Especi-ally, perianth and ovary was effective in shoot formation from flower organ expants. T-he optimum concentration of sucrose for shoots and roots formation of Iris ensata explants was 3 and 6%, respectively.

Somatic embryogenesis from the axillary meristems of peanut (Arachis hypogaea L.)

  • Singh, Shweta;Hazra, Sulekha
    • Plant Biotechnology Reports
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    • v.3 no.4
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    • pp.333-340
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    • 2009
  • Developmental anomalies in the plumule meristem of peanut (Arachis hypogaea L.) somatic embryos resulted in poor shoot differentiation and reduced plant recovery. Existing meristems with caulogenic potential have never been tested for embryogenesis in peanut. The present experiment was designed to test the mature zygotic embryo axis derived plumule with three meristems for somatic embryogenesis. Embryogenic masses and embryos developed from the caulogenic meristems in the axils. Exposure of 2 weeks in primary medium with $90.5{\mu}M$ 2,4-D suppressed the shoot tip differentiation temporarily which then regained the ability to form the shoot on withdrawal of 2,4-D. Exposure of 4 weeks in primary medium with $90.5{\mu}M$ 2,4-D suppressed the shoot tip differentiation irreversibly. No shoot formation was noted from the tips in any of the cultures which were in secondary medium with $13.6{\mu}M$ 2,4-D. Development of somatic embryos directly from axillary meristems was confirmed histologically. Conversion frequency of these embryos was 11%. Thus, in this report, we describe a method to obtain somatic embryos from the determined organogenic buds of the axillary meristem, by culturing the nodal explant vertically on embryo induction medium. It also displays the possibility of obtaining both embryogenic and organogenic potential in two parts of the same explant simultaneously. The possibility of extending this approach for genetic transformation in in vivo system through direct DNA delivery or Agrobacterium injection in meristems can also be explored. Using Agrobacterium rhizogenes, we have demonstrated the possibility of gene transfer in the axillary meristems of seed-derived plumule explant.

Effect of silver nitrate(AgNO3) and polyamines on shoot organogenesis and plant regeneration of Lycopersicon esculentum cultivar, Micro-Tom (질산은과 polyamines이 미니토마토, Micro-Tom 신초 기관발생과 식물체 재분화에 미치는 영향)

  • Kim, Yong-Ho;Park, Cheol-Ho;Park, Sang-Un
    • Journal of Plant Biotechnology
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    • v.29 no.1
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    • pp.25-29
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    • 2002
  • The study was carried out to establish an efficient protocol for shoot organogenesis and plant regeneration from stem explant cultures of Lycopersicon esculentum cv. Micro-Tom. The regenerated shoots obtained from stem explant cultures on solid MS medium containing the different concentrations of BAP. The highest number of shoots (5.3) per explant and shoot growth (0.7 cm) was obtained on MS medium containing 4.0 mg/L BA. The additions of AgNO$_3$ and putrescine substantially improved the shoot regeneration frequency, at the optimal concentration of 7 mg/L and 50 mg/L respectively. The regenerated shoots (about 1 cm) were normal and could be easily rooted with 0.1 mg/L IBA treatment. The rooted plants were hardened and transferred to vermiculite with a 92% survival rate where they grew normally.

In vitro micropropagation of water hyacinth (Eichhornia crassipes)

  • Suh, Eun-Jung;Park, Byoung-Mo;Han, Bong-Hee
    • Journal of Plant Biotechnology
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    • v.37 no.4
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    • pp.505-510
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    • 2010
  • This study was conducted to refine a micropropagation method of water hyacinth (Eichhornia crassipes) in vitro. When young shoots were cultured on media with various concentrations of BA or TDZ alone, LS medium containing $5.0\;mgl^{-1}$ BA was found favorable for shoot proliferation from young shoots with a mean of 4.2 shoots. Using BA together with IAA, more shoots were obtained on LS medium containing $5.0\;mgl^{-1}$ BA and $1.0\;mgl^{-1}$ IAA with a mean of 5.7 shoots. In liquid medium, number of shoots and fresh weight per explant increased significantly. The best shoot proliferation and increasing of fresh weight were achieved on LS liquid medium containing $5.0\;mgl^{-1}$ BA and $1.0\;mgl^{-1}$ IAA with 6.9 shoots and more than 4,000 mg fresh weight. Of the different concentrations of LS salt, double strength of LS medium provided the highest shoot proliferation with 7.3 shoots, and fresh weight with 5,539 mg per explant. Shoot proliferation on LS medium containing $50\;gl^{-1}$ sucrose had better results with 8.7 shoots and 5,979 mg per explant in fresh weight than other conditions. In conclusion, the optimal level for shoot proliferation and biomass increase of water hyacinth was attained with the application of the double strength of LS medium containing $5.0\;mgl^{-1}$ BA, $1.0\;mgl^{-1}$ IAA and $50\;gl^{-1}$ sucrose.

Effect of kinds of medium and plant growth regulators for plantlets regeneration by bud culture in Disporum sessile (윤판나물(Disporum sessile) 아배양에 의한 식물체 재분화에 영향하는 배지 종류 및 생장조절물질 효과)

  • Lee, Na-Nyum;Kim, Ji-Ah;Kim, Tae Dong;Kim, Yong Wook
    • Journal of Plant Biotechnology
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    • v.44 no.1
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    • pp.42-48
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    • 2017
  • This study was conducted to establish the optimal condition for in vitro propagation of Disporum sessile. In the experiments with different kinds of media (MS, B5 and WPM) and explant types for shoot induction and elongation, the highest number of shoot inductions (2.5/explant) was shown when the axillary bud explants were cultured on MS medium without plant growth regulators (PGRs). The best shoot elongation (7.2 cm) was also obtained when the apical buds were cultured on MS medium. The effect of BA pulse treatments with in vitro shoots was also examined. The highest in shoot induction (2.29/explant) and elongation (7.28 cm) was observed when the shoots from axillary buds were cultured on the media without PGRs. However, it decreased with increasing duration of BA pulse-treatment. The highest rooting rate (100%) and number of root inductions (21.3/explant) were achieved with 1.0 mg/L indole-3-butyric acid (IBA) treatment, whereas no differences were observed by different shoot types. The regenerated in vitro plantlets were hardened and successfully established in soil.

Effects of Growth Regulators on Plant Regeneration from the Cotyledon Explant in Oriental Melon (Cucumis melo L.) (참외 (Cucumis melo L.)자엽절편으로부터 식물체 재분화에 미치는 생장조절물질의 영향)

  • 문정길;추병길;두홍수;권태호;양문식;류점호
    • Korean Journal of Plant Tissue Culture
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    • v.27 no.1
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    • pp.1-6
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    • 2000
  • This study was carried out to find the effects of plant growth regulators on callus formation, rooting and shooting from cotyledon explant in oriental me]on. Various combinations of 0.1 mg/L auxins (IAA, NAA) and 0.5, 1.0. 1.5, 2.0 mg/L cytokinins (BA, kinetin, zeatin) were treated to the MS basal medium, respectively. Callus was induced mort effectively as 2,437.0 mg (FW)/explant in MS medium supplemented with 0.1 mg/L NAA and 2.0 mg/L BA, but that was non-embryogenic callus as colored yellow white and broke easily. Root was induced most effectively at a frequency of 98.0% in MS medium supplemented with 0.1 mg/L NAA and 0-5 mg/L kinetin. Shoots formed on cut part of vein at a frequency of 98.0% in MS medium supplemented with 0.1 mg/L IAA and 2.0 mg/L BA, that were multiple shoots. in case of its concentration, BA and lower concentration of IAA and NAA (0.01 and 0.05 mg/L). respectively. shooting ratio was not increased. The result of treatment with BA 0-5 mg/L and IAA 0.1 mg/L, callus induced at a week, and shoot start to form multiple shoots about 3 weeks after inoculation. After 2 times subculture as 2 weeks intervals, divided shoots rooted and developed into intact plantlets at 10 weeks and then that grown normally on pots after acclimatization.

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