• Title/Summary/Keyword: regenerated shoots

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더덕의 체세포배로부터 식물체 재생과 사포닌 함량 변화 (Plant Regeneration and Saponin Contents in Codonopsis lanceolata L.)

  • 최명석;최필선
    • 한국약용작물학회지
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    • 제7권4호
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    • pp.275-281
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    • 1999
  • 더덕 (Codonopsis lanceolata)의 자엽기의 체세포배 는 생장조절제 를 함유하지 않은 MS고체배지에 배양하였을 때 완전한 식물체로 분화되었다. 체세포배로부터 줄기 재분화에는 2% sucrose가, 뿌리의 분화에는 8% sucrose가 가장 좋았다. 줄기의 생장은 2% sucrose가, 뿌리의 생장은 4% sucrose 농도에서 가장 좋았다. 자연산 더덕뿌리, 재분화 개체의 뿌리, 배발생 캘러스, 비배발생 캘러스에서의 saponin계 물질의 함량을 HPLC로 조사한 결과 각 조직간의 물질 함량은 커다란 차이를 보였으며, 고농도의 sucrose 함유 배지에서 재분화된 뿌리의 saponin계 물질의 함량은 자연산뿌리와 비슷하였다. 세포분화가 진행됨에 따라 saponin계 물질의 함량은 급격히 감소하였다. 이러한 결과는 식물의 유용물질 생산에 효율적으로 활용될 수 있을 것이다.

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Plant Regeneration via Organogenesis from Seed Explants in Red Pepper (Capsicum annuum L.)

  • Lee, Kwang-Woong
    • Journal of Plant Biology
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    • 제39권3호
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    • pp.167-172
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    • 1996
  • Efficient plant regeneration has been achieved via organogenesis in the red pepper plant (Capsicum annum L.). Shoots were induced from seed explants of cultivar 'Friendship' on Murashige and Skoog's (MS) basal medium supplemetned with; NAA or IAA, and BAP or zeatin. Seed explants on the medium supplemented with 0.1-0.3 mg/L IAA and 2-5 mg/L zeatin for 2 weeks vigorously formed normal shoots in more than 90% of the explants. When these were transferred to MS medium containing 0.5-1.0 mg/L GA, 90-100% of the shoots have elongated within 1-2 weeks. The elongated shoots rooted in media supplemented with 0.3 mg/L NAA. It was revealed that this method is very rapid and efficient regeneration system for red pepper and regenerated plants can be obtained after only 5-6 weeks of culture.

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Adventitious Shoots Regeneration from Seed Explants of Xanthoceras sorbifolium

  • Hyunseok Lee
    • 한국자원식물학회:학술대회논문집
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    • 한국자원식물학회 2020년도 추계국제학술대회
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    • pp.58-58
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    • 2020
  • Xanthoceras sorbifolium Bunge (yellowhorn) is a woody tree in the soapberry family, Sapindaceae, native to northern China. This species has been identified as a major woody bioenergy plant for bio-diesel production because of high oil content in seed. But the flowers do not bear fruit well while the many flowers blooming. This study was performed to regenerate in vitro plantlet using adventitious shoot formation. To establish the protocol of plant regeneration, adventitious shoots formation rate in the culture of cotyledon of immature zygotic embryos was 68.6% in 1/2 MS medium with 0.5 mg l-1 BA and 3% sucrose (w/v). In the culture of cotyledons of mature zygotic embryos, induction of adventitious shoots was needed to contain high sucrose in pre-culture medium and the frequency of shoot induction was 64.4%. Multiple shoots were induced in 0.5 mg l-1 TDZ, and rooting of shoot was induced 4.0 mg l-1 IBA. Flow cytometry analysis revealed that all the regenerated plantlets were diploid.

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옥수수 전위유전자 Ac 및 Ds의 2배체종 감자 Genome 내로의 도입 (Introduction of Maize Transposable Elements, Ac and Ds into the Genome of a Diploid Potato Species)

  • 김화영;임용표
    • 식물조직배양학회지
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    • 제27권1호
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    • pp.39-45
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    • 2000
  • 전위유전자 표지를 이용한 2배체 야생근연종 감자의 유용 유전자 cloning 체계를 개발하기 위하여 옥수수의 진위유전자 Ac를 조작하여 전위효소는 생산하나 이동은 불가능하도록 제작된 immobilized Ac (iAc)와 자체 이동은 불가능하나 iAc의 작용에 의해 이동이 가능한 Ds를 Agrobacterium tumefaciens를 이용한 형질전환에 의해 2배체 근연종 감자 (Solanum tuberosum Group Phureja) 계통 1.22에 도입하였다. iAc및 Ds 삽입 binary vector들을 보유하는Agrobacterium 계통들로 형질전환 처리된 1.22 기내배양신초의 잎과 줄기 절편의 경우 50mg/L 의 kanamycin이 첨가된 배지에서도 캘러스를 형성하였으며, 잎 절편으로부터 재분화신초가 획득되었다. 형질전환 처리되지 않은 1.22 신초는 100mg/L의 kanamycin이 첨가된 배지에서 전혀 발근이 되지 않았으나, 형질전환 처리에 의해 획득된 재분화 신초는 동일 배지에서 발근이 되었다. iAc와 Ds의 염기배열에 대해 특이적인 oligonucleotide primer들을 이용하여 형질전환 처리 획득 식물들로부터 추출된 DNA에 대한 PCR분석을 실시한 결과, 사용된 primer들의 iAc 와 Ds의 염기배열에 있어서의 위치에 의해 예상되는 크기의 DNA들이 형성되어 iAc 와 Ds의 1.22 genome내 도입이 확인되었다.

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Plant Regeneration from Leaf Explants of Kalanchoe daigremontiana Hamet & Perrier

  • ;Kim, Teh-Ryung;In, Jun-Gyo;Yang, Deok-Chun;Choi, Kwan-Sam
    • 한국약용작물학회지
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    • 제14권5호
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    • pp.293-298
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    • 2006
  • Optimum culture conditions for high frequency plant regeneration from leaf explants of Kalanchoe daigremontiana Hamet &Perrier were established. Shoot regeneration was achieved from leaf explant cultures using MS medium supplemented with indole-3-acetic acid (IAA) and thidiazuron (TDZ) or benzyladenine (BA). Percent regeneration was influenced by plant growth regulators and source of explants. MS medium supplemented with TDZ (1.0 mg/l) and IAA (0.4 mg/l) was the most effective, providing shoot regeneration for 76.7 % of ex vitro leaf explants associated with a high number of shoots per explant (9.5 mean shoots per explant), whereas 100% shoot regeneration associated with 12.4 shoots per explant occurred from in vitro leaf explants on the same medium. Clusters of shoots were multiplied and elongated on MS medium containing several concentrations of BA. MS medium supplemented with 0.25 mg/l BA was proved as the most effective shoot elongation medium. Elongated shoots (2-3 cm) were rooted at 100% on half-strength MS medium. Rooted plantlets were then transferred to potting soil. Regenerated plants were established in the soil with 90% success.

In vitro Selection of Acifluorfen-tolerant Solanum ptycanthum and Phenotypic Variation in Regenerated Plants

  • Yu, Chang-Yeon;Lim, Jeong-Dae;Kim, Myong-Jo;Kang, Won-Hee;Hyun, Tae-Kyoung
    • 한국약용작물학회지
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    • 제10권4호
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    • pp.263-268
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    • 2002
  • Acifluorfen-tolerant callus lines of Solanum ptycanthum were isolated by stepwise selection. Growth of the unselected line was completely inhibited at 0.5 uM. while some selected lines grew at 8 uM acifluorfen. Twenty-two of twenty-five acifluorfen-tolerant callus lines regenerated shoots. Many of the regenerated somaclones were variants, differing in leaf shape, leaf color, number of flower parts, flower color, and fertility. The acifluorfen tolerant S. ptycanthum callus lines differed.

돌나물의 식물체 분화에 미치는 Glutamine과 AgNO3의 영향 (Effects of glutamine and AgNO3 on plant regeneration of Sedum sarmentosum)

  • 안정호;김현순;이승엽
    • Journal of Plant Biotechnology
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    • 제36권1호
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    • pp.18-22
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    • 2009
  • 돌나물 (Sedum sarmentosum)의 형질전환 및 유전자원 기내보존을 위한 효율적인 식물체 분화 체계를 확립하기 위하여, 잎절편 유래의 켈러스로부터 식물체 분화에 미치는 glutamine과 $AgNO_3$의 영향과 분화 식물체의 순화 후 생존율을 조사하였다. 캘러스 유도는 3 mg/L 2,4-D와 1 mg/L BA, 식물체 분화는 0.2 mg/L NAA와 3.0 mg/L BA를 각각 첨가한 MS배지를 사용하였다. 금산 수집종에서 glutamine 첨가는 50일째에 100%의 식물체 분화를 보였으며, 350mg/L glutamine 첨가시 켈러스당 17.6개의 가장 많은 식물체가 분화되었다. $AgNO_3$ 첨가배지에서도 금산 수집종은 100% 식물체 분화를 보였으며, $15{\mu}M$ 첨가배지에서 캘러스당 16.1개의 식물체가 분화되어 가장 양호하였다. 완주 수집종은 glutamine 첨가에 관계없이 식물체가 전혀 분화되지 않았고, $5-15{\mu}M$ $AgNO_3$ 첨가배지에서만 소수의 식물체가 분화되었다. 분화 식물체는 생장조절제를 첨가하지 않은 MS 배지로 옮겨 shoot생장과 발근을 유도한 다음, vermiculite와 perlite를 혼합한 인공토양에서 순화시킨 결과 88-100%의 생존율을 보였다. 생존 식물체들은 강모래:밭흙 (1:1, v/v)을 혼합한 토양에 정식한 후에도 양호한 생장을 보였으며, 모식물과 형태적으로 차이가 없었다.

액체배지 첨가에 의한 Anthurium andreanum ′Atlanta′의 기내생육 촉진 (Growth Stimulation of In Vitro Shoots by the Post-supplying of Liquid Medium in Anthurium andreanum ′Atlanta′)

  • 한봉희;구대희
    • Journal of Plant Biotechnology
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    • 제30권2호
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    • pp.185-188
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    • 2003
  • In order to enhance shoot elongation and rooting of Anthurium andreanum 'Atlanta' in vitro, 15mL of liquid media containing various concentrations of activated charcoal, sucrose and MS salts were added in same vessels after small shoots were induced from the calli on mudium supplemented with 10.0mg/L BA and 0.1mg/L 2.4-D. The post-supplying of 15mL liquid medium containing MS macro and micro elements, 30g/L sucrose and 5.0∼10.0g/L activated charcoal was significantly stimulated the shoot elongation and rooting of regenerated shoots from calli. The medium addition was also resulted in the enhanced soil survival, elongation and rooting of plantlets in cultural soil mixed with perlite and vermiculite(1 : 1)

철 저장단백질 관련 Ferritin 유전자를 이용한 상추의 형질전환 (Transformation of Lettuce (Lactuce sativa L.) Using Iron Storage Protein Ferritin Gene)

  • 김성하;노일섭;최장선;강권규
    • 식물조직배양학회지
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    • 제28권3호
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    • pp.147-151
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    • 2001
  • Explants of Lactuce sativa cultivar, chungchima, were co-cultivated with Agrobacterium tumefaciences LBA4404, EHA101 strains containing nptll gene and ferritin gene encoding iron storage protein from soybean for transformation. Through initial selection of regenerated explants by culturing on a kanamycin and carbenicillin containing MS medium, multiple shoots were obtained after 2 months of culture. For a complementary step of selection, putative transgenic shoots were transferred to 1/2 MS basal medium supplemented with 100 mg/L kanamycin and 500 mg/L carbenicillin. The selected shoots were tested with PCR analysis using nptll, ferritin specific primers whether ferritin gene was introduced to genome of the plants. These results confirmed that produced the specific PCR bands in the putative transgenic lines. Additionally the Northern blot showed that transcripts of ferritin gene were detected in mature leaf of the transgenic lines. These results suggest that ferritin gene be successfully integrated and transcribed in the putative transgenic lettuce plants.

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Efficient transformation of Actinidia arguta by reducing the strength of basal salts in the medium to alleviate callus browning

  • Han, Meili;Gleave, Andrew P.;Wang, Tianchi
    • Plant Biotechnology Reports
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    • 제4권2호
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    • pp.129-138
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    • 2010
  • An efficient transformation system for high-throughput functional genomic studies of kiwifruit has been developed to overcome the problem of necrosis in Actinidia arguta explants. The system uses Agrobacterium tumefaciens strain EHA105 harbouring the binary vector pART27-10 to inoculate leaf strips. The vector contains neomycin phosphotransferase (nptII) and ${\beta}$-glucuronidase (GUS) (uidA) genes. A range of light intensities and different strengths of Murashige and Skoog (MS) basal salt media was used to overcome the problem of browning and/or necrosis of explants and calli. Callus browning was significantly reduced, resulting in regenerated adventitious shoots when the MS basal salt concentration in the culture medium was reduced to half-strength at low light intensity ($3.4\;{\mu}mol\;m^{-2}\;s^{-1}$) conditions. Inoculated leaf strips produced putative transformed shoots of Actinidia arguta on half-MS basal salt medium supplemented with 3.0 $mg\;l^{-1}$ zeatin, 0.5 $mg\;l^{-1}$ 6-benzyladenine, 0.05 $mg\;l^{-1}$ naphthalene acetic acid, 150 $mg\;l^{-1}$ kanamycin and 300 $mg\;l^{-1}$ $Timentin^{(R)}$. All regenerated plantlets were deemed putativ transgenic by histochemical GUS assay and polymerase chain-reaction analysis.