• Title/Summary/Keyword: callus cultures

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High Frequency Production of Doubled-haploid Plants by Colchicine Application in Anther Cultures of Rice (벼 약배양에서 콜히친이 식물체 배수성에 미치는 영양)

  • 권용삼;이형규;박규환;손재근
    • Korean Journal of Plant Tissue Culture
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    • v.28 no.1
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    • pp.37-40
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    • 2001
  • To optimize the in uitro chromosome doubling procedure in anther cultures of rice, anthers were cultured on callus induction medium with 0.001 to 0.1 mg/L colchicine for 30 days. The addition of colchicine slightly reduced callus formation and plant regeneration in comparison to the colchicine-free medium (control medium). This reduction was greater with higher concentration colchicine. Microspore-derived rice plants in control medium were found to be mainly haploid (50 to 58.8%) and doubled-haploid (31 to 40%) in anther culture of 3 Japonica and 1 Tonsil type cultivars. The application of 0.001 mg/L colchicine was increased to 54.3∼60.0% in the frequency of fertile doubled-haploid plants. These results indicate that the addition of colchicine to the callus induction medium is an efficient means to obtain doubled-haploid plants in anther cultures of rice.

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High Frequency Somatic Embryogenesis and Plant Regeneration in Seedling Explant Cultures of Melon (Cucumis melo L.) (멜론(Cucumis melo L.) 유묘 절편으로부터 고빈도의 체세포배발생과 식물체 재분화)

  • 최필선;소웅영;조덕이;유장렬
    • Korean Journal of Plant Tissue Culture
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    • v.21 no.1
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    • pp.1-6
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    • 1994
  • Cotyledonary and hypocotyl explants of melon seedlings were cultured on Murashige and Skoog's (MS) medium supplemented with various concentrations of 2,4-Dichlorophenoxyacetic acid (2,4-D) and benzyladenine (B.A).Up to 22% of cotyledonary explants and 7%, of hypocotyl explants, respectively: Produced somatic embryos through intervening two types of calli: bright yellow compact (BYC) callus and pale-yellow compact (PYC) callus. BYC callus was capable of producing somatic embryos at initial culture, but it became necrotic as subrulhues proceeded. In contrast UC callus was incapable of producing somatic embryos during initial culture (first 6 weeks), but it became bright-yellow friable (BYF) callus with forming a few globular embryos after 2 months of subculture, indicating that the callus turned embryogenic. The embryogenic capacity of BYF maintained for over one year when the callus was sucultured at 4-week interval. Upon transfer onto MS basal medium the callus gave rise to numerous somatic embryos and subsequently converted to plantlets. Plantlets were transplanted to potting soil and grown to maturity in the phyotron.

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Effect of Macro-nutrients for the Salidroside Production from Callus Cultures of Rhodiola sachalinesis A. Bor (참돌꽃의 캘러스로부터 salidroside 생산에 미치는 배지성분의 영향)

  • Lee, Jae-Seung;Choi, Hye-Jin;Hwang, Baik;Park, Hyoung-Jae;Ahn, Jun-Cheul;Hwang, Sung-Jin
    • Korean Journal of Medicinal Crop Science
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    • v.15 no.4
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    • pp.246-250
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    • 2007
  • Rhodiola sachalinensis A. Bor is a Chinese herb containing the natural compound salidroside, which has been known to possess medical properties such as enhancing bodies' ability to survive in adverse environments and extending human life. To improve a metabolite production from Rhodiola sachalinensis callus cultures, the concentrations of macro nutrients were investigated. To investigate the salidroside production in Rhodiola sachalinensis callus cultures, we analyzed salidroside content in each callus which was cultured in 30 $m{\ell}$ of $2\;{\times}\;B_5$ liquid medium for 4 weeks. The optimal concentrations of macro nutrients for salidroside production ($67.96\;{\pm}\;3.41\;mg/{\ell}$) were found to be 99 mM $KNO_3$, 1 mM $(NH_4)_2SO_4$, 2 mM $NaH_2PO_4{\cdot}2H_2O$, 2 mM $CaCl_2{\cdot}2H_2O$ and 1 mM $MgSO_4{\cdot}7H_2O$. From these results, we determined the more upgraded culture condition for industrial production of salidroside.

Improvement of Catechin Productivity in Callus Cultures of Camellia sinensis Leaves (차나무 잎의 캘러스 배양을 통한 카테킨류의 생산성 개선)

  • Oh, Soon-Ja;Koh, Seok-Chan
    • Korean Journal of Plant Resources
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    • v.18 no.2
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    • pp.351-358
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    • 2005
  • The effects of thiamine-HCl or polyvinylpyrrolidone (PVP) on growth and catechin contents of calli from Camellia sinensis leaves were investigated to improve catechin productivity in callus cultures on the selective medium. The growth of calli was great on the proliferation medium (the MS medium with 0.2 mg/L 2,4-D and 1.0 mg/L TDZ) supplemented with $20{\sim}30\;mg/L$ thiamine-HCl. Although the caffeine content was low in calli compared to young leaves, the total content of catechins was high in calli. Particularly,(-)-epicatechin (EC) which not detected in young loaves was also detected in calli on the propagation medium containing thiamine-HCl or PVP. In addition, the (-)-epicatechingallate (ECG) content was significantly higher in calli than in young leaves. In conclusion, the proliferation medium supplemented with 30 mg/L thiamine-HCl as a vitamin source seemed to be optimal condition for the growth and catechin production in callus culture.

Scopolamine Production in Suspension Cultures of Tumor Calli from Datura metel L. (흰독말풀(Datura metel L.)종양 캘러스의 현탁배양으로부터 Scopolamine 생성)

  • 이수경;윤길영;김용해;양덕조
    • Korean Journal of Plant Tissue Culture
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    • v.27 no.3
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    • pp.203-211
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    • 2000
  • In this study to produce large-scale scopolamine we were examined in the tumor calli of Datura metel L. induced by Agrobacterium tumefaciens $Ery{101}$. The growth and scopolamine contents of tumor calli were higher under light condition than in dark. The optimum condition of growth and scopolamine production were fluence rate of 16 $\mu$mol $m^{-2}s^{-1}$, spectra of red light region and 16 hour light periods on 50 mL SH liquid medium in 4 weeks culture. To increase of the scopolamine contents in tumor calli, the optimum concentration of nitrogen source were 1.8 mM NH$_4$+ and 40 mM NO$_3$. The optimum elicitor concentration for production of scopolamine were 10 mg/L chitosan and 15 mg/L yeast extract. The effect of precursors were good at the concentration of 0.2 mM tropine and 0.3 mM tropic acid, respectively. In order to increase of growth and scopolamine contents. we induced mutant from Datura metel L. tumor callus. Mutants of tumor calli were obtained by 3 Krad, 4 Krad and 6 Krad of ${60}^Cor-ray$. Among them, 3 Krad tumor callus was excellent on the growth and teratoma induction. The 4 Krad tumor callus was negligible for both growth and teratoma induction. But the 6 Krad tumor callus was the best in growth and teratoma induction. The formation of the mutant calli can be enhanced through hormonal combination of 1 mg/L 2,4-dichlorophenoxyacetic acid and 0.5 mg/L benzyladenine. We carry out selection mutant tumor calli for high content tropane alkaloid and suspension cultures for scopolamine production.

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Gamma Radiation-Induced Changes of Antioxidant Enzymes in Callus Cultures of Cassava(Manihot esculenta Crantz) (감마선에 의한 카사바 (Manihot esculenta Crantz) 배양세포의 항산화효소 활성 변화)

  • 이행순;유순희;권석윤;김재성;곽상수
    • Korean Journal of Plant Tissue Culture
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    • v.26 no.1
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    • pp.53-58
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    • 1999
  • The gamma radiation-induced changes of antioxidant enzymes such as superoxide dismutase (SOD), peroxidase (POD) and catalase (CAT) in callus cultures of cassava (Manihot esculenta Crantz) selected as a high yield of cell line for SOD were investigated. In normal cultures, the cell growth reached a maximum at 30 days after subculture (DAS), followed by a rapid decrease with further cultures. The SOD and POD specific activities (units/mg protein) showed the highest at the immediately after subculture and subsequently decreased to 20 DAS, and then increased to 30 DAS, whereas the CAT activity showed the lowest at just after subculture, and it continuously increased from 15 DAS to 30 DAS, showing a good correlation with the cell growth. Irradiation of gamma-ray of 50 and 70 Gy on 7 DAS inhibited significantly the cell growth by 50% and 80% at 14 days after treatment (DAT), respectively. In the cells irradiated with 70 Gy, SOD and POD specific activities increased by 4 and 2.5 folds at 14 DAT, respectively, whereas CAT activity was not affected. The results indicate that SOD and POD may be involved in the antioxidative mechanism in relation to oxidative stresses induced by subcultures and by gamma radiation in callus cultures of cassava.

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Effect of Fungal Elicitor and Heavy Metals on the Production of Flavonol Glycosides in Cell Cultures of Ginkgo biloba

  • KIM, MIN SOO;CHUL KIM;DO HYUN JO;YEON WOO RYU
    • Journal of Microbiology and Biotechnology
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    • v.9 no.5
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    • pp.661-667
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    • 1999
  • The effect of fungal elicitor and heavy metal salts on the production of flavonol glycosides in cell cultures of Ginkgo biloba was investigated. Among the fungi tested, Trichoderma longibrachiatum ATCC 52326 was found to be the most efficient in the production of flavonol glycosides. Kaempferol production from the elicited callus increased ten-fold as compared to the unelicited callus, while quercetin concentration of elicited cells was nine-fold higher than that of uneliceited cells in suspension cultures. The maximum quercetin concentration of 0.362㎎/l was obtained in 1.25㎎/l of the homogenate elicitor. Among the heavy metal salts tested, CuSO₄ showed a significant effect on quercetin accumulation, reaching to the concentration of 0.526 ㎎/l. Quercetin concentration increased to a maximum of l2-fold in response to CuSO₄ treatment as compared to that of untreated cells. The phenylalanine ammonia-lyase (PAL) activity and flavonol glycosides production simultaneously increased for 5 days of culture after fungal elicitor feeding, and their contents showed the same proportional patterns during the culture period. In contrast, PAL activity of cell cultures treated with CuSO₄ was almost constant during the culture period, although quercetin production increased remarkably.

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Plant Regeneration and Somatic Embryo Formation from Root-Derived Callus of Rice (벼 뿌리조직 유래의 캘러스로부터 체세포배 형성과 식물체 재분화)

  • 손재근;김경민;김종수
    • Korean Journal of Plant Tissue Culture
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    • v.22 no.3
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    • pp.143-148
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    • 1995
  • The competence of callus formation and plant regeneration from root derived callus was higher in japonica cultivars than those of Tongil-type cultivars of rice. A japonica type cultivars Yeongdeogbyeo, showed the highest capacity (13%) for plant regeneration from root calli of 6 cultivars tested. The callus induced from seed and root tissues maintained higher capacity for plant regeneration during 7 passages of subculture on N$_{6}$ solid media at 2-week intervals. The maximum frequency (2 x 10$^{5}$ mL) of round cells and their cell colonies showed about 24 days after suspension culture of root-derived callus in N$_{6}$ medium with lmg/L 2,4-D, 300mg/L casein hydrolysate, 10mM L-proline, 20g/L sucrose and 30g/L sorbitol. The frequency of somatic embryo formation in suspension cultures of root-derived callus increased with prolonged advance of subculture time from 30 to 90 days, but their regenerative capacities decreased.

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High-frequency plant regeneration from transgenic rice expressing Arabidopsis thaliana Bax Inhibitor (AtBI-1) tissue cultures

  • Cho, A-Ra;Lee, Dong-Kil;Kim, Kyung-Min
    • Journal of Plant Biotechnology
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    • v.42 no.2
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    • pp.83-87
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    • 2015
  • Genetic transformation was affected by material of explant, age of callus, and medium of regeneration. Two rice seed cultivars (Ilpum and Baekjinju) and mediums were investigated in this study for enhancing regeneration of transgenic rice expressed AtBI-1 gene encoding the Arabidopsis thaliana Bax inhibitor. Regeneration rate of Ilpum rice transformant in gelrite of 5 and 8 g were 27.4% and 18.0%, respectively. In Baekjinju, regeneration rate of transformant was 5.4% and 4.3% in 5 and 8 g gelrite, respectively. The highest number of transformant plant in this study was regenerated from Ilpum cultivar on MS medium (30.4%) and was applied for the subsequent experiment. The callus regeneration rate of transformant were 40.7% in callus infection of up-side, it was higher regeneration then in the down-side (3.9%). The regeneration rate of callus of 25 days and 35 days were 14.7% and 38.6%, respectively. The most important application of this work is in genetic transformation of rice, particularly for improvement transgenic plant tissue culture protocol with high frequency of plant regeneration.

Plant Regeneration from Leaf and Internode Segment Cultures of Boxthorn (Lycium chinense Mill.) (구기자나무의 잎과 마디절편체 배양에 의한 식물체 재생)

  • 김동찬;정해준;민병훈;양덕춘
    • Korean Journal of Plant Tissue Culture
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    • v.28 no.6
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    • pp.329-333
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    • 2001
  • Callus and shoot formation from medicinal crop, Lycium chinense Mill. cv. 'Cheongyang', as influenced by various media, explant sources and plant growth regulators were investigated. The rate of shoots formation, number of shoots, and fresh weight of shoots were the best on MS medium followed by B$_{5}$, WPH, and SH. Callus induction was more effective in leaf than internode segments, and was the best on MS medium containing 0.5 mg/L NAA with 0.2 mg/L BA. Effects of plant growth regulators in shoot formation were more effective in BA than TDA combined with NAA. Shoot formation from callus induced in leaf and internode segments was the best on MS medium containing 0.01 mg/L NAA with 0.2 mg/L BA.

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