• Title/Summary/Keyword: MG/MS

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In vitro shoot propagation of Ranunculus kazusensis Makino, an endangered aquatic plant (멸종위기 수생식물인 매화마름(Ranunculus kazusensis Makino)의 기내 신초 증식)

  • Park, Min Wan;Ryu, Shi Hyun;Nam, Su Hwan;Bae, Kee Hwa
    • Journal of Plant Biotechnology
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    • v.44 no.3
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    • pp.325-329
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    • 2017
  • A micropropagation method via callus for Ranunculus kazusensis Makino, an endangered species, was established. When stem segments were cultured on MS media supplemented with 1.0 mg/L IAA, NAA, IBA and 2,4-D, the highest frequency of callus induction was achieved on MS medium supplemented with 1.0 mg/L NAA. Multiple shoot per explant was obtained, the MS medium containing 1.0 mg/L BA and 0.5 mg/L NAA. Additionally, effect of activated charcoal (AC) and sucrose on shoot growth in in vitro culture were examined. The most suitable conditions for shoot growth after 4 weeks of culture were the MS medium with AC and sucrose. This in vitro propagation protocol will be valuable for conservation and mass propagation of this endangered plant.

Effects of Plant Growth Regulators, Medium Salt Strength and Nitrogen Ratio on Cell Culture of Gymmema sylvestre (식물생장조절물질, 무기물 농도 및 질소원 비율이 Gymmma sylvestre 세포 배양에 미치는 영향)

  • Lee, Eun-Jung;Han, Eun-Joo;Paek, Kee-Yoeup
    • Journal of Plant Biotechnology
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    • v.33 no.2
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    • pp.105-110
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    • 2006
  • This study was carried out to investigate the effects of plant growth regulators, medium salt strength and nitrogen ratio on cell culture of Gymnema sylvestre. Cell growth was inhibited by 2,4-D higher than 1.0 mg L$^{-1}$, but not by kinetin lower than 0.5 mg L$^{-1}$. Maximal cell growth was obtained at 1.0 mg L$^{-1}$ 2,4-D and 0.1 mg L$^{-1}$ kinetin. Cell growth was greatest at 1x MS medium but high strength of MS medium inhibited cell growth due to low water potential in the medium. In $NH_4^+:NO_3^-$ ratio of 0:60 (i.e. 0.0 mM $^NH_4^+$ and 60.0 mM $NO_3^-$), cells growth was highest but cells were smaller and whiter compared with those in other $NH_4^+:NO_3^-$ ratio. Reduced cell growth was observed with continuous culture. These results suggested that optimal cell culture of G. sylvestre could be achieved with 1x MS medium with 20:40 ratio of $NH_4^+:NO_3^-$ supplemented with 1.0 mg L$^{-1}$ 2,4-D and 0.1 mg L$^{-1}$ kinetin.

Effect of explant parts and medium components on in vitro regeneration in Osmunda cinnamomea var. forkiensis (꿩고비(Osmunda cinnamomea var. forkiensis) 기내 포자체 재생에 영향을 미치는 배양부위와 배지구성물질)

  • Kwon, Hyuk Joon;Shin, So Lim;Lee, Cheol Hee;Kim, Soo-Young
    • Journal of Plant Biotechnology
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    • v.44 no.4
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    • pp.448-453
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    • 2017
  • This study was carried out to find culture materials (explant parts) and medium components (medium type, sucrose and $NaH_2PO_4$ concentration) for in vitro propagation of Osmunda cinnamomea var. forkiensis sporophyte. The results of study: chopped segments of leaf blades, stipes, rhizomes and roots were cultured on a 1/2MS medium supplemented with 0.1% activated charcoal. Among these explant types, only the rhizome segments produced young sporophyte, regenerating vigorously on a 1/8MS medium. Adjusting the sucrose concentration to 2% and supplement to $50mg{\cdot}L^{-1}\;NaH_2PO_4$ in the 1/8MS medium proved to be more efficient for plant regeneration. Consequently, the addition of 0.1% activated charcoal to a modified 1/8MS medium (2% sucrose, $50mg{\cdot}L^{-1}\;NaH_2PO_4$, pH 5.8 and 0.8% agar) yielded the highest sporophyte regeneration.

Development of LC-MS/MS Quantitation Method for Ethoxyquin in Fishery Products (수산물 중 에톡시퀸의 LC-MS/MS 정량분석법 개발)

  • Shin, Dasom;Chae, Young-Sik;Kang, Hui-Seung;Lee, Soo-Bin;Cho, Yoon-Jae;Cheon, So-Young;Jeong, Jiyoon;Rhee, Gyu-Seek
    • Journal of Food Hygiene and Safety
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    • v.31 no.6
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    • pp.432-438
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    • 2016
  • Ethoxyquin (EQ, 1,2-dihydro-6-ethoxy-2,2,4-trimethyl-quinoline) is quinoline-based antioxidant used in the animal feed and food industry to protect the raw materials and final products against oxidation. In recent years the use of synthetic antioxidants in fishmeal ingredients carry-over to farmed fish fillets has received increasing attention in food safety. This study was conducted to develop an analytical method to determine EQ in aquatic products. The analytes were confirmed and quantified via liquid chromatography-tandem mass spectrometry (LC-MS/MS) in the positive ion mode using multiple reaction monitoring (MRM). The sample was extracted with 1 N HCl (in case of flatfish extracted with 1 N HCl containing 10% acetonitrile). Then, solid phase extraction (SPE) was used for the cleanup. Standard calibration curves presented linearity with the correlation coefficient ($r^2$) > 0.99, analyzed at 0.005-0.2 mg/kg concentration. The developed method was validated according to the Codex Alimentarius Commission (CAC) guideline. The limits of quantitation for EQ were 0.01 mg/kg. Average recoveries ranged from 81.3% to 107%. The repeatability of measurements, expressed as the coefficient of variation (CV, %), was below 10%. The analytical method was characterized with high accuracy and acceptable sensitivity to meet CODEX guideline requirements and would be applicable to analyze the EQ residue in aquatic products.

Anomalous somatic embryos formation and plant regeneration from the cultures of immature embryos of Camellia japonica L. (동백나무 미숙배 배양으로부터 비정상 체세포배 형성과 식물체 재생)

  • Choi, Jong-Hye;Kwon, Suk-Yoon;Choi, Pil-Son
    • Journal of Plant Biotechnology
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    • v.38 no.4
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    • pp.258-262
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    • 2011
  • Embryogenic callus was induced from the cultures of immature embryos of Camellia japonica L. on Murashige & Skoog's (MS) solid medium supplemented with 1 mg/L 2,4-dichlorophenoxy acetic acid (2,4-D), and then the embryogenic callus was proliferated on same medium for 4 weeks over. The embryogenic callus was sub-cultured on MS basal medium without 2,4-D to produce coyledonary stage of somatic embryo. The frequency (%) of somatic embryogenesis was 25.1%, and the majority of somatic embryos formed had a abnormal morphology with cupshaped cotyledon (48.3%), one cotyledon (12.6%), three cotyledons (9.4%), four cotyledons (1.9%), whereas was only normal morphology with two cotyledon (27.5%). When the somatic embryos with normal or abnormal cotyledons transfer to MS basal medium or $\frac{1}{2}$ MS medium with/or without plant growth regulators ($GA_3$, IBA) for regeneration, the frequency (%) of two-cotyledon embryos regenerated into plantlets was higher 11.1% than one cotyledon (0.0~8.3 %), three cotyledons (0.0~5.8%), four cotyledons (0.0%), cup-shaped (0.3~4.2%). These results demonstrated that the anomalous cotyledons of somatic embryos could caused to decrease the rate of plant regeneration.

Development of Analytical Method for Colistin in Fish and Shrimp using Liquid Chromatography Mass Spectrometry (LC-MS/MS를 이용한 수산물 중 콜리스틴 분석법 개발)

  • Shin, Dasom;Kang, Hui-Seung;Lee, Soo-Bin;Cho, Yoon-Jae;Cheon, So-Young;Jeong, Jiyoon;Rhee, Gyu-Seek
    • Journal of Food Hygiene and Safety
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    • v.31 no.5
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    • pp.319-326
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    • 2016
  • Colistin is a last resort antimicrobial agent against multi-drug resistant Gram-negative bacteria. This study was conducted to develop an analytical method to determine colistin in fish and shrimp. The analytes were confirmed and quantified via liquid chromatography-tandem mass spectrometry (LC-MS/MS) in the positive ion mode using multiple reaction monitoring (MRM). The sample was extracted with acidified 5% methanol (containing 0.5% formic acid). Then, solid phase extraction (SPE) was used for cleanup. Matrix-matched calibration curves were linear over the calibration ranges (0.05-1.2 mg/kg) for all the analytes into blank sample with $r^2$ > 0.99. All the values fulfilled the criteria requested by the Codex guidelines. Average recoveries ranged from 85.9% to 107.9%. The repeatability of measurements, expressed as the coefficient of variation (CV, %), was less than 15%. The limit of detection (LOD) was 0.02 mg/kg, and the limit of quantitation (LOQ) was 0.05 mg/kg. This improved method showed higher accuracy and acceptable sensitivity to meet the CAC guideline requirements and is applicable for the analysis of residual colistin (A+B) in fish and shrimp.

Induction and growth of adventitious roots and bioreactor culture in Codonopsis lanceolata (더덕 (Codonopsis lanceolata)의 부정근 유도 및 생장에 미치는 배양조건과 생물반응기 배양)

  • Ahn, Chang-Ho;Bae, Kee-Hwa;Yi, Jae-Seon;Choi, Yong-Eui
    • Journal of Plant Biotechnology
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    • v.35 no.2
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    • pp.155-161
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    • 2008
  • This paper reported the establishment of mass production system of adventitious roots of Codonopsis lanceolata through shake flask and bioreactor culture. Induction of adventitious roots was started from the explants of leaf, stem and root on 1/2 strength Murashing and Skoog (MS) solid medium. Stem segments were the best explants for induction of adventitious roots compared to leaf and root segments. Among the different auxins tested (NAA, IBA and IAA), number of adventitious root per explant was highest on solid medium with 1.0 mg/L IBA, and produced $9.9{\pm}1.2$ roots per explant. However, growth of adventitious roots was fast in the presence of IBA at low concentration (0.1 mg/L). In shake flask culture, maximum production of adventitious roots (fresh weight) was obtained in half-strength MS medium compared to full-strength and one-third MS medium. When the adventitious roots produced in shake-flask culture were transferred to 5 L air-lift bioreactor, 16 times of fresh weight increase was gained after one month of culture. These results indicate that this protocol for the production of C. lanceolata adventitious roots can be applied to large scale culture for practical application.

Rapid Micropropagation by Axillary Buds Cultures of Smilax china

  • Song, Hyun-Jin;Sim, Seon-Jeong;Jeong, Mi-Jin;Heo, Chang-Mi;Kim, Hak-Gon;Jeong, Gwon-Yong;Heo, Su-Yeoung;Choi, Yong-Weon;Park, Geun-Hye;Yang, Jae-Kyung;Moon, Hyun-Shik;Choi, Myung-Suk
    • Journal of agriculture & life science
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    • v.44 no.6
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    • pp.39-44
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    • 2010
  • An efficient method for the rapid propagation of Smilax china from axillary buds was established. Plants with thick leafage were selected from Korea native S. china population. Axillary buds of S. china collected from selected plant and were cultured in various culture media (2MS, MS, 1/2MS, WPM, B5 and SH medium). Shoot was induced from axillary bud on MS basal medium after 4 weeks of culture. 1/2MS medium showed a higher growth rate than those of the others, while the lowest shoot growth was obtained in 2MS medium. Among the sucrose concentrations, 5% sucrose was the optimum level for shoots growth from axillay buds. Among cytokinins, $0.5mgL^{-1}$ 6-benzylaminopurine (BAP) treatment showed the best performance on shoot multiplication, yielding average shoot multiplication forming about 2.4. Rooting was induced directly near the base of the shoot on 1/2MS medium containing with three-auxins ${\alpha}-napthalene$ acetic acid (NAA), indole acetic acid (IAA) and ${\beta}-indolebutyric$ acid (IBA) (0.5 and $1.0mgL^{-1}$). The $1.0mgL^{-1}$ IBA treatments induced earliest rooting with maximum of root number and root growth. These rooted plantlets were successfully transferred to pots for 4 weeks hardening process, and were transferred to soil with above 90% survival rate.

Comparison of chrysanthemum cultivars based on direct shoot regeneration rates in tissue culture

  • Han, Bong-Hee;Lee, Su-Young;Park, Byoung-Mo
    • Journal of Plant Biotechnology
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    • v.36 no.3
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    • pp.275-280
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    • 2009
  • Direct shoot regeneration from leaf or internode or petiole segments was conducted in 33 cultivars of chrysanthemum. Shoot regeneration rates varied according to cultivars, culture media, and explant types. The high shoot regeneration rate of more than 70% in 15 cultivars (‘Pink Pangpang’, ‘Orange Memory’, ‘Relance’, ‘Zinba’, ‘Beakma’, ‘Innocence’, ‘Sunny Pangpang’, ‘Euro Yellow’, ‘Dublin’, ‘Boramae’, ‘Peak’, ‘Euro White’, ‘Vesuvio White’, ‘Linneker Salmon’ and ‘Pink Pride’) and 2 ones (‘Forward’ and ‘Agason’) was obtained from the segments of leaves and internodes, respectively, cultured on MS medium containing 1.0 mg-$L^{-1}$ BAP, 0.5 mg-$L^{-1}$ IAA and 30 g-$L^{-1}$ sucrose. That in 6 cultivars (‘Shuhonochikara’, ‘Hakunosen’, ‘Whitney Pangpang’, ‘Plaisir D’Amour’, ‘Grace’ and ‘Kumsu’) was observed from the segments of leaves or internodes cultured on 1/2 MS medium 1.0 mg-$L^{-1}$ BAP, 0.5 mg-$L^{-1}$ IAA and 15 g-$L^{-1}$ sucrose In case of 3 cultivars (‘Ilweol’, ‘Puma White’ and ‘Sharon’), when internode explants excised from mother plants, which were pre-cultured on MS medium containing 2 g-$L^{-1}$ activated charcoal and 30 g-$L^{-1}$ sucrose for two months in the dark, and cultured on MS medium containing 1.0 mg-$L^{-1}$ BAP, 0.5 mg-$L^{-1}$ IAA and 30 g-$L^{-1}$ sucrose, that was shown. Seven cultivars including ‘Puma Yellow’, ‘Argus’, ‘Yes Morning’, ‘Whiparam’, ‘Hakunohikari’, ‘Charming Eye’ and ‘Moon light’ requires more improved culture conditions. Tissues with the highest shoot regeneration rate were in descending order, leaf, petiole, and internode segments.

Selection of the fittest varieties of chrysanthemum (Dendranthema grandiflorum Kitamura) and set of culture condition for efficient transformation (형질전환 효율성 향상을 위한 국화 최적 품종 선발 및 배지.배양조건설정)

  • Kang, Chan-Ho;Yun, Seung-Jung;Han, Bum-So;Han, So-Gon;Kown, Sung-Hwan;Song, Young-Ju;Jang, Mi-Hyang
    • Journal of Plant Biotechnology
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    • v.36 no.4
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    • pp.373-383
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    • 2009
  • To set efficient transformation system in chrysanthemum, thirty-four chrysanthemum (Dendranthema grandiflorum Kitamura) varieties were collected and cultured for shoot regeneration. Five varieties, ‘Shuho-no-chikara', ‘Zinba', ‘Baekma', ‘Pink pride' and ‘Keumsu' of them were selected, because they had a high shoot regeneration efficiency. MS medium containing 1.0 mg/L NAA and BA respectively was very adequate for shoot regeneration in those varieties. MS medium with 3.0 mg/L NAA and 1.0 mg/L kinetin in ‘Shuho-no-chikara' and the medium with 0.5 mg/L NAA and 3.0 mg/L BA in ‘Keumsu' were also suitable for shoot regeneration. The most efficient callus induction and shoot regeneration were obtained on MS medium. Shoot regeneration was enhanced more than 8% on MS medium with 0.3% phytagel and 10-15 mg/L putrescine. The best cultural material for shoot regeneration was stem. When stem was used as a culture material, shoot regeneration rate was increased more than 26% and the days to shoot regeneration was shortened about 14 days.