• Title/Summary/Keyword: callus development

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Increase of isoflavones in soybean callus by Agrobacterium-mediated transformation

  • Jiang, Nan;Jeon, Eun-Hee;Pak, Jung-Hun;Ha, Tae-Joung;Baek, In-Youl;Jung, Woo-Suk;Lee, Jai-Heon;Kim, Doh-Hoon;Choi, Hong-Kyu;Cui, Zheng;Chung, Young-Soo
    • Plant Biotechnology Reports
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    • v.4 no.4
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    • pp.253-260
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    • 2010
  • Plant secondary metabolites have always been a focus of study due to their important roles in human medicine and nutrition. We transferred the isoflavone synthase (IFS) gene into soybean [Glycine max (L.) Merr.] using the Agrobacterium-mediated transformation method in an attempt to produce transformed soybean plants which produced increased levels of the secondary metabolite, isoflavone. Although the trial to produce transgenic plant failed due to unestablished hygromycin selection, transformed callus cell lines were obtained. The induction rate and degree of callus were similar among the three cultivars tested, but light illumination positively influenced the frequency of callus formation, resulting in a callus induction rate of 74% for Kwangan, 67% for Sojin, and 73% for Duyou. Following seven to eight subcultures on selection media, the isoflavone content of the transformed callus lines were analyzed by high-performance liquid chromatography. The total amount of isoflavone in the transformed callus cell lines was three- to sixfold higher than that in control callus or seeds. Given the many positive effects of isoflavone on human health, it may be possible to adapt our transformed callus lines for industrialization through an alternative cell culture system to produce high concentrations of isoflavones.

Improvement of Shoot Regeneration from Scutella-Derived Callus in Rice

  • Kim, Yong-Wook;Cho, Joon-Hyeong;Lee, Jang-Yong
    • KOREAN JOURNAL OF CROP SCIENCE
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    • v.49 no.1
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    • pp.52-60
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    • 2004
  • The optimized in vitro culture system was investigated for improvement of regeneration efficiencies by observing the responses of scutella-derived callus of Korean rice (Oryza sativa L.). Large variations of callus induction (43.9-93.9%) and shoot regeneration (0-88.7%) were observed among the rice cultivars depending on medium. However, shoot regeneration was significantly improved by selected utilization of basal medium, growth regulators, and carbon sources. N6 basal medium was more efficient for embryogenic callus induction than MS or LS basal medium, while MS was superior to N6 for shoot regeneration. The calli of highly regenerative cultivars grew faster and showed higher rates of green tissue formation (GT) and shoot regeneration (SR) and lower rate of callus browning (CB) than those of recalcitrant cultivars. Although a higher level of kinetin stimulated the GT and SR in highly regenerative cultivars, $10\textrm{mgL}^{-1}$ kinetin generally suppressed the GT and SR, while CB was accelerated compared to $2\textrm{mgL}^{-1}$ kinetin. Additional benefits of sorbitol combined with maltose (or sucrose) under $5\textrm{mgL}^{-1}$ kinetin were certainly confirmed on regeneration efficiencies compared to sucrose alone as carbon source and osmotic regulator. This combination showed high rate of GT and SR with multiple shoots while low rate of CB. With MSRK5SM-Pr medium ($5\textrm{mgL}^{-1}$ kinetin, 3% sorbitol, 2% maltose, $500\textrm{mgL}^{-1}$ proline), the regeneration efficiencies of total 17 out of 24 cultivars were practically improved 160% on average compared to MSRK2S ($2\textrm{mgL}^{-1}$ kinetin, 3% sucrose) control medium. Especially, the medium was most effective to the cultivars showing a medium level of regenerability such as Daesanbyeo and Dongjinbyeo and Suwon477, enhancing efficiencies more than 300-600% compared to MSRK2S medium.

Evaluation of Alcea rosea L. Callus Extract as a Natural Cosmetic Ingredient (접시꽃 캘러스 추출물의 천연화장품 원료로서의 효능 평가)

  • Lee, Gibok;Yeom, Areum;Won, Kim Dong;Park, Chang-Min;Joung, Min-Seok;Lee, Gi Yong;Jeong, Cheol-seung
    • Journal of the Society of Cosmetic Scientists of Korea
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    • v.44 no.3
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    • pp.295-302
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    • 2018
  • In this study, we investigated the biological effects of Alcea rosea L. callus extract for the development of natural cosmetics ingredients. The antioxidant activities of A. rosea L. callus extract was measured through DPPH, ABTS and FRAP assay. As a result, A. rosea L. callus extract were found to have a strong antioxidant ability in a dose dependent manner. In addition, A. rosea L. callus effectively reduced the intracellular oxidative stress induced by AAPH at a concentration of 10 mg/mL. In a tyrosinase activity assay, we found that A. rosea L. callus extract reduced tyrosinase activity by 51% at 10 mg/mL. Based on these results, A. rosea L. callus extract is considered as a promising natural ingredients for cosmetics with antioxidant and whitening functions.

In vitro Tissue Culture of Aloe arborescens Mill

  • Rha, Eui-Shik;Kim, Hyun-Soon;Lee, Seung-Yeob
    • Plant Resources
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    • v.1 no.2
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    • pp.109-112
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    • 1998
  • Aloe in vitro culture was attempted to induce callus and regeneration ability from different explant sources onto MS medium with 0.5mg/l NAA plus 1.0mg/l BA. Anthers that no developed any callus and plant regeneration, while only four out of 274 filament explants induced calli at cut edge without regenerated plants. Twenty ovary explants regenerated four direct plantlets without via callus from the base of epidermal tissues. Regenerated plants on the root tip gave 2n=14 of chromosome numbers.

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Formation and Production of Ascorbate Oxidase by Cucumber Tissue Cultured Cells (오이조직 배양세포에 의한 Ascorbate Oxidase 생성 및 생산)

  • Lee, Jong-Hwa;Chung, Ho-Kwon;Shinmyo, Atsuhiko;Lim, Bun-Sam
    • Microbiology and Biotechnology Letters
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    • v.21 no.4
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    • pp.329-335
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    • 1993
  • Ascorbate oxidase activity in various cucumber tissue extracts was highest in young fruit peeling. Cucumber callus was induced from young fruit peeling and callus cell lines were selected for more than 7 months, which porduced high levels of ascorbate oxidase and had a high growth rate. Induction of callus was optimized with Linsmaier-Skoog(LS) medium at 25$^{\circ}C$ in dark phase. Ascorbate oxidase activity reached a maximum at 5 days after transfer to LS basal liquid-medium ant then declined. The enzyme activity in callus cells was stimulated by addition of 10${\mu}$M $CuSO_4$ in the early logarithmic phase of growth. And also, adding 10${\mu}$M $CuSO_4$ at 3rd day 7th day of culture period, ascorbate oxidase activity in callus cells was maintained to high level. Maximum yield of ascorbate oxidase was found at the 25th day by flask shaking culture, but three-fold of ascorbate oxidase activity was obtained at the 16th day by jar fermentation.

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Identification of mitochondrial mutant (NADH-dehydrogenase) using PCR method and regeneration of mutants from Zea mays (PCR 기법을 사용한 옥수수 미토콘드리아 변이체 (NADH-dehydrogenase)의 선별과 재분화)

  • 설인환
    • Journal of Life Science
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    • v.8 no.1
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    • pp.8-13
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    • 1998
  • The maize mitochondrial mutant (NCS2) is derived from homologous recombination between genes encoding NADH dehydrogenase subunit 4 and subunit 6. Plants from mitochondria mutants exhibited severe related growth and development including dwarfism and striping on the leaves. Aborted embryos from NCS2 mutants have been rescued and cultured on the N6 medium supplemented with 2,4-D 1 mg/l. Most calli from NCS2 aborted embryos showed slow growing pattern at first stage. However, upon continuous culturing them on the medium, those were segregated into mutant and normal callus lines. These segregations could be detected by using PCR method with three primers. Such segregation seems to be resulted from the preferential growth of normal cells over the mutant cells on the normal culture condition. Therefore, this method can be used for determining rate of indirect cytoplasmic segregation by estimating amplified band intensities. When NCS2 mutant callus lines cultured on regeneration medium, no adventitious shoot induction was observed. However, callus lines with more mitochondria induced adventitious shoots. These studies suggest that mitochondria NADH-dehydrogenase for electron transport in the inner membrane of mitochondria is essential for the differentiation and development of plants.

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Genetic Improvement for the Low Salinity-Tolerant Porphyra Sp. by Cell Culture Technique I. Tissue Culture of Porphyra yezoensis foma narawaensis (세포배양기술에 의한 김의 내저염성 품종개량 I. 큰방사무늬김의 조직배양)

  • Hong Yang Ki;Sohn Chul Hyun;Chang Jung Won
    • Journal of Aquaculture
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    • v.2 no.1
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    • pp.1-7
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    • 1989
  • Axenic tissue culture of a marine red algae Porphyra yezoensis foma narawaensis was established for the vegetative propagation of tissues as a seed stock and for the development of a low salinity-tolerant cell line. Callus tissues have been induced from the vegetative area of blade away from the hold fast when grown on PES-agar medium. The brownish red fragile callus was maintained under fluorescent light of ca. 2000 lux with 12 : 12 hr L : D at $16^{\circ}C$. Amounts of carbohydrate and protein was determined against the weight of callus. Optimum temperature of the callus growth was $14^{\circ}C\~18^{\circ}C$. Optimun concentration of sodium chloride was $2.0\%$ for the callus growth in PES-agar medium.

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Development of a Robot for Automation of a Callus Inoculation (식물조직배양 자동화를 위한 로봇개발 - 엔드이펙터 및 시스템의 성능시험 -)

  • Chung, Suk-Hyun;No, Dae-Hyun
    • Journal of Bio-Environment Control
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    • v.18 no.2
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    • pp.87-94
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    • 2009
  • This study was conducted to develop an automation system of inoculation processing of a lily callus. The results are summarized as followings: The end-effector was manufactured as suction and machine type. And these end-effectors can separate the callus from the mediums and divide the separated callus and then inoculate the divided callus to new mediums. Using the machine type end-effect0r, the results of the experiment showed the success rate in the division process was 100% while the separation and inoculation process was 92%. To develop the automation controller of inoculation process, the system was developed to control an external device and the manipulator. The data communication program between a robot and a personal computer was also developed using CAsyncsocket and Ethernet Interface.

Effect of cefotaxime on reduction of contamination for callus tissues in calla 'Gagsi' (Cefotaxime 처리를 통한 칼라 기내 식물체의 오염 감소 효과)

  • Lee, Sang Hee;Kim, Young Jin;Yang, Hwan Rae;Kim, Jong Bo
    • The Journal of the Convergence on Culture Technology
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    • v.5 no.1
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    • pp.409-412
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    • 2019
  • We investigated the development of a micropropagation protocol for multiplication of calla 'Gagsi' by using shoots as explant. The callus was induced on Murashige and Skoog (MS) basal medium containing cefotaxime antibiotics (25, 50, 100 mg/L). Also, MS basal medium with NAA 0.5 mg/L and BA 1.0 mg/L was used. The callus induction and browning rates were compared by treatment supplemented cefotaxime 25, 50 and 100 mg/L in basal MS medium. The callus induction rate was 10.5 % and browning rate was also, 10.5 % on the MS containing 25 mg/L. In the MNB containing cefotaxime, the callus induction rate was 34.5 % and browning rate was 27.0 %. The cefotaxime experiment has been widely used in previous studies. It is thought that it will help establish the mass multiplication system by positively affecting the growth and browning reduction of calla plants.

Effects of 5-azacytidine, a DNA methylation inhibitor, on embryogenic callus formation and shoot regeneration from rice mature seeds (벼 성숙종자로부터 배상체 캘러스 형성 및 식물체 재분화에 DNA methylation 억제제인 5-azacytidine의 영향)

  • Lee, Yeon-Hee;Lee, Jung-Sook;Kim, Soo-Yun;Sohn, Seong-Han;Kim, Dool-Yi;Yoon, In-Sun;Kweon, Soon-Jong;Suh, Seok-Chul
    • Journal of Plant Biotechnology
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    • v.35 no.2
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    • pp.133-140
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    • 2008
  • The modification of DNA and histone plays an important role for gene expression in plant development. The objective of this research is to observe the effects of methylation on the gene expression during dedifferentiation from rice mature seeds to callus and differentiation from callus to shoots. The embryogenic callus with ability to shoot regeneration was not induced on the N6A medium supplemented with 5-azacytidine and abnormal callus with brown color was formed. When the normal rice callus was placed on the regeneration MSRA medium supplemented with 5-azacytidine, the shoot regeneration was inhibited. The results showed that 5-azacytidine, DNA demethylating agent, had negative effects on normal embryogenic callus formation and shoot regeneration. This suggested that DNA methylation of some genes was required for normal cell dedifferentiation and differentiation in tissue culture. The microarray and $GeneFishig^{TM}$ DEG screening were used to observe the gene transcript profile in callus induction and regeneration on N6A (N6 medium + 5-azaC) and MSRA (MS regeneration medium + 5-azaC). Subsets of genes were up-regulated or down-regulated in response to 5-azaC treatments. The genes related with epigenetic regulation, electron transport, nucleic acid metabolism and response to stress were up and down regulated. The different expression of some genes (germin like protein etc.) during callus induction and shoot regeneration was confirmed using RT-PCR and northern blot analysis.