• Title/Summary/Keyword: Explants

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High Frequency Induction of Multiple Shoots from Nodal Explants of Vitex negundo L. Using Sodium Sulphate

  • Chandramu C.;Rao D. Manohar;Reddy V. Dashavantha
    • Journal of Plant Biotechnology
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    • v.5 no.2
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    • pp.107-113
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    • 2003
  • The effect of sodium sulphate on shoot induction and multiple shoot formation from nodal explants of Vitex negundo L. was tested on Murashige and Skoog's (MS) medium fortified with different auxins, cytokinins and sucrose. Highest percentage $(97.78\%)$ of explants for shoot induction and multiple shoot (20.68/explant) production were observed in the combination treatment of $N^6-Benzyl$ adenine (BA) $(17.80\;{\mu}M/L)$, ${\alpha}-Naphthalene$ acetic acid (NAA) $(2.15\;{\mu}M/L)$ and $5\%$ sucrose supplemented with 100 mg/L sodium sulphate. In vitro raised shoots were rooted on the half-strength MS medium fortified with different concentrations of NAA, Indole-3-acetic acid (IAA), and Indole-3-butyric acid (IBA) alone and in combinations. Among the treatments, $4.90\;{\mu}M/L$ of IBA was found most effective $(95.56\%)$ in inducing roots. The rooted plantlets were shifted to glasshouse for acclimatization and later transferred to the field with cent percent survival. Furthermore, in vitro flowering was observed in the shoots cultured on MS medium supplemented with BA $(8.90\;{mu}M/L)$ and NAA $(1.61\;{\mu}M/L)$.

Development of an efficient protocol for high-frequency regeneration system in Hibiscus syriacus L.

  • Seo, Sang-Gyu;Ryu, Sun-Hee;Zhou, Yang;Kim, Sun-Hyung
    • Journal of Plant Biotechnology
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    • v.44 no.2
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    • pp.164-170
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    • 2017
  • A rapid and efficient in vitro regeneration system was established for Hibiscus syriacus L. The successful regeneration protocol employs induction of shoot organogenesis on leaf, petiole, and root explants. Among the various plant growth regulators evaluated, thidiazuron (TDZ) was the most effective for inducing rapid shoot formation. Most efficient shoot regeneration frequency was obtained from Murashige and Skoog (MS) media containing 0.01 mg/L TDZ. Regeneration efficiency was highest in the roots, and lowest in the leaves. A combination of 0.01 mg/L TDZ with benzyladenine (BAP) markedly improved the frequency of shoot differentiation from the root (up to 98%) and petiole (up to 88%) explants. Furthermore, leaf and petiole explants showed the highest frequency of shoot induction in half-strength MS media containing 0.01 mg/L TDZ and 1.0 mg/L BAP, while root explants formed the greatest number of shoots when 0.01 mg/L TDZ and 0.1 mg/L BAP were added to half-strength MS media. Although the frequency of shoot differentiation from leaf explants was only 50%, the leaf is considered the most efficient plant organ for use in tissue culture because leaves are easier to obtain than roots and petioles. Our findings show that various organs of H. syriacus can be used for plant regeneration, and the protocol developed in this study may be applicable in the horticulture industry.

High Frequency Somatic Embryogenesis and Plant Regeneration in Seedling Explant Cultures of Melon (Cucumis melo L.) (멜론(Cucumis melo L.) 유묘 절편으로부터 고빈도의 체세포배발생과 식물체 재분화)

  • 최필선;소웅영;조덕이;유장렬
    • Korean Journal of Plant Tissue Culture
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    • v.21 no.1
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    • pp.1-6
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    • 1994
  • Cotyledonary and hypocotyl explants of melon seedlings were cultured on Murashige and Skoog's (MS) medium supplemented with various concentrations of 2,4-Dichlorophenoxyacetic acid (2,4-D) and benzyladenine (B.A).Up to 22% of cotyledonary explants and 7%, of hypocotyl explants, respectively: Produced somatic embryos through intervening two types of calli: bright yellow compact (BYC) callus and pale-yellow compact (PYC) callus. BYC callus was capable of producing somatic embryos at initial culture, but it became necrotic as subrulhues proceeded. In contrast UC callus was incapable of producing somatic embryos during initial culture (first 6 weeks), but it became bright-yellow friable (BYF) callus with forming a few globular embryos after 2 months of subculture, indicating that the callus turned embryogenic. The embryogenic capacity of BYF maintained for over one year when the callus was sucultured at 4-week interval. Upon transfer onto MS basal medium the callus gave rise to numerous somatic embryos and subsequently converted to plantlets. Plantlets were transplanted to potting soil and grown to maturity in the phyotron.

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High Frequency Bulblet Formation in Scale and Stem Thin Cell Layer Explant Cultures of Lilium Oriental Hybrids (오리엔탈 백합의 인편 및 줄기의 박판 세포층 절편으로부터 고빈도 자구형성)

  • Oh, Seung-Cheol;Chung, Myung-Hee;Kim, Suk-Won;Liu, Jang-Ryol
    • Journal of Plant Biotechnology
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    • v.30 no.3
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    • pp.251-255
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    • 2003
  • An efficient system for in vitro bulblet formation of Lilium oriental hybrids(cvs). Casa Blanca and Siberia is described. Transverse thin cell layer(tTCL)(1mm thick) explants of 'Casa Blanca' formed bulblets at a frequency of 97.7% when cultured on Murashige and Skoog (MS) medium supplemented with 1mg/L 2,4 dichlorophenoxyacetic acid(2,4-D) (On average 15.6 bulblets were formed per explant). The frequency of bulblet formation was drastically reduced when the explant ghickness was thinner than 1 mm. Explants from the outermost layer of bulb scale produced greater frequency of bulblet formation than middle or innermost layer. Among auxins supplemented to culture medium at 1 mg/L, 2,4-D led to greater frequency of bulblet formation on explants than dicamba, picamba, or phenylacetic acid(PAA). tTCL explants from the middle region of the outermost layer bulb scale yielded greater frequency of bulblet formation than the upper or lower region. tTCL stem explants of 'Siberia' formed bulblets at a frequency of 95.3% when cultured on MS medium with 1 mg/L 2,4-D(On average 9.1 bulblets were formed per explant). The system estabilished in this study will be useful for in vitro rapid propagation and genetic transformation of Lilium Oriental hybrids.

Micorpropagation of Corylopsis coreana by Thidiazuron Treatment (Thidiazuron 처리에 의한 히어리나무의 기내번식)

  • Kang, Hyo-Jin;Moon, Heung-Kyu;Yi, Jae-Seon
    • Journal of Plant Biotechnology
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    • v.30 no.3
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    • pp.263-267
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    • 2003
  • This study was conducted to evaluate the effect of thidazuron(TDZ) on shoot proliferation and growth from axillary buds of 20-years-old Corylopsis coreana. Shoots proliferation was effectively achieved on WPM(Woody Plant Medium) supplemented with 0.03∼0.1mg/L TDZ. The highest shoot number(6.5$\pm$0.7) was obtained on 0.1mg/L TDZ treatment. On the TDZ medium shoots formed as clusters less than 1cm in height and therefore needed to subculture on GA$_{3}$ containing medium to induce elongation. In consecutive cultures, phenolic compounds were excreted at the proximal part of the explants and inhibited growth of the explants. Growth inhibition by the compounds was overcome using liquid and paper bridge culture system. About 60% of the elongated shoots rooted on half- strength MS medium containing IBA. Generally, IBA was mire effective on in vitro rooting than NAA with optimal range of 0.5mg/L to 1.0mg/L. Rooted plantlets were transferred in an artificial soil(vermculite) and acclimatized in high humidity greenhouse condition. Survival rate differed greatly depending on rooting types of the explants. Two types of rooting were observed. The first type was direct rooting from the explants. The second type was callus formation followed by rooting from the callus. The explants showing the 1st type rooting survived can be multiplicated in vitro by TDZ treatment followed by elongation with GA$_{3}$ and rooting with IBA.

Plantlet Regeneration by Tissue Cultures of Cyclamen persicum Mill. (시클라멘 (Cyclamen persicum Mill.)의 조직배양에 의한 식물체 재분화)

  • 은종선;김영선;한상권
    • Korean Journal of Plant Tissue Culture
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    • v.27 no.6
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    • pp.479-484
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    • 2000
  • This study was conducted to examine the effects of explant sources and plant growth regulators on mass propagation of Cyclamen persicum. Tuber, cotyledon, and petiole tissues were cultured on MS medium supplemented with different concentrations and combinations of auxins and cytokinins. Shoots were not induced from calli on cotyledon and petiole explants cultured on MS medium containing various concentrations of 2,4-D or NAA. However, multiple shoots were formed directly from tuber explants cultured on the medium containing 0.5 and 1.0 mg/L 2,4-D or NAA. In MS medium with cytokinin alone, TDZ was more effective in shoot formation than BA or kinetin in all explants. The combinations of NAA and BA was found to be most effective in shoot formation from tuber, cotyledon and petiole explants. Especially, shoots were formed from all the tuber explants on the medium containing 0.5 mg/L of NAA and BA. Hormonal effects on root formation were examined by subculturing single shoots on MS medium containing NAA or IBA. The medium with 0.5 mg/L IBA was most effective in root induction and subsequent plantlet regeneration.

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Effect of Cinnamomum Cassia on Cartilage Protection in Rabbit and Human Articular Cartilage

  • Baek, Yong-Hyeon;Huh, Jeong-Eun;Lee, Jae-Dong;Choi, Do-Young;Park, Dong-Suk
    • The Journal of Korean Medicine
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    • v.28 no.4
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    • pp.148-157
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    • 2007
  • Background & Objective: Articular cartilage is a potential target for drugs designed to inhibit the activity of matrix metalloproteinases (MMPs) to stop or slow the destruction of the proteoglycanand collagen in the cartilage extracellular matrix. The purpose of this study was to investigate the effects of Cinnamomum cassia in inhibiting the release of glycosaminoglycan (GAG), the degradation of collagen, and MMP activity in rabbit and human articular cartilage explants. Methods: The cartilage-protective effects of Cinnamomum cassia were evaluated by using glycosaminoglycan degradation assay, collagen degradation assay, colorimetric analysis of MMP activity, measurement of lactate dehydrogenase activity and histological analysis in rabbit cartilage explants culture. Results: Interleukin-1a (IL-1a) rapidly induced GAG, but collagen was much less readily released from cartilage explants. Cinnamomum cassia significantly inhibited GAG and collagen release in a concentration-dependent manner. Cinnamomum cassia dose-dependently inhibited MMP-1, MMP-3 and MMP-13 activities from IL-1a-treated cartilage explants culture when tested at concentrations ranging from 0.02 to 1 mg/ml. Conclusion : These results indicate that Cinnamomum cassia inhibits the degradation of proteoglycan and collagen through the down regulation of MMP-1, MMP-3 and MMP-13 activities of IL-1a-stimulated rabbit and human articular cartilage explants.

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High-frequency shoot regeneration from leaf explants through organogenesis in bitter melon (Momordica charantia L.)

  • Thiruvengadam, Muthu;Rekha, K.T.;Yang, Chang-Hsien;Jayabalan, Narayanasamypillai;Chung, Ill-Min
    • Plant Biotechnology Reports
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    • v.4 no.4
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    • pp.321-328
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    • 2010
  • An efficient protocol for in vitro organogenesis was achieved from callus-derived immature and mature leaf explants of Momordica charantia, a very important vegetable and medicinal plant. Calluses were induced from immature leaf explants excised from in vitro (15-day-old seedlings) mature leaf explants of vivo plants (45 days old). The explants were grown on Murashige and Skoog (MS) medium with Gamborg (B5) vitamins containing 30 g $1^{-1}$ sucrose, 2.2 g $1^{-1}$ Gelrite, and 7.7 lM naphthalene acetic acid (NAA) with 2.2 ${\mu}M$ thidiazuron (TDZ). Regeneration of adventitious shoots from callus (30-40 shoots per explant) was achieved on MS medium containing 5.5 ${\mu}M$ TDZ, 2.2 ${\mu}M$ NAA, and 3.3 ${\mu}M$ silver nitrate ($AgNO_3$). The shoots (1.0 cm length) were excised from callus and elongated in MS medium fortified with 3.5 ${\mu}M$ gibberellic acid ($GA_3$). The elongated shoots were rooted in MS medium supplemented with 4.0 ${\mu}M$ indole 3-butyric acid (IBA). Rooted plants were acclimatized in the greenhouse and subsequently established in soil with a survival rate of 90%. This protocol yielded an average of 40 plants per leaf explant with a culture period of 98 days.

High Frequency Organogenesis and Plant Regeneration in Tissue Cultures of Lettuce Seedling Explants (상추 유식물체 절편의 조직배양에 의한 고빈도 기관발생과 식물체 재분화)

  • Jung, Min;Woo, Je-Wook;Jung, Won-Joong;Yoo, Jang-Ryul
    • Korean Journal of Plant Tissue Culture
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    • v.26 no.3
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    • pp.219-222
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    • 1999
  • To induce adventitious buds, hypocotyl and cotyledonary explants from 7 to 10 day-old seedlings of lettuce (Lactuca sativa L.: two Japanese cultivars of crisphead lettuce and four Korean cultivars of leaf lettuce) were cultured or Murashige and Skoog (MS) and Schenk and Hildebrandt (SH) media supplemented with BA and NAA in the light for five weeks. Cotyledonary explants produced adventitious shoots at greater frequencies than hypocotyl explants. MS medium was more favorable to adventitious shoot formation than HS medium. Combination of 0.5 mg/L BA and 0.1 to 1 mg/L HPh in MS medium led to the greatest frequency (86%) in adventitious shoot formation. Creator than 95% of shoots excised from explants were rooted when cultured on MS basal medium.

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The development of herbicide-resistant maize: stable Agrobacterium-mediated transformation of maize using explants of type II embryogenic calli

  • Kim, Hyun A.;Utomo, Setyo Dwi;Kwon, Suk Yoon;Min, Sung Ran;Kim, Jin Seog;Yoo, Han Sang;Choi, Pil Son
    • Plant Biotechnology Reports
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    • v.3 no.4
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    • pp.277-283
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    • 2009
  • One of the limitations to conducting maize Agrobacterium-mediated transformation using explants of immature zygotic embryos routinely is the availability of the explants. To produce immature embryos routinely and continuously requires a well-equipped greenhouse and laborious artificial pollination. To overcome this limitation, an Agrobacterium-mediated transformation system using explants of type II embryogenic calli was developed. Once the type II embryogenic calli are produced, they can be subcultured and/or proliferated conveniently. The objectives of this study were to demonstrate a stable Agrobacterium-mediated transformation of maize using explants of type II embryonic calli and to evaluate the efficiency of the protocol in order to develop herbicide-resistant maize. The type II embryogenic calli were inoculated with Agrobacterium tumefaciens strain C58C1 carrying binary vector pTF102, and then were subsequently cultured on the following media: co-cultivation medium for 1 day, delay medium for 7 days, selection medium for $4{\times}14$ days, regeneration medium, and finally on germination medium. The T-DNA of the vector carried two cassettes (Ubi promoter-EPSPs ORF-nos and 35S promoter-bar ORF-nos). The EPSPs conferred resistance to glyphosate and bar conferred resistance to phosphinothricin. The confirmation of stable transformation and the efficiency of transformation was based on the resistance to phosphinothricin indicated by the growth of putative transgenic calli on selection medium amended with $4mg\;1^{-1}$ phosphinothricin, northern blot analysis of bar gene, and leaf painting assay for detection of bar gene-based herbicide resistance. Northern blot analysis and leaf painting assay confirmed the expression of bar transgenes in the $R_1$ generation. The average transformation efficiency was 0.60%. Based on northern blot analysis and leaf painting assay, line 31 was selected as an elite line of maize resistant to herbicide.