• Title/Summary/Keyword: 캘러스 형성

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Plant Regeneration and Protein Analysis from Cadmium Resistant Callus of Tobacco (Nicotiana tabacum cv. BY4) (담배 (Nicotiana tabacum cv. BY4)카드뮴 저항성 캘러스로부터 식물체 재생과 단백질 분석)

  • 오승철;소웅영;조덕이;양덕춘
    • Korean Journal of Plant Tissue Culture
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    • v.28 no.1
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    • pp.7-13
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    • 2001
  • Calli were induced from diploid and haploid tobacco after 4 weeks and maintained on MS medium with combination of 2.0 mg/L 2,4-D,0.1 mg/L BAP and 2.0 mg/L kinetin. Suspension cells were screened through 65 $\mu$m-nylon mesh and 100 $\mu$m-mesh, then they were smeared on selection medium combined with cadmium and PFP by using the low melting agarose of 0.8%. After 30days smeared cultures of the medium the cell was treated with 500 $\mu$M and 1000 $\mu$M to select the resistant cell line were selected. Plant regeneration was induced from the selected cell lines on medium with 0.5, 1.5, 2.0 mg/L BAP and on media with combination of auxin and BAP under 500 $\mu$M and 1000 $\mu$M cadmium. At this time, plant regeneration was achived on cadmium free medium. In case of haploid, occurred from the cell line which is selected in medium with cadmium and PFP. In case of diploid regeneration occurred is in the medium with cadmium alone. The plantlet regenerated from cadmium resistant calli grew well in cadmium 500 $\mu$M. Protein pattern of leaf, root, stem of regenerated plants was analyzed. The quantum was 6.5188 ug/mg.fr.wt in the leaf of plant, 5.3611 ug/mg.fr.wt in the stem, 3.0213 ug/mg.fr.wt in the root. On the other hand, 5.9652 ug/mg.fr.wt. in the leaf of control, 3.5974 ug/mg.fr.wt in the stem of the control, 4.3766 ug/mg.fr.wt. in the root of the control. The one dimension bends regenerated from cadmium resistant calli resistant to cadmium in leaf were 49 involving 198.7KD etc. Disappeared were 4 involving 160.5KD etc, The protein bends were combinized were 3 involving 83.4KD etc. The bends resistant to cadmium stress in stem were 41 involving 4.3KD etc. Disappeared were 5 involving 114.8KD etc. The protein bends combinized were 6 involving 128.7KD etc. The bends which had the resistance to cadmium stress in root is 27 in volving 166,9KD etc. The bends which disappeared were 198.7KD etc. There were 5 involving 83.4KD etc.

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Effect of IAA and Zeatin Riboside on Plantlet Induction from Leaf Disks of Solanum tuberosum L. and Variation of Regenerated Plants (IAA와 Zeatin Riboside가 감자의 엽절편체로부터의 식물체 유기 및 재분화개체의 변이에 미치는 영향)

  • Park, Young-Doo;Boe, Arthur A.
    • Horticultural Science & Technology
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    • v.19 no.4
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    • pp.459-464
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    • 2001
  • Leaf disks from cultivar 'Kennebec' and one selection line (ND 860-2) were cultured on Murashige-Skoog medium with various combinations of indole acetic acid (IAA) and zeatin riboside. Shoots, roots and callus were induced at various combinations of plant growth regulator levels. The medium containing $3.5mg{\cdot}L^{-1}$ IAA and $4.0mg{\cdot}L^{-1}$ zeatin riboside produced the most plantlets. Rooted regenerants were grown in the greenhouse. The growth of regenerated plants obtained from the MS medium supplemented with $7.0mg{\cdot}L^{-1}$ IAA and $3.0mg{\cdot}L^{-1}$ zeatin riboside was significantly greater than those grown from nodal expalnts. In ND 860-2, a leaf chimera with chlorophyll deficient (light yellow) sectors was found in plants regenerated fiom leaf disks (grown on MS medium supplemented with $3.5mg{\cdot}L^{-1}$ IAA and $3.0mg{\cdot}L^{-1}$ zeatin riboside) but not in plants grown from nodal explants. The phenotypic variability was also observed in the tuber number, size and weight.

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Efficient Light Treatment for Graft-take and Early Growth of Grafted Tomato Seedlings (토마토 접목시 활착과 순화에 효과적인 광 처리 방법 구명)

  • Kim, Sung Eun;Lee, Moon Haeng;Kim, Young Shik
    • Journal of Bio-Environment Control
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    • v.22 no.4
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    • pp.322-327
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    • 2013
  • This research was conducted to elucidate the appropriate light environment right after grafting to produce vigorous cherry tomato seedlings. Tomato plants were grafted and then treated in 4 different ways: to keep in the natural light (Non), to cover the grafted stem part with aluminum foil to make only that part dark (Part), to put the grafted seedlings in the acclimation room for two (Day-2) or four days (Day-4) to make the whole seedlings in the dark condition. Tube grafting method was used for grafting, in which silicon tube of 1.5mm in diameter was used. The survival rate was the maximum in the treatment Day-2. The SPAD value, seedling quality and yield of $1^{st}$ and $2^{nd}$ cluster were the best in the treatment Part. The treatment Part needs cost more than other treatments but is more economic thanks to higher yield. Therefore it was concluded to be economically feasible to make the grafted stem part dark right after grafting in case of cherry tomato.

Effects of a variety of treatments affecting Chinese cabbage protoplast culture, and plant regeneration from protoplast-derived callus (배추 원형질체 배양에 미치는 다양한 처리의 효과와 원형질체 유래 캘러스로부터 신초 재분화)

  • Han, Jeung-Sul;Yoon, Moo-Kyeong;Jeong, Mi-Hye
    • Journal of Plant Biotechnology
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    • v.35 no.3
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    • pp.235-243
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    • 2008
  • Here we describe a procedure for Chinese cabbage protoplast culture and effects of various treatments. Chinese cabbage protoplasts were isolated from different parts of young seedlings as using an enzyme mixture, of which yield was maximized in seven hours around after digestion. The highest rate of initial cell division followed by micro-callus formation was obtained in the medium with 1.0 mg/L 2,4-D, 0.5 mg/L NAA, and 1.0 mg/L BA when the cotyledon-derived protoplasts were cultured. Initiation of cell division and micro-callus proliferation significantly depended upon Chinese cabbage genotype under a same culture circumstances. The micro-calli developed from cotyledon tissue of Norang-Bom cultivar successfully grew toward callus colonies on the solidified medium with 0.2 mg/L zeatin and 0.1 mM spermidine. The callus colonies generated de novo shoots at the maximum frequency of 4.3% on the medium with 5.0 mg/L BA and 1.0 mg/L NM. Our results might be helpful for further studies to enhance the regeneration efficiency in Chinese cabbage protoplast culture.

Improvement of Cultural Efficiency Using DNA Markers in Anther and Seed Culture of Rice (DNA marker를 이용한 벼의 조직배양 효율 재선)

  • Kim Kyung-Min
    • Journal of Life Science
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    • v.16 no.3 s.76
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    • pp.527-533
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    • 2006
  • The purpose of this study is to improve the culturability of a indica type rice cultivar, IR 36, using DNA marker associated with the ability of plant regeneration in anther and seed culture. The varietal difference of ability of callus formation and plant regeneration was investigated in anther and seed culture of 8 rice cultivars. Three japonica rice cultivars showed to have better culturability than those of tongil and indica type genotypes. But two indica/japonica lines, 'MGRI 079' and 'MGRI 036', which were selected to have good culturability in previous study showed the highest regenerability (20%) in anther culture of 8 rice cultivars. Thirty four $BC_2F_4$ lines were selected by marker screening using RZ400 for 100 $BC_2F_4$ lines derived from a cross $'MGRI\;079/IR\;36^{^*3}'$. The frequency of callus formation of 30 $BC_2F_4$ lines was higher than those of 'IR 36' in anther culture of the selected $BC_2F_4$ lines. The ability of plant regeneration of 15 lines was higher than that of 'IR 36' in the seed culture of 34 $BC_2F_4$ lines. A promising line, $BC_2F_4-28$, was selected to have better culturability in the anther and seed culture of the $BC_2F_4$ lines. The heading date and grain shape of the $BC_2F_4-28$ was similar to 'IR 36'. In seed culture of 50 $BC_4F_3$ lines derived from a rice cross $'MGRI\;079/IR\;36^{^*5}'$, 11 lines including $BC_4F_3-3$ showed to have higher regenerability compared with 'IR 36'. The highest frequency of plant regeneration (11%) was obtained from $BC_4F_3-46$ in seed culture of the $BC_4F_3$ lines.

Characterization of Arabidopsis Histidine Kinase 3 and Proteomic Analysis of Its Mutant (애기장대 histidine kinase 3 (AHK3)의 특성과 결손돌연변이체인 ahk3의 프로테옴 분석)

  • Liang Ying-Shi;Cha Joon-Yung;Ermawati Netty;Jung Min-Hee;Lee Kon-Ho;Son Dae-Young
    • Journal of Life Science
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    • v.16 no.3 s.76
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    • pp.447-453
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    • 2006
  • Histidine kinase plays important roles in signal transduction in plant. We characterized the function of Arabidopsis histidine kinase 3 (AHK3) and analyzed the expression patterns of genes and proteins in its mutant ahk3 by trans-zeatin (t-zeatin). The ahk3 exhibited decreased sensitivity to t-zeatin during callus formation, seedling growth, and leaf senescence. From proteomic analysis of ahk3, eukaryotic translation initiation factor 5A-2, auxin binding glutathione S-transferase, and NDPK1 were identified not to be induced by t-zeatin, when compared to the wild-type. In addition, the expression levels of ARR4 and ARR16 among A-type response regulators (ARRs) markedly decreased in ahk3 by t-zeatin treatment. These results suggest that AHK3 plays an important role in cytokinin signaling and the proteins identified from proteomic analysis and specific ARRs, ARR4 and ARR16 may be directly or indirectly associated in AHK3-mediated cytokinin signaling.

Plant Regeneration and in vitro Tuber Enlargement from Callus in Pinellia ternata(Thunb.) Breit (반하(半夏) 캘러스로부터 식물체(植物體) 재생(再生)과 기내(器內) 괴경(塊莖) 생장(生長) 유도(誘導))

  • Kim, Tai-Soo;Park, Moon-Soo;Park, Ho-Ki;Kim, Sun;Jang, Yeong-Sun
    • Korean Journal of Medicinal Crop Science
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    • v.2 no.3
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    • pp.246-250
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    • 1994
  • This study carried out to induce plant regeneration and callus formation from leaflet in MS medium with 2.4-D, NAL and IAA for in vitro growth of tuber, Kinetin and BA were used for plant regeneration. $NH_4NO_3$ and $KNO_3$ as a nitrogen source and $(NH_4)_2SO_4$, as a sulfate source were tested for in vitro growth of tuber. The resutls ars as fellows : 1. For the callus formation from leaflet and differentiation potency of organ, 2.4-D was more effective than IAA in MS medium under $26^{\circ}C$ and light condition of 8 hours a day. 2. For the plant regeneration from callus, MS medium with $2.0\;mg/{\ell}$ BA was most effective under $26^{\circ}C$ and light condition of $16{\sim}24$ hours a day. 3. For the in vitro growth of tuber, $KNO_3$ by $3.0g/{\ell}$ in MS medium was effective. This condition enhanced the growth of tuber 2.5 times compared with that in MS medium with $2.0\;mg/{\ell}$ BA.

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Plant Regeneration from the Segments of Petioles of Cacalia firma (병풍쌈의 엽병 조직 절편으로부터 식물체 재분화)

  • Choi, Soo-Wan;Lim, Soon;Park, Wan-Geun;Choi, Yong-Eui
    • Korean Journal of Plant Resources
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    • v.24 no.5
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    • pp.483-488
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    • 2011
  • Cacalia firma recently has been used increasingly as leaf vegetables but endangered in natural forest. In this work, we established the plant regeneration via adventitious shoot formation from petiole segments of seedling and in vitro plantlets. Wounding of seed coats and $GA_3$ treatments were effective to induce in vitro germination of seeds, whereas, seed did not germinate at all without these treatment. When cotyledon, leaf, petiole, and root segments of seedling were cultured on medium with 2 $mg{\cdot}L^{-1}$ benzyl adenine (BA) and 0.5 $mg{\cdot}L^{-1}$ naphthaleneacetic acid (NAA), petiole segments showed highest number of shoots per explant among the other segments. Among the various kinds of cytokinins, BA, isopentyl adenine (2-ip), kinetin, zeatin, thidiazuron (TDZ), TDZ and BA treatments were effective to induce high frequency of adventitious shoot formation from petiole segments of in vitro propagated plants. NAA stimulated the frequency of adventitious shoot formation but not for number of adventitious shoots per explants compared to TDZ or BA treatment alone. Most of adventitious shoots were developed directly from surfaces of explants. Adventitious shoots were transferred on medium with IBA for root formation, thereafter the plantlets were successfully transferred to soil.

Development of transgenic strawberry plants expressing monellin, a sweet protein (감미단백질 모넬린 발현 딸기 형질전환 식물체 개발)

  • Min, Sung Ran;Ko, Suk Min;Lyu, Jae Il;Park, Ji Hyun;Yi, So Young;Lee, In-Ha;Kim, Hyun Sook;Kim, Tae Il;Choi, Pil Son;Jeong, Won-Joong;Kim, Suk Weon;Kim, Jonghyun;Liu, Jang R.
    • Journal of Plant Biotechnology
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    • v.42 no.3
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    • pp.180-185
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    • 2015
  • Leaf discs from 'Yeobong' and 'Maehyang' strawberry plants were used as explants for transformation. The Agrobacterium tumefaciens strain EHA105 harboring the monellin gene under the control of the CaMV 35S promoter was used in co-cultivation experiments. The frequencies of callus formation and plant regeneration from leaf explants after co-cultivation in 'Yeobong' were higher than those of 'Maehyang'. These transgenic plants showed normal growth patterns and flowering. PCR and Southern hybridization confirmed that 1 to 2 copies of the monellin gene were integrated into genome of the transgenic strawberry plants. Northern blot analysis confirm that the transcripts were expressed in transgenic strawberry plants. Although long-term subcultured transgenic strawberry plants showed a phenomenon to escape the transgene, the transformation system established in this study provides new opportunities for genetic improvement of strawberry plants.

Introduction of Bean Chitinase Gene into Korean Ginseng by Agrobaterium tumefaciens (Agrobacterium tumefaciens에 의한 강낭콩 키틴가수분해효소 유전자의 고려인삼으로의 도입)

  • 이행순;권석윤;백경희;김석원;이광웅;유장렬
    • Korean Journal of Plant Tissue Culture
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    • v.22 no.2
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    • pp.95-99
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    • 1995
  • We have previously established a system for plant regeneration through somatic embryogenesis and Agrobacterium-mediated transformation of Korean ginseng. In this study to produce a fungus-resistant plant, we introduced a bean chitinase gene into ginseng using the transformation system. A binary vector pChi/748 was constructed by introducing the bean basic chitinase gene into EcoRI site of pGA748 which carries the CaMV 35S promoter governing the introduced gene and neomycin phosphotransferase II(NPT-II)gene as a positive selection marker. Cotyledonary explants were cocultured with A. tumefaciens strain LBA4404 harboring the binary vertor pChi/748 for 48 h, and transferred to MS medium supplemented with l mg/L2,4-D,0.1mg/L kinetin, 100 mg/L kanamycin, and 500mg/L carbenicillin. Kanamycin-resistant calli were formed on the cut surface of cotyledonary explants after one month of culture, and subsequently they gave rise to somatic embryos. Upon transfer onto medium containing 1 mg/L each of BA and GA$_3$, most of them converted to plantlets after 5 weeks of culture. The genomic DNA of eight kanamycin-resistant regenerants was subjected to polymerase chain reaction (PCR) using two specific 21-mer oligonucleotides derived from the chitinase gene. PCR-Southern blot analysis confirmed that the chitinase gene was incorporated into six out of the eight regenerants..

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