• Title/Summary/Keyword: in vitro bud culture

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A STUDY ON CHONDROGENIC POTENTIAL IN MANDIBULAR AND LIMB BUD MESENCHYMAL CELLS OF HUMAN EMBRYOS : A POSSIBLE ROLE OF PROTEIN KINASE C

  • Kook, Yoon-Ah;Kim, Eun-Cheol;Kim, Sang-Cheol
    • The korean journal of orthodontics
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    • v.26 no.6
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    • pp.667-676
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    • 1996
  • We have examined the in vitro stage-related chondrogenic potential of human mandibular and limb bud mesenchyme cells using micromass culture. Our results indicate that limb bud mesenchyme cells as early as stage 16 by Carnegie system (37 days), well before the initiation of in vivo chondrogenesis, have chondrogenic potential which is expressed in micromass culture. These results are correlated with stage-related chondrogenic potential of human limb bud in vivo as a result of Alcian blue staining. The proliferation of chondrogenic cells increased in the first 3 days after culture and then decreased. These results were correlated with the cell cycle analysis of which the number of $G_0^1/G_1$ phase increased markedly after 3 days of culture, while the percentage of cells in S phase was decreased. On the other hand, it was rarely differentiated in the mandible. We examined the effects of two PKC modulators such as phorbol 12-myristate 13-acetate (PMA), a potent activator of PKC, and staurosporine (STSN), an inhibitor of PKC. PMA inhibited the chondrogenesis, whereas STSN promoted the chondrogenesis in a dose dependent manner. In addition, PMA exerted no inhibitory effect when the cells were pretreated for 24 h with STSN, implying that the chondrogenic events might be settled at an early step in vitro and FKC may act as a negative modulator. Collectively, these results demonstrate, for the first time, the stage-related chondrogenic potential of human mandibular and limb bud mesenchyme cells and the role of PKC during chondrogenesis in vitro & in vivo.

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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.

Micropropagation via Axillary Bud Induction of Eucalyptus pellita (액아유도에 의한 Eucalyptus pellita의 기내번식)

  • Moon, Heung-Kyu;Kim, Ji-Ah;Lee, Hyun-Shin;Kang, Ho-Duck
    • Journal of Plant Biotechnology
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    • v.30 no.3
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    • pp.269-273
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    • 2003
  • In order to develop an efficient micropropagation protocal for Eucalyptus pellita, on in vitro culture system has been was established by inducing axillary buds from greenhouse stock materials. Among 6different media tested, DKW medium was the best ot induce bast induce both shoot proliferation and growth. Average number of proliferated shoots of 403per explant was obtained at the concentration of 0.1mg/LBA. Most of the stem materials excreted phenolic compounds at the proximal part of the explant and caused darking of the media. Therefore, it was necessary to transfer frequently to a fresh medium and/or to add activated charcoal at the concentration of 0.02%(w/v). Generally on vitro roots were formed easily on 1/2DKW medium with NAA treatment. All the explants rooted at the medium containing 0.2mg/L NAA and displayed vigorous root growth in vitro culture conditions. After transferred to an artificial soil mixture (peatmoss: vermiculrite: perlite, 1:1:1, v/v/v) in the greenhouse, most rooted plantlets survived well without any morphological abnormalities. The results show that the species can be micropropagated effectively by the application of axillary bud culture system.

Effect of explant's position and culture method on shoot proliferation and micro-cuttings for a rare and endangered species, Abeliophyllum distichum Nakai (희귀 및 멸종위기 식물 미선나무(Abeliophyllum distichum Nakai)의 절편위치 및 치상방법에 따른 기내증식 및 미세삽목)

  • Lee, Na Nyum;Kim, Ji-Ah;Kim, Yong-Wook;Choi, Yong Eui;Moon, Heung Kyu
    • Journal of Plant Biotechnology
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    • v.42 no.3
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    • pp.228-234
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    • 2015
  • Using either the apical or axillary bud of the endangered species Abeliophyllum distichum Nakai, we tested the effect of bud position and culture method on shoot proliferation and rooting. In shoot proliferation, the axillary bud explant was more effective than the apical bud and the effect was fostered by BA treatment, whereas no differences were observed in shoot elongation by the explant position. Spontaneous rooting was observed in the MS basal medium and resulted in conspicuous differences in the explant position : more than 80% in apical bud explant and 28% in axillary bud explant was achieved, respectively. The positional effects were also observed in BA pre-treatments: generally vertical culture method appeared to be better in shoot proliferation, growth, and rooting than that of the horizontal culture method regardless of the BA pre-treatment duration. The highest shoot multiplication was achieved through the vertical culture method with axillary bud explant, whereas the best shoot elongation and rooting was obtained using the vertical culture method with the apical bud explant. Apical bud explant was superior to axillary bud explant in ex vitro micro-cuttings and revealed a significant difference in shoot growth and root development. The above results suggest that explant position and culture method influence the efficiency of micropropagation for a rare and endangered plant Abeliophyllum distichum.

Variation of the Regenerated Plantlets from in Vitro Culture of Neoregeria carorinae 'Tricolor' and in Vivo Growth of Regenerated Plantlets (네오레게리아 기내배양시 변이발생과 기외 생육)

  • 정향영;한봉희;신학기;김의영
    • Korean Journal of Plant Tissue Culture
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    • v.22 no.5
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    • pp.273-276
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    • 1995
  • In vitro propagation of Neoregeria carorinae 'Tricolor' was achieved by using immature flowers and lateral buds, and the plantlets from tissue culture were transplanted and cultivated in greenhouse. The picking times of explants to decrease disappearance of stripes, and in vivo the growth and flowering of regenerated plantlets as influenced by in vivo healed nun were investigated. The normal plantlet were obtained at a frequency of 67%, in the culture of immature flowers picked at 4 weeks after flower bud differentiation, while all leaf stripes disappeared in the culture of immature flowers picked 1 and 5 weeks after flower bud differentiation. In vivo growth of plantlet from immature flower buds was better than those from lateral buds, and the flowering of 27.8% showed in the greenhouse culture of plantlet from immature culture, but the plantlets from lateral buds did not flower at all. The plantlets rooted on the medium with 0.5 mg/L IBA were the most favorable in green house culture, and the kinds and concentrations of auxin in vitro did not have any influence on variation of plane cultured in greenhouse.

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In vitro propagation and multiple shoot induction of Rhodiola rosea L. by axillary bud culture (홍경천(Rhodiola rosea L.)의 액아배양을 통한 다신초 유도 및 기내 대량증식)

  • Bae, Kee-Hwa;Ko, Myung-Suk;Kim, Nam-Young;Song, Jae-Mo;Song, Gwan-Pil
    • Journal of Plant Biotechnology
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    • v.39 no.2
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    • pp.114-120
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    • 2012
  • An efficient in vitro propagation was established by using axillary bud explants of roseroot (Rhodiola rosea L.), which has been known as a medicinal plant in East Asia. Among various media tested, MS medium supplemented with 1.0 mg/L BA and 1.0 mg/L $GA_3$ was found to be the best for multiple shoot formation (15 axillary shoots per axillary bud). In addition 1/2MS medium containing 50 g/L sucrose was best for shoot elongation (7.8 cm) and increasing total chlorophyll contents (8.64 mg/g) best. Maximum number of roots (17.7 roots per explant) was observed on the medium without plant growth regulators. Propagated plants were successfully acclimatized to ex vitro conditions, with a survival frequency of 97% after 12 weeks. Most rooted shoots grew well and produced viable seeds when grown in vitro culture conditions. Therefore, R. rosea can be effectively propagated in vitro by the system we developed in this study.

Micropropagation of Echinosophora koreensis Nakai, a Korean Endemic Species in Danger, Using Axillary Buds

  • Hyunseok Lee
    • Proceedings of the Plant Resources Society of Korea Conference
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    • 2020.12a
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    • pp.60-60
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    • 2020
  • To establish in vitro axillary bud culture conditions of Echinosophora koreensis Nakai, one of Korean endemic endangered species famous for beautiful flowers, we tested the influence of plant growth regulators (PGRs) in shooting and rooting stage from in vitro plants. In shoot multiplication, addition of 6-benzylaminopurine (BA) to the media induced 2.5 to 3 shoots per bud during 4 weeks of culture. And media including 0.5 mg L-1 thidiazuron (TDZ) produced 3 to 4 shoots per bud. However, zeatin and isopentenyl adenine (2-ip) were not successful to increase shoot number, and the combination treatments of BA with other PGRs were also not effective. Shoots were smaller than 2 cm in length, in most of the treatments. In rooting, naphthalene acetic acid (NAA) treatments in the range of 0.5 to 4.0 mg L-1 appeared to increase rooting rate by 10% to 60% approximately when compared with the control but roots developed with callus clusters. Indole butyric acid (IBA) addition had little effect on rooting (below 10%), while some roots were longer than in NAA treatments and some shoots were longer on high IBA concentrations (4.0 to 8.0 mg L-1). It is suggested that micropropagation is a highly applicable and promising to multiplication and conservation of rare and endangered endemic species.

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In vitro propagation of endangered species, Hylotelephium ussuriense (Kom.) H. Ohba (멸종위기종 둥근잎꿩의비름 (Hylotelephium ussuriense (Kom.) H. Ohba)의 기 내 증식)

  • Bae, Kee-Hwa;Yoo, Kyoung-Hwa;Kim, Ji-Ah;Yoon, Eui-Soo
    • Journal of Plant Biotechnology
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    • v.41 no.1
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    • pp.38-43
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    • 2014
  • To establish the system of in vitro plant regeneration, the different explants (stem with axillary bud and stem without axillary bud) of Hylotelephium ussuriense were cultured on the Murashige and Skoog's medium containing 6-benzylaminopurine (BA) and indolebutyric acid (IBA). The adventitious shoot induction was more effective in the stem with axillary bud explants than the stem without axillary bud explants, and was the best on MS medium containing 3.0 mg/L BA and 0.01 mg/L IBA. Frequency of plantlet growth was not significantly treated on MS and sucrose. Total chlorophyll contents under ventilation treatment were higher than those in control (non-ventilation). This in vitro propagation protocol will be useful for conservation and mass propagation of this endangered plant.

Effects of Plant Growth Regulators on in vitro Propagation of Echinosophora koreensis Nakai

  • Yi, Jae-Seon;Lee, Hyunseok;An, Chanhoon
    • Journal of Forest and Environmental Science
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    • v.29 no.4
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    • pp.275-281
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    • 2013
  • To establish in vitro nodal culture conditions of Echinosophora koreensis Nakai, one of rare and endangered species famous for beautiful flowers in the Korean Peninsula, the influence of plant growth regulators (PGRs) on shooting and rooting from in vitro shoots was investigated. In shoot multiplication, addition of 6-benzylaminopurine (BA) to the half-strength Driver and Kuniyuki's media in the range of 2.22 to 8.88 ${\mu}M $induced 2.5 to 2.7 shoots per axillary bud; and addition of 2.27 ${\mu}M $ thidiazuron (TDZ) produced 3.2 shoots, during 4 weeks of culture, while zeatin and isopentenyl adenine (2ip) were not effective on shoot multiplication as observed from several combination treatments of BA with other PGRs. Shoots established were smaller than 2 cm in length, in most of the treatments. while in BA 8.88 ${\mu}M $ treatment more than 30% of shoots were longer than 2 cm and shorter than 4 cm. In rooting, naphthalene acetic acid (NAA) from 5.37 to 21.48 ${\mu}M $ showed the rooting rate from 40.0 to 62.5%. Indole butyric acid (IBA) addition had little effect on rooting (<10%), although some roots in IBA-containing media were longer than those in NAA. Micropropagation from axillary buds of nodular explants was applicable and promising to multiplication and conservation of Echinosophora koreensis Nakai.

Study on the Propagation System and the Photosynthetic Rate of Chrysantemum zawadskii H. (약용자원식물 구절초의 고소득화를 위한 번식체계 확립 및 재분화 식물체의 광합성 능력증대 I. 구절초의 기내배양 및 재분화 식물체의 RAPD 분석)

  • 김정률
    • Korean Journal of Plant Resources
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    • v.11 no.1
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    • pp.1-8
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    • 1998
  • This study was conducted to establish mass propagation system from the axillary bud culture of chrysanthemum zawadskii H. which was used as material of medicinal plants. Shoot egeneration was better on MS medium with NAA and BA. The optimum concentraions of growth regulator for shoot regeneration differed depending on accessionsof C. Zawadskii. Shoot regeneration in Keungucheolcho was better on MS Medium with NAA 0.01mg/1 and BA 0.1mg/1 while Hyangrobonggucheocho was better with NAA 0.1mg/1and BA 0.3mg/1. Addition of NAA into medium was effective for induction of root from shoots regenerated. Shoot multiplcation was more effective when 10mg/1 spermine was added into medium than when other polyamines were treated ino medium . Randomly and specifically amplified polymorphic DAC banding patterns based on polymerase chain reaction (PCR) analysis were used to assess the genetic variation of plants regenerated from in vitro culture.

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