• Title/Summary/Keyword: cotyledon culture

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Changes of RNA and Protein During Callus Induction and Plant Regeneration from Perilla frutescens (들깨로부터 Callus의 유기와 재분화에 따른 단백질 및 RNA의 변화)

  • 정상훈;양선경;김현경;정대수;조영수;김도훈
    • Journal of Life Science
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    • v.9 no.1
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    • pp.29-34
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    • 1999
  • Cotyledon and hypocotyl explants of perilla were cultured on MS medium containing a combined concentration of BA(0.5, 1.0 and 1.5mg/$\ell$) and NAA(0.1, 0.5 and 2.0mg/$\ell$) in order to regenerate the explant and induce the callus. The best regeneration of the explant and induction of the callus were observed in a combined concenteration of 0.5mg/$\ell$ of BA and 0.5mg/$\ell$ NAA both in cotyledon and hypocotyl explants. In cotyledon explants, rooting was achieved upon transferring shoots to MS medium containing 0.5mg/$\ell$ of BA and 0.1mg/$\ell$ of NAA. We also investigated the change of protein and RNA content on developmental stage of callus and plant regeneration of perilla. Protein content was increased but RNA content was decreased as the culture period increases. The banding pattern of polypeptide revealed that both 30KD and 45KD polypeptides were obvious in cotyledon obtained from pre-culture explants, but only 30KD polypeptide was further getting obvious as the culture period increases.

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Plantlet Regeneration Cotyledon and Petiole Cultures of Cyclamen Persicum Mill (시클라멘(Cyclamen Persicum Mill.)의 자엽과 엽병배양에 의한 식물체 재분화)

  • 은종선;고정애;김영선
    • Korean Journal of Plant Tissue Culture
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    • v.22 no.4
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    • pp.212-216
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    • 1995
  • These experiments were carried out to examine the effect of explant sources and plant growth regulators on callus induction and plantlet differentiation. Cotyledon and petiole explants of Cyclamen persium were cultured on MS medium supplemented with different concentrations of auxins and cytokinins. Cotyledon cultured on medium containing 2, 4-D and kinetin did not form callus or shoots. But when calli induced from petiole explants on medium with 1.0 mg/L 2, 4D and 1.0 mg/L kinetin were subcultured on the same medium the formation of shoots from calli occurred after 150 days of culture. The combination of NAA and BA were more effective than that of 2, 4 D and kinetin in the formation of shoots from calli, cotyledon culture was most effective on medium with 0.2 mg/L NAA and 0.5 mg/L BA. Shoots excised from calli were rooted on medium with 1.0 mg/L NAA. The plantlets were subsequently transplanted to potting soil.

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Effects of Date and Growth Regulators on the Culture of' Immature Zygotic Embryos of North American Ginseng

  • Hovius, Marilyn H. Y.;Saxena, Praveen K.;Proctor, John T. A.
    • Journal of Ginseng Research
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    • v.31 no.1
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    • pp.14-22
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    • 2007
  • As the zygotic embryo of North American ginseng (Panax quinquefolius L.) matured during stratification over 203 days it grew from 0.75 to 5.2 mm. Embryo excision and culturing on media containing different concentrations of two growth regulators, gibberellic acid ($GA_3$, 1 to 10 ${\mu}M$) and benzyladenine (BA, 1 to 5 ${\mu}M$), during stratification, showed that shoot and root number and the shoot, root and cotyledon length increased with increased stratification time. Gibberellic acid was the more effective growth regulator for increasing shoot and root number and shoot, root and cotyledon lengths. Immature embryos (stratified for up to 63 days) needed growth regulators for further development. Cultures on $GA_3$ at the last culture date (stratified for 203 days) when embryos were mature, produced multiple shoots but there was no effect of $GA_3$ concentration. Benzyladenine inhibited shoot and root growth regardless of embryo stratification. Growth regulators had little effect on cotyledon length of mature embryos. Embryos cultured on $GA_3$ combined with BA were green on all culture dates whereas greening in the control and BA treatments increased with culture date. The BA treatments induced 100% swelling of the embryos on the final culture date while in the control and $GA_3$ treatments there was no swelling. There was little or no curling in the control and BA treatments and a linear decrease in curling with culture date in the $GA_3$ and $GA_3$ + BA treatments.

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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Origin of Direct Somatic Embryos from Cultureed Cotyledon Segments of korean Ginseng (Panax ginseng C.A. Meyer) (한국 인삼 (Panax ginseng C.A. Meyer)의 자엽절편 배양으로부터 형성되는 체세포배의 기원)

  • 최용의;소웅영
    • Korean Journal of Plant Tissue Culture
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    • v.21 no.3
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    • pp.177-182
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    • 1994
  • Cotyledon segments of korean ginseng produced somatic embryos when cultured on MS basal medium, whereas plumule or excised axis explants did not. histological examination revealed that the cells in proximal region of cotyledon turned meristematic and densely cytoplasmic was composed of smaller and more densely cytiplasmic cells than the subepidermal cells. however, in the case both epidermis and subepidermal cells were almost the same in size and cytoplasmic density, the embryo originated from multiple cells.

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Plant Regeneration from Turnip (Brassica rapa ssp. rapifera) Organs

  • Gendaram Sarantuya;Bae Chang-Hyu
    • Plant Resources
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    • v.8 no.3
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    • pp.286-292
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    • 2005
  • Shoot induction system was developed in the recalcitrant plant species, Brassica rapa ssp. rapifera by using optimum selection of profit organ, phytohormone combination, seedling age and kind of culture container. Out of in vitro cultured leaf segment, petiole, hypocotyl, and cotyledon with petiole, only cotyledon with petiole derived from 4 day-old seedlings induced multiple shoot. The optimum combination of auxin and cytokinin for the multiple shoot induction was MS medium containing 5mg/L BA and 0.5mg/L NAA. The major factors for multiple shoot propagation were part of plant organ, age of seedling, and ratio of auxin and cytokinin. In addition, shoot regeneration was promoted in the 100ml Erlenmeyer flask compared with the $90mm{\times}20mm$ Petri-dish. The induced shoots formed roots easy on MS medium containing 0.1mg/L IBA and the whole plants were successfully cultivated in soil.

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Structural Characteristics of Vascular Tissue in Carrot Seedlings with Anomalous Cotyledon Developed from Somatic Embryos (당근의 체세포배로부터 발생한 이상자엽 유식물의 유관속 조직의 구조적 특성)

  • 홍성식;소웅영
    • Korean Journal of Plant Tissue Culture
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    • v.25 no.1
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    • pp.69-74
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    • 1998
  • The somatic embryos of Daucus carota L. cv. Hongshim developed in MS basal medium far 4 weeks had varied number of cotyledons. Palisade and spongy parenchyma of cotyledon were not clearly discriminated in all seedlings developed from the embryos. No independent existence of collateral vascular bundle was observed in all seedlings with various types of cotyledon ; instead, vascular bundles were either interconnected or partially connected with one another. Most of the cotyledonary bases on hypocotyl showed short cylinder structure which encircle plumule. The vascular tissues of cotyledonary bases and nodes of seedlings with jar-shaped or 1 cotyledon were connected in ring forms, showing the pattern of ectophloic shiphonostele, and similar ring form structure was also found in the vascular arrangement of 5 cotyledon seedlings. The vascular bundles of seedlings with 2, 3 and 4 cotyledons in many cases had independently arranged within the cotyledonary bases and nodes, showing the pattern of eustele. In hypocotyl, tetrarch or hexarch xylems prevailed in seedlings with jar-shaped cotyledon or 1 and 5 cotyledon; tetrarch xylems prevailed in 2 cotyledon seedlings; and triarch xylems prevailed in 3 cotyledon seedlings. In most of seedlings, cortex vascular bundles were dispensed in the region from cotyledonary node to hypocotyl, but double vascular bundles were also observed occasionally. In roots, diarch xylems were observed in most of seedlings with 2 cotyledons, triarch xylems in half of seedlings with 3 cotyledons, and diarch xylems in most of the remaining seedlings with the occasional occurrences of tetrarch xylems.

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Changes in the Contents and Composition of Dietary Fiber during the Growth of Soybean Sprout (콩나물 성장중에 식이섬유 함량과 조성의 변화)

  • 이꽃임
    • Journal of Nutrition and Health
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    • v.29 no.10
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    • pp.1142-1149
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    • 1996
  • This study was conducted to determine changes in th contents and composition of dietary fiber during the growth of soybean sprout. Soybean was soaked in water at $25^{\circ}C$ for 2hrs and cultivated at 2$0^{\circ}C$ for 7 days under dark condition. The soybean sprouts were divided into cotyledon and axis and sampled every 24hrs. The analysis methodlogies used were Van Soest's NDF, AOAC's ADF and lignin and Prosky's IDF, SDF, TDF. The weight of 100 sprouts increased gradually from 20.26g to 90.12g during the growth periods. The weight increased to 344.9% of the original weight. The germination rate was 100% after soaking at $25^{\circ}C$ for 2hrs. Root length increased gradualy from 0.6cm at 1st day to 17.2cm at 7th day. The crude ash and crude fat contents showed no significant change in the cotyledon and axis. The crude protein contents increased in the cotyledon and axis, whereas the total carbohydrate content didn't have general tendency. The insoluble dietary fiber(IDF), soluble dietary fiber(SDF) and total dietary fiber(TDF) contents of cotyledon were no significantly different from 20.01%, 1.45%, 21.46% at 1st day to 22.75%, 2.07%, 24.82% at 7th day on dry basis. In axis those contents increased from 23.19%, 1.97%, 25.16% at 1st day to 32.78%, 3.02%, 35.80% at 7th day, respectively. The neutral detergent fiber(NDF) contents of cotyledon and axis increased from 4.35% to 6.39% and from 6.44% to 26.60% respectively on dry basis. The acid detergent fiber (ADF) contents of cotyledon and axis increased from 2.84% to 4.91% and from 2.5% to 4.7%, but there were no significantly different in the hemicellulose and lignin contents on dry basis. The hemicellulose and lignin contents of axis increased with culture periods from 1.70% to 4.41% and from 0.20% to 2.11%, respectively. The cellulose contents increased from 4.54% to 20.35% on dry basis.

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Changes of Protein and Lipid During Callus Induction and Plant Regeneration from Perilla frutescens (들깨 조직으로부터 callus 유기에 따른 지질 및 단백질 조성의 변화)

  • 김현경;김도훈;정순재;남재성;정대수
    • Journal of Life Science
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    • v.11 no.3
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    • pp.254-258
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    • 2001
  • The biochemical change during regeneration of perilla callus were investigated by comparing total protein and lipid contents, protein band pattern in SDS-PAGE, and fatty acid composition in the calli cultured for various period(0, 1, 3, 5 and 6 weeks) Calli were induced from cotyledon and hypocotyl explants of peplants of perilla on perilla on MS medium containing BA(0.5 mg/L) and NAA(0.5mg/L). The protein contents reached the peak at 3 weeks after induction of calli, and then was decreased. Total lipid contents was decreased as the culture period increased. The band pattern of polypeptides showed that 30KD and 45KD polypeptides and 22KD and 45KD polypetides were major proteins in the cotyledon and hypocotyl explants, respectively. However increase of culture period only 30KD protein was highly accumulated.

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Establishment of Efficient Regeneration System Through In Vitro Culture of Lettuce (Lactuca sativa)

  • Kim, Young-Sook;Kwon, Tea-Ho
    • Plant Resources
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    • v.2 no.1
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    • pp.16-21
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    • 1999
  • An efficient regeneration system was established by using in vitro plantlets of germinated seedlings from different cultivars of lettuce (Lactuca sativa cv. Chongchima, Chongchuckmyun, Jeokchima, Jeokchuckmyun). Shoot formation were observed from all cultivars on MS medium supplemented with 0.1 mg/L NAA and 0.5 mg/L BA. In all cultivars, when cotyledon was cultured, the number of shoot per explant was more greater than that hypocotyl and leaf disc were cultured. Shoot formation rate (91.7%) was high in a cotyledon culture of cultivar, Chongchukmyun. The growth of multiple shoots derived from the cultivar, Chongchukmyun, was most effective on medium containing 0.5 mg/L BA and 1.0 mg/L GA$_3$. When shoots were transferred on MS medium without plant growth regulators, roots were effectively differentiated. Rooted plantlets were acclimated on pots for further propagation.

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