• Title/Summary/Keyword: Plant Cell Culture

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식물세포의 관류배양을 위한 초음파 분리기의 개발

  • Gu, Yeong-Han;Jo, Gyu-Heon
    • 한국생물공학회:학술대회논문집
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    • 2000.11a
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    • pp.402-404
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    • 2000
  • In this study, we have developed an ultrasonic separation system for plant cells and its operating conditions in terms of voltage, flow rate and concentration were examined. For plant cell, the operation of ultrasonic separator highly depended on concentration of cells. Holding capacity highly depended on flow rate in chamber. Optimum voltage was 30V in high density culture

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Chromosome Aberration in Suspension Culture of Ginseng(Panax ginseng C. A. Meyer) Callus (인삼 캘러스 현탁배양에 있어서의 염색체 이상)

  • Park, Jong-Bum
    • Journal of Environmental Science International
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    • v.15 no.12
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    • pp.1193-1197
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    • 2006
  • This study was to examine the variations of chromosome number and the ranges of variety in the suspension culture of ginseng (Panax ginseng C. A. Meyer) callus cell, and the effect of plant hormones for the chromosome aberration. Plant hormones added with MS medium in the suspension culture were 2,4-D, kinetin, and 2,4-D+kinetin and concentration of the plant hormones were $1000{\mu}M$ and $0.1\;{\mu}M$ respectively. As a result of these experiment the following conclusion has been obtained. Media contained with 2,4-D+kinetin in $10{\mu}M$ concentration was very effective in the suspension culture result from 26.4% mitosis frequency, and found the various variation of chromosome number. Variety of chromosome number was diversed ($9\sim110$), espicially frequency of hypohaploid and hyperhaploid cells were very higher than hyperdiploid cells. In this experiments, it is suggested that $10{\mu}M$ 2,4-D+kinetin added with medium in the suspension culture of ginseng callus was effect in the variations of chromosome number.

Enhancement of Growth and Secondary Metabolite Biosynthesis: Effect of Elicitors Derived from Plants and Insects

  • Jeong Gwi-Taek;Park Don-Hee
    • Biotechnology and Bioprocess Engineering:BBE
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    • v.10 no.1
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    • pp.73-77
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    • 2005
  • Plant-derived natural products have been and will continue to be valuable sources. Elicitors have been employed to modify cell metabolism in order to enhance the productivity of useful metabolites in plant cell/tissue cultures. In this study, several elicitors were used to improve the productivity of useful metabolites and to reduce culture time for archiving high concentration in P. ginseng hairy root cultures. The addition of chitosan, chitosan oligosaccharide and alginate oligosaccharide to the culture of P. ginseng hairy roots caused growth to be inhibited with the increase in elicitor concentration. The usage of the chitosan elicitor and D-glucosamine caused a slight decrease in hairy root growth, whereas total ginseng saponin accumulated slightly with the increase in elicitor concentration. When gel beads were added to the culture medium at the initial period, hairy root growth was enhanced. The maximum growth was 1.35 times higher than that of the control at $1\%$ (w/v). Total ginseng saponin content decreased due to the addition of alginate beads. This would result in consistent diffusion of lower levels of calcium ions during the culture period that promotes biomass growth.

Polyploidy and Acclimatization Rate of Broccoli Plane Derived from Anther Culture (葯培養 由來 녹색꽃양배추(Broccoli)의 幼植物 馴化 및 培數性 調査)

  • 이광식;박재복;안춘희;윤여중
    • Korean Journal of Plant Tissue Culture
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    • v.25 no.5
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    • pp.315-318
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    • 1998
  • This study was conducted to clarify the acclimatization rate and ploidy level of anther culture-derived plants in broccoli. The acclimatization rate from 71.4 to 100% was obtained from 7 varieties in 2 years. It was possible to identify the ploidy of the Plants obtained through anther culture by measuring the number of chloroplast in the guard cell. The average numbers of chloroplasts per guard cell in haploid diploid, and tetraploid were 8.5, 13.5 and 18.5, respectively. The regenerated plants could be classified based on these results into 47.1-51.3% of haploids, 47.9∼51.7%, diploid, and 0.8∼l.2% of tetraploids.

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Histological Observation of Embryogenic and Non-embryogenic Callus in Long-term Subculture of Wild Viola (Viola patrinii DC.) (흰제비꽃 배양세포에 있어서 분화세포와 미분화세포 조직의 비교 관찰)

  • 정용모;손병구;이재헌;서정해;정정한;권오창
    • Korean Journal of Plant Tissue Culture
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    • v.27 no.3
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    • pp.233-238
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    • 2000
  • To obtain a basic information of the development of Genus Viola, morphological and histological observation of in vitro calli and cells in Viola culture cells were investigated. There were two callus types obtained by long term subculture of wild viola (Viola partrinii DC. ) petiole callus. One was friable callus - soft and pale green in color and small cells in size, and the other was compact callus - compact and deep bluish green in color, large cells in size. In scanning electron microscopic observation, friable callus was composed of voculated cell around small. cell clump, while compact callus was composed of cells filled with protoplasm Somatic embryogenesis was observed from suspension culture of the compact callus.

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Isolation and culture of protoplasts from leaf tissue of Capsicum annnum var. accumnatum Fingerh and C. frutescensL. [Syn. C. minimum Roxb.] (Bird chilli)

  • Lee, Kue-Jae;Lee, Wang-Hyu
    • Proceedings of the Plant Resources Society of Korea Conference
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    • 2003.10a
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    • pp.50-58
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    • 2003
  • Isolation and culture of leaf protoplasts from two chilli cultivars (Capsicum annuum var. accumnatum and Bird chilli) were developed to enhance selection process in the somatic hybridization programmes. In order to isolate the protoplasts from leaves of these two chilli cultivars different incubation periods (3, 5 and 10 hours) were tested with combinations of enzyme mixtures containing cellulase and macerozyme. Leaves were incubated on three enzyme mixtures (2% cellulase + 0.4% macerozyme, 1% cellulase + 0.2% macerozyme and 0.5% cellulase + 0.1 % macerozyme in 13% mannitol) at 251oC in the dark. Three hours of incubation using 2% cellulase and 0.4% macerozyme was the best for the protoplast isolation of both chilli cultivars tested. The yield was 5 ${\times}$ 108protoplasts/ml/ g leaf tissue in both chilli varieties. It was found that in the mixed nurse method using Nagata and Takebe (NT) medium supplemented with 1.0mg/12,4-D, NAA and BAP with 0.5M mannitol and 1.2% Sea Plaque agarose is the best medium for protoplast culture. Protoplasts of Capsicum annum var. accumnatum were alive for 14 days forming cell walls and initiating cell division.

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STUDIES ON THE TISSUE CULTURE OF PANAX GINSENG

  • Harn C
    • Proceedings of the Ginseng society Conference
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    • 1974.09a
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    • pp.9-22
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    • 1974
  • Unlike the tissue culture in animals and human being, in higher plants various parts of the plant are cultured for varied purposes, and they are named variously depending on which parts are used as explants or what purposes they are cultured for. Followings are some of the names of culture used frequently: organ culture, tissue culture, callus culture, single cell culture, meristem culture, mericlone culture, ovary culture, ovule culture, embryo culture, endosperm culture, anther culture, pollen culture, protoplast culture, etc.. As the names of the culture indicate, in some kinds of culture the explants used for culture are actually not tissues, but organs, single cells, or protoplasts. It seems, however, convenient to call all of the above-mentioned cultures grossly as tissue culture. Several kinds of tissue culture were attempted using Panax ginseng as material and some of the results were summarized below. 1. Callus culture After dormancy of the sed was broken, whole embryo or parts (hypocotyl, cotyledon and epicotyl) of partly grown embryo were cultured in the media supplemented with growth regulators. Rapid swelling occurred in a few weeks, but most of the swelling was observed only in the basal part of epicotyl, changes in the other parts of embryo appearing in much later stages. The swelling or increase in size, however, was resulted not from the divisions of cells, but from the mere expansion of cell. Real calli were formed about two months after inoculation of explants. Callus tissues developed from cortex, pith, and vascular bundle in the cases of hypo- and epicotyl, from mesophyl tissue in the case of cotyledon. Shoots developed more easily from cotyledons regardless of whether they are detached from or attached to the embryo proper. 2. Culture in the Knudson C medium When cotyledons, detached from or attached to the embryo proper, were cultured in the growth regulator-free Knudson C medium comprision only several kinds of mineral compounds and sucrose, shoot primordium or callus developed profusely and finally plantlets were produced directly from shoot primordium or indirectly through callus. In this medium epidermal cells as well as mesophyl cells of the cotyledon became meristematic and divided, changing into multinucleate cells or multicellular bodies, developing eventually into either shoot primordia or calli. 3. Anther culture Anthers were cultured in the media supplemented with various growth regulators applied singly or in combinations. Callus was formed mostly in the connective tissue of anther. Cells of anther wall layers changed in appearance, but no division occurred. Microspores of all stages in development were not changed, ruling out the possibility that microspore-originated callus might be formed. 4. Isolation of protoplast Protoplasts were isolated from young root, leaf, and epicotyl, using 0.7M D-mannitols as osmoticum and using macerozyme and cellulase respectively for maceration and digestion of the cell wall. Production in large number of naked intact protoplast was rather difficult as compared with other plant species. Fusion of protoplasts occurred infrequently mainly due to the fewer number of naked protoplasts in the solution.

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Antibody Production in Plant Cell Cultures

  • Lee, James M.
    • Proceedings of the Botanical Society of Korea Conference
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    • 1995.06a
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    • pp.67-78
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    • 1995
  • Monoclonal antibodies (MoAbs) are a highly diversified class of proteins with major research and commercial applications such as diagnostics and therapeutics. Currently, the dominant method for producing MoAbs is through the hybridoma technique. However, this technique is slow, tedious, labor intensive, and expensive. The production of MoAbs in cultured transgenic plant cells can offer some advantages over that in the over that in the mammalian systems. The media to cultivate plant cells are well defined and inexpensive. Contamination by bacteria or fungi is easily monitored in plant tissue cultures. Furthermore, these contaminants are usually not potent pathogens to human beings. In our interdisciplinary research efforts, heavy chain monoclonal antibody (HC MAb) was inserted into Ti plasmid vector and transferred into A. tumefaciens for the transformation in tobacco cells. It was found that 76% of the transformants produced HC MAb. The presence of HC MAb in the cell membrane fraction indicated that the signal peptide was functional and efficient. The change of the HC MAb concentration during a batch culture followed a similar trend as dry cell concentration, indicating that the production of HC MAb was growth related. The long-term repeated subcultures of 11 cell lines showed that there was no obvious trend of neither the decrease nor the increase of the productivity with the repeated subcultures.

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A Study on the Cytotoxic Effects of Several Plant Extracts on the Cell viability and Cell Adhesion Activity in Cultured NIH3T3 Fibroblast (몇 가지 식물추출물이 배양 NIH3T3 섬유모세포의 세포생존율과 세포부착률에 미치는 세포독성에 관한 연구)

  • Rim, Yo-Sup;Song, Won-Seob;Seo, Young-Mi;Park, Seung-Taeck;Kim, Shin-Moo
    • Korean Journal of Clinical Laboratory Science
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    • v.42 no.3
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    • pp.116-124
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    • 2010
  • This study was aimed to clerify the cytotoxicity of some plant extracts such as Hosta longissima HONDA (HL), Hemerocallis fulva var. Kwanso REGL (HFVK), Hemerocallis fulva L (HF), Macrocapium officinale NAKAI (MO) and Mentha canadensis var. piperascens HARA (MCVP), the cultured NIH3T3 fibroblasts were treated with 25, 50, 100, 150 and $200{\mu}g/mL$ of five kinds of plant extracts for 48 hours, respectively. The cytotoxicity of plant extracts was measured by MTT and NR assays for the cell viability, and XTT assay for the cell adhesion activity. In this study, HL, MO and FHVK extracts showed the range of midtoxic-non toxic by the criteria of chemical cytotoxicity. While, the HF and MCVP extracts showed midtoxic. In the extract cytotoxicity, HL, MO and FHVK extracts showed non-toxic by the criteria of extract cytotoxicity. While, HF extract was determined as lower-toxic. In the responsive sensitivity of each plant extract on colorimetric assays, HF extract was sensitive to mitochondrial enzyme by MTT assay, lysosomal enzyme by NR assay and mitochondrial nucleus by XTT assay. While, MCVP extract was sensitive to mitochondrial enzyme by MTT assay and lysosomal enzyme by NR assay than other assays. While, HL, HFVK and MO extracts were most sensitive to NR assay. Cell culture is one of useful materials in the screening of cytotoxic and recovary effect on the putative chemical agents or plant extract. And also, colorimetric assay is regarded as very useful tools for quantitative measurement of cytotoxic effect on plant extracts in vitro.

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