• Title/Summary/Keyword: Callus & shoot induction

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Efficient Plant Regeneration from Shoot Tip and Young Leaf in Rhodiola sachalinensis A. Bor.

  • Chi, Hyung-Joon;Yoon, Jae-Ho;Yang, Deok-Chun;Song, Won-Seob
    • Plant Resources
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    • v.6 no.3
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    • pp.233-241
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    • 2003
  • The shoot tip and young leaf of Rhodiola sachalinensis were cultured to invest the plant growth regulator condition for callus induction, shoot and root regeneration. When the shoot tip was sterilized in 2.0% of NaOCl for 20min., the contamination rate was the lowest. And the survival rate of the culture material was good in carbenicillin 500mg/L treatment group. Callus was obtained from shoot tip and young leaf segments. NAA 0.1-1.0mg/L and 2,4-D 0.1-0.5mg/L alone treatment were shown to have a good response on callus induction from shoot tip culture. In the case of young leaf culture, NAA and 2,4-D 0.1-0.5mg/L alone treatment were good in callus induction. In culturing shoot tip NAA 0.5mg/L and BA 0.5mg/L, NAA1.0mg/L and BA 0.lmg/L combination treatment was good in shoot regeneration. The regenerated shoots were rooted on MS medium supplemented with NAA and BA combination treatment. Especially, NAA 1.0mg/L and BA 0.1mg/L combination treatment was effective for root regeneration.

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Micropropagation by Leaf and Meristem Cultures of Pelargonium citrosa Van leenen (구문초 (Pelargonium citrosa Van leenen)의 잎과 정분열조직배양에 의한 미세증식)

  • 은종선;고정애;김영선;김명준
    • Korean Journal of Plant Tissue Culture
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    • v.21 no.4
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    • pp.247-252
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    • 1994
  • The effects of explant sources, plant growth regulators on callus induction and plantlet differentiation from leaf blade, petiole, and meristem tissue of Pelalgonium citrosa were investigated under illumination or in dark condition Leaf blade explants cultured on Murashige and Skoog's medium containing 2,4-D and kinetin did not form callus or organ. But those cultured on medium with NAA and BA produced callcus and shoots. Dark condition was more effective than light condition to callus induction and showed that some of shoot were differentiated directly from leaf blade explane. Callus proliferated vigorously on meristem tissue after 7 days of culture, and multiple shoots were obtained Sum callus on medium with 0.5 mg/L NAA and BA. Roots formed readily from about 80% of the shoots cultured on medium with 1.0 mg/L NAA. Regenerated plantlets regenerated had phenotypically normal leaves and roots.

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An Improved Method of Organogenesis from Cotyledon Callus of Acacia sinuata (Lour.) Merr. using Thidiazuron

  • Shahzad Anwar;Ahmad Naseem;Anis Mohammad
    • Journal of Plant Biotechnology
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    • v.8 no.1
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    • pp.15-19
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    • 2006
  • An efficient protocol for in vitro multiple shoot bud induction and plant regeneration from mature green cotyledon derived callus tissues of Acacia sinuata has been developed. Callus formation occurs at all the concentrations of thidiazuron (TDZ) in Murashige and Skoog's (MS) medium, but 0.6 ${\mu}M$ proved to be the best with maximum callus formation frequency. Supplementation of TDZ in combination with indole-acetic acid (IAA) in MS media accelerates shoot bud organogenesis in differentiating callus tissues with 60-70% conversion of shoot buds into shoot Most efficient shoot organogenesis was recorded when TDZ induced calli were subcultured at different concentrations of 6-benzyla-denine (BA). Optimum shoot bud induction and plant regeneration from callus was achieved when 0.6 ${\mu}M$ (TDZ) induced calli were subcultured at 3.0 ${\mu}M$ (BA) where $16.6{\pm}0.74$ shoots/unit callus on obtained. Rooting in in vitro differentiated shoots was achieved when transferred to medium containing different concentration of indole-3-butyric acid (IBA) in full & half strength MS medium. The well rooted plantlets were hardened and transferred to net house with 90% survival rate.

Multiple shoot induction and callus regeneration in Sarcostemma brevistigma Wight & Arnott, a rare medicinal plant

  • Thomas, T. Dennis;Shankar, Surabhi
    • Plant Biotechnology Reports
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    • v.3 no.1
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    • pp.67-74
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    • 2009
  • An efficient micropropagation protocol based on multiple shoot induction and callus regeneration has been standardized in Sarcostemma brevistigma, a rare medicinal plant. The nodal cuttings were cultured on MS medium supplemented with BA ($0.5-8{\mu}M$) or Kn ($0.5-8{\mu}M$) alone or in combination with NAA ($0.5-1.5{\mu}M$). Maximum multiple shoot induction was observed on MS medium supplemented with $4{\mu}M$ BA. On this medium, 100% cultures responded with an average number of 11.3 shoots per explant. However, the average shoot length was limited to only 0.9 cm on this medium. The addition of $1{\mu}M$ NAA along with $4{\mu}M$ BA gave rise to an average number of 10.9 shoots with an average shoot length of 1.8 cm. Luxuriantly growing callus was obtained on MS medium supplemented with BA ($5{\mu}M$) and 2,4-D ($2{\mu}M$). The callus was subcultured on MS medium supplemented with BA ($2-15{\mu}M$) or Kn ($2-15{\mu}M$) alone or in combination with NAA ($0.5-2{\mu}M$) for shoot organogenesis. Optimum callus regeneration was obtained on MS medium supplemented with $10{\mu}M$ BA and $1{\mu}M$ NAA. On this medium, 100% cultures responded with an average number of 13.4 shoots per culture. The shoots obtained via multiple shoot induction and organogenesis were rooted on half-strength MS medium supplemented with NAA ($1-7{\mu}M$) or IBA ($1-7{\mu}M$). IBA was better than NAA in terms of both the percentage of cultures that responded and the average number of roots per explant. The rooted shoots were successfully transplanted to soil with 86% success. This standardized protocol will help to conserve this rare medicinal plant.

The Effects of Plant Growth Substances on the Callus Induction and Multiple Shoot Formation of Korean Lawngrass( Zoysia japonica Steud.) (식물생장조절제 처리가 들잔디의 Callus유기 및 Multiple Shoots형성에 미치는 영향)

  • 심재성;김동찬;서병기
    • Asian Journal of Turfgrass Science
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    • v.8 no.3
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    • pp.137-147
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    • 1994
  • We have established a high-frequency plant regeneration system via organogenesis from mature seed of Korean lawngrass(Zoysia japonica Steud.). The effects of 2,4-dichiorophenoxy acetic acid (2,4-D), 6-furfuryl amino purine (kinetin), $\alpha$-naphthaiene acetic acid (NAA), N6-benzyl amino pu-rine (BAP), and casein hydrolysate (CR) on cailus induction and multiple shoot formation on ex-posure to light were evaluated. Callus produced on the Murashige and Skoog (MS) medium containing 2,4-D and kinetin had high organogenesis potency. A single addition of 1.0 mg $L-^1$ 2,4-D significantly induced callus. Also, 1.0 mg L-$^1$ 2,4-D, with the addition of 0.1 mg $L-^1$ kinetin highly enlianced callus induction. The trend of cailus induction was also found on mediurn containing 0.1 mg $L-^1$ BAP with 1.0 mg $L-^1$2,4-D, and 1 g $L-^1$ CR with the addition of 1.0 mg $L-^1$ 2,4-D. However, NAA was no effective on callus formation. The growth of root was significantly high in the presence of 0.1 mg $L-^1$ kinetin compared to other concentrations. Over 2 mg $L-^1$ kinetin highly lengthened roots. Fresh weight of plantlet was highest on medium containing 0.1 mg $L-^1$ 2,4-D. Also, on medium containing 0.1 mg $L-^1$ BAP, fresh weight of piantlet was highly enhanced. BAP was significantly effective on multiple shoot formation, particularly when 2.0 mg $L-^1$ was added with 0.1 mg $L-^1$ 2,4-D. Callus induction and multiple shoot formation were achieved on MS basal medium containing 1.0 g $L-^1$ CH.

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Somatic embryogenesis and in vitro plant regeneration from various explants of the halophyte Leymus chinensis (Trin.)

  • Sun, Yan Lin;Hong, Soon-Kwan
    • Journal of Plant Biotechnology
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    • v.36 no.3
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    • pp.236-243
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    • 2009
  • The halophyte Leymus chinensis (Trin.) is a perennial rhizome grass (tribe Gramineae) that is widely distributed throughout China, Mongolia and Siberia. This study was conducted to investigate an optimal condition for plant regeneration from mature seeds, leaf base segments, and root segments in L. chinensis. Plant growth regulators affecting embryogenic callus induction and plant regeneration were investigated by four-factor-three-level [L9 (34)] orthogonal test in this study. The effects of explants types (mature seeds, leaf base segments and root segments), callus types, medium types were examined in this study. Wild type (WT) and Jisheng No. 1 plants (JS) were used for primary callus induction. A clear explants difference was seen during callus induction; mature seeds were considered as the preferred explants; and the highest frequency of callus induction was obtained in Medium 6 using mature seeds as explants in WT. Plant regeneration ability was evaluated by frequencies of green callus forming, shooting, rooting, and shooting with roots. Effect of α-naphthalene acetic acid (NAA) on shoot regeneration was remarkable with the highest frequency of 70.8% in WT after 2-month culture. The medium with 0.2- 0.5 mg/L NAA was found to have the highest shoot induction. All regenerated shoots were successfully rooted when transferred on half-strength Murashige and Skoog (MS) basal medium. The acclimatized plantlets were grown to mature with flowering and seeds setting in green house conditions.

Changes of Cllus Induction, Betacyanins and Protein Contents from Cotyledons of Sugar Beet(Beta vulgaris L.) (사탕무(Beta vulgaris L.) 의 자엽소 배양에 의한 callus 유기 적색색소 및 단백질 \ulcorner량의 변화)

  • Kim, Hyoun-Kyoung;Kim, Doh-Hoon;Chung, Dae-Soo;Park, Hyeon-Jin
    • Journal of Life Science
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    • v.7 no.4
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    • pp.270-275
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    • 1997
  • In order to produce betacyanins from sugar beet(Beta vulagris L.) in vitro, callus induction, shoot formation, root formation, betacyanin contents and protein contents determined from callus which was induced cotyledons of sugar beet seedling under an addition of NAA and BAP on the MS medium. The results were summarized as follows; The combination 3.0mg/l NAA and 1.0mg/l BAP treatment showed the most high callus induction rate, betacyanin and protein contents. The combination NAA and BAP treatments were not shoot formation, but BAP treatments showed high root formation rate. But high concentrations of BAP have not shown root formation.

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Effects of Plant Growth Regulators on in vitro Propagation of Cymbidium kanran and Cymbidium hybrida (한란 및 심비디움의 기내 증식에 미치는 생장조절물질의 효과)

  • Kim, Hak-Yoon;Kwon, Soom-Tae
    • Current Research on Agriculture and Life Sciences
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    • v.18
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    • pp.1-7
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    • 2000
  • This study was carried out to determine the effects of plant growth regulators on organogenesis from Cymbidium kanran and Cymbidium hybrida. Optimal rhizome formation from Cymbidium kanran was obtained on MS medium with 10 ppm kinetin+2 ppm NAA. and optimal protocorm formation from Cymbidium hybrida was obtained on MS medium with 10 ppm kinetin+0.05 ppm NAA. However, in this study the optimal media for the callus induction from both explants was not identified. Optimal shoot induction from rhizome of Cymbidium kanran was obtained on MS medium with 10 ppm BA+2 ppm NAA and 5 ppm BA+2 ppm NAA. Optimal shoot induction from protocorm of Cymbidium hybrida was obtained on MS medium with 10 ppm kinetin+2 ppm NAA.

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Improvement of Shoot Regeneration from Scutella-Derived Callus in Rice

  • Kim, Yong-Wook;Cho, Joon-Hyeong;Lee, Jang-Yong
    • KOREAN JOURNAL OF CROP SCIENCE
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    • v.49 no.1
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    • pp.52-60
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    • 2004
  • The optimized in vitro culture system was investigated for improvement of regeneration efficiencies by observing the responses of scutella-derived callus of Korean rice (Oryza sativa L.). Large variations of callus induction (43.9-93.9%) and shoot regeneration (0-88.7%) were observed among the rice cultivars depending on medium. However, shoot regeneration was significantly improved by selected utilization of basal medium, growth regulators, and carbon sources. N6 basal medium was more efficient for embryogenic callus induction than MS or LS basal medium, while MS was superior to N6 for shoot regeneration. The calli of highly regenerative cultivars grew faster and showed higher rates of green tissue formation (GT) and shoot regeneration (SR) and lower rate of callus browning (CB) than those of recalcitrant cultivars. Although a higher level of kinetin stimulated the GT and SR in highly regenerative cultivars, $10\textrm{mgL}^{-1}$ kinetin generally suppressed the GT and SR, while CB was accelerated compared to $2\textrm{mgL}^{-1}$ kinetin. Additional benefits of sorbitol combined with maltose (or sucrose) under $5\textrm{mgL}^{-1}$ kinetin were certainly confirmed on regeneration efficiencies compared to sucrose alone as carbon source and osmotic regulator. This combination showed high rate of GT and SR with multiple shoots while low rate of CB. With MSRK5SM-Pr medium ($5\textrm{mgL}^{-1}$ kinetin, 3% sorbitol, 2% maltose, $500\textrm{mgL}^{-1}$ proline), the regeneration efficiencies of total 17 out of 24 cultivars were practically improved 160% on average compared to MSRK2S ($2\textrm{mgL}^{-1}$ kinetin, 3% sucrose) control medium. Especially, the medium was most effective to the cultivars showing a medium level of regenerability such as Daesanbyeo and Dongjinbyeo and Suwon477, enhancing efficiencies more than 300-600% compared to MSRK2S medium.

Callus Induction and Plant Regeneration from Stolon in Zoysiagrass (한국잔디류에서 포복경 배양을 통한 캘러스 유기와 재분화에 관한 연구)

  • 김종보;박순정;김두환
    • Asian Journal of Turfgrass Science
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    • v.11 no.4
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    • pp.311-320
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    • 1997
  • This study was carried out to induce and maintain callus from 59 zoysiagrass lines, to know the effective disinfestation method for zoysiagrass stolon as explant and the difference in the response of callus induction among 59 lines, and to investigate the effect of medium, growth regulators, light, temperature, stolon part and internode position on callus induction and emhryogenic callus(E.C.) formation. The treatment of 0.lmg/L $HgCl_2$for 15 min resulted in no contamination and the highest callus induction(46.6%). Callus was induced from the 59 zoysiagrass lines. The callus growth of Z. japonica and Z. sinica was generally better than Z. matrella Ten cell lines whose callus and stolon grow fast in culture and in field, respectively were selected to he used for breeding. Callus induction was the most effective at 2.0mg /L of both 2, 4-D and picloram in MS medium. MS medium was the best for callus induction and growth while LS medium was the best for embryogenic callus and shoot formation. Callus induction and growth was better at 28, 31$^{\circ}C$. than 25$^{\circ}C$. and dark condition was better than light condition in MS me-dium containing 2mg/L 2,4-D. While callus induction was better with node part as explant than with internode part, callus growth and embryogenic callus formation was better with internode part. In 'Japonica 1', the first internode was the most effective in callus induction, but third internode was the best in '$M_2$ X $S_2$'.

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