• Title/Summary/Keyword: K-L transformation

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Plantlet Regeneration via Somatic Embryogenesis from Hypocotyls of Common Buckwheat (Fagopyrum esculentum Moench.)

  • Kwon, Soo-Jeong;Han, Myong-Hae;Huh, Yoon-Sun;Roy, Swapan Kumar;Lee, Chul-Won;Woo, Sun-Hee
    • KOREAN JOURNAL OF CROP SCIENCE
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    • v.58 no.4
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    • pp.331-335
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    • 2013
  • Buckwheat sprout is used as vegetable, and also flour for making noodles, and so on. Currently, information about tissue culture in buckwheat is limited and restricted to micro-propagation. We carried out somatic embryogenesis and plant regeneration using hypocotyl segments as explant of the cultivated buckwheat species, Fagopyrum esculentum which differs from existing studies in the growth regulator combinations used. Maximum callus regeneration was induced on MS medium containing 2,4-dichlorophenoxyacetic acid (2,4-D) $2.0mg{\cdot}L^{-1}$, benzyladenine (BA) $1.0mg{\cdot}L^{-1}$ and 3% sucrose. Friable callus was transferred to solidified MS media containing BA ($1.0mg{\cdot}L^{-1}$) with various concentrations of 2,4-dichlorophenoxyacetic acid for the induction of embryogenesis. The optimum concentrations of growth regulators (for regeneration of plantlet) were indole-3-acetic acid ($2.0mg{\cdot}L^{-1}$), Kinetin ($1.0mg{\cdot}L^{-1}$), BA ($1.0mg{\cdot}L^{-1}$). Only 2,4-D did not show any significant effect on callus induction or embryogenesis. Regeneration of embryonic callus varied from 5% to 20%. Whole plants were obtained at high frequencies when the embryogenic calli with somatic embryos and organized shoot primordia were transferred to MS media with 3% sucrose. The main objective of this research was to develop an efficient protocol for plant regeneration for common buckwheat, and to apply in future for genetic transformation.

Enhancement of artemisinin content by constitutive expression of the HMG-CoA reductase gene in high-yielding strain of Artemisia annua L.

  • Nafis, Tazyeen;Akmal, Mohd.;Ram, Mauji;Alam, Pravej;Ahlawat, Seema;Mohd, Anis;Abdin, Malik Zainul
    • Plant Biotechnology Reports
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    • v.5 no.1
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    • pp.53-60
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    • 2011
  • Artemisinin is effective against both chloroquine-resistant and -sensitive strains of Plasmodium species. However, the low yield of artemisinin from cultivated and wild plants is a serious limitation to the commercialization of this drug. Optimization of artemisinin yield either in vivo or in vitro is therefore highly desirable. To this end, we have overexpressed the 3-hydroxy-3-methyl-glutaryl-coenzyme A reductase (HMGR) gene (hmgr) from Catharanthus roseus L. in Artemisia annua L. and analyzed its influence on artemisinin content. PCR and Southern blot analyses revealed that the transgenic plants showed stable integration of the foreign hmgr gene. The reverse transcriptase-PCR results suggested that the hmgr was expressed at the transcriptional level in transgenic lines of Artemisia annua L., while the high-performance liquid chromatography analysis showed that artemisinin content was significantly increased in a number of the transgenic lines. Artemisinin content in one of the A. annua transgenic lines was 38.9% higher than that in non-transgenic plants, and HMGR enzyme activity in transgenic A. annua L. was also higher than that in the non-transgenic lines.

Numerical Simulation of Irregular Wave Transformation due to Wave-induced Current over a Submerged Elliptic Shoal (수중타원형 천퇴상 불규칙파의 파랑쇄파류에 의한 변형 수치모의)

  • Choi, Jun-Woo;Baek, Un-Il;Yoon, Sung-Bum
    • Journal of Korean Society of Coastal and Ocean Engineers
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    • v.19 no.6
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    • pp.565-573
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    • 2007
  • The effect of wave and current interactions on irregular wave transformation over a submerged elliptic shoal is investigated based on numerical simulations of the Vincent and Briggs experiment [Vincent, C.L., Briggs, M.J., 1989. Refraction-diffraction of irregular waves over a mound. Journal of Waterway, Port, Coastal and Ocean Engineering, 115(2), pp. 269-284]. The numerical simulations are conducted by a combination of REF/DIF S(a wave model) and SHORECIRC(a current model) and a time dependent phase-resolving wavecurrent model, FUNWAVE. In the simulations, the breaking-induced currents defocus waves behind the shoal and bring on a wave shadow zone that shows relatively low wave height distributions. The computed results of the combined model system agree better with the measurements than the computed results obtained by neglecting wave-current interaction do. In addition, the results of FUNWAVE show a good agreement with the measurements. The agreement indicates that it is necessary to take into account the effect of breaking-induced current on wave refraction when wave-breaking occurs over a submerged shoal.

Transformation of Populus Species by an Agrobacterium Binary Vector System (Agrobacterium Binary Vector에 의한 포플러 형질전환(形質轉換)을 위한 기초연구(基礎研究))

  • Chun, Young Woo;Klopfenstein, Ned B.;McNabb, Harold S. Jr.;Hall, Richard B.
    • Journal of Korean Society of Forest Science
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    • v.77 no.2
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    • pp.199-207
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    • 1988
  • Three clones of Populus alba ${\times}$ P. grandidentata have been tested for susceptibility to Agrobacterium tumefaciens strains A281 and A348. We determined the optimum concentration of kanamycin sulfate for effective selection of leaf disc-derived, transgenic tissues transformed using Agrobacterium binary vector pGA472 containing a neomycin phosphotransferase gene (NPT-II) which confers kanamycin resistance. Of the wild type Ti plasmids contained by the two Agrobacterium strains, pTiBo542 of strain A281 appears to be best suited to serve as a helper plasmid for binary vector systems. A relatively low concentration (10mg/l) of kanamycin sulfate inhibited adventitious shoot initiation from leaf discs on regeneration medium. Transformed kanamycin-resistant calli were obtained by culturing Agrobacterium inoculated leaf discs on selective regeneration medium. The transformed kanamycin-resistant calli continued to grow on regeneration media supplemented with kanamycin sulfate to levels of 50 and 200mg/l. The growth of non-co-cultivated control calli was severely inhibited on regeneration medium containing 50mg/l kanamycin sulfate.

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Transformation of Endocrine Disrupting Chemicals (EDCs) by Manganese(IV) Oxide (망간산화물을 이용한 내분비계장애물질의 변환에 관한 연구)

  • Lee, Seung-Hwan;Choi, Yong-Ju;Chung, Jae-Shik;Nam, Taek-Woo;Kim, Young-Jin;Nam, Kyoung-Phile
    • Journal of Soil and Groundwater Environment
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    • v.14 no.1
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    • pp.44-50
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    • 2009
  • The occurrence of endocrine disrupting compounds (EDCs), chemicals that interfere with human hormone system, are increasing in the freshwater, waste water and subsurface as well. In this study, we determined the reactivity of three EDCs in the presence of birnessite. In aqueous phase, bisphenol A, 2,4-dichlorophenol and 17${\beta}$-estradiol, which possesses phenoxy-OH, were very rapidly transformed by birnessite: up to 99% of initial concentrations (50 mg/L for bisphenol A, 100mg/L for 2,4-dichlorophenol, and 1.5mg/L for 17${\beta}$-estradiol) were destroyed within 60 minutes. Especially, bisphenol A was the most reactive chemical, disappearing by 99% in a few minutes. The reaction occurred on the surface of birnessite, showing a linear increase of first-order kinetic constants with the increase of the surface area of birnessite. In soil slurry phase, the reactivity of birnessiteto EDCs was faster than in aqueous phase probably due to the cross coupling reaction of phenoxy radicals with soil organic matter. Considering the rapid transformation of the EDCs in the both phases, this oxidative cross coupling reaction mediated by birnessite would be an effective solution for the remediation of EDCs in environmental media, especially in soil.

In vitro Propagation of Arbuscular Mycorrhizal Fungi using Ri t-DNA Transformed Carrot Roots (Ri t-DNA로 형질전환된 당근 뿌리를 이용한 Arbuscular 균근균의 기내증식)

  • Cho, Ja Yong;Sohn, Bo-Kyoon;Lee, Hyo-Yeon;Chung, Soon-Ju
    • Horticultural Science & Technology
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    • v.18 no.6
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    • pp.802-807
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    • 2000
  • This study was conducted to propagate the arbuscular mycorrhizal fungi in vitro using the hairy root of carrot transformed by Agrobacterium rhizogenes with Ri t-DNA. Mycorrhizal spores and roots in sudangrass plants were wet-sieved, surface-sterilized and inoculated onto the hairy root of carrot on the Modified Strullu & Romand (MSR) medium. The mycorrhizal spores of Glomus sp. propagated in vitro for 12 weeks was about $50{\mu}m$, and the shapes of spores were round or elliptic. Spores were formed mainly at the middle of the hyphae. Number of mycorrhizal spores propagated using dual culture of the transformed carrot roots and the mycorrhizal inoculum for 12 weeks were about 1,200 per plates.

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Effect of Surface Sterilization Method on Agrobacterium-mediated Transformation of Field-grown Zoysiagrass Stolon (포장생육 잔디 포복경을 이용한 잔디 형질전환에 있어서 살균방법의 영향)

  • Ahn, Na-Young;Alam, Iftekhar;Kim, Yong-Goo;Bae, Eun-Ji;Lee, Kwang-Soo;Lee, Byung-Hyun
    • Journal of The Korean Society of Grassland and Forage Science
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    • v.33 no.2
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    • pp.100-104
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    • 2013
  • Zoysiagrass (Zoysia japonica Steud.) is an important forage and turfgrass that spreads by stolons and rhizomes. Zoysiagrass stolon can be used directly for Agrobacterium-mediated genetic transformation by exploiting the potential of direct shoot formation. However, surface sterilization of field-grown stolons is difficult and remains to be explored. We developed an effective surface sterilization and culture method using the stolon explant for infection with Agrobacterium tumefaciens. Among various treatments, sequential disinfection in 30% bleach for 15 min followed by 0.1% mercuric chloride for 25 min resulted in the highest number of clean stolons. The efficacy of mercuric chloride was increased under vacuum conditions by incubating at 800 mbar for 5 min. The inclusion of 2.5 mg/l amphotericin B further prevents fungal growth in in vitro cultures. This protocol would speed up the development of transgenic plants by utilizing field-grown stolon nodes.

Oxidative Transformation of 1-Naphthol Using Manganese Oxide (망간산화물을 이용한 1-Naphthol의 산화 제거 연구)

  • Lim, Dong-Min;Kang, Ki-Hoon;Shin, Hyun-Sang
    • Journal of Korean Society of Environmental Engineers
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    • v.28 no.5
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    • pp.535-542
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    • 2006
  • In this study, removals of 1-naphthol by oxidative-coupling reaction using birnessite, one of natural Mn oxides present in soil, was investigated in various experimental conditions(reaction time, Mn oxide loadings, pH, etc). Removal efficiency of 1-naphthol by birnessite was high in all the experimental conditions, and UV-vis. and mass spectrometric analyses on the supernatant after reaction confirmed that the reaction products were oligomers formed by oxidative-coupling reaction. Pseudo-first order rate constants, f, for the oxidative transformation of 1-naphthol by birnessite was derived from the kinetic experiments under various amount of birnessite loadings, and using the observed pseudo-first order rate constants with respect to birnessite loadings, surface area-normalized specific rate constant, $k_{surf}$ was also determined to be $9.31{\times}10^{-4}(L/m^2{\cdot}min)$ for 1-naphthol. In addition, the oxidative transformation of 1-naphthol was found to be dependent on solution pH, and the pseudo-first order rate constants were increased from 0.129 at pH 10 to 0.187 at pH 4.

Algorithm for the L1-Regression Estimation with High Breakdown Point (L1-회귀추정량의 붕괴점 향상을 위한 알고리즘)

  • Kim, Bu-Yong
    • Communications for Statistical Applications and Methods
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    • v.17 no.4
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    • pp.541-550
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    • 2010
  • The $L_1$-regression estimator is susceptible to the leverage points, even though it is highly robust to the vertical outliers. This article is concerned with the improvement of robustness of the $L_1$-estimator. To improve its robustness, in terms of the breakdown point, we attempt to dampen the influence of the leverage points by means of reducing the weights corresponding to the leverage points. In addition the algorithm employs the linear scaling transformation technique, for higher computational efficiency with the large data sets, to solve the linear programming problem of $L_1$-estimation. Monte Carlo simulation results indicate that the proposed algorithm yields $L_1$-estimates which are robust to the leverage points as well as the vertical outliers.

Comparison of the Cell Surface Barrier and Enzymatic Modification System in Brevibacterium flavum and B. Lactofermentum

  • Jang Ki-Hyo;Britz Margaret L.
    • Biotechnology and Bioprocess Engineering:BBE
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    • v.10 no.3
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    • pp.225-229
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    • 2005
  • To investigate impediments to plasmid transformation in Brevibacterium flavum BF4 and B. lactofermentum BL1, cell surface barriers were determined by measuring growth inhibition whilst enzymatic barriers were determined by comparing DNA methylation properties. B. lactofermentum was more sensitive to growth inhibition by glycine than B. flavum. Release of cellular proteins during sonication was more rapid for B. lactofermentum than for B. flavum. Plasmid DNA (pCSL 17) isolated from B. flavum transformed recipient $McrBC^+$ strains of Escherichia coli with lower efficiency than $McrBC^-$. McrBC digestion of this DNA confirmed that B. flavum contain methylated cytidines in the target sequence of McrBc sequences but B. lactofermentum contained a different methylation pattern. DNA derived from the B. lactofermentum transformed recipient $EcoKR^+$ strains of E. coli with lower efficiency than $EcoKR^-$, indicating the presence of methylated adenosines in the target sequence of EcoK sequences. The present data describe the differences in the physical and enzymatic barriers between two species of corynebacteria and also provide some insight into the successful foreign gene expression in corynebacteria.