• Title/Summary/Keyword: lCP-MS

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Plant regeneration from suspension-cultured cell clusters of Arabidopsis thaliana (애기장대(Arabidopsis thaliana)의 현탁배양세포괴로부터 식물체 재분화)

  • 김명덕;김준철;진창덕;임창진;한태진
    • Korean Journal of Plant Tissue Culture
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    • v.25 no.3
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    • pp.195-200
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    • 1998
  • Callus induction from leaf and stem explants of Arabidopsis thaliana was successfully obtained when leaf explants were cultured on MS medium containing 2.0 mg/L 2,4-D in the dark and also, when stem explants were cultured on CP medium containing 0.5 mg/L 2,4-D and 0.1 mg/L BAP. Explant-derived sliced calli were suspension-subcultured every week in CP liquid medium with 0.5 mg/L 2,4-D and 0.1 mg/L BAP in the dark, and shoot-forming cell clusters of nodular, pale yellow and knobby type were selected after 7-8 weeks of culture. Shoots were initiated from the green spots of the selected shoot forming calli cultured on MS regeneration medium containing 0.05 mg/L IAA, 7.0 mg/L 2-iP and 30 g/L sucrose under continous illumination for four weeks. Shoot regeneration frequency (calli regenerating at least one shoot) was more than 50%. For plant regeneration, excised shoots were trnasferred to hormone free medium for root initiation after 4 weeks of culture. The regenerants were bolting after 2 weeks of culture and formed in vitro flowering buds within bracts after 4 weeks of culture.

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Plant Regeneration from Protoplasts of Suspension Cultured Cells in Arabidopsis thaliana (애기장대(Arabidopsis thaliana) 현탁배양세포의 원형질체로부터 식물체 재분화)

  • 김명덕;김준철;진창덕;임창진;한태진
    • Korean Journal of Plant Tissue Culture
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    • v.27 no.2
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    • pp.125-131
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    • 2000
  • Protoplasts of Arabidopsis thaliana were easily isolated from the shoot-forming (SF) suspension-cultured cell clusters with 4 hours-shaking condition (40 rpm) on CPD enzyme solution containing 1% cellulase R-10, 0.25% pectolyase Y-23 and 0.5% driselase. Protoplasts were cultured on liquid KAO medium supplemented with 1 mg/L 2,4-D, 0.5 mg/L kinetin, 200 mg/L spermidine and 68 g/L glucose. Also, protoplasts were cultured on 0.2 $\mu$M membrane filter placed onto CP solid medium containing the suspension cells as feeder cells in the dark at $25^{\circ}C$ for 4 weeks. Protoplast-derived-SF calli were cultured on MS medium containing 0.05 mg/L IAA, 7 mg/L 2 ip and 30 g/L sucrose under the continuous illumination for four weeks. The frequency of shoot formation was about 60%. The regenerants were transferred into potting soil to grow mature plants. The regenerants formed the silques with seeds after 8 weeks of cultures.

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Shoot Proliferation and Plant Regeneration from Suspension-Cultured Cells of Dianthus gratianopol (패랭이꽃속 Dianthus gratianopol의 현탁배양세포로부터 Shoot 증식과 식물체 재분화)

  • Kim Joon-Chul
    • Journal of Plant Biotechnology
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    • v.32 no.4
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    • pp.301-306
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    • 2005
  • Conditions for efficient organogenesis and plant regeneration from Dianthus gratianopol suspension cultured cells were established. Shoot-forming calli of glossy surface, pale green and knobby type were selected from leaf explant-derived calli and were suspension-subcultured every week in CP liquid medium with 1.0 mg/L 2,4-D and 0.5 mg/L BAP. Combinations of 1.0 mg/L 2,4-D and 0.5 mg/L BAP, and 1.5 mg/L 2,4-D and 0.5 mg/L BAP were effective for the induction of regenerative callus from the suspension cultured cell clusters. Multiple shoot primordia were initiated from the green spots of these regenerative callus and formed shoots on MS medium with 1.0 mg/L TDZ and 0.5 mg/L PAA. Shoot regeneration frequency (calli regenerating at least one shoot) was about 87%. For plant regeneration, proliferated shoots were excised and transferred to MS medium with 0.1 mg/L NAA for root initiation after 9 weeks of culture. The regenerants were potted in soil and formed the flowering buds and petals. Also, adventitious shoots were formed from the excised green shoot primordia of regenerative callus and these shoots proliferated successfully and regenerated to whole plants.

Optimization of Embryogenic Callus Induction and Plant Regeneration in Orchid Coelogyne cristata

  • Naing, Aung Htay;Lim, Ki-Byung
    • Horticultural Science & Technology
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    • v.29 no.3
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    • pp.260-266
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    • 2011
  • An efficient protocol was established for high frequency somatic embryogenesis through a callus culture of Coelogyne cristata. The best frequency of callusing was obtained from a PLB segment (3-5 mm) cultured on MS medium supplemented with coconut water (CW) and a combination of both 3 $mg{\cdot}L^{-1}$ of 2,4-D and BA. When the calli were sub-cultured on the MS medium without any PGRs, the average number of somatic embryos were higher than those with PGRs treatment. NAA is the most critical factor among PGRs, which dramatically hindered for the formation of a somatic embryo. The efficacy of the addition of coconut powder (CP) for somatic embryogenesis was almost the same in all treatments. However, the number of somatic embryos formed distinctly depended on age of the callus. The somatic embryos converted into healthy plants with well-developed shoots on the same medium. Plantlets showed the best responses of root and shoot growth when transferred to $\frac{1}{2}$ MS medium containing 1.5 $g{\cdot}L^{-1}$ of activated charcoal. All plants with above 3.0-cm-high were successfully acclimatized in the greenhouse.

Efficient Plant Regeneration from Mesophyll Protoplast of Arabidopsis thaliana and Morphological Characterization of Regenerants (애기장대 (Arabidopsis thaliana)의 엽육원형질체로부터 효율적인 식물체 재분화와 이들의 형태적 특성)

  • 김명덕;김준철;진창덕;임창진;한태진
    • Korean Journal of Plant Tissue Culture
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    • v.26 no.2
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    • pp.127-132
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    • 1999
  • Protoplasts were isolated from the leaf mesophyll tissue of in vitro 4-weeks-old Arabidopsis thaliana and cultured in MS liquid medium supplemented with 2.0 mg/L NAA, 0.5 mg/L BAP and 9% mannitol in the dark at $25^{\circ}C$. When protoplast-derived microcolonies were dehydrated, the frequency of callus induction enhanced approximately 7-fold higher compared with non-dehydrated microcolonies in CP medium. Fifty callus lines were selected from dehydrated microcolonies. Shoots were efficiently initiated from the green spots of the selected shoot forming calli cultured on MS regeneration medium supplemented with 0.05 mg/L IAA, 7.0 mg/L 2-iP and 30 g/L sucrose under continous illumination for 4 weeks. Shoot regeneration frequencies (calli regenerating at least one shoot) were 3.5%~56%. Histological observations of shoot forming callus revealed that tracheary elements initiated from inner compact cells, and that meristemoids developed to shoot primordia and shoots. Roots were induced from these regenerating shoots on MS medium without phytohormones. These regenerants were successfully transplanted into potting soil. Morphological characterization of 50 protoplast-derived plants showed that the frequency of normal type was 78%.

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A Study of Analytical Method for Trace Metal Ions in Whole Blood and Urine by Inductively Coupled Plasma-Mass Spectrometry using Solid-Liquid Extraction Technique (유도결합 플라스마-질량분석법과 고체-액체 추출법을 이용한 혈액 및 소변중 미량금속의 분석에 관한 연구)

  • Lee, Won;Hur, Young-Hoe;Park, Kyung-Su
    • Analytical Science and Technology
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    • v.11 no.4
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    • pp.281-291
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    • 1998
  • An analytical method for the simultaneous measurement of trace Cu, Sn, and Bi in blood and urine has been investigated by Inductively Coupled Plasma-Mass Spectrometry (ICP-MS). Microwave oven was used for the pretreatment of blood samples using nitric acid and hydrogen peroxide in a closedvessel digestion system with 1 mL whole blood for 8 minutes. Amberlite IRC-718 resin was used as a solid phase in solid-liquid extraction technique for the removal of matrix interferences such as Na, S, P, and other polyatomic ion species. Detection limits for Cu, Sn, and Bi by this method were 0.000375 ng/mL, 0.000297 ng/mL, and 0.000174 ng/mL, respectively. Recoveries of 99.1% for Cu, 102.5% for Sn, and 98.4% for Bi were obtained for the standard spiked NIST SRM 955a blood sample. The developed method was applied for whole real blood and urine samples.

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Assessment of Physicochemical Characteristics among Different Types of Pale Ale Beer (Pale ale류에 속하는 맥주 신제품의 이화학적 특성 평가)

  • Kim, Ki Hwa;Park, Sue Jee;Kim, Jee Eun;Dong, Hyemin;Park, In Seon;Lee, JaeHwan;Hyun, So Yang;Noh, Bong Soo
    • Korean Journal of Food Science and Technology
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    • v.45 no.2
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    • pp.142-147
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    • 2013
  • This study was conducted to evaluate and compare new beer (NB) with market beers, e.g., New castle brown ale (NC), Victoria bitter (VB), and Coopers pale ale (CP) using physicochemical parameters. In addition, pattern recognition analyses were carried out using an electronic nose based on mass spectrometry (MS-E nose) and an electronic tongue (E-tongue) for differentiation of the different types of beer. The measured alcohol content of NB was 4.37%. NB was not significantly different compared with other types of beer with regard to bitterness unit, color, and polyphenol content (p<0.05). On the basis of the flavor pattern determined by the MS-E nose, NB was separated by DF1 (first score from discriminant function analysis), while NC, VB, and CP were located in the same group. The result of the E-tongue showed that the different samples could be clearly discriminated; NB was less sour. It was suggested that the discriminant function analysis (DFA) given by the MS-E nose and E-tongue could be used for evaluations during new product development. Furthermore, because of its simplicity, it might be possible to use the validated method for the evaluation of beer.

Sample Pretreatment for the Determination of Metal Impurities in Silicon Wafer (실리콘 웨이퍼 중의 금속 불순물 분석을 위한 시료 전처리)

  • Chung, H.Y.;Kim, Y. H.;Yoo, H.D.;Lee, S.H.
    • Journal of the Korean Chemical Society
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    • v.43 no.4
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    • pp.412-417
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    • 1999
  • The analytical results obtained by microwave digestion and acid digestion methods for sample pretreatment to determine metal impurities in silicon wafer by inductively coupled plasma-mass spectrometry (ICP-MS) were compared. In order to decompose the silicon wafer, a mixed solution of $HNO_3$ and HF was added to the sample and the metal elements were determined after removing the silicon matrix by evaporating silicon in the form of Si-F. The recovery percentages of Ni,Cr and Fe were found to be 95∼106% for both microwave digestion and acid digestion methods. The recovery percentage of Cu obtained by the acid digestion method was higher than that obtained by the microwave digestion method. For Zn, however, the microwave digestion method gave better result than the acid digestion method. Fe was added to a silicon wafer using a spin coater. The concentration of Fe in this sample was determined by lCP-MS, and the same results were obtained in the two pretreatment methods.

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Isolation and Purification of Berberine in Cortex Phellodendri by Centrifugal Partition Chromatography (Centrifugal Partition Chromatography에 의한 황백으로부터 Berberine의 분리 및 정제)

  • Kim, Jung-Bae;Bang, Byung-Ho
    • The Korean Journal of Food And Nutrition
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    • v.27 no.3
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    • pp.532-537
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    • 2014
  • Cortex Phellodendri (CP) is derived from the dried bark of Phellodendron amurense. It has been widely used as a drug in traditional Korea medicine for treating diarrhea, jaundice, swelling pains in the knees and feet, urinary tract infections, and infections of the body surface. Many analytical methods have been used to study oriental herbal medicines, such as thin-layer chromatography, column liquid chromatography, and high performance liquid chromatography (HPLC). In this study, preparative centrifugal partition chromatography (CPC) was successfully carried out in order to separate pure compounds from a CP methanol extract. The optimum two-phase CPC solvent system was composed of n-butanol: acetic acid: water (4:1:5 v/v/v). The flow rate of the mobile phase was 3 mL/min in ascending mode with rotation at 1,000 rpm. The CPC-separated fraction and purification procedures were carried out by preparatory HPLC. The $^1H$ NMR spectrum revealed that the resonances at ${\delta}$ 4.10 and 4.20 ppm corresponded to three protons ($-OCH_3$), whereas those at ${\delta}$ 6.10 ppm corresponded to two protons ($-OCH_2O-$). Further, two aromatic protons (H-11 and H-12) conveys a doublet-doublet pattern. The H-11 doublet and H-12 doublet appear at ${\delta}$ 7.98 and 8.11, respectively. The $^{13}C$ NMR. spectrum showed a tetrasubstituted with a methylenedioxy group at C2 and C3, and two methoxy groups at C9 and C10. The chemical structure of the berberine was identified by $^1H$, $^{13}C$-nuclear magnetic resonance and electrospray ionization-mass spectroscopy spectral data analysis.

Development of Antibiotics Marker-free Potato Having Resistance Against Two Herbicides (두 가지 제초제에 대하여 저항성을 가지는 항생제 마커-프리 형질전환 감자 육성)

  • Fang, Yi-Lan;Kim, Jin-Seog;Gong, Su;Mo, Hwang-Suk;Min, Seok-Ki;Kwon, Suk-Yoon;Li, Kui-Hua;Lim, Hak-Tae
    • Journal of Plant Biotechnology
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    • v.34 no.3
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    • pp.253-261
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    • 2007
  • This study was conducted to develop an antibiotics marker-free potato (Solanum tuberosum L., cv. Taedong valley) plant having resistance against two herbicides. Agrobacterium tumefaciens strain EHA105, harboring a binary vector plasmid pCAMBIA3300 containing bar gene under the control of a promoter CaMV35S and linked CP4-EPSPS genes driven by CaMV35S promoter, was used in the current study. The leaf segments of newly bred potato variety (cv. Taedong Valley) was co-cultured with Agrobacterium. Then, the regenerated individual shoots were excised and transferred to potato multiplication medium supplemented with 0.5 mg/L phosphinothricin. The shoots were rooted in MS medium without hormone and obtained putative transgenic plant E3-6. Integration of target genes into the E3-6 plant and their expression was confirmed by PCR, Southern analysis, and ELISA test. The tissue necrosis test on young leaf blade and shikimic acid accumulation test using the tissue of E3-6 plant were conducted to investigate the resistance to glufosinate-ammonium and glyphosate, respectively. The transgenic plants (E3-6) simultaneously showed a high resistance to both herbicides. The same results were surely obtained also in the whole plants foliar-treated with alone or mixture of two herbicides, glufosinate-ammonium and glyphosate.