• Title/Summary/Keyword: MG/MS

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Production of Novel Flavonoids in Cell Cultures of Cudrania tricuspidata (꾸지뽕나무 (Cudrania tricuspidata)세포배양에 의한 신규 Flavonoids 생산)

  • 최명석;곽상수;유장렬;이인경;유익동
    • Korean Journal of Plant Tissue Culture
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    • v.28 no.3
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    • pp.159-164
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    • 2001
  • To produce novel bioactive flavonoids, Gericudranin A and Gericudranin B, a cell culture system of Cudrania tricuspidata including callus induction and optimization of culture conditions was established. Friable calli were efficiently induced from the hypocotyl segments of seedlings on B5 medium supplemented with 1.0 mg/L NAA, 0.1 mg/L kinetin and 3% sucrose. Several factors were optimized for the Gericudranin production and the cell growth in suspension cultures. Low level of basal salt medium (1/8 MS), 1.0 mg/L IAA and 0.1 mg/L zeatin, and high level of sucrose (5%) were effective for the production of Gericudranins, whereas WPM with 1.0 mg/L NAA, 0.1 mg/L zeatin, and 5% sucrose were more effective for the cell growth. When cells were cultured on MS liquid medium supplemented with 1.0 mg/L IBA, about 2200 $\mu\textrm{g}$/g dry wt of Gericudranin A could be produced. The level might be about 10 times of the native inner bark. About 2350 $\mu\textrm{g}$/g dry wt of Gericudranin B was also produced on MS liquid medium with 5% sucrose, 1.0 mg/L NAA, 0.1 mg/L kinetin. The content was estimated about 3 times of the level of native inner bark.

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Plant Regeneration of Geranium (Pelargonium graveolense) Callus and Changes of Peroxidase Isozyme Pattern (제라늄(Pelargonium graveolense) 캘러스의 재분화 및 peroxidase isozyme 발현패턴 변화)

  • Lee, Seok-Hyun;Lee, Mi-Young
    • Applied Biological Chemistry
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    • v.43 no.3
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    • pp.184-189
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    • 2000
  • Callus was induced from the petioles of scented-geranium (Pelargonium graveolense) in MS medium containing various concentrations of plant growth regulators. The highest frequency of more than 70% of callus was induced in 2 mg/l NAA and 0.5 mg/l BAP or 2 mg/l 2,4-D and 0.5 mg/l BAP combined treatment, while not in 2,4-D, NAA or BAP alone. When the callus was transferred to the MS medium containing 0.05 mg/l NAA and 0.5 mg/l BAP, were highest intensity of shoot formation, 14 shoots/callus, was induced after 5 weeks. The highest rooting was observed on hormone-free rooting media from the regenerated shoots after 3 weeks, indicating that the regeneration from geranium callus might be possible by changing the hormone ratios. Peroxidase (POD) specific activities in callus induced from 2 mg/l NAA and 0.5 mg/l BAP were higher than those of 2 mg/l 2,4-D and 0.5 mg/l BAP callus during the entire culture periods. POD isozyme C3 was the main cathodic POD isozyme expressed in NAA and BAP callus, while C1 was the main in 2,4-D and BAP callus. However, anodic POD isozymes, A1, A2 and A3 were expressed with similar activities in both hormone combinations.

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Transformation of Korean Ginseng (Panax ginseng C.A. Meyer) with Salt Toleranc SAL1 Gene (염류내성관련 SAL1 유전자에 의한 인삼 형질전환)

  • In, Jun-Gyo;Yang, Deok-Chun
    • Korean Journal of Medicinal Crop Science
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    • v.13 no.1
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    • pp.57-62
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    • 2005
  • Salt-tolerant transgenic Panax ginseng plants were produced by introducing the SAL1 geue (3'(2'), 5'-bis-phosphate nucleotidase) that confers tolerance to the salts through Agrobacterium tumefaciens co-cultivation. Cotyledon explants of immature ginseng zygotic embryos cultured on Murashige and Skoog medium lacking growth regulators formed somatic embryos directly with below 10%, but the 74% tranformation rate were observed at the treatment of phytohormone with 1.0 mg/l 2,4-D and 0.5 mg/l kinetin. Somatic embryos were initially cultured on MS medium supplemented with 250 mg/l cefotaxime for 3 weeks and subsequently subcultured five times to a medium containing 100 mg/l kanamycin and 250 mg/l cefotaxime. Upon development into the cotyledonary stage, these somatic embryos were transferred to on the medium containing 50 mg/l kanamycin and 10 mg/l gibberellic acid to induce germination and strong selection. Integration of the transgene into the plants was confirmed by polymerase chain reaction with specific primers. The ginseng transformants with well-developed shoots and roots were successfully acclimatized in a greenhouse when they were planted in soil.

유전자총 및 Agrobacterium을 이용한 Bentgrass의 형질전환

  • 임용우;김기용;정영수
    • Proceedings of the Korean Society of Grassland Science Conference
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    • 1999.06a
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    • pp.74-74
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    • 1999
  • Bentgrass의 형질전환식물 생산을 위하여 유전자총 (Gene-gun) 및 Agrobacterium기법을 이용하여 형질전환을 시도한 결과를 요약하면 다음과 같다. 1. 캘러스의 유도 및 증식 : Bentgrass (Agrostis palustris)의 종자를 MS-5 배지 (MS 기본배지, 2,4-D 5mg/L, casein hydrolysate 2g 포함)에 치상하여 캘러스를 유도하였다. 유도된 캘러스는 증식을 위하여 MS-3 배지 (MS기본배지, 2,4-D 3mg/L, casein hydrolysate 2g 포함)에서 2주 간격으로 subculture 하였다.(중략)

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Plant Regeneration via Multiple Shoots Formation from Sucker Explants of Rubus fruticosus L. (블랙베리(Rubus fruticosus L.)의 맹아절편체로부터 다경유도를 통한 식물체 재분화)

  • Shin Jeong-Sun;Sim Ock-Kyeong;Lee Jong-Chon;Cho Han-Jik;Kim Ee-Yup;Lee Kang-Seop
    • Korean Journal of Plant Resources
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    • v.18 no.3
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    • pp.456-461
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    • 2005
  • This study was carried out to induce plant regeneration via shoot formation from sucker explants of Rubus fruticosus L. To induce adventitious shoots, sucker explants were sterilized in $1.2\%$ NaOCl solution, and cultured on the MS solid medium supplemented with kinetin (0.5, 1.0, 3.0 mg/L) and BA (0.5, 1.0, 3.0 mg/L), respectively. As above, to induce adventitious shoots, sucker explants were cultured on the MS solid medium supplemented with IBA (0, 0.1, 1.0 mg/L) and BA (0, 0.1, 1.0, 2.0 mg/L). After 4 weeks of culture, the highest frquency $(100\%)$ of shoot formation from sucker explants was obtained from the medium with 1.0 mg/L BA. The highest shoot number per explant from in vitro shoot explants was 5.3. After 10 weeks of culture, the number of shoot per explant was increased. The highest frequency $(85\%)$ of root formation was obtained at 0.5 mg/L glycine medium, when the explant with shoot were cultured on the MS medium containing glycine at various concentrations from 0 to 2.0 mg/L. The survival rate of the plantlets after transfer to plastic pots containing sand, soil, and vermiculite (1:1:1, vol.) was $95\%$. The results indicate that micropropagation procedure can be applied for an efficient mass propagation of Rubus fruticosus.

Determination of Post-harvest Fungicide in Citrus Fruits Using LC-MS (LC-MS를 이용한 감귤류의 수확 후 처리 살균제 분석)

  • Choi, Su-Jeong;Kim, Eun-Jeong;Lee, Jae-In;Cho, In-Soon;Park, Won-Hee;Hwang, In-Sook;Kim, Moo-Sang;Kim, Gun-Hee
    • Korean Journal of Food Science and Technology
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    • v.45 no.4
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    • pp.409-415
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    • 2013
  • Post-harvest fungicide residue was measured in citrus fruits. Samples were collected from local markets in Seoul and analyzed using liquid chromatography coupled with mass spectrometry (LC-MS). LC-MS results were validated for the assay of pesticides by using linearity, accuracy, precision, and limits of detection and quantification. The linearity in the concentration ranged from 0.005 to 2.0 mg/kg ($R^2$ >0.999). Sample recoveries ranged from 80.2 to 98.3% with relative standard deviations below 4.0% for spiking levels from 0.01 to 1.0 mg/kg. The limits of detection ranged between 0.002 and 0.008 mg/kg, and the limits of quantification ranged between 0.006 and 0.027 mg/kg. The highest residue levels for carbendazim, thiabendazole, imazalil, and azoxystrobin in citrus fruits were 0.541, 0.958, 0.721, and 0.052 mg/kg, respectively. The pesticide residues found in citrus fruits were blow maximum residue limits (MRLs) and are not a serious public health problem.

Quantitative Analysis of the Seven Marker Components in Asarum sieboldii using the LC-MS/MS and GC-MS (LC-MS/MS와 GC-MS를 이용한 세신 추출물 중 7종 성분의 함량분석)

  • Seo, Chang-Seob;Shin, Hyeun-Kyoo
    • Korean Journal of Pharmacognosy
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    • v.44 no.4
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    • pp.350-361
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    • 2013
  • Asarum sieboldii has been used for treatment of fever, pain, common cold, and chronic sinusitis in Korea. In this study, we performed quantification analysis of seven major constituents including aristolochic acid I, aristolochic acid II, ${\alpha}$-asarone, ${\beta}$-asarone, elemicin, methyl eugenol, and safrole in the 70% ethanol extract of Asarum sieboldii and its solvent fractions, n-hexane, ethylacetate, n-butanol, and water ones using a ultra-performance liquid chromatography-electrospray ionization-mass spectrometer(UPLC-ESI-MS) and gas chromatography-mass spectrometer(GC-MS). Regression equations of seven components were acquired with $r^2$ values >0.99. The values of limit of detection(LOD) and quantification(LOQ) were 0.1-3.9 ng/mL and 0.3-11.7 mg/mL, respectively. The amount of the seven compounds in Asarum sieboldii were not detected -143.66 mg/g. The established LC-MS/MS and GC-MS methods will be helpful to improve quality control of Asarum sieboldii.

Callus Formation and Plant Regeneration from Immature Embryos of Eleutherococcus senticosus (가시오갈피 미숙배 배양으로부터 Callus 형성 및 식물체 재분화)

  • Yu, Chang-Yeon;Kim, Jae-Kwang;Ahn, Sang-Deuk
    • Korean Journal of Medicinal Crop Science
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    • v.5 no.1
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    • pp.49-55
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    • 1997
  • This study was conducted to establish mass propagation system from the tissue culture using immature embryos in Eleutherococcus senticosus. Immature embryos from seeds were removed under the microscope and placed on MS, $MSB_5\;and\;B_5$ medium containing several plant growth regulators. While the calli were well formed on media containing 2 mg/l of 2, 4-D, 2 mg/l of 2, 4-D and 0.7 mg/l of TDZ, shoot regeneration was better on MS medium with combinations of high concentrations of TDZ and low concentrations of 2, 4-D. Treatment of 2, 4-D alone was better than treatment of TDZ alone in callus induction, but plant regeneration was reversed. The results of callus formation and shoot regeneration on $MSB_5\;and\;B_5$ media were similar to those of MS media. The rate of callus formation was nearly 100% when 2, 4-D was added to $B_5$, medium on concentration of 2 mg/l or 0.7 mg/l. TDZ showed very significant effect on the formation of multiple shoots.

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Appropriate in Vitro Culture Conditions of Growing Medium for New Ever-bearing Strawberry 'Goha' (사계성 신품종 딸기 '고하'의 기내배양을 위한 배지의 적정 조건)

  • Lee, Jong-Nam;Kim, Hye-Jin;Kim, Ki-Deog;Kwon, Young-Seok;Im, Ju-Sung;Yeoung, Young-Rok;Lim, Hak-Tae
    • Horticultural Science & Technology
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    • v.28 no.6
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    • pp.1051-1056
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    • 2010
  • This study was carried out to determine suitable in vitro culture conditions of new ever-bearing strawberry ($Fragaria$ $ananassa$ Duch.), 'Goha'. Four-week old plantlets which were derived from the meristem culture were used in this study. Three different culture media including MS, Gamborg B5 and White medium were used for the plant culture and the medium concentrations were at the 5 levels of $1/3{\times}$, $1/2{\times}$, $1{\times}$, $2{\times}$ and $3{\times}$. Sucrose content ranged at four levels of 1, 3, 5 and 8% (w/v). Crown diameter on the MS medium was thicker (2.1 mm) than in other media. Fresh weight on the MS medium was 482 mg, which was heavier than 88 mg or 260 mg of Gamborg B5 or White medium, respectively. Particularly, 1/2MS medium was found to have higher growth rate than these in other treatments (fresh weight, dry weight and D/F rate). Shoot length in the treatment of 1% sucrose concentration was 3.6 cm which was the longest. Shoot length was in inverse proportion to the increasing concentration of sucrose. Fresh weight was increased up to 3% sucrose concentration, but decreased above 5% sucrose concentration. From the results, we found that the best condition for in vitro culture of new ever-bearing strawberry 'Goha' was 1/2MS medium supplemented with 1% sucrose concentration.

Herbicide Resistant Cabbage (Brassica oleracea ssp. capitata) Plants by Agrobacterium-mediated Transformation

  • Lee, Yeon-Hee;Lee, Seung-Bum;Suh, Suk-Chul;Byun, Myung-Ok;Kim, Ho-Il
    • Journal of Plant Biotechnology
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    • v.2 no.1
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    • pp.35-41
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    • 2000
  • Transgenic cabbage (Brassica oleracea ssp. capitata) plants resistant to the commercial herbicide Bast $a^{R}$ were obtained by Agrobacterium tumefaciens - mediated transformation. Hypocotyl segments of in vitro grown plants were infected with Agrobacterium tumefaciens LBA 4404 harboring plasmid pMOG6-Bar which contains hpt and bar genes. Explants were cultured on callus induction medium (MS basal medium + 1 mg/L NAA + 2 mg/L BA + 2 mg/L AgN $O_3$+ 100 mg/L carbenicillin + 250 mg/L cefotaxime) supplemented with 15 mg/L hygromycin. Hygromycin resistant calluses were transferred to shoot regeneration medium (MS basal medium + 0.1 mg/L NAA + 2 mg/L BA + 3% sucrose + 2 mg/L AgN $O_3$+ 15 mg/L hygromycin + 250 mg/L cefotaxime + 100 mg/L carbenicillin). In order to induce roots, elongated shoots were placed on the MS medium without plant growth regulators and hygromycin. Southern blot analysis of several putative transgenic plants indicated that one to five intact copies of Apt and bar genes were incorporated into the genome. Expression of bar gene was confirmed by Northern blot analysis and by herbicide resistant phenotype. Seed progeny from self-pollinated transformants expressed the herbicide resistance and showed Mendelian segregation of the introduced gene.e.

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