• Title/Summary/Keyword: Callus Induction

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Transformation of Birdsfoot trefoil by BcHSP17.6 Gene using Agrobacterium tumefaciens (BcHSP17.6 유전자 도입에 의한 버즈풋 트레포일의 형질전환)

  • 김기용;성병렬;임용우;최기준;임영철;장요순;정의수;김원호;김종근
    • Journal of The Korean Society of Grassland and Forage Science
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    • v.21 no.3
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    • pp.145-150
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    • 2001
  • This study was conducted to obtain the transformed birdsfoot trefoil (Lotus corniculatus L.) plants with BcHSP17.6 gene using Agrobacterium turnefaciens LBA4404 and we confirmed transformed gene from the regenerated birdsfoot trefoil plants. The expression vector, pBKH4 vector, harboring BcHSP17.6 gene was used for production of transgenic birdsfoot trefoil plants. The callus of birdsfoot trefoil was cocultivated with Agrobacteriurn turnefaciens and transformed calli were selected on kanamycin-containing SH-kc medium to regenerate into plants. The transformed birdsfoot trefoil plants were produced 4 momths after cultivation on BOi2Y medium. The transgenic birdsfoot trefoil plants were analyzed by isolation of genomic DNA and genomic Southern hybridization using a -32P labelled BcHSPl7.6 fragments. (Key words : Birdsfoot trefoil, Transgenic plant. BcHSP17.6 gene, Callus induction, Plant regeneration)

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Callus induction and plant regeneration of Lychnis wilfordii (Regel) Maxim a critically endangered plant in Korea

  • Bae, Kee Hwa;Lee, Mi Hyun;Choi, Yong Eui;Yoon, Eui Soo
    • Journal of Plant Biotechnology
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    • v.41 no.1
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    • pp.33-37
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    • 2014
  • Lychnis wilfordii (Regel) Maxim is a rare and valued ornamental plant. Germination rate reached 46.6% when seeds were treated with $100mg{\cdot}l^{-1}$ GA (Gibberellic acid). The highest callus induction was observed in the leaf explants of the seedlings on MS medium containing specific concentrations of $0.5mg{\cdot}l^{-1}$ BA ($N^6$-benzyladenine) and $3.0mg{\cdot}l^{-1}$ NAA (a-naphthalene acetic acid). The adventitious shoot was formed in 97.3% of calli on 1/2 WPM (Woody Plant Medium) medium. Shoot elongation of in vitro propagated plantlets was no difference among various medium. The plantlets grew well after transferring to the pot. This in vitro propagation protocol should be useful for conservation of this endangered plant.

Induction of Callus and Culture of Protoplasts from Cotyledon-Originated Callus in Alnus hirsuta (물오리나무(Alnus hirsuta)의 칼루스 유도 및 자엽유래 칼루스로부터의 원형질체 배양)

  • 김형하
    • Journal of Plant Biology
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    • v.33 no.4
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    • pp.253-257
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    • 1990
  • Callus-inducing ability of Alnus hirsuta was examined by culturing various tissues (leaf, hypocotyl, cotyledon and seed) on NT (Nagata & Takebe) medium, supplemented with 2.5$\mu$M 2,4-D. Leaf-originated callus was cultured on media varying in auxin (IBA and NAA) and cytokinin (BAP) concentrations to examine the effects of auxin and cytokinin on callus growth. Maximum growth was obtained at 10 $\mu$M IBA+10$\mu$M BAP and 10$\mu$M NAA without cytokinin. Cell suspensions established from cotyledon-originated callus yielded viable protoplasts after incubation for 16-18 hours in an enzyme mixture (1% (w/v) Onozuka R-10 0.5% (w/v) Macerozyme, CPW salts and 13% (w/v) mannitol, pH 5.8). Protoplasts were cultured on NT medium, supplemented with glucose, hormones and coconut milk. After 6 weeks of culture, protoplasts sustained cell divisions to form microcallus, which showed various colors from red to white.

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Effect of gamma ray irradiation and ethyl methane sulphonate on in vitro mutagenesis of Citrullus colocynthis (L.) Schrad

  • Ramakrishna, D.;Chaitanya, G.;Suvarchala, V.;Shasthree, T.
    • Journal of Plant Biotechnology
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    • v.45 no.1
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    • pp.55-62
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    • 2018
  • In the present study in vitro mutagenesis was used to study the effect of gamma irradiation and EMS on callus induction, morphogenesis and production of multiple shoots from different explants of Citrullus colocynthis (L.) Schrad. Gamma radiations (5 kR to 20 kR) and certain chemicals have been effected on plant growth developments and changes of biochemical metabolisms in plants. Murashige and Skoog (MS) medium containing with auxins such as NAA, IAA, 2,4-D (0.5 ~ 2.0 mg/l), cytokinines BAP, kn TDZ, (0.5 ~ 2.5 mg/l), L-Glutamic acid (1 ~ 2 mg/l) and Coconut milk (10 ~ 20%). After 5 weeks on induction media, explants and callus (EC) were exposed to 5 kR, 10 kR, 15 kR and 20kR, of gamma radiation and treated with 1, 2, 3, 4 and 5 mM ethyl methane sulphonate (EMS) for 30 min. The highest percentage of callusing was observed (70%) stem irradiated with 5 kR and significantly decrease in fresh and dry weight of callus in the below 4 kR doses and above 20 kR doses, there was a progressive decrease in the fresh weight and dry weights when compared to control callus. Maximum percentage of plantlet regeneration (59%) was induced from callus exposed to 15 kR gamma irradiation on MS media fortified with 2.0 mg/l 2,4-D + 2.0 mg/l BAP + 2.0 mg/l L-glutamic acid. Increase in gamma irradiation dose above 15 kR and 5 mM EMS reduced regeneration capacity of callus. Doses higher than 20 kR and 7 mM EMS was lethal to micropropagated plants of Citurullus colocynthis.

Plant Regeneration from Leaf and Internode Segment Cultures of Boxthorn (Lycium chinense Mill.) (구기자나무의 잎과 마디절편체 배양에 의한 식물체 재생)

  • 김동찬;정해준;민병훈;양덕춘
    • Korean Journal of Plant Tissue Culture
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    • v.28 no.6
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    • pp.329-333
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    • 2001
  • Callus and shoot formation from medicinal crop, Lycium chinense Mill. cv. 'Cheongyang', as influenced by various media, explant sources and plant growth regulators were investigated. The rate of shoots formation, number of shoots, and fresh weight of shoots were the best on MS medium followed by B$_{5}$, WPH, and SH. Callus induction was more effective in leaf than internode segments, and was the best on MS medium containing 0.5 mg/L NAA with 0.2 mg/L BA. Effects of plant growth regulators in shoot formation were more effective in BA than TDA combined with NAA. Shoot formation from callus induced in leaf and internode segments was the best on MS medium containing 0.01 mg/L NAA with 0.2 mg/L BA.

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Plant Regeneration from Callus and Adventitious Root Segments of Pulsatilla Koreana Nakai

  • Jung, Su-Jin;Jeong, Jae-Hun;Yoon, Eui-Soo;Choi, Yong-Eui
    • Journal of Plant Biotechnology
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    • v.34 no.2
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    • pp.153-159
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    • 2007
  • Plant regeneration of Pulsatilla koreana was achieved via adventitious shoot formation indirectly from callus and directly from adventitious root segments. For the callus induction from leaf or petiole explants, combination of 2,4- dichlorophenoxyaceticacid (2,4-D) with $2.22\;{\mu}M$ 6-benzyladenine (BA) was effective. Adventitious shoot induction from callus was enhanced by the combined treatment with $0.1\;{\mu}M$ polyvinylpyrrolidone (PVP) compared to cytokinin treatment alone. Adventitious roots were induced from the petiole segments on 1/2 MS medium with $4.93\;{\mu}M$ IBA. High frequency direct adventitious shoot formation from the segments of adventitious roots was achieved on medium with $4.92\;{\mu}M$ 2-isopentenyladenine (2-ip). Elongated shoots were rooted on half-strength MS medium containing $5.71\;{\mu}M$ indole acetic acid (IAA). Regenerated plantlets with well-developed shoots and roots were successfully transferred to soil. This in vitro propagation protocol might be useful for mass propagation as well as conservation of this plant.

High-frequency Plant Regeneration from Mature Seed-derived Callus Cultures of Orchardgrass (오차드그래스 성숙종자로부터 캘러스 유도 및 고효율 식물체 재분화)

  • Lee, Sang-Hoon;Lee, Dong-Gi;Kim, Jin-Soo;Lee, Byung-Hyun
    • Journal of Plant Biotechnology
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    • v.30 no.4
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    • pp.341-346
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    • 2003
  • In an effort to optimize tissue culture conditions for genetic transformation of orchardgrass (Dactylis glomerata L.), an efficient and high-frequency plant regeneration system from seed-derived calli was established. Embryogenic calli induced on MS medium containing 3mg/L 2,4-D and 0.1mg/L BA had significantly improved regeneration ability. Plant regeneration rate was 92% when embryogenic calli were cultured on N6 medium supplemented with 1mg/L 2,4-D and 3mg/L BA. Among three kinds of medium, MS and N6 medium were optimal for embryogenic callus induction and plant regeneration, respectively. Ho difference in callus induction frequency was observed among four cultivars of orchardgrass, however, "Roughrider" cultivar showed higher regenerability with the frequency of 61%. Addition of maltose to the regeneration medium as a carbon source dramatically increased regeneration frequency up to 69%. A short tissue culture period and high-frequency regeneration system would be beneficial for molecular breeding of orchardgrass through genetic transformation.

Establishment of efficient Alstromeria callus induction system using node culture and various hormones (마디배양과 다양한 호르몬을 이용한 효율적인 알스트로메리아 캘러스 유도 시스템 체계 확립)

  • Yang, Hwan Rae;Lee, Sang Hee;Kim, Jong Bo
    • The Journal of the Convergence on Culture Technology
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    • v.5 no.1
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    • pp.413-416
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    • 2019
  • Alstroemeria (Alstroemeriaceae) is one of the most important cut flowers in international market. Especially, characteristics like long vase-life, various colors, tolerance to low temperature and a low energy requirement during cultivation have stimulated this success. Because of its characteristics such as low multiplication rates, time-consuming process and high risk of carrying viral disease, in vitro propagation techniques based on rhizome meristems culture have been developing nowadays. The callus induction has various cultivation sites compared with the direct plant generation method, and if the callus is maintained well, the plant differentiation can be performed simultaneously while maintaining the callus, so that it can be used for mass proliferation. In this study, we tested various hormones and cultivars for efficient callus induction. As a result of culturing between the nodes and the internodes, the callus began to be formed after 8 weeks, and the calli incidence in the nodes was higher than that between the internodes. Also, in the comparison of 2,4-D and picloram, the callus incidence rate was up to 2 times higher in the medium treated with 2,4-D. Using these results, it is thought that it will help establish the system of mass propagation system of Alstroemeria and cultivate new varieties.

Establishment of Efficient Callus Induction System with Picloram Hormone in Lily Plants (Picloram을 이용한 효율적인 백합 캘러스 유도 체계 확립)

  • Kim, Jong Bo
    • The Journal of the Convergence on Culture Technology
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    • v.7 no.3
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    • pp.605-608
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    • 2021
  • Lily is one of the most important 5 cut flowers in international flower market and lilies are distributed in Asia, Eurasia and North America. To develop a new lily cultivar, in addition to hybridization, mutation and selection methods, biotechnological techniques including tissue culture are also required. Establishment of tissue culture system is one of the requirement for the breeding program in Lily. Among many fields of plant tissue culture, establishment of regeneration system via embryogenic calluses are studied in many crops. In this study, research was carried out to decide the proper concentration of picloram which is used for the induction of embryogenic calluses. As a result, 3 different types of callused were observed after 3-4 weeks. They were CEC (compact embryogenic callus), FEC (friable embryogenic callus) and white callus type. 1.0 mg /l of picloram showed the best result for the production of embryogenic callus, however, due to its higher rate of browning in this concentration, 0.75 mg/l of picloram was selected as a proper concentration of picloram for the induction of CEC and FEC in Lily. These results can be contributed to the establishment of both regeneration system and mass propagation in lily in the future.