• Title/Summary/Keyword: embryogenic callus induction

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Chromosome Doubling of Allium wakegi Araki by In Vitro Cultures (기내배양에 의한 쪽파의 체세포 염색체 배가)

  • Yim, Sun-Hee;Ahn, Chang-Soon;Jeong, Chang-Nam;Han, Tae-Ho
    • Journal of Plant Biotechnology
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    • v.29 no.4
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    • pp.259-264
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    • 2002
  • Induction of embryogenic callus from Allium wakegi Araki explants was promoted on medium containing 2,4-D, and production of abnormal embryos from the embryogenic callus increased as 2,4-D concentration was raised. Shoot tip was found to be the best explant source for embryogenic callus formation followed in the order by bulb scale and leaf section. Medium containing 0.09 M sucrose was effective for embryogenic callus production. The regenerants from embryogenic callus on medium containing 2,4-D and BA at different concentrations was consisted of diploids, tetraploids and a few mixaploids of 2n+4n, and their chromosomal aberration rate ranged from 8.0 to 33.3%. Frequency of chromosomal aberrants was the highest (18.7%) in the regenerants obtained from bulb scale-derived embryogenic callus among others. Plant regeneration rate was high (33.5%) from the shoot tip-derived embryogenic callus and the frequency of chromosomal aberrants was very low (7.0%). The plantlets regenerated on medium containing 0.26 M sucrose resulted in high chromosomal aberrants. The regenerants on medium containing sucrose at 0.09∼0.20 M produced chromosomal aberrants at around 15.2∼16.6%.

Factors Affecting Callus Induction and Plant Regeneration from Mature Seed of Zoysiagrass (Zoysia japonica Steud.) (들잔디 성숙종자로부터 캘러스배양 및 식물체 재분화에 미치는 몇 가지 요인의 영향)

  • 이상훈;김범수;원성혜;조진기;김기용;박근제;성병렬;이효신;이병현
    • Journal of The Korean Society of Grassland and Forage Science
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    • v.24 no.1
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    • pp.29-36
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    • 2004
  • In an effort to optimize tissue culture responses of zoysiagrass(Zoysia japonica Steud.) for genetic transformation, factors affecting callus induction and plant regeneration were investigated. MS medium containing 3 mg/L 2,4-D was optimal for embryogenic callus induction from mature seed. The plant regeneration frequency of 73.3% was observed when embryogenic calli induced in this medium were transferred to N6 medium supplemented with 0.1 mg/L 2,4-D and 5 mg/L BA. Among several basic media, MS and N6 medium were optimal for callus induction and plant regeneration, respectively. Regenerated plants were grown normally when shoots transplanted to the soil. A rapid and efficient plant regeneration system established in this study will be useful for molecular breeding of turfgrass through genetic transformation.

Effects of Medium Supplements on Seed-derived Callus Culture of Italian Ryegrass (배지첨가물질이 이탈리안 라이그래스의 종자유래 캘러스 배양에 미치는 영향)

  • Woo, H.S.;Lee, B.H.
    • Journal of Animal Science and Technology
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    • v.46 no.2
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    • pp.243-250
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    • 2004
  • In an effort to optimize tissue culture responses of Italian ryegrass(Lolium multiflorum Lam.) for future genetic manipulations to improve forage characteristics, the effects of culture medium supplements on tissue culture responses were investigated with mature seeds of three cultivars, 'Jeanne', 'Florida-80' and 'Metro', as explant tissues. For all explants, MS medium containing 5mg/L 2,4-D was optimal for embryogenic callus induction from mature seed and had a strong effect on successive plant regeneration. The optimal concentration of dicamba for the induction of embryogenic callus from mature seeds was 7mg/L. The highest plant regeneration frequency was observed when embryogenic callus was transferred to N6 medium supplemented with 1mg/L 2,4-D and 5mg/L BA. Plant regeneration frequency of callus cultured in the dark was higher than that of cultured in the light. Casein hydrolysate and L-proline improved both in embryogenic callus induction from mature seeds and plant regeneration. High-frequency regeneration system established in this study will be useful for molecular breeding of Italian ryegrass through genetic transformation.

Improvement of Shoot Regeneration from Scutella-Derived Callus in Rice

  • Kim, Yong-Wook;Cho, Joon-Hyeong;Lee, Jang-Yong
    • KOREAN JOURNAL OF CROP SCIENCE
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    • v.49 no.1
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    • pp.52-60
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    • 2004
  • The optimized in vitro culture system was investigated for improvement of regeneration efficiencies by observing the responses of scutella-derived callus of Korean rice (Oryza sativa L.). Large variations of callus induction (43.9-93.9%) and shoot regeneration (0-88.7%) were observed among the rice cultivars depending on medium. However, shoot regeneration was significantly improved by selected utilization of basal medium, growth regulators, and carbon sources. N6 basal medium was more efficient for embryogenic callus induction than MS or LS basal medium, while MS was superior to N6 for shoot regeneration. The calli of highly regenerative cultivars grew faster and showed higher rates of green tissue formation (GT) and shoot regeneration (SR) and lower rate of callus browning (CB) than those of recalcitrant cultivars. Although a higher level of kinetin stimulated the GT and SR in highly regenerative cultivars, $10\textrm{mgL}^{-1}$ kinetin generally suppressed the GT and SR, while CB was accelerated compared to $2\textrm{mgL}^{-1}$ kinetin. Additional benefits of sorbitol combined with maltose (or sucrose) under $5\textrm{mgL}^{-1}$ kinetin were certainly confirmed on regeneration efficiencies compared to sucrose alone as carbon source and osmotic regulator. This combination showed high rate of GT and SR with multiple shoots while low rate of CB. With MSRK5SM-Pr medium ($5\textrm{mgL}^{-1}$ kinetin, 3% sorbitol, 2% maltose, $500\textrm{mgL}^{-1}$ proline), the regeneration efficiencies of total 17 out of 24 cultivars were practically improved 160% on average compared to MSRK2S ($2\textrm{mgL}^{-1}$ kinetin, 3% sucrose) control medium. Especially, the medium was most effective to the cultivars showing a medium level of regenerability such as Daesanbyeo and Dongjinbyeo and Suwon477, enhancing efficiencies more than 300-600% compared to MSRK2S medium.

Callus Induction and Plant Regeneration from Leaf Tissue. Culture of $\emph{Aralia elata}$ S. (두릅의 엽조직배양에 의한 Callus유기 및 식물체 재분화)

  • 장한호;박철호;조동하;신영범
    • KOREAN JOURNAL OF CROP SCIENCE
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    • v.38 no.4
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    • pp.366-370
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    • 1993
  • This study was conducted to determine the optimum culture conditions for inducing callus and regenerating plantlets from cultured leaf tissues of Aralia elata. Young leaf tissues(1cm) of A. elata plant were cultured on MS medium supplemented with 2, 4-D and Thidiazuron. Embryogenic callus was induced along the leaf veins, more efficiently on the medium containing 1.0mg /1 Thidiazuron in 4 weeks after culture initiation. Calli were subcultured to proliferate on MS media containing 2, 4-D, Dicamba, Picloram, and Thidiazuron. Callus was better proliferated on the medium containing Dicamba than on the others.. However, callus subcultured on the medium containing Thidiazuron was more embryogenic and light green-colored, of which some showed embryoid-like structure on the surface. Hormone-free medium was more efficient to regenerate plantlets than media supplemented with Kinetin, BA, and Thidiazuron.

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Embryogenic cell suspension culture and plant regeneration in zoysiagrass (Zoysia japonica Steud) (한국들잔디 배아세포의 부유배양과 식물체 재생)

  • Fang, Wenjuan;Han, Liebao;Qi, Chunhui;Li, Deying;Park, Tae-Yun
    • Asian Journal of Turfgrass Science
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    • v.23 no.2
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    • pp.345-352
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    • 2009
  • Zoysiagrass (Zoysia japonica Steud) is a warm season turfgrass species widely used for sports field and golf courses. Many cultivars are propagated through vegetative methods. This study was conducted to develop an optimum culture medium and culture conditions for embryogenic callus induction and plant regeneration, and to establish a cell suspension culture system for use in zoysiagrass breeding and propagation. The results indicated that adding $Cu^{++}$ at 2.5 mg $L^{-1}$ to the induction medium was optimum for callus induction. Increasing the numbers of sub-culture cycles improved the quality of calli. The optimum dosage for cell suspension culture ranged from 2.5 to 10 mL. The embryogenic callus suspension used in this study had a plant regeneration rate of 58%.

Factors Affecting Callus Culture and Plant Regeneration in Kentucky Bluegrass (켄터키 블루그래스에 있어서 캘러스 배양 및 식물체 재분화에 미치는 요인의 영향)

  • Lee, K.W.;Lee, S.H.;Lee, D.G.;Woo, H.S.;Kim, D.H.;Choi, M.S.;Won, S.H.;Seo, S.;Lee, B.H.
    • Journal of Animal Science and Technology
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    • v.47 no.6
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    • pp.1067-1074
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    • 2005
  • In order to optimize tissue culture conditions of Kentucky bluegrass(Poa pratensis L.), effects of culture medium supplements, media and cultivars on embryogenic callus induction and regeneration of plants were investigated. MS medium containing 3mg/L 2,4-D and 0.1mg/L BA was optimal for embryogenic callus induction from mature seeds. The highest plant regeneration frequency(57.7%) was observed when the embryogenic calli were cultured on N6 medium supplemented with 1mg/L 2,4-D and 3mg/L BA. Among several basic media, MS and N6 medium were optimal for callus induction and plant regeneration, respectively. Genotype was an important factor in plant regenerability. ‘Newport’ showed to have higher regeneration frequency of 53.4%. Regenerated plants were grown normally when shoots transplanted to the soil. A short tissue culture period and high-frequency regeneration system would be beneficial for molecular breeding of Kentucky bluegrass through genetic transformation.

Efficient Plant Regeneration from Alfalfa Callus by Osmotic Stress Treatment (알팔파 캘러스로부터 삼투압 스트레스 처리에 의한 효율적인 식물체 재분화)

  • Kim, J.S.;Lee, D.G.;Lee, S.H.;Woo, H.S.;Lee, B.H.
    • Journal of Animal Science and Technology
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    • v.46 no.5
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    • pp.879-886
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    • 2004
  • Effects of culture mediwn supplements and osmotic stress treatment on embryogenic callus induction and somatic embryogenesis were investigated in order to optimize tissue culture conditions of alfalfa(Medicago sativa L.). SH mediwn containing 5mgIL 2,4-D and 0.2mgIL kinetin was optimal for embryogenic callus induction from cotyledon tissue of alfalfa. Somatic embryos were formed when the embryogenic callus was cultured on SH mediwn supplemented with ImgIL 2,4-D and 2mgIL BA. Supplementation of 5mM L-proline and IgIL casein hydrolysate into the regeneration mediwn further increased plant regeneration frequency. Osmotic stress treatment of callus appeared to improve the frequency of somatic embryo formation, but the frequency of somatic embryo formation differed by the osmotic stress treatment using different osmotic stressors. The highest plant regeneration frequency of 30.7% was observed when embryogenic callus was treated with 0.7M sucrose for 18h. Efficient regeneration system established in this study will be useful for molecular breeding of alfalfa through genetic transformation.

High frequency plant regeneration from mature embryos of an elite barley cultivar (Hordeum vulgare L. cv Baegdong)

  • Lee, Kui-Jae;Wtpsk, Senarath;Lee, Wang-Hyu
    • Proceedings of the Plant Resources Society of Korea Conference
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    • 2003.10b
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    • pp.21-21
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    • 2003
  • An efficient plant regeneration system was developed for Hordeum vulgare L. cv Baegdong - an important Korean cultivar. The protocol was based on a series of experiments involving the sizes of mature embryos and the culture media. The embryo size is found to be critical for the establishment of embryogenic callus. Embryos of 1.1-1.5 mm size showed a much higher ability to produce embryogenic callus capable of regenerating green plants. The auxins picloram and dicamba proved effective in inducing callus from mature embryos. 2.5 mg $I^{-1}$ dicamba and 4.0 mg $I^{-1}$ picloram in Murashige and Skoog's (MS) medium was optimum for the induction of primary callus. The induced primary callus was loose and friable which ultimately developed into creamy white and compact callus after transferring into the fresh medium. Multiple shoots were induced in the MS medium supplemented with 6.0 g $I^{-1}$ maltose, 20 mg $I^{-1}$ sorbitol, 0.5 mg $I^{-1}$ 2,4-D and 1.0 mg $I^{-1}$ kinetin and the rate was 6.5 shoots per embryo. Regenerated plants were hardy and developed roots rapidly in the medium containing 0.2 $I^{-1}$ IBA. This efficient plant regeneration system provides a foundation for generating transgenic plants of this important barley cultivar.

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High frequency plant regeneration from mature embryos of an elite barley cultivar (Hordeum vulgare L. cv Baegdong)

  • Lee, Kui-Jae;Wtpsk, Senarath;Lee, Wang-Hyu
    • Proceedings of the Plant Resources Society of Korea Conference
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    • 2003.10a
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    • pp.59-67
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    • 2003
  • An efficient plant regeneration system was developed for Hordeum vulgare L. cv Baegdong - an important Korean cultivar. The protocol was based on a series of experiments involving the sizes of mature embryos and the culture media. The embryo size is found to be critical for the establishment of embryogenic callus. Embryos of 1.1-1.5 mm size showed a much higher ability to produce embryogenic callus capable of regenerating green plants. The auxins picloram and dicamba proved effective in inducing callus from mature embryos. $2.5\;m;I^{-1}$ dicamba and $4.0\;mg\;I^{-1}$ picloram in Murashige and Skoog's (MS) medium was optimum for the induction of primary callus. The induced primary callus was loose and friable which ultimately developed into creamy white and compact callus after transferring into the fresh medium. Multiple shoots were induced in the MS medium supplemented with $6.0\;g\;I^{-1}$ maltose, $20\;mg\;I^{-1}$ sorbitol, $0.5\;mg\;I^{-1}$ 2, 4-D and $1.0\;mg\;I^{-1}$ kinetin and the rate was 6.5 shoots per embryo. Regenerated plants were hardy and developed roots rapidly in the medium containing $0.2\;I^{-1}$ IBA. This efficient plant regeneration system provides a foundation for generating transgenic plants of this important barley cultivar.

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