• 제목/요약/키워드: two-cotyledon embryo

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당근의 체세포배로부터 발생한 경정부의 구조적 특성 (Structural Characteristics of Shoot Apex Developed from Somatic Embryos of Daucus carota L.)

  • 홍성식;소웅영
    • 한국자원식물학회지
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    • 제12권2호
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    • pp.133-138
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    • 1999
  • 당근(Daucus carota L. cv. Hongshim)의 접합자배와 체세포배의 경정부의 구조는 주사전자현미경과 종단면 관찰을 통하여 이루어졌다. 접합자배의 경정부는 도립된 보우트형이고, 2ㆍ3ㆍ4자엽 체세포배의 경정부는 각각 도립된 보우트형, 피라미드형 및 볼록한 다이아몬드형으로 나타났다. 접합자배의 경정부는 다소 작은 세포들로 구성되어 있고, 초층과 내체의 구별이 된다. 체세포배의 경정부는 다소 큰 세포들이 불규칙하게 배열되어 있다.

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당근의 배양세포로부터 형성된 체세포배의 다자엽 구조 (Multicotyledonary Structure of Somatic Embryos Formed from Cell Cultures of Daucus carota L.)

  • 소웅영
    • Journal of Plant Biology
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    • 제39권1호
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    • pp.71-77
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    • 1996
  • 배발생 캘러스의 선발은 1.0mg/L 2,4-D를 첨가한 MS 기본배지에서 배양한 당근 유식물의 배축절편으로부터 유도된 캘러스에서 이뤄졌다. 체세포배 발생을 유도하기 위해서 배발생 캘러스로부터 얻어진 세포괴는 2,4-D를 제거한 MS 배지에 옮겼다. 세포괴를 1.0mg/L 2,4-D 첨가배지에서 1주간 그리고 2,4-D 제거 배지에서 2주간 배양한 후에 2자엽배의 발생은 63%에 그쳤고 나머지는 1자엽 5%, 3자엽 21%, 4자엽 6%, 5자엽 5%, 6자엽 0.2% 및 나팔형 자엽 1%의 발생빈도를 나타냈다. 체세포배의 발아율은 다자엽 체세포배에서 보다 2자엽 배에서 높았다. 나팔형 자엽의 체세포배는 경엽부 발생없이 자엽과 뿌리가 다소 신장 확대되는데 그치는 등 정상적으로 발아되지 못했다. 해부학적인 관찰에서 체세포배의 뿌리와 환상 전형성층은 배축의 중간부위에서 분지되기 시작하고 자엽절을 거쳐서 자엽으로 이어지는데 배축에서 전형성층속은 자엽수와 같은 수였다. 1자엽 체세포배는 다자엽 체세포배의 각각의 자엽보다 크고 전형성층과 함께 말굽형 자엽을 가지고 있었다. 따라서 체세포배의 자엽구조는 전형성층 배열상태와 밀접한 관련이 있다는 것을 확인할 수 있었다.

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STUDIES ON THE TISSUE CULTURE OF PANAX GINSENG

  • Harn C
    • 고려인삼학회:학술대회논문집
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    • 고려인삼학회 1974년도 학술대회지
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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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Effect of 2,4-D on embryo formation and its morphology in anther culture of herbaceous peony (Paeonia lactiflora Pall.)

  • Park, Gyu-Hwan;Kim, Dai-Hee;Kim, Jin-Ho;Choi, Yong-Hwa;Oh, Jung-Youl;Kwon, Yong-Sham;Kim, Myung-Min
    • Journal of Life Science
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    • 제12권1호
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    • pp.19-21
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    • 2002
  • The pathway of embryos formed anther culture in herbaceous peony was influenced by addition of 2,4-D. MS medium with 2,4-Dichlorophenoxy acetic acid (2,4-D) alone did not arise direct embryogenesis, but was proliferate callus. Embryos through calli were produced on medium containing 0.2 mg/1 zeatin or without growth regulators. Direct embryogenesis was obtained from MS basal medium. However, after the anthers were cultured on medium with 0.1 mg/1 2,4-D, 3 g/1 AC, 30 g/1 sucrose, 2 g/1 gelrite for 40 days. Its efficiency (32.3 %) was markedly improved when anthers cultured on medium without 2,4-D. Embryo morphology was also affected by the 2,4-D used in medium. The induction of normal embryos with two cotyledons was higher in the embryos formed through direct embryogenesis than those formed callus. The embryos formed from calli were mainly showed abnormal embryo with one, three, four cotyledons or hors and bowling pin type.

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당근(Daucus carota L.)의 현탁배양을 통한 체세포배 발생에 미치는 카드뮴의 영향 (Effect of Cadmium on Somatic Embryogenesis from Cell Culture of Daucus carota L.)

  • 조덕이;신은경;소웅영
    • 식물조직배양학회지
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    • 제27권3호
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    • pp.227-232
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    • 2000
  • 당근 유식물체의 자엽을 절편으로 유도된 배형성능 세포를 여러가지 농도의 카드뮴 (0, 50. 100, 200. 500, 1000 $\mu$M)이 첨가된 MS 액체배지에 배양한 다음 MS 기본배지에 옮겨서 체세포배 발생을 유도했고 한편 카드뮴 처리구와 카드뮴이 처리되지 않은 대조구에서 체세포배를 얻었다. 현탁배양에서 정상적인 2개의 자엽을 갖는 체세포배 발생은 카드뮴 첨가 배지에서 감소하였으나 비정상적인 3개의 자엽을 갖는 체세포배가 많이 관찰되었다. 또한 비정상적인 1개, 3개 및 4개의 자엽을 갖는 체세포배보다 정상적인 2개의 자엽을 갖는 체세포배의 식물체 재생률이 가장 높았다. 카드뮴 처리구에서 발생된 체세포배의 발아율은 자엽수에 관계없이 대조구에서 발생된 체세포배의 발아율에 비해 감소하였다.

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동백나무 미숙배 배양으로부터 비정상 체세포배 형성과 식물체 재생 (Anomalous somatic embryos formation and plant regeneration from the cultures of immature embryos of Camellia japonica L.)

  • 최종혜;권석윤;최필선
    • Journal of Plant Biotechnology
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    • 제38권4호
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    • pp.258-262
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    • 2011
  • 동백의 배발생 캘러스는 1 mg/L 2,4-D가 첨가된 MS배지에서 미숙배 배양을 통해 유도하였으며, 동일배지에 계대 배양하여 4주 이상 증식시켰다. 배발생 캘러스로부터 자엽기 체세포배를 생산하기 위하여 2,4-D가 첨가되지 않은 MS기본배지 배양하였다. 동백 미숙배로부터 체세포배 발생율은 25.1%였으며, 형성된 대부분의 체세포배중에서 48.3%가 cup모양, 12.6%가 1개 자엽, 9.4%가 3개자엽, 1.9%가 4개 자엽을 갖는 비정상적인 형태의 체세포배였으며, 반면 2개의 자엽을 갖는 정상적인 체세포배는 27.5%였다. 정상 및 비정상형태의 체세포배를 MS기본배지, $\frac{1}{2}$MS배지, 0.1 mg/L $GA_3$가 첨가된 $\frac{1}{2}$MS배지 및 0.1 mg/L IBA와 $GA_3$가 조합 첨가된 $\frac{1}{2}$MS배지에 치상하였을때 2개의 자엽을 갖는 정상체세포배의 식물체 재생율이 11.1%로 0.0 ~ 8.3%를 나타내는 1개 자엽 (0.0 ~ 8.3%), 3개 자엽 (0.0 ~ 5.8%), 4개 자엽 (0.0%) 및 cup 모양 (0.3 ~ 4.2%) 보다 높았다. 이러한 결과로 볼 때 비정상 자엽을 갖는 체세포배 빈도가 높을수록 식물체 재생빈도가 감소될 수 있음을 알 수 있었다.

알팔파 캘러스로부터 형성된 이차체세포배의 형태가 유식물 발달에 미치는 영향 (Effect of Embryo Morphology on Plant Development in Secondary Somatic Embryogenesis of Alfalfa)

  • 원성혜;이병현;조진기
    • 한국초지조사료학회지
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    • 제19권4호
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    • pp.297-302
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    • 1999
  • 알파파의 재분화된 식물체로부터 다량의 이차체세포배를 유도하였으며, 이들 체세포배로부터 재분화되는 식물체의 획득빈도를 향상시키기 위하여, 캘러스에서 유래된 배의 형태 및 형태별 유식물 분화 양상에 관한 실험을 수행하였다. 2,4-D 농도에 따라 체세포배의 형성에 차이를 나타내었는데, 생장조절제가 첨가되지 않았거나, 2,4-D가 $0.1mg/{\ell}$ 첨가된 배지에서 형성된 배는 약 57% 이상이 2개의 자엽을 갖은 정상배였으며, 2,4-D 농도가 증가할수록 정상체세포배의 출현빈도는 감소하였고, 2,4-D $4mg/{\ell}$에서는 10%만이 정상배로 나타났다. 배의 형태에 따른 발아율 및 유식물 분화양상을 조사한 결과, 2개의 자엽을 갖는 정상배의 경우는 발아율이 85%로 가장 높았으며, 정상식물체로 발육되는 비율도 80%로 나타났다. 그러나 자엽이 1개 또는 3~4개인 배는 정상적으로 발육되는 식물체의 비율이 10% 이하로 매우 낮았으며, 자엽이 5개인 배와 나팔모양의 배는 정상적인 식물체로 발달하지 못하였다.

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Esterase isozyme patterns in developing plant regeneration from calli of citrus junos Sieb.

  • Lee, Hyun-Hwa;Lee, Sook-Young;Park, Min-Hee;Jang, Hyun-kyu;Kim, Hong-Sub
    • Plant Resources
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    • 제2권1호
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    • pp.1-9
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    • 1999
  • The callus from the hypocotyl region of immature embryo of Citrus junos Sieb. was efficiently induced in the $\frac{1}{2}$ MS medium containing 45uM BA after 8 weeks culture. The callus was developed into the two callus type, embryogenic callus and nonembryogenic callus, which can be distinguished by visual examination depending on color and appearence. In vitro regeneration of callus established efficiently in the hormone-free MS medium from the embryogenic callus. In order to investigate the physiological changes depending on the developmental stage of embryo, the embryo was formed in the MS medium. The embryogenic and nonembryogenic callus, and the various stages of the somatic embryo were examimed the changes of esterase activity, and their isozyme patterns as well. The protein content and esterase activities was gradually increased on the developmental stages of embryo. Total protein pattern were different by the SDS-PAGE and were appeared strong band of 23 KD in the torpedo stage. The pattern of the esterase isozyme was exhibited a difference between embryogenic callus and nonembryogenic callus. It was appered pI 6.0, 8.0, 8.2 in the embryogenic callus. Also the new band of pI 4.75 was appeared in the cotyledon. These results suggest that the changes of esterase activities and isozyme patterns are importent factor in the differentiation and development of citrus.

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Germination of Hemerocallis Seeds as Influenced by Seed Development and Temperature Treatments

  • Kim, Ji Hee;Suh, Jeung Keun;Lee, Ae Kyung
    • 원예과학기술지
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    • 제34권6호
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    • pp.830-839
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    • 2016
  • Germination of Hemerocallis seeds as influenced by cold stratification at $5^{\circ}C$ (CS) and maturity of seeds evaluated using X-ray imaging has not been well investigated. Seeds of H. lilioasphodelus, H. citrina, H. citrina 'April Flower', and H. minor collected from China and H. thunbergii collected from Korea were germinated at $20^{\circ}C$ without pre-temperature treatment, while H. hongdoensis, H. dumortieri, H. minor, and H. vespertina seeds were treated with CS. Harvesting 'Stella de Oro' capsules at 35-40 days after anthesis yielded mature seeds with well-developed embryo and cotyledons analyzed by X-ray images with a 92% germination in 17 days after sowing. Seeds of H. thunbergii and H. citrina germinated in less than 13 days without CS; two weeks of CS did not accelerate seed germination. Seeds of H. hongdoensis germinated in 24 days when seeds were stored at $25^{\circ}C$ without CS and in less than 27 days when cold stratified. Therefore, 'Stella de Oro' capsules should be harvested at 35-40 days after anthesis to harvest mature seeds. Cold stratification is not required to accelerate seed germination in the Hemerocallis taxa evaluated in this study.

산복사나무(Prunus davidiana FR.)의 성숙배와 유식물에 있어서 유관속계의 분화 (Differentiation of the Vascular System in Mature Embryo and Seedling of Prunus davidiana FR.)

  • 홍성식
    • Journal of Plant Biology
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    • 제28권4호
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    • pp.285-296
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    • 1985
  • To examine the differentiation and transition of vascular system in Prunus davidiana FR., the mature embyro and developing seedlings were embedded in paraplast and treated by clearing method. In mature embryo, the procambium was connected with the epicotyl-hypocotyl-radicle axis and cotyledons, whereas protophloem and protoxylem were restricted primarily to the mid-vein and two lateral veins of the cotyledonary base. With the onset of germination, protophloem and protoxylem were differentiated both acropetally and basipetally from the cotyledonary base. The first and second leaf traces appeared in the cotyledonary node, and then differentiated bidirectionally toward the epicotyl and the root tip. The 3rd to 6th leaf traces were connected with the cotyledonary traces in hypocotyl. At the part of the root tip, the xylem was a diarch. As the first and second leaf traces were superimposed at the middle part of the root, the diarch xylem was changed to a tetrarch. As the cotyledonary traces were diverged below the root base, the tetrarch xylem was changed to an octarch. It was suggested that the vascular system of the epicotyl might be superimposed on that of the cotyledon-hypocotyl-root during the formation of the primary vascular system of Prunus davidiana.

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