• Title/Summary/Keyword: cotyledon explants

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Relationship of Cotyledon Number with Procambium Differentiation in Somatic Embryogenesis of Codonopsis lanceolata L. (더덕의 체세포배발생에서 전형층분화와 자엽수의 관계)

  • Choi Pil-Son;Soh Woong-Young;Cho Duck-Yee;Liu Jang-Ryol
    • Journal of Plant Biotechnology
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    • v.32 no.2
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    • pp.135-138
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    • 2005
  • Embryogenic callus was obtained from cotyledonary explants of Codonopsis lanceloata on Murashige & Skoog's medium supplemented with 1 mg/L 2,4-D. Suspension cultures of the embryogenic calli were grown on a shaker at 100 strokes/min, and then the calli were subcultured for 2 weeks in 2,4-D-free medium to produce somatic embryo. In somatic embryos at the globular stage, cotyledon initials began to differentiate themselves in the near distal end of the procambial strand. Dicotyledons, tricotyledon, tetracotyledon and fused cotyledon were differentiated from the distal ends of two, three, four and circular procambial strands, respectively. Nearly circular procambial strand in lower hypocotyls were independently differentiated into two, three, four procambial tissues at cotyledonary node and cotyledons to form somatic embryos with dicotyledon, tricotyledon, tetracotyledon. If the distal subepidermal cells of globular embryo exclusively became cotyledon initials, the torpedo or cotyledonary embryo was characterized by somatic embryos with fused cotyledon.

Developmental and Structural Diversity of Regenerated Plants in Cell and Tissue Cultures (세포조직배양계에서 재생된 식물의 발생 및 형태학적 다양성)

  • 소웅영
    • Proceedings of the Botanical Society of Korea Conference
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    • 1993.07a
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    • pp.1-36
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    • 1993
  • It is possible to regenerate plants from calli, single cells and protoplasts of numerous species via organogenasis or embryogenesis in cell and tissue culture systems. Also such regeneration of plants can directly occur from cells of explants. However certain plant species has not been yet provided cultures suitable for plant regeneration from cells or tissues. For example, we have to confirm the regenerability of plant from cells before preparing transformed cells for application. Even more, it is very important to notice that regenerated plants in cell and tissue cultures often show structural abnormality. The mojority of those plants is functionally disordered and eventually cases degenerated. One of such examples is vitreous plants which are manifested mainly in the leaves and manifesteds to a lesser extent in the stems and roots. Regenerants in suspension cultures show more frequent vitrification than on gelled media so that relative humidity and water potential are the key factors involved in abnormal morphogenesis in vitro. The other is that somatic embryos formed in media containing BAP or high concentration of sucrose show frequently cotyledon aberrancy such as polycotyledon and born type cotyledon. The embryos with aberrant cotyledon of Codonopsis lanceolata could not germinate or regenerate into plants in many cases. In contrast, the polycotyledon embryos of Aralia cordata germinated in higher percentage than two cotyledonary embryos, but horn type cotyledonary embryos rarely germinated. The major cause of poor germination is the abnormal development of plumule apex meristem.

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Plant Regeneration from Cotyledon and Hypocotyl Tissues of Chinese Cabbage (배추의 자엽과 배축 절편체로부터의 식물체 재분화)

  • Kang, Byung-Kook;Lim, Chae-Wan;Chung, Kyu-Hwan;Park, Young-Doo
    • Horticultural Science & Technology
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    • v.19 no.3
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    • pp.315-319
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    • 2001
  • The study was carried out to develop a simple and efficient system to regenerate plants from cotyledon and hypocotyl tissues of Chinese cabbage (Brassica campestris L. ssp. pekinensis cv Seoul). Among the various combinations of naphthalene acetic acid (NAA) and 6-benzyladenine (BA) tested, the best shoot induction medium for cotyledon, with 2.67 shoots per explants, contained $2.0mg{\cdot}L^{-1}$ NAA, $1.0mg{\cdot}L^{-1}$ BA and $16.7mg{\cdot}L^{-1}$ $AgNO_3$. The shoot induction medium with $1.0mg{\cdot}L^{-1}$ NAA, $5.0mg{\cdot}L^{-1}$ BA and $16.7mg{\cdot}L^{-1}$ $AgNO_3$, was best for shoot induction from hypocotyl explants, with 1.87 shoots per explants. After shoot induction, regenerated shoots were excised and rooted on rooting medium. Rooted plantlets were then hardened in the high humidity growth chamber and transplanted to pots, and then grown in the greenhouse. Regenerated plants appeared phenotypically normal and there were no changes in chromosome number.

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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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Effects of Growth Regulators on the Formation of Somatic Embryo and Adventitious Bud from the Cotyledon of Korean Ginseng (Panax ginseng C.A. Meyer) (고려인삼(Panax ginseng C.A. Meyer)의 자엽으로부터 체세포배 및 부정아의 발생에 미치는 식물호르몬의 영향)

  • Yang Deok-Chun;Yoon Eui-Soo;Choi Kwang-Tae
    • Journal of Ginseng Research
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    • v.23 no.4
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    • pp.199-204
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    • 1999
  • Cotyledon explants of Korean ginseng (Panax ginseng C.A. Meyer), a perennial medicinal plant, produced direct somatic embryos at a high frequency on MS medium without growth regulators. Cytokinin highly suppressed the somatic embryogenesis but stimulated direct fomlation of adventitious buds. BAP was more effective than kinetin for the formation of adventitious bud. IBA combination with cytokinin enhanced the frequency of adventitious bud formation. The highest frequency of adventitious bud formation were $40\%$ at 0.05 mg/l IBA and 5 mg/l BAP. Adventitious buds were mainly formed near the distal portion of cotyledon, while somatic embryos were only formed near the proximal portion of cotyledon. Adventitious buds were covered with sheath similar to axillary buds in the zygotic embryos, and then leaf-like epicotyls were developed.

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Plant Regeneration from Cotyledon and Hypocotyl Culture in Apple Hybrid Seedling (P.16 $\times$ Malus prunifolia) (사과 P.16 $\times$ Malus prunifolia 교잡실생의 자엽 및 배축배양에 의한 식물체 재분화)

  • 김송남;오성도;김영숙
    • Korean Journal of Plant Tissue Culture
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    • v.27 no.3
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    • pp.191-195
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    • 2000
  • Adventitious shoots were induced from cotyledon and hypocotyl explants of apple hybrid seedlings (P.16$\times$ Malus prunifolia) on MS medium supplement with 2,4-D and various cytokinine (Kn. BA, TDZ) The shoot regeneration from the cotyledon culture was the highest on MS medium supplemented with 1.0 mg/L NAA and 2.0 mg/L BA. Whereas in case of hypocotyl culture, it was the highest on MS medium supplemented with 0.5 mg/L NAA and 0.5 mg/L BA. However, in the MS medium without BA there was no shoot regeneration. Hypocotyl culture seemed to be more effective than cotyledon culture in shoot regeneration. Specially, the top position of the hypocotyl found to be the best explant for shoot induction among the other segments of hypocotyls. Regenerated shoots were rooted on half-stength MS medium with 1.0 mg/L NAA. Above results suggest that Apple hybrid (P.16 $\times$ Malus prunifolia) can be multiplied via cotyledon or hypocotyl culture systems.

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Somatic embryogenesis and plant regeneration in zygotic embryo explant cultures of rugosa rose

  • Kim, Suk Weon;Oh, Myung Jin;Liu, Jang R.
    • Plant Biotechnology Reports
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    • v.3 no.3
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    • pp.199-203
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    • 2009
  • Rugosa rose (Rosa rugosa) is cultivated as a garden flower and an important genetic resource for the breeding of roses (R. hybrida). This study describes culture conditions for high frequency plant regeneration from zygotic embryo explants via somatic embryogenesis in rugosa rose. Mature zygotic embryo, cotyledon, and radicle explants formed embryogenic calluses at frequencies of 38, 6.7, and 8.8% when cultured on half-strength Murashige and Skoog medium (${\frac{1}{2}}MS$) supplemented with 2.26, 9.05, and $9.05{\mu}M$ 2,4-dichlorophenoxyacetic acid, respectively. Embryogenic calluses produced numerous somatic embryos, which then developed into plantlets on ${\frac{1}{2}}MS$ without growth regulators. Regenerated plantlets were grown to whole plants in a growth chamber.

Thidiazuron Induced High Frequency Adventitious Shoot Formation and Plant Regeneration in Capsicum annuum L.

  • VENKATAIAH PEDDABOINA;CHRISTOPHER THAMIDALA;SUBHASH KARAMPURI
    • Journal of Plant Biotechnology
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    • v.5 no.4
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    • pp.245-250
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    • 2003
  • An efficient procedure was developed for adventitious shoot bud induction and plantlet regeneration from various explants of the ten genotypes of Pepper (Capsicum annuum L.) using Thidiazuron (TDZ). Among various treatments at 1.0-3.0 mg/L TDZ Induced maximum number of adventitious shoots depending upon the explant type and genotype compared to other treatments. Among the explants tested, leaf induced maximum number of adventitious shoots than the cotyledons. TDZ-mediated organo-genesis was possible in 10 pepper cultivars, the extent of the response being genotype-dependent. Of the ten genotypes tested, C. annuum cvs CA960, $G_4$ and X-235 were produced maximum number of adventitious shoots and Sell was the least, and all other genotypes gave moderate response. Elongation of multiple shoots was observed on medium supplemented with SA (0.05 mg/L) in combination of IAA (0.05 mg/L). Differences in ability for in vitro shoot regeneration and elongation depend upon the variety and explant type. The elongated shoots were success. Fully rooted on MS medium containing at 1.0 mG/L IAA. Plantlets regenerated from different explants of ten genotypes were found to be diploid (2n=24) and were devoid of any chromosomal aberrations. Regenerated plants were successfully established in soil where 85-90% of them developed into morphologically normal and fertile plants.

Effects of different types and ages of explants and cytokinins on shoot regeneration in Brassica juncea L. (절편부위와 사이토키닌이 갓의 재분화에 미치는 영향)

  • Roh, Kyung Hee;Kwak, Bo-Kyung;Kim, Jong-Bum;Lee, Kyeong-Ryeol;Kim, Hyun Uk;Kim, Sun Hee;Kang, Han Chul
    • Journal of Plant Biotechnology
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    • v.40 no.2
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    • pp.72-78
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    • 2013
  • To establish an efficient protocol for plant regeneration of Brassica juncea L. Czern, the effects of explant types, explant ages and cytokinins on shoot regeneration were examined in this study. Shoot regeneration was markedly affected by the explant types used in the following order: cotyledon with petiole> hypocotyl> leaf with petiole> cotyledon> leaf. Five-day-old seedlings of cotyledon with petiole explants showed the maximum shoot regeneration frequency. Of the six cytokinins-6-${\gamma}$-${\gamma}$-Dimethylallylamino-purine (2-ip), 6-${\gamma}$-${\gamma}$-Dimethylallylamino-purine riboside (2-ip riboside), 6-Benzyl amino-purine (BAP), Thidiazuron (TDZ), Zeatin, Zeatin riboside-TDZ ($8{\mu}M$) was found to be the best cytokinin for shoot regeneration with the highest shoot induction frequency (80%) from cotyledon with petiole after 4 weeks. All the regenerated plantlets were developed well and they produced morphologically normal flowers.

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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