• Title/Summary/Keyword: callus development

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Effective Micropropagation of Pulsatilla cernua var. koreana through Apical Meristem Culture (할미꽃 정단 분열조직 배양을 통한 효율적 미세번식)

  • Ko, Jeong-Ae;Kim, Hyun-Soon
    • Korean Journal of Plant Resources
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    • v.21 no.5
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    • pp.362-367
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    • 2008
  • In order to investigate the effect of plant growth regulators on effective in vitro micropropagation, apical meristems of Pulsatilla cernua var. koreana were cultured on Murashige and Skoog's (MS) medium with 2,4-D, NAA, TDZ and BA. Media containing 2,4-D and kinetin, 2,4-D and TDZ, NAA and TDZ were not effective on callus induction. However, embryogenic or organogenic callus was obtained on media containing NAA and BA. Especially, on MS medium with 0.5mg/L NAA and 1.0mg/L BA was optimal for a high frequency (62%) of shoot or shoot bud obtained from callus. Callus proliferation, shoot multiplication and elongation were significantly increased by adding 10% coconut water on MS media with 0.5mg/L NAA and 1.0mg/L BA. Repeated subculturing of in vitro grown shoots resulted in propagation rate of 12.9 shoots per explant every 30 days. Root formation from the adventitious shoots was not easily achieved. However, roots were only produced through callus on MS medium with 2.0mg/L NAA alone or 0.5mg/L NAA and 1.0mg/L BA. These roots were used materials for callus and shoot production repetitively.

In vitro seed germination and callus formation on flower bud of Korean mistletoe ( Viscum album L. var. cololatum [Kom.] Ohwi) (겨우살이 종자 발아 및 화아 배양에 의한 캘러스 형성)

  • Kim, Suk-Weon;Ko, Suk-Min;Liu, Jang-R.
    • Journal of Plant Biotechnology
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    • v.35 no.1
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    • pp.47-53
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    • 2008
  • Effects of growth regulators and culture conditions on seed germination, haustorium development, and callus formation of Korean mistletoe (Viscum album var. coloratum (Kom.) Ohwi) were described. Histological examination showed that seed of V. album contained one or two zygotic embryos with rod shape, and actively dividing cells were mainly distributed in radicle region rather than cotyledon of zygotic embryo. The most significant factor for seed germination and haustorium development of V. album was the requirement of the light. Various growth regulators examined in this study failed to substitute the effect of the light on seed germination. The frequency of callus formation was highest at 27.3% when flower buds were cultured onto B5 medium containing $0.1\;mgl^{-1}$ IAA. Explants from other organs were recalcitrant in forming calluses. Culture conditions described in this study could be applied for production of useful metabolites and multiplication of V. album in future.

New embryogenesis from atypical bodies and plant regeneration from long-term subcultured embryogenic callus in rose (장기간 계대배양 된 장미 배발생 캘러스로부터 식물체 재분화 및 비정형체로부터 새로운 배발생캘러스 재생)

  • Lee, Su Young;Do, Kyoung Ran;Cheon, Kyeong-Seong;Kim, Won Hee;Kwon, O Hyeon;Lee, Hye Jin
    • Journal of Plant Biotechnology
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    • v.41 no.2
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    • pp.89-93
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    • 2014
  • Long-term subcultured rose embryogenic calluses, which had been maintained for more than 5 to 6 years since the first embryogenesis from calluses induced from in vitro roots of rose, were identified as potential material for the development of transgenic plants. The first embryogenic calluses from 'Sweet Yellow' and two breeding lines (KR056002 and KR056006) were obtained in 2007 and 2009, respectively. Subsequently, we found that plants regenerated from long-term embryogenic calluses (LEC). Whereas the LEC from 'Sweet Yellow' takes 3 to 4 months to regenerate plants, those of the two breeding lines take 4 to 5 months. This period of time is the same as that taken for plants to regenerate from the first embryogenic callus. New embryogenesis was observed from atypical bodies (ABs) that appeared during the process of long-term subculture. We found that it is possible to use the AB as a material for new embryogenesis.

Callus Induction and Embryogenesis Through Pollen Culture in Paeonia albiflora PALL (작약의 화분배양에 의한 캘러스 및 배발생)

  • 김영숙;이병기
    • Korean Journal of Plant Tissue Culture
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    • v.22 no.1
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    • pp.13-17
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    • 1995
  • In order to induce haploid plant through pollen culture, pollens of Paeonia albiflora were cultured on MS liquid medium The development of micospore through pollen culture was examined The effect of low temperature (5$^{\circ}C$, 10 days) pretreatment on callus induction and embryogenesis in pollen culture was not evident Calli derived from pollen gave rise to globular embryos when transferred onto solid medium containing 0.5 mg/, 2,4-L. The effect of low temperature pretreatment and medium. combination to pollen viability was unrecognized. Pollen viability was reduced as the culture proceeded.

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Growth Regulators and Colchicine Treatments for Embryo Culture Efficiency in Barley (보리 배배양 효율증진을 위한 생장조절제와 콜히친처리 효과)

  • Bong Yeon, Kim
    • KOREAN JOURNAL OF CROP SCIENCE
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    • v.40 no.6
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    • pp.757-767
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    • 1995
  • This experiment was done to determine the optimum concentration of IAA for root development in plants regenerated from the callus culture of barley embryos. Two concentrations of 2,4-D, 3ppm and 5ppm selected as an optimum among five different concentrations in the previous experiment were used for callus induction and proliferation in this experiment. For callus induction, 3ppm of 2,4-D produced 35.6% in immature embryos and 4.4% in mature embryos, while 5ppm gave 33.8% in immature and 5.6% in mature embryos. Out of 320 immature embryos cultured, 111 embryos were induced to calli and 684 plants were produced from them, while only 16 embryos were induced to calli from 320 mature embryos and 92 plants were restored. The rates of callusing and plant regeneration were 34.7%, 214% in immature embryos and 5.0%, 28.7% in mature embryos, respectively. The average root lengths and root numbers of plants restored from callus at five different IAA concentrations of 0ppm, Ippm, 5ppm, l0ppm and 30ppm were 7.9mm, 3.6; 18.4mm, 5.2; 16.1mm, 3.9; 8.5mm, 3.5 and 6.4mm, 3.4, while plants directly obtained from mature embryos were 14.8mm, 4.9; 4.9mm, 3.6; 4.3mm, 3.1; 3.6mm, 2.6 and 3.2mm, 2.1, respectively. Therefore, 1ppm gave the best result for the root. promotion in callus, whle 0ppm, a control, gave the largest root developmemt in embryos. High concentration of lAA(30ppm) in callus and any exogeneous supplement of lAA in embryos negatively affected to the root lengths and root numbers. Genotypic effect was also observed in given four varieties, Bruce, Klages, Olbori and Albori. For chromosome doubling, when 0.1% colchicine was applied on 428 plants under three different conditions such as air circulation, temperatures and growth stages, 319 plants of doubled haploids were obtained so that the rate was 74.5%

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Development of Somatic Embryos from Stem Segments and Anthers in Kalanchoe daigremontiana (칼란코에의 줄기 切片 및 葯 培養으로부터 體細胞胚의 發生)

  • 이강섭
    • Korean Journal of Plant Resources
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    • v.10 no.1
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    • pp.86-93
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    • 1997
  • In order to induce somatic embryogenesis from the stem explants and anther of Kalanchoe daigremontiana, the explants were cultured on MS medium supplemented with auxin (2,4-D, IAA, NAA) and/or cytokinin (BAP) for 8 weeks. Callus from explants was induced most efficiently on MS medium containing. 2.0mg/L NAA and 0.2mg/L BAP. Somatic embryogenesis in stem callus was formed by transfering embryogenic callus from induction media containing growth regulators to medium without growth regulators and then to the medium containing auxin and cytokinin (0.1 mg/L IAA and 1.0mg/L BAP). Callus formation occurred actively in the anthers at early uninucleate stage, and by low temperature pretreatment at $4^{\circ}C$ for 3days. Somatic embryogenesis from the anther callus was induced on MS medium containing 1.0mg/L NAA and 1.0mg/L BAP, 2.0mg/L NAA and 0.2mg/L BAP. The tetraploid of 5.4% was obtained among plants regenerated from anthers.

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Mass Production of Calla Lily(Zantedeschia spp. Southern Light) by the Immature Zygotic Embryo Culture (유색칼라(Zantedeschia spp. Southern Light) 미숙배 배양에 의한 다량증식)

  • 고정애;최소라;김현순
    • Korean Journal of Plant Resources
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    • v.16 no.2
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    • pp.160-167
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    • 2003
  • In order to investigate the effects of developmental stage of embryos and plant growth regulators on mass production of Zantedeschia spp. Southern Light, immature zygotic embryos of Zantedeschia spp. Southern Light were cultured on Murashige and Skoog(1962) basal media or containing 2,4-D, NAA and BA. Globular embryos did not grow on any of the 2,4-D, NAA and BA combinations. The most suitable stage of immature zygotic embryo culture on the induction callus and multiple shoot was at early cotyledonary embryo stage, and at this stage of embryos were germinated up to 87.5%. The whitish watery callus and yellowish compact nodular callus produced on all 2,4-D, NAA and BA media. The best combination for inducing embryogenic callus was 0.5 mgL NAA and 1.0 mg/L BA. Whitish watery calli have been subcultured for more than 8 months and have retained their producing ability, Plant regeneration was only obtained by direct shoot development and yellowish compact nodular calli. Abundant plantlets were regenerated from cotyledonary stage of embryo culture on MS medium supplemented with 0.5 mg/L NAA and 1.0 mg/L BA. Supplementation of the media with 10% coconut water showed as the best concentration for plant differentiation from direct developed of shoots. The number of regenerated plants from one embryo could be seperated 25-35s plantlets. All yellowish compact callus-derived plantlets were transferred to pots containing a mixture of vermiculite, perlite and sand(1:1;1 v/v) and 100% of divided plantlets were phenotypically normal.

In vitro Callus and Somatic Embryo Induction of Six Hosta Species Native to Korea

  • Choi, Han;Lee, Seung Youn;Ryu, Sun Hee;Yoon, Sae Mi;Kim, Sang Yong;Lee, Jong Suk;Yang, Jong Cheol
    • Proceedings of the Plant Resources Society of Korea Conference
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    • 2018.10a
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    • pp.80-80
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    • 2018
  • Hosta is a genus of the family Asparagaceae and distributed in East Asia. There are six Hosta species (Hosta capitata (Koidz.) Nakai, H. clausa Nakai, H. jonesii M.G.Chung, H. minor (Baker) Nakai, H. venusta F.Maek., and H. yingeri S.B.Jones) native to Korea and among them, four species (H. minor, H. jonesii, H. venusta and H. yingeri) are endemic to the Korea peninsula. Hosta is generally propagated by seed, crown division or tissue culture. However, tissue culture is a more efficient method to mass proliferation, a new cultivar development and disease-free plantlet production in a limit time. Hence, we conducted this study to evaluate the influence of various plant growth regulators (PGRs) treatments on the induction of callus and somatic embryo of the six Hosta species. Leaf, petiole and root were used to select optimum tissue culture explants. Petiole explants thus only were used for callus induction and somatic embryogenesis with TDZ (0.1, 0.5 or 1.0mg/L) and NAA (0.1 or 0.5 mg/L) combinations. After 12 weeks of culture, the highest rate of somatic embryogenesis was achieved on modificated MS medium containing 1.0 mg/L TDZ and 0.1 mg/L NAA in H. capitata and H. minor (15.5%, respectively), 0.1 or 0.5 mg/L TDZ and 0.1 mg/L NAA in H. jonesii (22.2%), 1.0 mg/L TDZ and 0.5 mg/L NAA in H. yingeri (26.7%), and 0.1 mg/L TDZ and 0.5 mg/L NAA in H. venusta (53.3%). H. clausa showed very low effect on somatic embryogenesis by PGRs; 2.2%. There was interspecies difference to PGRs respond for callus and somatic embryo induction. Regenerated multiple shoots and plantlet of H. minor, H. jonesii, H. venusta and H. yingeri were obtained via somatic embryogenesis.

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REDIFFERENTIATION FROM TISSUE CULTURE AND ISOLATION OF VIABLE PROTOPLASTS IN PANAX GINSENG C.A. MEYER (고려인삼의 조직배양에 의한 기관형성과 원형질체배양에 관한 연구)

  • Choi Kwang-Tae;Yang Deok-Chun;Kim Nam-Won;Ahn In-Ok
    • Proceedings of the Ginseng society Conference
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    • 1984.09a
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    • pp.1-11
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    • 1984
  • Ginseng cotyledon calli were cultured on 1/2MS media supplemented with combination of various growth regulators to induce more embryoids and plantlets in a short period. And tissues of ginseng root and calli were also incubated under various factors or conditions to establish methods for the isolation of viable protoplasts in Panax ginseng C.A. Meyer. The calli derived from cotyledon produced numerous embryoids in 1/2MS media containing 0.5mg/$\ell$ 2,4-D and 0.5mg/$\ell$ kinetin after 2 months' culture. But only shoot formation was less frequent. Further development of these embryoids occurred on 1/2MS medium supplemented with the same concentration of BA and GA. Viable protoplasts were isolated from the root tissue and callus of ginseng. The specific conditions for the isolation of viable protoplasts were required of ginseng materials, root tissue and callus, being processed. For the production of viable protoplasts from 1-year old ginseng root tissue, an enzyme mixture of $2\%$ cellulase 'Ono-zuka' and $0.5\%$ macerozyme, an enzyme solution pH of 5.2 to 5.8, a 7- to 8- hour incubation period at $28{\pm}1^{\circ}C$, and 0.9M mannitol as osmoticum in the cell enzyme mixture were optimum, while the treatments with an enzyme mixture of $2\%$ cellulase 'Onozuka', $2\%$ macerozyme and $1\%$ driselase, and 25-hour incubation period at $28{\pm}1^{\circ}C$, were more efficient for the production of viable protoplasts from ginseng callus.

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Development of Herbicide Resistant Plant Through Plant Tissue Culture (제초제(除草劑) Butachlor 및 Simetryne에 저항성(抵抗性)인 식물체(植物體) 선발육성(選拔育成))

  • Kim, K.U.;Kim, S.H.
    • Korean Journal of Weed Science
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    • v.7 no.2
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    • pp.200-207
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    • 1987
  • This study was conducted to select and develop herbicide resistant plant through tissue culture. Growth response of seedlings and callis of various rice varieties with Echinochloa species was assessed under the treatment of various rates of butachlor [N-(butoxy methyl)-2-chloro-2', 6'-diethyl acetamide] and simetryne [2,4-bis(ethyl amino)-6-methyl thio-1,3,5-triazine]. Further, succinate dehydrogenase activity was determined in herbicide treated callus to characterize different response of plants to herbicide. Rice variety like Sangpung showed relative resistance in both callus and seedling states against butachlor, indicating maintenance of resistance. However, in the simetryne treatment, the similar response was not observed in callus and seedling state, although there was a great different response among plant materials against simetryne. Rice variety which exhibited resistance in callus and seedling states showed low succinate dehydrogenase inhibition index. Succinate dehydrogenase inhibition index can be used as an important marker characters to differentiate varietal response of plant to herbicide. Rice plant was differentiated from butachlor and simetryne tolerant callus treated at $2.5{\times}10^{-5}$ M is growing under the growth chamber and can be used for resistant source.

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