• 제목/요약/키워드: embryogenic calli

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Somatic Embryogenesis and Plant Regeneration from Stem Tissues of Orostachys japonicus A. Berger

  • Song, Min-Jung;Park, Young-Goo
    • Journal of Plant Biotechnology
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    • 제34권3호
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    • pp.181-187
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    • 2007
  • Orostachys japonicus A. Berger is a Perennial herbaceous plant which has been traditionally used as an anti-inflammatory agent to treat hepatitis and as an anticancer agent. The objective of this study was 1) to establish and proliferate in vitro plant of O. japonicus 2) to induce indirect somatic embryogenesis from O. japonicus. General calli and embryogenic calli in all ranges of 2,4-D and BA combination, were induced and were best at 22% (embryogenic cell) in 5.0 mg/L 2,4-D and 0.5 mg/L BA combination. Embryogenic cell line was maintained by subculture at 2 week intervals and transferred to solid and liquid medium for embryo formation. In solid medium culture, globular and heart shaped embryos were observed in MS medium containing 5.0 mg/L 2,4-D and 0.5 mg/L BA combination. The number of embryos was 6.5 per 0.5 g cell, and then the immature embryos transferred to MS basal medium for embryo development. In a suspension culture of embryogenic cells, globular and heart shaped embryos were emerged in MS medium supplemented with 3.0 mg/L 2,4-D and 0.3 mg/L BA combination after 10 days of incubation. The embryo formation rate was about 33% by suspension culture. The ratio of embryo germination was 60.9%, on the other side, the root formation rate was 74.3% in 1/2 MS continuously.

인삼근 캘루스의 체세포 배아 발생과 관련한 IAA 및 IAAsp의 수준에 관한 연구 (STUDIES ON THE LEVELS OF INDOLE-3-ACETIC ACID (IAA) AND INDOLE-3-ACETYL-L-ASPARATE(IAAsp)IN RELATION TO SOMATIC EMBRYOGENESIS OF CALLI DERIVED FROM GINSENG (PANAX GINSENG C.A. MEYER) ROOTS)

  • Chen Kai-hsien;Hsing Yue-ie;Chen Shuh-chun;Chang Wei-chin
    • 고려인삼학회:학술대회논문집
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    • 고려인삼학회 1984년도 학술대회지
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    • pp.45-48
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    • 1984
  • 인삼근에서 유래된 캘루스중 배분화 유조직과 배분화가 되지 않는 유조직에 있어서 Indol-3-acetic acid (IAA) 및 Indole-3-acetyl-L-aspartate(IAAsp) 함량을 Ion-pair reverse phase HPLC 기법을 이용하여 분석 비교하였다. 배분화 유조직의 IAA 및 IAAsp 함량은 배분화가 불가능한 유조직에 비해 각각 7 및 18배 함량이 높았다. 배분화가 되지 않은 캘루스로부터 얻은 추출물의 HPLC 프로파일상의 인돌 화합물 피크 부근에 특이한 미확인 피크가 나타났다. 이와 같은 분명한 차이는 배상체 발생 잠재력이 높은 유조직을 얻기 위한 배양조직을 선별하기 위한 확실한 파라메타로 이용될 수 있을 것이다.

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Selection and Characterization of S-Aminoethyl-L-Cysteine Resistant Plants from Gamma-ray Irradiated Embryogenic Callus in Sweet potato

  • Lee In-Sok;Kim Dong-Sub;Hong Chang-Pyo;Kang Si-Yong;Song Hi-Sup;Lee Sang-Jae;Lim Yong-Pyo;Lee Young-Il
    • Journal of Plant Biotechnology
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    • 제5권4호
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    • pp.233-238
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    • 2003
  • Sweet potato cells derived from Yulmi were isolated from embryogenic callus and irradiated with 50 Gy dose. Resistant cells were selected on a MS medium containing 1.0 mM S-aminoethyl-L-cysteine (AEC). This level of AEC approximately inhibits non-selected wild type cells. The callus resistant to this analog of lysine was subcultured for 30 days in absence of AEC to proliferate. The three resistant calli (AR-1, AR-2 and AR-3) with better growth were divvied into 0.5~1mm diameter and placed on MS medium with 0, 0.4, 0.6, 0.8 and 1.0 mM AEC. There are considerable growth difference between control callus and AEC resistant callus on the AEC-medium. The selected calli were placed on the hormone-free medium for regeneration. Three plantlets, five plantlets and six plantlets were recovered from AR-1, AR-2 and AR-3 calli, respectively. Each two regenerants in AR-1, AR-2 and AR3 were randomly selected for RAPD and SDS PAGE analysis. RAPD polymorph isms between Yulmi and AEC resistant plant from irradiated calli were detected in several Wako primers. Also, it was identified that two AEC resistant plants had higher protein than the original variety Yulmi.

원형질체 내 Plasmid Electroporation에 의한 벼 배발생세포의 형질전환 1. 벼의 Electroporation 원형질체로부터 식물체 재분화 (Transformation of Rice Embryogenic Cells by Electroporation Mediated Plasmid Uptake into Protoplasts 1. Plant Regeneration from Electroporated Protoplasts of Rice)

  • 김명덕;최성진김준철
    • KSBB Journal
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    • 제10권1호
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    • pp.23-29
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    • 1995
  • 발아된 벼(Oryza sativa L. cv. Nakdong)의 하배 축에서 callus를 유도하고, 현탁배양한 세포에서 원 형질체를 분리하여 MS 배지에서 재분화를 수행하였다. Callus는 하배축으로부터 $2.5mg/{\ell}2$,4-0를 첨가한 LS 배지에서 최고 65%의 치상효율로 유도되었으며, 형태척으로 선별 가능한 embryogenic callus 는 $2mg/{\ell}2$,4-D, O.2mgj P kinetin과 $0.1mg/{\ell} GA_3$ 가 첨가된 AA, 배지에서 현탁배양하였다. 4개월 이상 현탁배양한 세포로부터 분리한 원형질체는 voltage를 400V jern와 Imsec 통안 electropora­tion했을 때 $2.5mg/{\ell}2$,4-D, $0.1mg/{\ell}$kinetin과 lOmM proline이 첨가된 PCM 배지에서 1.1%의 치 상효율을 보였다. 형성된 microcalli는 LS2.5 배지 에 치상된 feeder cell에 $0.2\mu\textrm{m}$membrane filter를 얹은 위에 옮겨 암소에서 2주 동안 배양한 후, $30{\pm}/3{\mu}E$.m^{-2}S^{-1}의 형광하에서 2주 통안 배양했을 때 2mm 직경의 노란색 callus를 형성하였다. 형성된 callus를 재분화 배지에 치상하였을 때, 녹점 green spot)에서 shoot 형성과정을 거쳐 완전한 식물체로 분화되었으며 재분화율은 1l~33%였다.

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Insilico Analysis for Expressed Sequence Tags from Embryogenic Callus and Flower Buds of Panax ginseng C. A. Meyer

  • Sathiyamoorthy, Subramaniyam;In, Jun-Gyo;Lee, Byum-Soo;Kwon, Woo-Seang;Yang, Dong-Uk;Kim, Ju-Han;Yang, Deok-Chun
    • Journal of Ginseng Research
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    • 제35권1호
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    • pp.21-30
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    • 2011
  • Panax ginseng root has been used as a major source of ginsenoside throughout the history of oriental medicine. In recent years, scientists have found that all of its biomass, including embryogenic calli and flower buds can contain similar active ingredients with pharmacological functions. In this study, transcriptome analyses were used to identify different gene expressions from embryogenic calli and fl ower buds. In total, 6,226 expressed sequence tags (ESTs) were obtained from cDNA libraries of P. ginseng. Insilico analysis was conducted to annotate the putative sequences using gene ontology functional analysis, Kyoto Encyclopedia of Genes and Genomes orthology biochemical analysis, and interproscan protein functional domain analysis. From the obtained results, genes responsible for growth, pathogenicity, pigments, ginsenoside pathway, and development were discussed. Almost 83.3% of the EST sequence was annotated using one-dimensional insilico analysis.

Plastid Transformation of Soybean Suspension Cultures

  • Zhang, Xing-Hai;Archie R.Portis. Jr.;Jack M.Widholm
    • Journal of Plant Biotechnology
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    • 제3권1호
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    • pp.39-44
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    • 2001
  • Plastid transformation was attempted with soybean [Glycine max (L.) Merr.] leaves and photoautotrophic and embryogenic cultures by particle bombardment using the transforming vector pZVII that carries the coding sequences for both subunits of Chlamydomonas reinhardtii Rubisco and a spectinomycin resistance gene (aadA). Spectinomycin resistant calli were selected from the bombarded leaves but the transgene was not present, indicating that the resistance was due to mutations. The Chlamydomonas rbcL and rbcS genes were shown to be site-specifically integrated into the plastid genome of the embryogenic cells with a very low transformation efficiency. None of the transformed embryogenic lines survived the plant regeneration process so no whole plants were recovered. This result does indicate that it should be possible to insert genes into the plastid genome of the important crop soybean if the overall methods are improved.

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Micropropagation of Medicinal Woody Eleutherococcus pedunculus via Somatic Embryogenesis

  • Choi, Yong Eui
    • Journal of Forest and Environmental Science
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    • 제23권1호
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    • pp.5-9
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    • 2007
  • Zygotic embryos just after harvest of seeds were immature globular to heart stage. Maturation of zygotic embryos rapidly proceed when zygotic embryos together with small excised parts of endosperm were cultured on 1/3-strength MS solid medium with 2% sucrose, and the zygotic embryos were germinated within two months. Embryogenic callus was formed from the excised segments of germinating zygotic embryos of Eleutherococcus pedunclus on Murashige and Skoog (MS) medium with $4.5{\mu}M$ 2,4-D. The embryogenic callus formation occurred at a low frequency (less than 7%) from hypocotyl segments. The embryogenic calli were maintained on the same medium as primary medium. High frequency somatic embryogenesis was obtained after the cells were transferred to medium lacking 2,4-D. Cotyledonary embryos were germinated and converted into plantlets on medium with $20{\mu}M$ $GA_3$. Embryogenic callus and somatic embryos were produced spontaneously on the surfaces of roots and/or hypocotyls of plantlets. The frequency of embryogenic callus formation was 85% in roots and 34% in hypocotyls. Therefore maintain of cell lines performed very easily. Plantlets with developed epicotyls at more than 3 cm acclimatized at high frequency (89%). While plantlets with small epicotyls (less than 1 cm) were acclimatized at low rate (32%). The soil survived plantlets produced new sprouts after over wintering in the field.

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벼 현탁배양을 통하여 분리된 원형질체로부터 식물체 재분화 (Plant Regeneration from Protoplasts Isolated through Embryogenic Cell suspension Culture in Rice)

  • 정병균
    • Journal of Plant Biology
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    • 제36권3호
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    • pp.211-218
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    • 1993
  • Plant regeneration was accomplished from protoplast culture of rice (Oryza sativa L. cv. Taebaeg). Embryogenic callus was induced from mature seed on MS medium containing 5 mM proline, 2.5 mg/L 2,4-D, 30 g/L sucrose in the dark at 28$^{\circ}C$ and used to establish embryogenic cell suspension culture. Suspension cells were subcultured every one week in N6 medium supplemented with 5 mM proline, 200 mg/L casein hydrolysate, 2.5 mg/L 2,4-D and amino acids of AA medium. Suspension cultures were composed of cells that were densely cytoplasmic, potentially embryogenic and were at least maintained for more than 6 months in liquid medium. Protoplasts were isolated from fast-growing suspension culture cells and cultured in a slightly modified KpR medium by mixed nurse culture. Isolated protoplasts began to divide within 5~7 days and thereafter, protoplast-derived calli were sequentially transferred to callus proliferating medium that soft agar MS medium contained 2 mg/L 2,4-D and produced distinct embryogenic cells. Microcolonies were then transferred to solid medium which consisted of MS medium containing 5 mg/L kinetin, 1 mg/L NAA, 1 mg/L ABA, 30 g/L sucrose and 10 g/L sorbitol under fluorescent light. Mulitple shoots of 4~5 per callus emerged and were transferred to hormone-free MS medium for root initiation. Thereafter, The plantlets were transferred to pots of soil to mature in the culture room.

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