• Title/Summary/Keyword: shoot culture

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Development of Cryopreservation System using Shoot-Apex in Yam (Dioscorea batatas)

  • Shin Jong-Hee;Kang Dong-Kyoon;Bae Jeong-Suk;Lee Bong-Ho;Sohn Jae-Keun
    • Journal of Plant Biotechnology
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    • v.8 no.1
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    • pp.43-50
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    • 2006
  • The goal of this research was to develop an efficient cryopreservation protocol for germplasms of yam (Diosorea batatas), that were cultivated in Korea. Comparative studies with four other cryogenic techniques and subsequent experiments for shoot regrowth were conducted. in vitro-grown shoot-apices of the D. batatas were successfully cryopreserved by encapsulation-dehydration. The maximum survival of shoot-apices could be achieved when the precultured (with 0.3 M of sucrose for one day) and encapsulated (with a 3%(w/v) Na-alginate solution) apices were dehydrated for $3.5{\sim}4\;h$ prior to direct immersion in LN (liquid nitrogen). The frequency of regrowth rate of cryopreserved apices was not decreased during 3-month storage period. The thawing method markedly affected survival of the cryopreserved apices, and thawing at $40^{\circ}C$ for 3 min produced the best results. When cryopreserved apices were post-cultured on the post-culture medium (MS), supplemented with $0.2mgl^{-1}$ of BA ($N_6$-benzyladenine) and $0.2mgl^{-1}$ of kinetin, they showed direct shooting without callusing.

Plant Regeneration from Leaf and Internode Segment Cultures of Boxthorn (Lycium chinense Mill.) (구기자나무의 잎과 마디절편체 배양에 의한 식물체 재생)

  • 김동찬;정해준;민병훈;양덕춘
    • Korean Journal of Plant Tissue Culture
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    • v.28 no.6
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    • pp.329-333
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    • 2001
  • Callus and shoot formation from medicinal crop, Lycium chinense Mill. cv. 'Cheongyang', as influenced by various media, explant sources and plant growth regulators were investigated. The rate of shoots formation, number of shoots, and fresh weight of shoots were the best on MS medium followed by B$_{5}$, WPH, and SH. Callus induction was more effective in leaf than internode segments, and was the best on MS medium containing 0.5 mg/L NAA with 0.2 mg/L BA. Effects of plant growth regulators in shoot formation were more effective in BA than TDA combined with NAA. Shoot formation from callus induced in leaf and internode segments was the best on MS medium containing 0.01 mg/L NAA with 0.2 mg/L BA.

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Effect of cytokinin on adventitious shoot formation and plant regeneration from explants of Pulsatilla koreana NAKAI (할미 꽃 (Pulsatilla koreana NAKAI) 식물 절편체로부터 부정아 유도에 미치는 cytokinin의 영향 및 식물체 재 분화)

  • Liam, Yu-Ji;Iin, Guan-Zhe;Kim, Won-Bae;Yoo, Dong-Lim;Zhao, Xiao-Mei
    • Journal of Plant Biotechnology
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    • v.36 no.2
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    • pp.170-173
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    • 2009
  • Leaf and petiole explants of Pulsatilla koreana NAKAI were cultured on MS medium supplemented with various concentrations of zeatin, kinetin or BAP combined with 0.05 mg/L IAA. After 6 weeks of culture, effects of cytokinin on adventitious shoot formation from explants were investigated. The highest frequency of shoot formation was obtained when petiole explants were cultured on medium with 0.5 mg/L zeatin and 0.05 mg/L IAA. Regenerated shoot were transferred on to root induction medium. The best root formation was observed at 1/2 MS medium with 1.5 mg/L NAA. Rooted plantlets were transplanted to a mixture of perlit and soil (1:3), where they were successfully acclimatized.

Organogenesis and Plant Regeneration in Tissue Cultures of Pelargonium aridum (Ligularia) and P. zonale(Ciconium) (Pelargonium aridum과 P. zonale의 기관발생에 의한 식물체 재분화)

  • 유순남
    • Korean Journal of Plant Tissue Culture
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    • v.22 no.6
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    • pp.307-310
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    • 1995
  • Cotyledon and epicotyl explants of P. aridum and P. zonale formed calli when cultured on MS medium supplemented with 2 mg/L NAA and 0.2 mg/L BA. Calli were subcultured on the same medium Upon transfer to MS medium with 0.1 to 0.5mg/L NAA and 0.25 to 2mg/L BA for P. aridum 0.1 to 0.5mg/L NAA and 1 to 2mg/L BA for P. zonale subcultured calli gave rise to the greatest number of shoots (0.78 shoot for P. aridum and 0.65 shoot per explant for P. zonale, respectively).Most shoots produced roots when cultured on 1/2MS basal medium. The regenerates were transferred to potting soil and grown to materity in a greenhouse.

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In vitro Propagation of Trichosanthes kirilowii Max. (조직배양에 의한 하늘타리의 증식)

  • Hwang, Sung-Jin;Hwang, Baik
    • Korean Journal of Medicinal Crop Science
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    • v.9 no.1
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    • pp.15-20
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    • 2001
  • Adventitious shoot buds, without an intervening callus phase, were induced from stem explants of Trichosanthes kirilowii on MS medium supplemented with 2 mg/L BA. The culture medium, carbon source as well as plant growth regulators were found to influence further shoot multiplication and eventual shoot growth. A maximum number of shoots was obtained on a MS medium containing 0.5 mg/L BA and 3% sucrose. Shoot produced in vitro were rooted on half-strength phytagelled 1/2MS medium prior to transfer to green house condition.

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In vitro Propagation using Shoot Tip Culture in Gold Tree[Dendropanax morbifera $L_{EV}]$. (황칠나무의 경정배양에 의한 기내번식)

  • 최성규;윤경원
    • KOREAN JOURNAL OF CROP SCIENCE
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    • v.46 no.6
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    • pp.464-467
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    • 2001
  • In order to establish a in vitro propagation system for gold tree[Dendropanax morbifera $L_{EV}$], the effects of auxins and cytokinins on shoot multiplication and rooting were investigated. Germination rate was the best in MS medium. The fresh weight and number of shoot were the best on the medium containing 0.1 or 1.0 mg/l BAP and 0.5 or 1.0 mg/l NAA. Shoots were successfully rooted in MS medium with 1.0 mg/l NAA. Roots were easily formed by the addition of auxins, especially 0.1 or 1.0 mg/l BAP.P.

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Highly efficient production of transgenic Scoparia dulcis L. mediated by Agrobacterium tumefaciens: plant regeneration via shoot organogenesis

  • Aileni, Mahender;Abbagani, Sadanandam;Zhang, Peng
    • Plant Biotechnology Reports
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    • v.5 no.2
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    • pp.147-156
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    • 2011
  • Efficient Agrobacterium-mediated genetic transformation of Scoparia dulcis L. was developed using Agrobacterium tumefaciens strain LBA4404 harboring the binary vector pCAMBIA1301 with ${\beta}$-glucuronidase (GUS) (uidA) and hygromycin phosphotransferase (hpt) genes. Two-day precultured leaf segments of in vitro shoot culture were found to be suitable for cocultivation with the Agrobacterium strain, and acetosyringone was able to promote the transformation process. After selection on shoot organogenesis medium with appropriate concentrations of hygromycin and carbenicillin, adventitious shoots were developed on elongation medium by twice subculturing under the same selection scheme. The elongated hygromycin-resistant shoots were subsequently rooted on the MS medium supplemented with $1mg\;l^{-1}$ indole-3-butyric acid and $15mg\;l^{-1}$ hygromycin. Successful transformation was confirmed by PCR analysis using uidA- and hpt-specific primers and monitored by histochemical assay for ${\beta}$-GUS activity during shoot organogenesis. Integration of hpt gene into the genome of transgenic plants was also verified by Southern blot analysis. High transformation efficiency at a rate of 54.6% with an average of $3.9{\pm}0.39$ transgenic plantlets per explant was achieved in the present transformation system. It took only 2-3 months from seed germination to positive transformants transplanted to soil. Therefore, an efficient and fast genetic transformation system was developed for S. dulcis using an Agrobacterium-mediated approach and plant regeneration via shoot organogenesis, which provides a useful platform for future genetic engineering studies in this medicinally important plant.

The Effect of NAA and BA on Polar-Regeneration of Shoot in Hybrid Poplar (NAA와 BA가 hybrid poplar(p. nigra var. betulifolla X P.trichocarpa)의 극성 분화에 미치는 영향)

  • 김용재;김명원강영희
    • KSBB Journal
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    • v.7 no.2
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    • pp.85-91
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    • 1992
  • The role of NAA(1-Naphthaleneacetic acid) and BA(6-Benzyladenine) in the adventitious shoot regeneratlon from Populus leaf segments and changes in the pattern of RNA and protein synthesis were investigated. The adventitious shoot regeneration octured at the basal cut end of Populus leaf segments. This process was effected by many factors, including wounding culture conditions, light and plant growth regulators etc. The highest adventitious shoot regeneration frequency was obtained at $0.01mg/\ell$ NAA with $0.2mg/\ell$ BA. In this condition adventitious shoot starved to regenerate on the 13th day of oullure. The most optimal hormone composition for RNA and protein synthesis was $0.01mg/\ell$ NAA with $0.2mg/\ell$ BA. The content of RNA and protein was greater at the proximal part. In the course of adventitious shoot regeneration, the proteins associated with polar-regeneration appeared at the proximal part of populus leaf segment.

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Effects of BA and 2, 4-D on Shoot and Root Formation from Petiole Segments of Strawberry Plant in In Vitro Culture (딸기의 엽병조직(葉柄組織)에서 BA와 2, 4-D가 Shoot 및 Root의 분화(分化)에 미치는 영향(影響))

  • Lee, Young Bok;Campbell, William F.
    • Korean Journal of Agricultural Science
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    • v.9 no.2
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    • pp.461-466
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    • 1982
  • Lower and upper petiole segments of strawberry plants (Fragaria${\times}$ananassa) were cultured on Murashige and Skoog (MS) medium containing 2mg/liter Benzylamino purine (BA), 0.02mg/liter 2,4-dichlorophenoxyacetic acid (2,4-D), or their coinbination. Of the lower petiole segments, 55% or more were differentiated on the media containing $2mg/{\ell}\;BA$ or $2mg/{\ell}\;BA+0.02mg/{\ell}\;2,4-D$. There was no significant difference in results be tween these treatments. BA did not influence root formation. No shoots we re initiated from the upper petiole segments. Root formation was stimulated by the $0.02mg/{\ell}\;2,4-D$ medium, whereas it inhibited shoot formation. Upper petiole segments produced more roots than did lower petiole segments.

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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.