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Effect of Embryo Maturity and Medium on Callus Formation and Plant Regeneration from Immature Embryo of Eleutherococcus senticosus (가시오갈피 미숙배로부터 Callus 형성 및 식물체 재분화에 미치는 배의 성숙정도 및 배지의 효과)

  • 유창연
    • Korean Journal of Plant Resources
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    • v.10 no.2
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    • pp.122-127
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    • 1997
  • This study was conducted to establish mass propagation system from the tissue culture using immature embroys in Eleutherococus senticosus. Immature embroys from seeds were removed under the microscope and placed on modified SH and WPM medium containing several plant growth regulators. The calli were well formed on media containing 1mg/l of 2,4-D on modified SH medium and 1mg/l of 2,4-D and 3mg/l of TDZ on WPM medium. Shoot regeneration was better on modified SH or WPM medium with combination of high concentration of TDZ and low concentration of 2,4-D. Treatment of 2,4-D alone was better than treatment of TDZ alone in callus induction on modified SH medium but plant regeneration reversed. Treatment of 2.4-D and TDZ combination was better than treatment of 2,4-D alone in callus induction on WPM medium. The results of callus formation and shoot regeneration on WPM media differed to those of SH media. The rate of callus formation was nearly 83% when 2,4-D was added to SH medium on concentration of 1mg/l. The rate of callus formation was nearly 38% when combination treatment of 2,4-D 1mg/l and TDZ 3mg/l was added to WPM medium. Also, plant regeneration differed depending on the mature degree of immature embryo.

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Development of Cryopreservation Protocols through Droplet-vitrification and its Application to Vegetatively Propagated Crop Germplasm (영양체 유전자원의 작은방울-유리화법에 의한 초저온동결보존 실용화기술개발)

  • Kim, Haeng-Hoon;Yi, Jung-Yoon;No, Na-Young;Cho, Gyu-Taek;Yoon, Mun-Sup;Baek, Hyung-Jin;Kim, Chung-Kon
    • Proceedings of the Plant Resources Society of Korea Conference
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    • 2010.05a
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    • pp.12-12
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    • 2010
  • We developed droplet-vitrification protocol, a combination of droplet-freezing and solution-based vitrification, and applied to germplasm collections of garlic, potato, lily as well as cell lines, including hairy roots, somatic embryos. To establish a garlic cryobank, four Korean garlic field collections at Danyang, Suwon, Mokpo and Namhae were cryopreserved last five years. The protocol applied consisted of preculture for 3-4 days at $10^{\circ}C$ on solid MS medium with 0.3M sucrose, loading for 40 min in liquid medium with 35% PVS3, dehydration with PVS3 for 150 min, cooling in $5{\mu}l$ droplets of PVS3 placed on aluminum foil strips by dipping these strips in liquid nitrogen, warming them by plunging the foil strips into pre-heated($40^{\circ}C$) 0.8M sucrose solution for 30s. A total of over 900 accessions of garlic were stored in liquid nitrogen for long-term conservation using unripe inflorescences, cloves or bulbils. Twelve alternative plant vitrification solutions were designed by modifying cryoprotectant concentrations from the original PVS2 and PVS3. The results suggest that PVS2-based vitrification solutions with increased glycerol and sucrose and/or decreased DMSO and EG concentrations can be applied for medium size explants which are tolerant to chemical toxicity and moderately sensitive to osmotic stress. PVS3 and variants can be used widely when samples are heterogeneous, of large size and/or very sensitive to chemical toxicity and tolerant to osmotic stress.

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Production and Process Monitoring of 5-Aminolevulinic Acid (ALA) by Recombinant E. coli I. Characteristics of ALA Production (유전자 재조합 대장균에 의한 5-Aminolevulinic Acid (ALA)의 생산 및 공정 모니터링 I. ALA의 생산 특성)

  • 이종일;정상윤;서국화;한경아;조성효;백경환
    • KSBB Journal
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    • v.19 no.1
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    • pp.17-26
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    • 2004
  • In this study the extracellular production of 5-aminolevulinic aicd (ALA) by recombinant E. coli BL2l (DE3) pLysS harboring the plasmid pFLS45 are investigated. Optimum concentrations of succinic acid and glycine for cell growth and ALA production were found to be 30 mM and 15 mM, respectively. Levulinic acid (LA) as an inhibitor of ALAD was added to the culture medium in the end of exponential cell growth phase and its optimum concentration was 30 mM. Growth of recombinant E. coli BL2l (DE3) pLysS (pFLS45) was largely dependent upon the pH value of culture medium. When the pH of culture medium was in the range of 6.0 and 6.5, high cell mass and ALA production were obtained. IPTG induction for the expression of the fusion gene did not enhance the production of ALA. Recombinant cell grew at 30't faster than at 37$^{\circ}C$, but ALA productivity was lower than at 37$^{\circ}C$. Repeated addition of glycine, succinic acid, and LA increased the production of ALA and the inhibition of intracellular ALA dehydratase activity, with up to 1.3 g/L ALA having been produced in the cultivation.

Aberration Retrieval Algorithm of Optical Pickups Using the Extended Nijboer-Zernike Approach (확장된 네이보어-제르니케 방법에 의한 광픽업의 파면수차 복원 알고리즘)

  • Jun, Jae-Chul;Chung, Ki-Soo;Lee, Gun-Kee
    • Journal of the Institute of Convergence Signal Processing
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    • v.11 no.1
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    • pp.32-40
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    • 2010
  • In this work, the method of acquiring the pupil function of optical system is proposed. The wavefront aberration and the intensity distribution of pupil can be analysed with the pupil function. This system can be adopted to the manufacturing line of optical pickup directly and also has good performance to analysing various property of optical instrument. It is one kind of inverse problem to get pupil functions by 3D beam data. The extended Nijboer-Zernike(ENZ) approach recently proposed by Netherlands research group is adopted to accompany to solve these inverse problem. The ENZ approach is one of a aberration retrieval method for which numerous approaches are available. But this approach is new in the sense that it use the highly efficient representation of pupil functions by means of their Zernike coefficients. These coefficients are estimated by using matching procedure in the focal region the theoretical 3D intensity distribution and measured 3D intensity distribution. The algorithm that can be applied more general circumstance such as high-numerical aperture instrument is developed by modifying original ENZ approach. By these scheme, MS windows based GUI program is developed and the good performance is verified with generated 3D beam data.

A comparative analysis of volatile organic compound levels in field samples between different gas chromatographic approaches (분석기법의 차이에 따른 현장시료의 VOC 분석결과 비교연구: 분석오차의 발생 양상과 원인)

  • Ahn, Ji-Won;Pandey, Sudhir Kumar;Kim, Ki-Hyun
    • Analytical Science and Technology
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    • v.23 no.5
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    • pp.465-476
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    • 2010
  • In this study, a number of volatile organic compounds (VOCs) including benzene, toluene, p-xylene, styrene, and methyl ethyl ketone were analyzed from samples collected in ambient air and under the field conditions. These samples were analyzed independently by two different set-ups for VOC analyses, i.e., between [1] gas chromatography/flame ionization detector with tube sampling - (F-T system) and [2] gas chromatography/mass spectrometer with bag sampling (M-B system). The analytical results derived by both systems showed fairly similar patterns in relative sense but with moderately large differences in absolute sense. The results of M-B system were high relative to F-T system with the F-T/M-B ratio below 1. If the relative biases of the two measurement techniques are derived in terms of percent difference (PD) in concentration values, the results were generally above 35% on average. A student t-test was applied to investigate the statistical significance of those differences between the systems. The results of both analytical systems were different at 95% confidence level for toluene, p-xylene, styrene, and methyl ethyl ketone (P < 0.043). However, F-T and M-B systems showed strong correlations for toluene and p-xylene. The observed bias is explained in large part by such factors as the differences in standard phases used for each system and the chemical loss inside the bag sampler.

Trace level analysis of Pb in plasma by inductively coupled plasma/mass spectrometry (유도결합플라즈마 질량분석법을 이용한 혈장 중 극미량 납 분석)

  • Lee, Sung-Bae;Yang, Jeong-Sun;Choi, Sung-Bong;Shin, Ho-Sang
    • Analytical Science and Technology
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    • v.25 no.3
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    • pp.190-196
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    • 2012
  • The human exposure of lead has usually detected the amount of lead in the whole blood, however, this method has a shortcoming to give the information on the short-term exposure to lead. In that sense, it is desirable to estimates the level of lead in plasma to draw the chronic bio-marker of lead exposure even though it is difficult to measure lead of several ng/L. An inductively coupled plasma-mass spectrometry (ICP-MS) method was developed for determining lead in plasma as the chronic bio-marker of lead of workers. To minimize the contamination of lead from the environment, we constructed class 1,000 clean room and compared the amount of floating dust before and after the operation of the clean room. The limit of detection (LOD) and the limit of quantification (LOQ) of lead in fetal bovine serum were 4.3 ng/L and 12.2 ng/L by NIOSH method (statistical calculation method) and 7.0 ng/L and 22.1 ng/L by signal/noise ratio, respectively. The accuracy was in a range of 92.3-101.3%, and the precision of the assay was less than 4% in the samples spiked in the concentration of 20 ng/L and 2,000 ng/L. The method was simple, reproducible and sensitive enough to permit reliable analysis of lead to the ng/L level in plasma and/or serum. The method was also useful for the biological monitoring of chronic exposure to lead.

Interaction of Forskolin with the Effect of $N^6-cyclopentyladenosine$ on Norepinephrine Release in Rat Hippocampus (흰쥐 해마에서 Norepinephrine 유리에 미치는 $N^6-cyclopentyladenosine$ 및 Forskolin의 영향)

  • Choi Bong-Kyu;Kim Do-Kyung;Son Yong;Yang Ue-Jong
    • The Korean Journal of Physiology and Pharmacology
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    • v.1 no.3
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    • pp.225-231
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    • 1997
  • As it has been reported that the depolarization-induced norepinephrine (NE) release is modulated by activation of presynaptic $A_1-adenosine$ heteroreceptor and various lines of evidence indicate the involvement of adenylate cyclase system in $A_1-adenosine$ post-receptor mechanism in hippocampus, it was attempted to delineate the role of adenylate cyclase system in the $A_1-receptor-mediated$ control of NE release in this study. Slices from rat hippocampus were equilibrated with $[^3H]-NE$ and the release of the labelled products was evoked by electrical stimulation.(3 Hz, $5Vcm^{-1}$, 2 ms, rectangular pulses). The influence of various agents on the evoked tritium-outflow was investigated. $N^6-Cyclopentyladenosine$ (CPA), a specific $A_1-adenosine$ receptor agonist, in concentrations Tanging from 0.1 to $10{\mu}M$ decreased the $[^3H]-NE$ release in a dose-dependent mauler without any change of basal rate of release. 8-Cyclopentyl-1,3-dipropylxanthine (DPCPX, $2{\mu}M$), a selective $A_1-receptor$ antagonist, inhibited the CPA effect. The responses to N-ethylmaleimide $(3&10{\mu}M)$, a SH-alkylating agent of G-protein, were characterized by increments of the evoked NE-release and the CPA effects were completely abolished by NEM pretreatment. Forskolin, a specific adenylate cyclase activator, in concentrations ranging from 0.1 to $30{\mu}M$ increased the evoked and basal rate of NE release in a dose-dependent manner and the CPA effects were inhibited by forskolin pretreatment. Rolipram $(1&10{\mu}M)$, a phosphodiesterase inhibitor, did not affect the evoked NE release but reduced the CPA effect. And 8-bromo-cAMP $(100&300{\mu}M)$, a membrane permeable cAMP analogue inhibited the CPA effect significantly. These results suggest that the $A_1-adenosine$ heteroreceptor plays an important role in NE-release via nucleotide-binding protein $G_i$ in the rat hippocampus and that the adenylate cyclase system might be participated in this process.

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In vitro Culture of Adventitious Roots from Dioscorea nipponica Makino for the Production of Steroidal Saponins (부채마 (Dioscorea nipponica Makino)의 부정근 배양조건과 Steroidal Saponin의 기내생산)

  • An Jung-Hee;Son Kun-Ho;Sohn Ho-Yong;Kwon Soon-Tae
    • Journal of Plant Biotechnology
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    • v.32 no.3
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    • pp.217-223
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    • 2005
  • Effects of growth regulators on growth of adventitious roots and accumulation of steroidal saponins, such as dioscin, prosapogenin A and prosapogenin C, in cultures of Dioscorea nipponica were determined. The maximum growth of adventitious roots was observed in MS medium supplemented with 30 g/L sucrose and 1.0 mg/L NAA. Addition of BA in combination with NAA appeared to be no effective in the growth of adventitious roots. Among the twenty different adventitious roots formed from different seeds, strain No. 10 was selected based on production ability of dioscin, and its stability through the successive liquid culture. During the first 4 weeks of incubation, contents of steroidal saponins in adventitious roots were negligible but the contents were markedly increased at 5 weeks of incubation. Dioscin and prosapogenin C content in IBA-treated adventitious roots were significantly higher than those in NAA-treated roots. However, content of prosapogenin A was not significantly different among NAA or IBA level. Results provide that liquid culture of adventitious roots of D. nipponica have a potential for mass production of dioscin including prosapogenin A and prosapogenin C.

Use of Paromomycin as a Selectable Marker for the Transformation of Chinese Cabbage (배추의 형질전환용 선발항생제로서 Paromomycin의 이용)

  • Cho, Mi-Ae;Min, Sung-Ran;Ko, Suck-Min;Liu, Jang-Ryol;Lee, Jun-Haeng;Choi, Pil-Son
    • Journal of Plant Biotechnology
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    • v.33 no.4
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    • pp.271-276
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    • 2006
  • Hypocotyl explants of Chinese cabbage (us. 'Jeong Sang' and 'Seoul') produced adventitious shoots on Murashige and Skoog (MS) basal medium supplemented with 4mg/L $AgNO_3$, 5 mg/L acetosyringone, 4 mg/L 6-benzyladenine and 3mg/L alpha-naphthaleneacetic acid (SI) after cocoultivation with strains of Agrobacterium tumefaciens (LBA4404) harboring the pCAMBIA1301 and the $_PPTN290$ containing hygromycin-resistance gene and paromomycin-resistance gene as a selectable marker genes, respectively. There was a significant difference in the frequency of transgenic plants depending on antibiotics and cultivars used. Paromomycin was better than hygromycin, and cultivar 'Jeong-sang' was higher than 'c.v. Seoul' in the frequency of transgenic plants. In particular, the highest frequency (0.70%) of transgenic plants was obtained from selection medium (SI) containing 100mg/L paromomycin in c.v., 'Jeong-sang' GUS positive response were obtained 9 plants and 3 plants from the cultivars, 'Jeong-sang' and 'Seoul', respectively. They were grown to maturity in a greenhouse and normally produced $T_1$ seeds. GUS histochemical assay for progeny $(T_1)$ revealed that the transgenes were expressed in the plant genome.

A Study on the External Fault Performance Evaluation of Grid-Connected Power Conditioning System for Residential Fuel Cell System (가정용 연료전지시스템 계통연계형 전력변환장치의 외부사고 성능평가에 관한 연구)

  • Lee, Jung-Woon;Seo, Won-Seok;Kim, Young-Gyu
    • 한국신재생에너지학회:학술대회논문집
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    • 2009.11a
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    • pp.131-131
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    • 2009
  • 최근 환경규제가 강화되면서 친환경적인 전력생산 요구 등의 외부환경 변화에 따른 우리나라의 전력구조가 기존의 중압 집중형 발전을 탈피한 분산전원 발전에 대한 관심이 증대되고 있는 실정이다. 특히, 분산전원으로서의 전력생산은 공급의 안전성, 경쟁력 및 에너지의 지속성 등이 요구되어지는데, 재생에너지가 가지고 있는 에너지 지속성의 한계 및 설치의 제약성을 탈피할 수 있는 시스템으로 현재 연료전지시스템이 가장 근접해 있는 실정이다. 즉, 도시가스 인프라가 우수하고 인구조밀 지역이 많은 우리나라의 특성상 각 가정 및 건물에 쉽게 설치하고 공급의 안전성을 갖는 건물용 연료전지는 최근 가장 각광받고 있는 분산전원 시스템 중의 하나이다. 올해부터 모니터링사업의 일환으로 수용가에 설치 될 연료전지 시스템이 얼마나 안정적으로 전기와 열을 각 가정에 공급하고 시스템의 안전성을 확보하는 가는 건물용 연료전지의 분산전원으로서의 가능성 및 국민의 수용성을 증대시키는 중요한 역할을 할 것이다. 연료전지시스템은 상용전력과 연계되어 있기 때문에 시스템의 안정성 뿐만 아니라 상용전력의 변화에 대응하여 안정적인 운전을 하는지에 대한 평가가 필수적이다. 이에 따라 본 연구에서는 가정용 연료전지시스템의 성능 및 안전성평가의 일환으로 계통연계형 전력변환장치의 성능 및 안전성을 평가 하고자 한다. 연료전지 검사를 위한 계통연계형 전력변환장치의 시험평가 항목으로는 크게 정상특성성능시험, 보호기능성능시험, 과도응답특성성능시험 및 외부사고성능시험 등으로 나뉘어진다. 본 연구에서는 외부사고 성능시험 항목들인 출력측 단락시험, 계통전압 순간정전?순간강하시험 및 부하차단 시험 등을 통하여 외부사고에 대한 성능 및 안전성을 평가하였다. 외부사고 성능시험의 주 목적은 시스템의 이상 운전이 아니라 외부의 영향에 따른 시스템의 안전성 및 전력품질을 평가한다. 출력측 단락시험을 수행하기 위해서 전력변환장치를 정격 출력 전압, 정격 출력 주파수 및 정격 출력에서 운전한 후, 교류 전원장치는 단락 전류를 검출하여, 사고 발생 후 0.3초 이내에 개방하도록 설정하였다. 여기서, 단락 저항 Rsc를 정격 전류의 10배 이상에 해당하는 부하와 같은 값으로 설정하였다. 스위치 SWSC를 폐로하여 단락 상태를 만들며, 이 때 전력변환장치의 출력전류와 차단 또는 정지 시간을 측정하였다. 실험 결과에 대한 판정기준은 단락전류를 검출하여 0.5초 이내에 개폐기 개방 또는 게이트 블록 기능이 동작하여 시스템을 안정하게 정지시키고 시스템 어떤 부위에도 손상이 없어야 한다. 실험 결과 파워컨디셔너의 출력전류 및 차단 또는 정지된시간이 40ms로 나타났고, 출력전류의 파형도 매우 안정함을 볼 수 있었다. 이와 같이 모든 실험을 수행한 결과 외부사고에 대하여 시스템이 안전하게 정지하는 등 연료전지 시스템의 안전성을 확인하였다.

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