• Title/Summary/Keyword: MG/MS

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Determination of Ametryn Residue in Agricultural Commodities Using HPLC-UVD/MS (HPLC-UVD/MS를 이용한 농산물 중 ametryn의 분석)

  • Lee, Su-Jin;Kim, Young-Hak;Song, Lee-Seul;Choung, Myoung-Gun
    • The Korean Journal of Pesticide Science
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    • v.15 no.2
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    • pp.125-133
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    • 2011
  • Ametryn is used in USA, China, and Japan, but not introduced in Korea yet. So, MRL (Maximum Residue Level), and analytical method of ametryn were not establishment in Korea. Therefore, this experiment was conducted to establish a determination method for ametryn residue in crops using HPLC-UVD/MS. Ametryn residue was extracted with acetone from representative samples of five raw products which comprised hulled rice, soybean, apple, green pepper, and Chinese cabbage. The extract was diluted with saline water, and dichloromethane partition was followed to recover ametryn from the aqueous phase. Florisil column chromatography was additionally employed for final clean up of the extract. The ametryn was quantitated by HPLC with UVD, using a Tosoh ODS 120T ($4.6{\times}250$ mm) column. The crops were fortified with ametryn at 2 levels per crop. Mean recovery ratio were ranged from 83.7% for a 0.2 mg/kg in soybean to 91.1% for a 1.0 mg/kg in hulled rice. The coefficients of variation were ranged from 1.2% for a 1.0 mg/kg in hulled rice to 3.6% for a 1.0 mg/kg in soybean. Quantitative limit of amatryn was 0.02 mg/kg in representative 5 crop samples. A LC/MS with selected-ion monitoring was also provided to confirm the suspected residue. Therefore, this analytical method was reproducible and sensitive enough to determine the residue of ametryne in agricultural commodities.

Efficient plant regeneration from immature embryo cultures of Jatropha curcas, a biodiesel plant

  • Varshney, Alok;Johnson, T. Sudhakar
    • Plant Biotechnology Reports
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    • v.4 no.2
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    • pp.139-148
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    • 2010
  • Jatropha curcas L. (Physic nut) is a commercially important non-edible oil seed crop known for its use as an alternate source of biodiesel. In order to investigate the morphogenic potential of immature embryo, explants from four developmental stages were cultured on medium supplemented with combinations of auxins and cytokinins. It was found that the size of embryo is critical for the establishment of callus. Immature embryos (1.1-1.5 cm) obtained from the fruits 6 weeks after pollination showed a good response of morphogenic callus induction (85.7%) and subsequent plant regeneration (70%) with the maximum number of plantlets (4.7/explant) on Murashige and Skoog's (MS) medium supplemented with IBA (0.5 $mg\;l^{-1}$) and BA (1.0 $mg\;l^{-1}$). The above medium when supplemented with growth adjuvants such as 100 $mg\;l^{-1}$ casein hydrolysate + 200 $mg\;l^{-1}$ L-glutamine + 8.0 $mg\;l^{-1}$ $CuSO_4$ resulted in an even higher frequency of callus induction (100%). Plant regeneration (90%) with the maximum number of plantlets (10/explant) was achieved on MS medium supplemented with 500 $mg\;l^{-1}$ polyvinyl pyrrolidone + 30 $mg\;l^{-1}$ citric acid + 1 $mg\;l^{-1}$ BA + 0.5 $mg\;l^{-1}$ Kn + 0.25 $mg\;l^{-1}$ IBA. It was observed that plantlet regeneration could occur either through organogenesis of morphogenic callus or via multiplication of pre-existing meristem in immature embryos. The age of immature embryos and addition of a combination of growth adjuvants to the culture medium appear to be critical for obtaining high regeneration rates. Well-developed shoots rooted on half-halfstrength MS medium supplemented with 0.5 $mg\;l^{-1}$ IBA and 342 $mg\;l^{-1}$ trehalose. The rooted plants after acclimatization were successfully transferred to the field in different agro-climatic zones in India. This protocol has been successfully evaluated on five elite lines of J. curcas.

Agrobacterium Mediated Transformation from Callus Pretreated with Particle Bombardment in Lilium lancifolium Thunb. (Particle Bombardment에 의해 전처리 된 참나리(Lilium lancifolium Thunb.) 캘러스의 Agrobacterium tumefaciens을 통한 형질전환)

  • Nam, Sang-Wook;Kim, Hei-Young
    • Journal of Plant Biotechnology
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    • v.31 no.1
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    • pp.13-17
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    • 2004
  • To improve transformation efficiency, the callus of Lilium lancifolium Thunb. were bombarded by particles coated with pIG 121 Hm which include NPT II and GUS genes, and then cocultivated with Agrobacterium tumefaciens EHA101 which contain pIG121Hm binary vector, carrying neomycin phosphotransferase (NPT II) and $\beta$-Glucuronidase (GUS) genes. Three days after cocultivation with Agrobacterium tumefaciens and particle bombardment, the callus clusters were transferred to MS medium containing 1mg/L 2,4-D, 0.1mg/L BAP, 100mg/L kanamycin and 200mg/L carbenicillin. Four weeks after transfer to the selection medium, GUS expression was detected and PCR analysis revealed the presence of NPT II fragment of the expected size (700 bp) in the transformed callus. The GUS expression from Agrobacterium-mediated transformants after particle bombardment increased to over 3-folds compared with that of callus cocultivated with Agrobacterium tumefaciens without particle bombardment. The callus clusters containing kanamycin resistant gene were transferred to MS medium containing 1mg/L NAA and 1mg/L BAP. Somatic embryos were developed in four weeks and microbulbs expressing GUS were formed.

Influence of Growth Regulators and Potassium Humate on in Vitro Multiplication of Apple Rootstock M.26 (생장조절제 및 Potassium Humate가 사과대목 M.26 기내 증식에 미치는 영향)

  • 임학태;용영록;송융남;한교필;김종화
    • Korean Journal of Plant Tissue Culture
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    • v.21 no.3
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    • pp.131-135
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    • 1994
  • This experiment was designed to improve the in vitro production system of apple rootstock M.26 as being influenced by the growth regulators, TDZ, BA, IAA, IBA, zeatin, and GA$_3$. Different levels of potassium humate (KH), known as cytokinin and auxin-like substance, were also supplemented to the MS basal medium along with IBA 0.6 mg/L to find out it effect on root formation in apple rootstock M.26. ID initiate and establish the in vitro multiplication of shoots byway of meristem culture, MS medium added with zeatin 1.0 mg/L was found to be the most suitable, showing the 100% of survival rate of shoot tips. A combination of thidiazuron (TDZ) 0.2 mg/L and NAA 0.5 mg/L promoted the shoot proliferation when shoot tips were used as explants. MS basal medium plus IBA 0.6 mg/L was very effective for root induction, but an addition of potassium humate (250 mg/L) to the medium containing IBA 0.6 mg/L stimulated the induction and proliferation of the rook by far the better.

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Transformation of Arabidopsis gamma-Tocopherol Methyltransferase into Lettuce (Lactuca sativa L.) (애기장대 gamma-Tocopherol Methyltransferase 유전자를 이용한 상추의 형질전환)

  • 김명준;백소현;유남희;윤성중
    • Korean Journal of Plant Tissue Culture
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    • v.27 no.6
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    • pp.435-439
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    • 2000
  • Explants of lettuce (Lactuca sativa L.) were cocultured with A. tumefaciens LBA 4404 harboring ${\gamma}$-tocopherol methyltransferase (${\gamma}$-TMT) gene from Arabidopsis thaliana. These explants were transferred to MS medium supplemented with 50 mg/L kanamycin, 500 mg/L carbenicillin, 0.1 mg/L NAA and 0.5 mg/L BA. After 4 weeks, kanamycin resistant shoots were obtained from the explants on the selection medium. The putative transgenic shoots were transferred to rooting MS medium supplemented with 50 mg/L kanamycin and 250 mg/L carbenicillin. Stable incorporation of the Arabidopsis ${\gamma}$-TMT cDNA into lettuce genomic DNA was confirmed by PCR and Southern analysis. HPLC analysis showed that $\alpha$- to ${\gamma}$-tocopherol ratio increased over four fold in a transgenic lettuce line indicating successful expression of the transgenic Arabidopsis ${\gamma}$-TMT in lettuce.

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Effects of Growth Regulators on Shoot Differentiation and Bulblet Formation in Shoot-Tip and Bulb-Scale Cultures of Lilium longiflorum (백합 경단 및 인편배양으로부터 유식물체 분화 및 자구형성에 미치는 생장조절제의 영향)

  • 이은모;정해준;민병훈;이영복
    • Korean Journal of Plant Tissue Culture
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    • v.22 no.2
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    • pp.83-87
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    • 1995
  • Regulation of organ differentiation by growth regulators was investigated through the shoot-tip and bulb-scale cultures of Lilium longiflorum (cv Georgia). When shoot tips were placed on MS medium supplemented with 0.1 mg/L NAA alone or 0.1 mg/L NAA and 0.1 mg/L BA, axillary shoots were proliferated. Root diffentiation and growth were stimulated on the basal medium. Although growth regulation did not seem to be necessary when bulb scales were used as explants for shoot differentiation, its differentiation was promoted vigorously by 0.2 mg/L NAA, but suppressed by BA. Bulblets were formed from bulb-scale-derived plantlets cultured on MS medium supplemented with 0.1 mg/L IBA. And more bulblets were formed from the plantlet in MS medium supplying 0.2 mg/L NAA with 6% than3% sucrose

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Changes of RNA and Protein During Callus Induction and Plant Regeneration from Perilla frutescens (들깨로부터 Callus의 유기와 재분화에 따른 단백질 및 RNA의 변화)

  • 정상훈;양선경;김현경;정대수;조영수;김도훈
    • Journal of Life Science
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    • v.9 no.1
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    • pp.29-34
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    • 1999
  • Cotyledon and hypocotyl explants of perilla were cultured on MS medium containing a combined concentration of BA(0.5, 1.0 and 1.5mg/$\ell$) and NAA(0.1, 0.5 and 2.0mg/$\ell$) in order to regenerate the explant and induce the callus. The best regeneration of the explant and induction of the callus were observed in a combined concenteration of 0.5mg/$\ell$ of BA and 0.5mg/$\ell$ NAA both in cotyledon and hypocotyl explants. In cotyledon explants, rooting was achieved upon transferring shoots to MS medium containing 0.5mg/$\ell$ of BA and 0.1mg/$\ell$ of NAA. We also investigated the change of protein and RNA content on developmental stage of callus and plant regeneration of perilla. Protein content was increased but RNA content was decreased as the culture period increases. The banding pattern of polypeptide revealed that both 30KD and 45KD polypeptides were obvious in cotyledon obtained from pre-culture explants, but only 30KD polypeptide was further getting obvious as the culture period increases.

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Agrobacterium-mediated Transformation of PAT into Platycodon grandiflorum A. De. candolle (Agrobacterium을 이용한 Phosphinothricin Acetyl Transferase의 도라지로의 형질전환)

  • Park, Jae-Seong;Kim, Ik-Hwan;Hong, Eui-Yon;Yun, Tae;Lee, Cheol-Hee;Jeong, Jae-Hun;Yang, Deok-Chun;Yun, Jong-Sun
    • Korean Journal of Medicinal Crop Science
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    • v.15 no.4
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    • pp.285-290
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    • 2007
  • This study was conducted to introduce phosphinothricin acetyl transferase (PAT) gene, resistant to basta which was non-selective herbicide, into balloon flower (Platycodon grandiflorum A. De. candolle). Seeds were germinated on MS medium, and 10-day-old immature cotyledon explants and 30-day-old leaf explants were cocultured with Agrobacterium tumefaciens strain MP 90 (pBinSyn) on 1/10 MS medium for 48 hours in the dark at $25^{\circ}C$. The cultures were transferred for selection of kanamycin-resistant shoots to the MS medium supplemented with 0.2 $mg/{\ell}$ NAA, 1.0 $mg/{\ell}$ BA, 3% sucrose, 100 $mg/{\ell}$ kanamycin, 500 $mg/{\ell}$ carbenicillin. Shoots were obtained from 10-day-old immature cotyledon explants after 4 weeks of culture. The shoots were subcultured twice every 4 weeks on the same medium for growth of transgenic shoots. Successful transformation was confirmed by histochemical GUS assay, PCR analysis, RT-PCR analysis, 10 $mg/{\ell}$ phosphinothricin treatment and 0.3% basta spray. The basta-resistant transgenic plants flowered normally.

Effects of glutamine and AgNO3 on plant regeneration of Sedum sarmentosum (돌나물의 식물체 분화에 미치는 Glutamine과 AgNO3의 영향)

  • Ahn, Jeong-Ho;Kim, Hyun-Soon;Lee, Seung-Yeob
    • Journal of Plant Biotechnology
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    • v.36 no.1
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    • pp.18-22
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    • 2009
  • This work was conducted to establish an efficient plant regeneration system for genetic transformation and the in vitro conservation of Sedum sarmentosum genetic resources. Effects of glutamine and $AgNO_3$ on plant regeneration between two genotypes were investigated using MS media supplemented with 0.2 mg/L NM and 3.0 mg/L BA. Calluses were formed on leaf explants placed on MS solid media supplemented with 3 mg/L 2,4-D and 1 mg/L BA. Calluses of Keumsan local strain produced shoots at a frequency of up to 100% after 50 days of culture on medium supplemented with glutamine. The highest number of shoots per callus was 17.6 at 350 mg/L glutamine. However, calluses of Wanju local strain gave rise to no shoots under the same culture conditions. Likewise, calluses of Keumsan local strain produced shoots at a frequency of up to 100% after 50 days of culture on medium supplemented with $AgNO_3$ whereas Wanju local strain sporadically produced shoots. The highest number of shoots per callus of Keumsan local stain was 16.1 at $15{\mu}M$ $AgNO_3$. Regenerated shoots were subcultured on hormone-free MS medium for rooting and shoot growth, and then 3-5 cm high plantlets were transplanted to the artificial soils comprising vermiculite and perlite, where they survived at a frequency of 88-100%. After being transplanted into upland soil:sand (1:1, v/v) in a greenhouse, regenerated plants showed a morphologically normal growth.

Optimization of apical tip culture condition for In Vitro propagation of 'Gisela 5' dwarf cherry rootstock (양앵두 왜성대목 'Gisela 5'의 기내번식을 위한 정단배양조건의 최적화)

  • Xu, Junping;Kang, In-Kyu;Kim, Chang Kil;Han, Jeung-Sul;Choi, Cheol
    • Journal of Plant Biotechnology
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    • v.42 no.1
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    • pp.49-54
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    • 2015
  • Based on the results in this study, here we propose a systematic micropropagation process for 'Gisela 5' that is one of the important dwarfing cherry rootstocks. When the apical tips detached from newly developed shoot in spring season were cultured on the half strength MS media with 0.5 mg/L IBA and 0.5 ~ 1.0 mg/L BA, the cultures scored the highest acquisition rate at 90% for normal shoot with vigorous growth and without hyperhydricity. As next step, the young shoots maintained in vitro well multiplied on the full strength MS medium supplemented with 0.5 mg/L IBA and 0.5 mg/L BA, in which multiplication rate was approximately nine-fold. Given the half strength MS medium containing 2.0 mg/L IBA, each transplanted shoot further developed robust roots. Finally, the plantlets were easily acclimatized in the compost consisted of vermiculite, perlite, and peatmoss in the proportion of 1:1:1. We expect that the results are useful for cherry cultivation and its rootstock production.