• Title/Summary/Keyword: Callus proliferation

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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.

Callus induction and plant regeneration of Iris dichotoma Pall. in endangered species

  • Bae, Kee-Hwa;Yoo, Kyoung-Hwa;Lee, Hak-Bong;Yoon, Eui-Soo
    • Journal of Plant Biotechnology
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    • v.39 no.3
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    • pp.182-188
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    • 2012
  • Iris dichotoma Pall. is an important endangered plant belonging to the family Iridaceae. A method was developed for the rapid micropropagation of I. dichotoma through plant regeneration from leaf, rhizome, and root explant-derived calli. Leaf, rhizome, and root segments were cultured on Murashige and Skoog (MS) medium supplemented with 2,4-dichlorophenoxy acetic acid (2,4-D; $0-3.0mg{\cdot}L^{-1}$) for callus induction. Callus production was highest at $1.0mg{\cdot}L^{-1}$ 2,4-D, where 73.8% and 45.5% of cultured rhizome and root cuttings, respectively, produced calli. The viable calli were maintained at an induced concentration of 2,4-D ($3.0mg{\cdot}L^{-1}$). They were then transferred to MS medium supplemented with various concentrations of 2,4-D ($0-3.0mg{\cdot}L^{-1}$) in combination with 6-benzyladenine (BA: 0, 1.0 and $3.0mg{\cdot}L^{-1}$) for adventitious shoot regeneration. The addition of a low concentration of 2,4-D into BA-containing medium significantly increased the frequency of shoot regeneration in leaf, rhizome, and root-derived calli. The highest number of adventitious shoots (26.4 per callus) formed at $0.5mg{\cdot}L^{-1}$ 2,4-D and 1.0 mg/l BA. For rooting of the shoots, half- strength MS medium supplemented with different concentrations of indole 3-butyric acid (IBA) $0-3.0mg{\cdot}L^{-1}$ was tested. The optimal results were observed using half-strength MS medium supplemented with $1.0mg{\cdot}L^{-1}$ IBA, on which 98% of the regenerated shoots developed roots with an average of 3.5 roots per shoot within 45 days. The plantlets raised in vitro were acclimatized and transferred to soil with 95% success. This in vitro propagation protocol will be useful for conservation and mass propagation of this endangered plant.

Effects of Growth Regulators on Plant Regeneration in Shoot-Tip-Derived Embryogenic Callus Cultures of Sweet Potato (Ipomoea batatas) (고구마 경단 유래 배발생 캘러스로부터 식물체 재분화에 미치는 생장조절제의 영향)

  • ;Shozo FUJIOKA
    • Korean Journal of Plant Tissue Culture
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    • v.21 no.5
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    • pp.281-286
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    • 1994
  • The hormonal regulation of organ differentiation was investigated in the tissue culture of sweet potato. Embryogenic callus was induced from shoot tips cultured on MS medium supplemented with 1 mg/L 2,4-D. When embryogenic callus was transferred to medium containing 0.1 mg/L GA$_4$, it proliferation was stimulated. The callus gave rise to plantlets when cultured on medium containing 0.1 mg/L BA. Addition of 0.1 mg/L jasmonic acid or 0.01 mg/L brassinolide to medium was effective for the development of healthy normal plantlets.

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Micropropagation through Callus Culture in Chinese Foxglove (Rehmannia glutinosa) (지황의 캘러스 배양에 의한 기내 대량증식)

  • 박충헌;성낙술;백기엽;이철희
    • Korean Journal of Plant Tissue Culture
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    • v.25 no.3
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    • pp.171-175
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    • 1998
  • Chinese foxglove (Rehmannia glutinosa) is receiving much attention as one of the principal medicinal crops and the crude drug damand expands rapidly.This study was conducted to obtain the basic breeding information of Chinese foxglove. Effects of supplemental plant growth regulators were investigated on leaf tissue for proliferation. 100% callus formation, 31% plantlet regeneration and 6% root differentiation were obtained by adding 0.5 mg/L NAA and 2.0 mg/L BA. 2,4-D and Zeatin treatment also resulted in 95% increase in callus formation, but shoot was not formed. During the subculture, callus propagation rate recorded 15.4% with 0.2 mg/L NAA and 1.0 mg/L BA and plant regeneration improved on MS medium supplemented with 0.2 mg/L NAA and 0.5 mg/L kinetin. The number of shoot formed ranged from 1.7 on WPM medium to 3.4 on MS medium with 0.1 mg/L NAA and 0.5 mg/L BA. Supplementation of 1.0 g/L activated charcoal improved the In vitro plant growth.

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Ultrastructural Characteristics of Developmental Stages During in vitro Regeneration in Citrus junos SIEB. (유자 (Citrus junos SIEB.) 의 발생단계에 따른 미세구조적 특성)

  • 박민희
    • Korean Journal of Plant Resources
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    • v.8 no.3
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    • pp.237-246
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    • 1995
  • In this study, the callus was induced and regenerated from the immature embryo and ultrastructural characteristics of developmental stages in Citrus junos SIEB, were investigated. The yellowish callus was induced by 5 to 6 week of culture of citrus. In proliferation callus after 6 weeks of culture, large vacuole was formed by fusion between adjacent small ones. In the non-embryogenic callus cultured for 12weeks, re-differentiated cells of callus showed the large nucleus with globular nucleus and amyloplast with large size of starches. In the embryogenic callus cltured for 14-16 weeks, the active exocytosis occurred in cells, secretory vesicles appeared on cell membrane and small particles from cytoplasm were released to intercelluar space. In the embryogenic callus cultured for 24 weeks, a sperical type of chloroplast bounded on cytoplasm by double membrane and typical grana was dispersed equally among matrix. In the normal plantlet after 26 weeks of culture, a lot of vessels and companion cells apperaed in the leaf cell of plantlet. In the normal plantlet after 30 weeks of culture, the immature leaf showed many small companion cells, sieve tubes and central vacuole. Also, the secondary vacuole protruded into the central vacuole and elongated chloroplasts near plasma membrane. In the matured plant habituated on the soil, palisada tissue composed of orderly arranged cells contained the nucleus in the center of the cell and large vacuoles on either side of the nucleus.

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An Experimental Study on the Effects of Hyperbaric Oxygen Therapy on the Healing of Mandibular Fracture of Streptozotocin-Induced Diabetic Rats. (고압산소요법이 당뇨백서 하악골 골절 치유에 미치는 영향에 관한 실험적 연구)

  • Sang, Jai-Woo
    • Maxillofacial Plastic and Reconstructive Surgery
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    • v.12 no.1
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    • pp.1-13
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    • 1990
  • The main objectives of this study was to observe the effects of hyperbaric oxygen therapy on the healing processes of mandibular fracture of streptozotocin-induced diabetic rats. Author used 60 rats (Sprague-Dawley Strain) deviding into control(30) and experimental groups(30). Complete fracture was produced on the left mandibular body of 60rats, rendered hyperbaric oxygen therapy (2 hrs. daily at 2.5 atm.) on experimental group and observed effects of hyperbaric oxygen therapy by microscopically. The obtained results were as follows; 1. Infiltration of inflammatory cells was no significant differences between the control and experimental group until 3rd week, but experimental group showed decreasing tendency after 4th week. 2. Severe proliferation of fibroblasts showed rather rapider in experimental group, at 2nd week, while at 3rd week in control group. 3. Osteoclasts appeared at 1st week in experimental group while at 3rd week in control group, and experimental group showed early bone resorption pattern. 4. Osteoblasts appeared at 1st week in experimental group while at 3rd week in control group, and experimental group showed prominent osteoblastic activity. 5. Moderate proliferation of capillary blood vessels showed in initial stage of experimental group while mild proliferation at 1-2nd week in control group. 6. Formation of cartilaginous callus showed at 4th week in experimental group, while at 6th week in control group. 7. Formation of bony callus showed mildly at 5th week, and moderately at 6th week in experimental group, while no appearance in control group, but complete bony union was not observed even in experimental group throughout this experiment.

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Micorpropagation of Corylopsis coreana by Thidiazuron Treatment (Thidiazuron 처리에 의한 히어리나무의 기내번식)

  • Kang, Hyo-Jin;Moon, Heung-Kyu;Yi, Jae-Seon
    • Journal of Plant Biotechnology
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    • v.30 no.3
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    • pp.263-267
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    • 2003
  • This study was conducted to evaluate the effect of thidazuron(TDZ) on shoot proliferation and growth from axillary buds of 20-years-old Corylopsis coreana. Shoots proliferation was effectively achieved on WPM(Woody Plant Medium) supplemented with 0.03∼0.1mg/L TDZ. The highest shoot number(6.5$\pm$0.7) was obtained on 0.1mg/L TDZ treatment. On the TDZ medium shoots formed as clusters less than 1cm in height and therefore needed to subculture on GA$_{3}$ containing medium to induce elongation. In consecutive cultures, phenolic compounds were excreted at the proximal part of the explants and inhibited growth of the explants. Growth inhibition by the compounds was overcome using liquid and paper bridge culture system. About 60% of the elongated shoots rooted on half- strength MS medium containing IBA. Generally, IBA was mire effective on in vitro rooting than NAA with optimal range of 0.5mg/L to 1.0mg/L. Rooted plantlets were transferred in an artificial soil(vermculite) and acclimatized in high humidity greenhouse condition. Survival rate differed greatly depending on rooting types of the explants. Two types of rooting were observed. The first type was direct rooting from the explants. The second type was callus formation followed by rooting from the callus. The explants showing the 1st type rooting survived can be multiplicated in vitro by TDZ treatment followed by elongation with GA$_{3}$ and rooting with IBA.

In Vitro Shoot Tip Culture of Pear 'Niitaka' as Related to Tree Vigor Sampling Time and Plant Growth Regulators (배 '신고'의 경정배양에 있어서 모수의 수세, 경정의 채취시기 및 생장조절제의 영향)

  • 이창후;김정선;김성복
    • Korean Journal of Plant Tissue Culture
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    • v.25 no.3
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    • pp.159-163
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    • 1998
  • In vitro shoot tip culture technique was established in pear (Pyrus pyrifolia 'Niitaka') as related to tree vigor, sampling time, and plant growth regulators and sucrose supplemented to medium. Shoot tips excised in June from the tree having medium-vigor developed good shoots. BA (1.0 and 2.0 mg/L) without NAA produced shoots suitable for proliferation, and NAA supplemented to medium resulted in poor shoot growth and excessive callus formation. BA of 2.0 mg/L combined with 0.01 mg/L NAA provided shoots suitable for rooting and sucrose of 30 g/L was recommended for proliferation medium. A fourth strength MS medium supplemented with 0.1 mg/L NAA produced plantlets in good quality of root number and root length.

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Selection of Glyphosate Resistant Cell Lines in Alfalfa (Medicago sativa L.) (알팔파의 Glyphosate 저항성 세포주 선발)

  • 류점호
    • Korean Journal of Plant Tissue Culture
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    • v.24 no.6
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    • pp.341-344
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    • 1997
  • Calli were induced from the cotyledon and the hypocotyl of alfalfa and the callus lines were tested for the resistance to glyphosate in the liquid medium containing 0.01-3.00 mM glyphosate. Some resistant cell lines were selected from the gradual increase of glyphosate concentration and the lines resistant to 10 mM glyphosate were analyzed with EPSPS activity. Vigorous callus proliferation from the cotyledon and the hypocotyl was observed from the MS medium containing 1 mg/L 2,4-D and 0.5 mg/L kinetin. The hypocotyl was thought to be better explant source for callus induction than the cotyledon. $ID_{50}$ (Inhibition Dosage of 50%) to glyphosate was between 0.1 mM and 0.2 mM level. A49-10G and A58-10G cell lines selected as resistant to 10 mM glyphosate had 8.0 and 9.1 fold increased EPSPS activity to those of the control lines, respectively.

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Effect of Antimitotic Agent Colchicine on In Vitro Regeneration of Watermelon

  • Jaskani Muhammad J.;Raza H.;Khan M. M.;Kwon Sung W.
    • Journal of Plant Biotechnology
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    • v.6 no.4
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    • pp.247-252
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    • 2004
  • In vitro cultures of watermelon were treated with antimitotic agent colchicine to induce ploidy alterations, particularly the induction of tetraploids. Explants cotyledon, embryonic end of seed, transverse sections of epicotyl and hypocotyl were cultured on MS media supplemented with BA ($1{\mu}M$) and colchicine ($0.01\%,\;0.05\%\;and\;0.1\%$). Explants were subcultured on colchicine free media after 4 and 7 days. Colchicine had negative effect on in vitro regeneration but this exhibited explants related response. However, hypocotyl section of seedlings induced maximum callus on $0.01\%$ colchicine. Shoot proliferation was more in cotyledon explants cultured on colchicine ($0.01\%$) for four days. Maximum root induction and root number were recorded in embryonic end explants. Overall, cotyledon and embryonic end explants, and low colchicine concentration ($0.01\%$) was found optimal in watermelon regeneration.