• Title/Summary/Keyword: Explants

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Plant regeneration and soil acclimatization through photoautotrophic culture from leaf explant of a rare species in Sedum tosaense Makino (희귀수종인 주걱비름(Sedum tosaense Makino)의 잎절편으로부터 기내 식물체 재분화 및 광독립배양을 통한 토양순화)

  • Ko, Myoung-Suk;Bae, Kee Hwa;Song, Gwanpil;So, In Sup
    • Journal of Plant Biotechnology
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    • v.40 no.2
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    • pp.79-87
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    • 2013
  • The aim of this study was to establish plant regeneration from leaf explants of Sedum tosaense Makino, which is globally rare and endangered species. The leaf explants of S. tosaense were cultured on the MS medium supplemented with different concentration of BA and NAA for callus induction. Callus induction was showed the highest (100%) on MS medium containing $2.0mg{\cdot}L^{-1}$ BA and $1.0mg{\cdot}L^{-1}$ NAA. The highest number of shoots were regenerated when callus were cultured on MS medium containing $2.0mg{\cdot}L^{-1}$ BA and $1.0mg{\cdot}L^{-1}$ NAA for 5 weeks. The axillary bud were cultured on the MS media supplemented with combination of BA and NAA for in vitro propagation. The highest number of adventitious shoot (7.9 per explants) formed at $1.0mg{\cdot}L^{-1}$ NAA and $2.0mg{\cdot}L^{-1}$ BA. For rooting, MS medium supplemented with or without $2.0g{\cdot}L^{-1}$ activated charcoal was tested. The optimal results were observed using MS medium supplemented with $2.0g{\cdot}L^{-1}$ activated charcoal, on which 85.7 (No. of root), 4.6 cm (length of root). 1,200 ppm $CO_2$ and 350 ppm $CO_2$ were supplied for make certain the effects of $CO_2$ on pre-acclimatization by photoautotrophic culture. 1,200 ppm $CO_2$ treatment was established higher than 350 ppm $CO_2$ treatment. Soil acclimatization of in vitro plantlets was the best in mixture soil consisted of peat moss and perlite with 100% survival rate and they showed the maximum growth.

Position Effect of Axillary Buds on Shoot Multiplication and Rooting in Bud Culture of Quercus acutissima (상수리나무 기내(器內) Axillary Bud의 치상부위(置床部位)에 따른 다경(多莖) 및 발근유도(發根誘道) 효과(効果))

  • Moon, Heung Kyu;Kim, Jae Hun;Park, Jae In
    • Journal of Korean Society of Forest Science
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    • v.76 no.4
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    • pp.370-375
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    • 1987
  • This study was conducted to examine the position effect of axillary buds on shoot multiplication and rooting with 1-year-old seedlings of Quercus acutissima. Shoot multiplication was greatly affected by axillary bud position: Productivity of multiple shoots were decreased in the order of top, mid and basal explants respectively. The best shoot multiplication (mean 6.1 shoots per explant) was obtained on WPM medium containing $1.0mg/{\ell}$ BAP and $0.1mg/{\ell}$ NAA using basal explant after 4-week-culture. Rooting was also greatly influenced by position. Its percentage was increased in the order of top, mid and basal explant respectively. Root initiation was better and more rapid on 1/2MS medium than GD medium. High rooting percentage (100%) was obtained on 1/2MS medium containing $0.2mg/{\ell}$ IBA after 15 days culture. Sucrose concentrations did not effect on rooting. However root development and shoot growth were greatly affected by them. Root was developed shortly on 1-2% levels and shoot growth was getting retarded, whereas both of them did not show significant difference at 3-6% levels. Rotting was decreased on 7-8% levels gradually, but shoot and leaf condition was better than any other concentrations. Survival rate of rooted explants in pot was varied according to the position of explants. Seedlings of top part were survived up to about 50% but most of mid and basal part seedlings did not survive over 4 weeks even in high humidity condition. Seedlings in pots showed normal growth over 10 months but most of them showed the condition of premature leaf shedding.

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Efficient plant regeneration through callus induction from the hypocotyl of Perilla frutescens L var. Dayu ('다유들깨'품종의 하배축에서 캘러스를 통한 고효율 식물재분화)

  • Ruyue Xu;Ji-Hi Son;Hong-Gyu Kang;Hyeon-Jin Sun;Hyo-Yeon Lee
    • Journal of Plant Biotechnology
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    • v.50
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    • pp.248-254
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    • 2023
  • This study was conducted to establish an efficient plant regeneration system in 'Dayu', a Korean variety of Perilla frutescens developed for seed oil production, in conjunction with the previously studied variety 'Namcheon'. The healthiest callus was formed on the hypocotyl explants cultured on a medium containing 0.1 mg/L NAA and 0.5 mg/L BA, outperforming the leaf and cotyledon samples. In both dark and long-day conditions, Dayu consistently exhibited significantly higher shoot regeneration rates compared with Namcheon. The highest shoot regeneration rates in Dayu were observed from the hypocotyl explants cultured on 0.1 mg/L NAA and 0.5 mg/L BA media, with shoot regeneration rates of 84.4% and 86.7% under dark and long-day conditions, respectively. Various combinations of plant growth regulators were tested to establish the optimal shoot regeneration conditions for Dayu hypocotyl explants. The results demonstrated that the highest shoot regeneration rate (90%) was achieved when 0.5 mg/L of BA was added to the medium without NAA. Among the regenerated shoots, 70.5% were normal plants, while 19.3% were abnormal. The addition of NAA or an increase in its concentration led to a higher occurrence of abnormal plants. After the regenerated shoots were transferred to 1/2 MS medium, roots were observed within 10-15 days. By day 30, they had developed into complete plants. The results obtained from the regeneration experiments with the perilla variety Dayu can valuably inform molecular breeding reliant on transformation techniques such as genome-editing and genetic modification technology.

Plant regeneration via callus culture of sea-milkwort (Glaux maritima L.) (갯봄맞이(Glaux maritima L.)의 캘루스 배양을 통한 식물체 재분화)

  • Han, Jeung-Sul
    • Journal of Plant Biotechnology
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    • v.44 no.2
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    • pp.171-177
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    • 2017
  • A callus-mediated regeneration protocol for sea-milkwort, an endangered coastal plant species in South Korea, is reported here. The explants of in vitro-plantlets generated from a node culture revealed distinguishable responses in callus induction depending on genotype, explant source, light condition, and 2,4-D concentration. Especially, continuous darkness exclusively facilitated callus induction from explants prior to other treatments. The calli initiated on the media with 2,4-D ranging from 0.1 mg/L to 3.0 mg/L in the dark vigorously proliferated when subcultured on the same media in continuous darkness. Given 1.0 mg/L zeatin in addition to darkness to the calli of the 'Pistachio' genotype, normal adventitious shoots were only regenerated from nodular structures that formed earlier from the calli at the frequency of 24.4 percent. Regenerated shoots easily grew into plantlets with roots and green color on a phytohormone-free MS medium under lighted condition, that were used for node culture as plant materials. Node culture effectively multiplied plantlets in accordance with protocol by Bae et al. (2016). Acclimatized plantlet clusters developed mature plant clusters under inland environment, followed by flowering the following April. Results were merged with node culture protocol suggested by Bae et al. (2016), which, as an in vitro propagation system for sea-milkwort, may contribute to natural habitat restoration.

Proteomic Responses of Diploid and Tetraploid Roots in Platycodon grandiflorum (2배체와 4배체 도라지의 단백질 발현양상 비교 분석)

  • Kim, Hye-Rim;Kwon, Soo-Jeong;Roy, Swapan Kumar;Cho, Seong-Woo;Kim, Hag-Hyun;Moon, Young-Ja;Boo, Hee-Ock;Woo, Sun-Hee
    • KOREAN JOURNAL OF CROP SCIENCE
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    • v.60 no.3
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    • pp.394-400
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    • 2015
  • The roots of Platycodon grandiflorum species either dried or fresh, are used as an ingredient in salads and traditional cuisine in Korea. To interpret the root proteins, a systematical and targeting analysis were carried out from diploid and tetraploid roots. Two dimensional gels stained with CBB, a total of 39 differential expressed proteins were identified from the diploid root under in vivo condition using image analysis by Progenesis Same Spot software. Out of total differential expressed spots, 39 differential expressed protein spots (${\geq}\;1.5$-fold) were analyzed using LTQ-FTICR mass spectrometry. Except two proteins, the rest of the identified proteins were confirmed as down-regulated such as Isocitrate dehydrogenase, Proteasome subunit alpha type-2-B. However, the most of the identified proteins from the explants were mainly associated with the oxidoreductase activity, nucleic acid binding, transferase activity and catalytic activity. The exclusive protein profile may provide insight clues for better understanding the characteristics of proteins and metabolic activity in various explants of the economically important medicinal plant Platycodon grandiflorum.

The Condition of Regeneration and Antibiotics Concentration for Gene Transformation of Zygotic Embryo in Panax ginseng C. A. Meyer (인삼(Panax ginseng C. A. Meyer)접합자 배의 형질전환을 위한 재분화 및 항생제 농도 조건)

  • 양덕춘;이은경;최원균;김무성
    • Korean Journal of Plant Resources
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    • v.16 no.1
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    • pp.74-81
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    • 2003
  • To obtain multi-shoot using zygotic embryos dissected from ginseng seed, the embryos were cultured on MS medium supplemented with CPA and BA. Effective multi-shoot induction was achieved on 0.5mg/ t CPA and 1.0mg/ t BA treatment. Among the various plant growth regulator treatment, MS basal medium with 1mg/ t 2,4-D and 0.5mg/ t kinetin was more competent and could be induced 4∼6 shoots per one embryo. Also, the best condition for pre-embryoid induction from ginseng cotyledon so as to ginseng transformation appeared to 1mg/ t 2,4-D and 0.5mg/ t kinetin treatment. The kanamycin level to select transformants varied greatly by different explant tyues. The petiole explants with leaf and embryo could survived up to 100$\mu\textrm{g}$ / ml kanamycin concentration where as petiole explants without leaf died all at the level. Conclusionally, our results suggest that optimum kanamycin concentration for ginseng transformation using somatic embryos is about 75∼100$\mu\textrm{g}$ / ml concentration.

Organ Induction by Combined Dose of bFGF and HGF in Animal Cap Assay of Early Xenopus laevis Embryos. (Xenopus laevis 초기 배의 동물극 분리배양에서 bFGF와 HGF 혼합처리에 의한 기관유도)

  • 진정효;윤춘식;이호선;박용욱;정선우
    • Journal of Life Science
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    • v.14 no.3
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    • pp.375-384
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    • 2004
  • Fibroblast growth factors (FGFs) are known to induce multiple functions in early development, including mesoderm formation, gastrulation movement and antero-posterior patterning. The induction of mesoderm from Xenopus presumptive ectoderm and the combination effect on inducing organs of bFGF(basic FGF) and HGF (Hepatocyte Growth Factor) were studied. Explants were cultured in the combined solution for 3 days to normal embryo arrive at St. 43. These effects on combined dose were examined by histological experiment and by immunohistochemical method. The concentrations of growth factors were tested in 0, 0.5, 1, 10 and also tested in 50 ng/ml of bFGF, and 0, 1, 10, 50 and 100ng/ml of HGF respectively. The synergistic effects were seen in the combined-dose of bFGF and HGF rather than in single dose. Various organs were differentiated and highest inducing effects were seen at the combined concentration of 1 ng/ml of bFGF and 10ng/ml of HGF, and at the concentration 10ng/ml of bFGF and 1 ng/ml of HGF. The bFGF induces various organs from cultured animal cap explants and the effects are time and dose-dependent. HGF is also a potent mitogen for renal tubular cells and for mature hepatocytes in primary culture. Eyes were developed in high percentage at the combined concentration of 1 and 10ng/ml of bFGF, and 1 and 10 ng/ml of HGF. From the induced eye and normal embryonic eye, RPE65 was commonly detected by monoclonal antibodies 40All and 25F5 and the localization of RPE65 was seen by AP reaction.

Multiple Shoot Formation of Gentiana axillariflora Leveille by in Vitro Culture (큰용담의 기내증식에서 multiple shoot의 유기)

  • Lim, Jung-Dae;Yu, Chang-Yeon
    • Korean Journal of Medicinal Crop Science
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    • v.8 no.1
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    • pp.41-48
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    • 2000
  • This study was aimed to proliferate Gentiana axillariflora Leveille which is one of the important medicinal and ornamental plants, by establishment of multiple shoot formation and embryogenesis through tissue culture technique. Callus was formed on MS (Murashige and Skoog) medium supplemented with 2,4-D, CPA, but not formed with BAP. The addition of 2,4-D 2 mg/ l into the medium was effective for callus formation and the rate of callus formation was about 90%. Somatic embryos were obtained on MS medium for two months. When callus was cultured on MS medium with combination treatment of 2,4-D 0.5 mg/ l and BAP 0.5 mg/ l, the number of embryo formed was better than that of other single or combination treatments and the total numbers of embryo a were 18.8 (number of total embryo/number of explants incubated = 753/40) at mean. Callus induction from stem and node explants was increased by addition of TDZ 2 mg/ l in the presence of 2,4-D 2 mg/ l, respectively. The best result about the differentiation of shoots was obtained on MS medium added BAP 2 mg/ l from node culture. Multiple shoots from shoot apex were induced on MS medium containing BAP 1 mg/ I and TDZ 1 mg/ l , BAP 2 mg/ l and TDZ 1 mg/ l. The number of multiple shoots per one explant was above seventy plants. It was the most effective regeneration system for rapid multiplication of Gentiana axillariflora Leveille.

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Establishment of Plant Regeneration from Apical Meristem of Sweetpotato (고구마 정단분열조직 유래 식물체 재분화 조건 확립)

  • Lee, Joon-Seol;Ahn, Young-Sup;Chung, Mi-Nam;Kim, Hag-Sin;Jeong, Byeong-Choon
    • KOREAN JOURNAL OF CROP SCIENCE
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    • v.51 no.spc1
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    • pp.233-236
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    • 2006
  • This study was conducted to investigate somatic embryogenesis capacity using callus derived from bud meristems in sweetpotato. Shoot apical meristem explants $(height:150{\mu}m;base:\;350{\mu}m)$were cultured on MS medium supplemented with 1 mg/L 2/4-D. Embryogenic callus were observed in five cultivars when their shoot apices were cultured on MS medium supplements with 1 mg/L 2,4-D. After 6 weeks of culture, greater than 80% of the survived explants produced embryogenic calli and the calli gave rise to somatic embryos at frequencies of 72% (Yulmi), 60% (Shinhwangmi), 78% (Geonmi), 70% (KoKei 14), 40% (Sinjami). The regenerated plants developed into whole plantlets after they were transferred onto the fresh hormon-free MS medium of 74% (Yulmi), 82% (Shinhwangmi), 86% (Geonmi), 74% (Kokei 14), 41% (Sinjami) respectively.

Effects of Light-emitting Diodes on In Vitro Growth of Virus-free Sweet Potato Plantlets (LED가 고구마 바이러스 무병묘의 기내 생장에 미치는 영향)

  • Yoo, Kyoung-Ran;Lee, Seung-Yeob
    • Horticultural Science & Technology
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    • v.35 no.4
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    • pp.490-498
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    • 2017
  • The in vitro growth of virus-free sweet potato [Ipomoea batatas (L.) Lam.] plantlets was investigated under different light sources: fluorescent lamp (control); red (660 nm), blue (460 nm), white light-emitting diodes (LED), and two mixtures of blue and red LED (R:B = 8:2, and 7:3). Single node explants (10 mm) of three cultivars ('Matnami', 'Shincheonmi', and 'Yeonhwangmi') were cultured on Murashige and Skoog medium supplemented with $0.2mg{\cdot}L^{-1}$ 6-benzyladenine for 4 weeks. Explants were exposed to $150{\pm}5{\mu}mol{\cdot}m^{-2}{\cdot}s^{-1}$ photosynthetic photon flux at a distance of 20 cm, constant temperature of $25^{\circ}C$, and under 16/8-h (day/night) photoperiod. Using the same method, the in vitro growth of 10 cultivars under red LED was also compared. After 3 weeks, vine length was highest in plantlets cultured under red LED, and lowest in plantlets cultured under blue LED. Fresh and dry weights were also greatest in plantlets cultured under red LED. Compared to the control, vine thickness was significantly higher in plantlets grown under white LED and the 7:3 R:B LED mixture. Significant differences were observed among the 10 cultivars grown under red LED. 'Matnami', 'Shincheonmi', and 'Shinhwangmi' all had excellent vine lengths, and fresh and dry weights. Compared to the control, vine elongation of sweet potato plantlets was most effective under red LED, and culture duration was about 1 week shorter.