• Title/Summary/Keyword: 식물체 재분화

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Identification of mitochondrial mutant (NADH-dehydrogenase) using PCR method and regeneration of mutants from Zea mays (PCR 기법을 사용한 옥수수 미토콘드리아 변이체 (NADH-dehydrogenase)의 선별과 재분화)

  • 설인환
    • Journal of Life Science
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    • v.8 no.1
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    • pp.8-13
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    • 1998
  • The maize mitochondrial mutant (NCS2) is derived from homologous recombination between genes encoding NADH dehydrogenase subunit 4 and subunit 6. Plants from mitochondria mutants exhibited severe related growth and development including dwarfism and striping on the leaves. Aborted embryos from NCS2 mutants have been rescued and cultured on the N6 medium supplemented with 2,4-D 1 mg/l. Most calli from NCS2 aborted embryos showed slow growing pattern at first stage. However, upon continuous culturing them on the medium, those were segregated into mutant and normal callus lines. These segregations could be detected by using PCR method with three primers. Such segregation seems to be resulted from the preferential growth of normal cells over the mutant cells on the normal culture condition. Therefore, this method can be used for determining rate of indirect cytoplasmic segregation by estimating amplified band intensities. When NCS2 mutant callus lines cultured on regeneration medium, no adventitious shoot induction was observed. However, callus lines with more mitochondria induced adventitious shoots. These studies suggest that mitochondria NADH-dehydrogenase for electron transport in the inner membrane of mitochondria is essential for the differentiation and development of plants.

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Plant Regeneration from Internode Explants of Actinidia arguta and its Histological Observation (다래(Actinidia arguta)의 절간절편체로부터 식물체 재분화 및 해부학적 관찰)

  • 김회경;김준철;진창덕
    • Korean Journal of Plant Tissue Culture
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    • v.24 no.5
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    • pp.263-268
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    • 1997
  • Effects of plant growth regulators and condition on shoot induction from internode explants of Actinidia arguta were investigated. Optimal condition for shoot induction was obtained when internode explants were cultured on MS medium containing 3.0 mg/L zeatin. The frequency showed about 62.5% under darks condition at 27$^{\circ}C$ after 6 weeks of culture. Regenerated shoots were rooted on WPM medium supplemented with 0.5 mg/L NAA and the regenerants were acclimated to an artificial soil with mixture (perlite : vermiculite = 1:1, v/v) for further growth and successfully transplanted to the soil in a highly humidified greenhouse. Histological observations revealed that meristemoids were formed from small and isodiametric cells, and then developed to shoot primordia.

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In vitro regeneration and the change of anatomical appearance in Poncirus trifoliata RAFIN. (탱자(Poncirus trifoliata RAFIN.)의 기내 재분화 및 조직학적 특성)

  • 박민희;이현화;장현규;이숙영;김홍섭
    • Korean Journal of Plant Resources
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    • v.12 no.2
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    • pp.107-119
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    • 1999
  • In this study, the induction regeneration of callus from immature embryo in trifoliata orange (Poncirus trifoliata RAFIN.) were accomplished. The embryogenic calli were induced from the immature embryo derived from seed when the calli were irradiated for 16hr at about 2,000 Lux in $\frac{1}{2}$ MS medium supplemented with 3% sucrose, and 44.4$\mu$M BA. Regeneration to whole plants was the most successful in MS medium containing 5.0$\mu$M BA. The yellowish callus was developed at 2 to 3 weeks of culture and the callus was changed from yellow to green at 5 to 6 weeks culture. In vitro regeneration was directly induced from embryogenic callus in MS medium containing 3% sucrose and 5.0$\mu$M BA. Multishoot was formed at 16 weeks culture. Moreover, when the root-formed plantlet was transplanted to soil, they grew to a whole plant. The compact cultured-cells were observed by light microscope after 4 weeks of cultivation and the embryogenic clumps were formed about the 5 weeks. At the same time, the neighboring cells were liquefied. In addition, differentiation of leaf and stem from the callus was observed after 12 weeks. The developed oil sacs and the profacicular cambium of the immature leaf were observed after 18 weeks. Therefore, we can see the considerable changes of cell arrangements according to the developmental stages of calli from trifoliata orange.

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Somatic embryogenesis and plant regeneration of Hovenia dulcis Thunb (헛개나무의 체세포배발생 및 식물체 재분화)

  • Eom, Seung-Hee;Shin, Dong-Yong;Lee, Hyeon-Yong;Kim, Myong-Jo;Kim, Jong-Dai;Choi, Won-Cheol;Heo, Kwon;Yu, Chang-Yeon
    • Korean Journal of Medicinal Crop Science
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    • v.10 no.1
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    • pp.41-45
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    • 2002
  • An efficient and reproducible procedure for the large scale propagation of Hovenia dulcis Thunb. is described. Shoot primodia emerging from the leaf surface was induced from MS medium supplemented with NAA. Stem cuttings were suitable explants for multiple shoot proliferation. They produced axillary shoots which branched repeatedly, yielding an average of 7 shoots per explants after 4 weeks in culture, when cultured on a woody plant medium (WPM) containing 0.1mg/l BA and 0.1mg/l NAA. Stem, leaf and root segments from axenic seedlings were used as explant source to induce somatic embryogenesis. A high frequency of somatic embryos were induced directly from leaf in MS medium with NAA, 2,4-D and in medium containing NAA, 2,4-D with BA. Somatic embryos were germinated in MS medium supplemented with 1mg/ l $GA_3$. Somatic embryos proliferated secondary somatic embryos rapidly after transfer to MS medium supplemented with 1mg/ l kinetin, 1mg/ l $GA_3$ and 2% dextrose.

Embryogenesis and plant regeneration of Panax ginseng Meyer via anther culture and ploidy assessment using flow cytometry (인삼 약 배양을 통한 배 발생과 식물 재분화 및 유세포 분석기를 이용한 배수성 검정)

  • Jung-Woo Lee;Kyong-Hwan Bang;Dong-Hwi Kim;Jang-Uk Kim;Young-Chang Kim;Ick-Hyun Jo
    • Journal of Plant Biotechnology
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    • v.50
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    • pp.19-26
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    • 2023
  • Korean ginseng (Panax ginseng Meyer) is an economically important plant because of it is rich in saponins. It is mainly cultivated in Asia, including Korea and China. Since ginseng requires a long breeding period due to juvenility, homozygote production techniques, such as anther culture, must be urgently established. In the present study, callus induction and embryogenesis through anther culture were observed in P. ginseng. Murashige and Skoog medium was used as the basal medium suitable for callus induction. When the medium was supplemented with 3% sucrose, the callus induction rate was high and the callus size was large. Cold pretreatment did not significantly affect callus induction and embryogenesis. Embryogenesis was the most efficient when the embryo-formation medium was supplemented with 1.0 or 3.0 mg/L 2,4-dichlorophenoxyacetic acid. Cultivar significantly affected anther culture efficiency. Specifically, 'Cheongseon' showed the highest embryo-formation efficiency, whereas no embryogenesis occurred in 'Sunun'. Ploidy assessment revealed the haploid status of the induced calli. Embryos derived from anther culture formed shoots upon transfer to germination medium, although no difference in ploidy was noted between the induced callus and control. Overall, the anther culture conditions established in the present study may contribute to the production of homozygous P. ginseng plants in the future.

Plant Regeneration from Immature Zygotic Embryos of Stewartia koreana Nakai via Somatic Embryogenesis (노각나무(Stewartia koreana Nakai)의 미숙배로부터 체세포배발생에 의한 식물체 재분화)

  • 최은경;박학봉;김광수;이용기
    • Korean Journal of Plant Tissue Culture
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    • v.22 no.2
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    • pp.77-81
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    • 1995
  • When cultured on MS medium supplemented with 0.5 mg/L NAA alone or 1.0 mg/L 2,4-D and 0.5 mg/L BA, immature zygotic embryos of Stewartia koreana formed embryogenic calli and somatic embryos. In investigate effect of sucrose concentration on somatic embryo development, embryogenic calli were transferred to MS basal medium containing 1.5,3, 6 or 9% sucrose. The greatest frequency of somatic embryos was obtained on medium containing 6% sucrose. However addition of 1.5 or 9% sucrose to medium inhibited somatic embryo germination and development into normal plantlet After 5 weeks of hardening culture on medium containing 6% sucrose, somatic embryos were transferred to half strangth MS medium supplemented with 0.1% charcol, wherein these embryo developed into the normal plantlets.

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Transformation of PAT gene into Lettuce (Lactuca sativa L.) using Agrobacterium tumefaciens (Agrobacterium tumefaciens를 이용한 상추 (Lactuca sativa L.)의 PAT유전자 형질전환)

  • 류정아;김창길;이현숙;최경배;양덕춘
    • Korean Journal of Plant Tissue Culture
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    • v.28 no.4
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    • pp.197-200
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    • 2001
  • Agrobacterium tumefaciens MP90 harboring PAT (phosphinothricin acetyltransferase) and NPTII-GUS gene were used for the genetic transformation of lettuce (Lactuca Sativa L.). Shoot regeneration from cotyledon explants were obtained from the MS medium supplemented with 0.1 mg.L$^{-1}$ NAA, 1.0 mg.L$^{-1}$ 2ip, 50 mg.L$^{-1}$ kanamycin and 500 mg.L$^{-1}$ carbenicillin after cocultivation with A. tumefaciens for 2 days. Kanamycin resistance test of transgenic plants indicated that the NPTII gene was integrated into the lettuce genome and was stably expressed. PCR and northern blot analysis indicated that bialaphos resistance gene (PAT) was stably integrated into the lettuce genome. The transgenic plant sprayed with Basta (1500x) remained healthy with continuous growth, while the control group exhibited fatality.

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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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Effect of Abscisic Acid, Kinds and Concentrations of Osmoticum on Somatic Embryo Induction, Germination and Plantlet Regeneration in Larix kaempferi (ABA 및 삼투압제 종류 및 농도에 따른 낙엽송 (Larix kaempferi) 체세포배 유도, 발아 및 식물체 재분화 효과)

  • Kim, Yong Wook
    • Journal of Korean Society of Forest Science
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    • v.100 no.4
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    • pp.693-697
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    • 2011
  • This study was conducted to examine effects of concentrations of abscisic acid (ABA) or /kinds of osmotica on induction of somatic embryos (SEs), germination and plantlet regeneration in Japanese larch (Larix kaempferi). In comparison of duration of culture, concentrations of ABA and osmoticum, the highest induction number (191/g tissue) of the SE was showed in $60{\mu}M$ ABA+0.2 M sucrose for 4 weeks culture. However, the lowest number (3.5~23.5) of SEs was induced from $4{\mu}M$ ABA+0.1 M sucrose, regardless of culture duration for SEs induction. In comparison of germination efficiency of SEs, the highest induction frequencies of cotyledon (90.9%), hypocotyl (95.8%) and root (96.5%), respectively, were obtained from the SEs that cultured from the treatment of $60{\mu}M$ ABA+0.2 M sucrose with 5 weeks culture. In contrast, the lowest germination response was showed in SEs that induced from the treatment of $4{\mu}M$ ABA+0.1 M sucrose. In comparison of effect of different kinds/concentrations of osmotica for germination and plantlet regeneration, the best response was obtained from the treatment of 0.2 M sucrose with induction of cotyledon (98.3%), hypocotyl (78.4%), root (57.5%) and plantlet regeneration (54.8%), respectively.

Plant Regeneration from Protoplasts of Suspension Cultured Cells in Arabidopsis thaliana (애기장대(Arabidopsis thaliana) 현탁배양세포의 원형질체로부터 식물체 재분화)

  • 김명덕;김준철;진창덕;임창진;한태진
    • Korean Journal of Plant Tissue Culture
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    • v.27 no.2
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    • pp.125-131
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    • 2000
  • Protoplasts of Arabidopsis thaliana were easily isolated from the shoot-forming (SF) suspension-cultured cell clusters with 4 hours-shaking condition (40 rpm) on CPD enzyme solution containing 1% cellulase R-10, 0.25% pectolyase Y-23 and 0.5% driselase. Protoplasts were cultured on liquid KAO medium supplemented with 1 mg/L 2,4-D, 0.5 mg/L kinetin, 200 mg/L spermidine and 68 g/L glucose. Also, protoplasts were cultured on 0.2 $\mu$M membrane filter placed onto CP solid medium containing the suspension cells as feeder cells in the dark at $25^{\circ}C$ for 4 weeks. Protoplast-derived-SF calli were cultured on MS medium containing 0.05 mg/L IAA, 7 mg/L 2 ip and 30 g/L sucrose under the continuous illumination for four weeks. The frequency of shoot formation was about 60%. The regenerants were transferred into potting soil to grow mature plants. The regenerants formed the silques with seeds after 8 weeks of cultures.

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