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Rapid Detection of Rifampicin Resistant M. tuberculosis by PCR-SSCP of rpoB Gene in Clinical Specimens (RpoB 유전자 PCR-SSCP법에 의한 임상검체내 Rifampicin 내성 결핵균의 신속진단)

  • Shim, Tae-Sun;Kim, Young-Whan;Lim, Chae-Man;Lee, Sang-Do;Koh, Youn-Suck;Kim, Woo-Sung;Kim, Dong-Soon;Kim, Won-Dong
    • Tuberculosis and Respiratory Diseases
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    • v.44 no.6
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    • pp.1245-1255
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    • 1997
  • Background : Rifampicin(RFP) is a key component of the antituberculous short-course chemotherapy and the RFP resistance is a marker of multi-drug resistant(MDR) tuberculosis. RPoB gene encodes the $\beta$-subunit of RNA polymerase of M. tuberculosis which is the target of RFP. And rpoB gene mutations are the cause of RFP resistance of M. tuberculosis. Although several reports showed that PCR-SSCP would be a rapid diagnostic method for identifying the RFP resistance, there were few reports Performed using direct, clinical specimens. So we Performed PCR-SSCP analysis of rpoB gene of M. tuberculosis in direct, clinical specimens. Methods : 75 clinical specimens were collected from patients at Asan Medical Center from June to August 1996. After PCR of IS 6110 fragments, 43 both AFB smear-positive and IS6110 fragment PCR-positive specimens were evaluated. The RFP susceptibility test was referred to the referral laboratory of the Korean Tuberculosis Institute. DNA was extracted by bead beater method. And heminested PCR was done using 0.1ul(1uCi) [$\alpha-^{32}P$]-dCTP. SSCP analysis was done using non-denaturating MDE gel electrophoresis. Results : The results of PCR of IS6110 fragments of M. tuberculosis were positive in 55(73%) cases of 75 AFB smear-positive clinical specimens. Of the 55 specimens, RFP susceptibility was confirmed in only 43 specimens. Of the 43 AFB smear-positive and IS6110 fragment-positive specimens, 29 were RFP susceptible and 14 were RFP resistant. All the RFP susceptible 29 strains showed the same mobility compared with that of RFP sensitive H37Rv in SSCP analysis of ropB gene. And all the other RFP resistant 13 strains showed the different mobility. In other words they showed 100% identical results between PCR-SSCP analysis and traditional susceptibility test. Conclusion : The PCR-sseP analysis of rpoB gene in direct clinical specimens could be used as a rapid diagnostic method for detecting RFP resistant M. tuberculosis.

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Growth of CdSe thin films using Hot Wall Epitaxy method and their photoelectrical characteristics (HWE방법에 의한 CdSe 박막 성장과 광전기적 특성)

  • Hong, K.J.;Lee, K.K.;Lee, S.Y.;You, S.H.;Shin, Y.J.;Suh, S.S.;Jeong, J.W.;Jeong, K.A.;Shin, Y.J.;Jeong, T.S.;Kim, T.S.;Moon, J.D.;Kim, H.S.
    • Journal of Sensor Science and Technology
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    • v.6 no.4
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    • pp.328-336
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    • 1997
  • The CdSe thin films were grown on the Si(100) wafers by a hot wall epitaxy method (HWE). The source and substrate temperature are $600^{\circ}C$ and $430^{\circ}C$ respectively. The crystalline structure of epilayers was investigated by double crystal X-ray diffraction(DCXD). Hall effect on the sample was measured by the van der Pauw method and studied on the carrier density and mobility dependence on temperature. From Hall data, the mobility was increased in the temperature range 30K to 150K by impurity scattering and decreased in the temperature range 150k to 293k by the lattice scattering. In order to explore the applicability as a photoconductive cell, we measured the sensitivity(${\gamma}$), the ratio of photocurrent to darkcurrent(pc/dc), maximum allowable power dissipation(MAPD), spectral response and response time. The results indicated that the photoconductive characteristic were the best for the samples annealed in Cu vapor compare with in Cd, Se, air and vacuum vapour. Then we obtained the sensitivity of 0.99, the value of pc/dc of $1.39{\times}10^{7}$, the MAPD of 335mW, and the rise and decay time of 10ms and 9.5ms, respectively.

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Effect of Different Cultivating Method on Growth, Yield in Rice Variety (재배방법의 차기가 수도 품종의 생육.수량에 미치는 영향)

  • Lee, J.S.;Lo, T.H.;Moon, C.S.;Suh, H.Y.;Park, K.D.;Jo, J.S.
    • KOREAN JOURNAL OF CROP SCIENCE
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    • v.14
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    • pp.159-164
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    • 1973
  • This study was carried out in the central part of Korea for three years from 1970 to 1972. Its aim was to investigate the variations of growth and yield of rice varieties of different cultivating methods (transplanting culture, direct sowing culture on irrigated paddy field, and direct sowing culture on non-irrigated paddy field. The results could be summarized as follows; 1. The number of days from sowing to heading by direct sowing culture on irrigated paddy field was remarkably shorter than transplanting culture. Direct sowing culture on non-irrigated paddy field was longer than transplanting culture. 2. The number of panicles showed direct sowing culture on irrigated paddy field was first, direct sowing culture on non-irrigated paddy field was second, and third transplanting culture. But the number of spikelets per panicle was first on the transplanting culture, second on the direct sowing culture on non-irrigated paddy field and third on the direct sowing culture on irrigated paddy field. 3. The weight of the brown rice showed that direct sowing culture on irrigated paddy field produced the highest yield among the cultivating method in all varieties. The transplanting culture increased more than the direct sowing culture on non-irrigated paddy field. The highly productive varieties were Sadominory, Akibale, Suwon 213-1 when the direct sowing culture on irrigated paddy field was used. 4. The correlation between the brown rice yield and the yield components were as follows; in each cultivating method, it showed highly significant positive correlation between the brown rice yield and the 1, 000 grain weight of hulled rice, and between the brown rice yield and the number of the spiklets per panicle in the transplanting culture, in the direct sowing culture On irrigated paddy field between the brown rice yield and the number of spikelets per panicle and between the brown rice yield and the number of panicles, in direct sowing culture on non-irrigated paddy field between the brown rice yield and number of panicles. 5. It showed highly significant negative correlation between the brown rice yield and the number of days from sowing to heading in the direct sowing culture on irrigated and non-irrigated paddy field, but it was showed very low negative correlation in the transplant culture.

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Primary Production and Nitrogen Regeneration by Macrozooplankton in the Kyunggi Bay, Yellow Sea (서해 경기만의 기초생산력 및 질소계 영양염의 재생산에 관한 연구)

  • Chung, Kyung Ho;Park, Yong Chul
    • 한국해양학회지
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    • v.23 no.4
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    • pp.194-206
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    • 1989
  • Seasonal variations of nutrients (ammonium, nitrite, nitrate, phosphate and silicate), primary productivity and ammonium regeneration rate of macrozooplankton were investigate to understand the relationship between nitrogen recycling and nitrogen requirement by phytoplankton from Feburuary 1986 to November 1987 in the Kyunggi Bay, shallow estuarine water of Yellow Sea. In general, nutrients increased during the winter and depleted during the spring and the early summer except temporally sharp increase after flood in September. Ammonium was prevalently generally found in high concentration throughout the study area and it occasionally raised N/P ratio in the range of 30 to 70 as in the freshwater environment. Daily net primary productivity ranged from 30.3 to 3580.0 mgC/$m^2$/d with an average of 883.9 mgC/$m^2$/d. Annual primary productivity was determined to be 320.0 gC/$m^2$/yr. Carbon assimilation number ranged from 2.9 to 19.4 mgC/mg chl-a/h which increased in the summer and decreased in the winter. Nitrogen requirement by phytoplankton ranged from 0.4 to 45.0 mg at-N/$m^2$/d and turnover time of inorganic nitrogen ranged from 2.4 in the late summer to 122.7 days in the winter. Nitrogen regeneration rate of mixed macrozooplankton determined by bottle incubation method ranged from 0.02 to 1.34 mg at-N $m^2$/d and it could contribute from 2.8 to 38.7% with an annual average of 14.9% of total nitrogen requirement by phytoplankton in this shallow estuarine environment.

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Ecosysteme de I′Etang de Berre (Mediterranee nord-occidentale) : Caracteres Generales Physiques, Chimiques et Biologiques

  • Kim, Ki-Tai
    • Korean Journal of Environmental Biology
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    • v.22 no.2
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    • pp.247-258
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    • 2004
  • Climatological, hydrological and planktonical research studies, measurements of primary production and photosynthetic efficiency from December 1976 to December 1978 have been carried out in two brackish lakes: Lake Etang de Berre and Lake Etang de Vaine located in the French Mediterranean coast, in the region of Carry-le-Rouet located on the north-west Mediterranean near Marseilles, and in fresh water inflows from 4 Rivers (Touloubre, Durance, Arc, Durancole) to Lake Etang de Berre. Physico-chemical parameters were measured for this study: water temperature, salinity, density, pH, alcalinity, dissolved oxygen (% saturation), phosphate, nitrate, nitrite, silicate etc. Diverse biological parameters were also studied: photosynthetic pigments, phaeopigments, specific composition and biomass of phytoplankton, primary pelagic production etc. Climatical factors were studied: air-temperature, solar-radiation, evaporation, direction (including strength) of winds, precipitation and freshwater volume of the four rivers. The changes in Lake ‘Etang de Berre’ ecosystem depend on the quality of the water in the Durance River, and on the effects of seawater near the entrance of the Caronte Canal. The water quality of the lake varies horizontally and vertically as a result of atmospheric phenomena, maritime currents and tides. The distribution of water temperatures is generally heterogeneous. Southeasterly winds and the Northeasterly Mistral wind are important in the origins of circulated and mixed water masses. These winds are both frequent and strong. They have, as a result, a great effect on the water environment of Lake Etang de Berre. In theory, the annual precipitation in this region is well over eight times the water mass of the lake. The water of the Durance River flows into Lake Etang de Berre through the EDF Canal, amounting to 90% of the precipitation. However, reduction of rainfall in dry seasons has a serious effect on the hydrological characteristics of the lake. The temperature in the winter is partially caused by the low temperature of fresh water, particularly that of the Durance River. The hydrological season of fresh and brackish water is about one month ahead of the hydrological season of sea water in its vicinity. The salinity of Lake Etang de Berre runs approximately 3$\textperthousand$, except at lower levels and near the entrance to the Caronte Canal. However, when the volume of the Durance River water is reduced in the summer and fall, the salinity rises to 15$\textperthousand$. In the lake, the ratio of fresh water to sea water is six to one (6:1). The large quantities of seston conveyed by rivers, particularly the Durance diversion, strongly reduce the transparency in the brackish waters. Although the amount of sunshine is also notable, transparency is slight because of the large amount of seston, carried chiefly by Tripton in the fresh water of the Durance River. Therefore, photosynthesis generally occurs only in the surface layer. The transparency progressively increases from freshwater to open seawater, as mineral particles sink to the bottom (about 1.7kg $m^{-2}a^{-1}$ on the average in brackish lakes). The concentration of dissolved oxygen and the rate of oxygen saturation in seawater (Carry-le-Rouet) ranged from 5.0 to 6.0 $m\ell$ㆍ.$1^{-1}$, and from 95 to 105%, respectively. The amount of dissolved oxygen in Etang de Berre oscillated between 2.9 and 268.3%. The monographs of phosphate, nitrate, nitrite and silicate were published as a part of a study on the ecology of phytoplankton in these environments. Horizontal and vertical distributions of these nutriments were studied in detail. The recent diversion of the Durance River into Lake Etang de Berre has effected a fundamental change in this formerly marine environment, which has had a great impact in its plankton populations. A total of 182 taxa were identified, including 111 Bacillariophyceae, 44 Chlorophyceae, and 15 Cyanophyceae. The most abundant species are small freshwater algae, mainly Chlorophyceae. The average density is about $10^{8}$ cells $1^{-1}$ in Lake Etang de Berre, and about double that amount in Lake Etang de Vaine. Differences in phytoplankton abundance and composition at the various stations or at various depths are slight. Cell biovolume V (equivalent to true biomass), plasma volume VP (‘useful’ biomass) and, simultaneously. the cell surface area S and S/V ratio through the measurement of cell dimensions were computed as the parameters of phytoplankton productivity and metabolism. Pigment concentrations are generally very high on account of phytoplankton blooms by Cyanophyceae, Chlorophyceae and Cryptophyceae. On the other hand, in freshwaters and marine waters, pigment concentrations are comparatively low and stable, showing slight annual variation. The variations of ATP concentration were closely related to those of chlorophyll a and phytoplankton blooms only in marine waters. The carbon uptake rates ranged between 38 and 1091 mg$Cm^{-2}d^{-1}$, with an average surface value of 256 mg; water-column carbon-uptake rates ranged between 240 and 2310 mg$Cm^{-2}d^{-1}$, with an average of 810, representing 290 mg$Cm^{-2}$, per year 45 000 tons per year of photosynthetized carbon for the whole lake. Gross photosynthetic production measured by the method of Ryther was studied over a 2-year period. The values obtained from marine water(Carry-le-Rouet) ranged from 23 to 2 337 mg$Cm^{-2}d^{-1}$, with a weighted average of 319, representing about 110 gCm$^{-2}$ per year. The values in brakish water (Etang de Berre) ranged from 14 to 1778 mg$Cm^{-2}d^{-1}$, with a weighted average of 682, representing 250 mg$Cm^{-2}$ per year and 38 400 tons per year of photosynthesized carbon for the whole lake.

Viabilities of Biopsied Mouse Embryos after Ultrarapid Refreezing and Thawing (미세조작된 생쥐수정란의 초급속 재동결융해 후 생존성)

  • 신상태;임준호;강만종;한용만;이경광
    • Korean Journal of Animal Reproduction
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    • v.20 no.2
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    • pp.207-214
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    • 1996
  • To examine the developmental capacity of manipulated embryos after ultrarapid refreezing and thawing, mouse embryos were biopsied at 4-cell stage, frozen twice at 4-cell and morula stages, respectively, and then transferred to rec-ipients. Single blastomeres were biopsied from 4-cell embryos by a modified aspiration method. Biopsied 4-cell embryos were equilibrated into freezing medium at room temperature for 2.5 min, loaded into 40 $\mu$I of freezing medium in 0.25 ml plastic straw and then directly immersed into liqiud nitrogen. Freezing medium for 4-cell embryos consisted of 4.0 M ethylene glycol and O.25 M sucrose in dPBS supplemented with 6 mg/lm BSA. Morulae were frozen into freezing medium containing 5.0 M glycerol instead of ethylene glycol. Thawing was conducted by agitating each straw in 3TC water for 20 sec. The c content of each straw was expelled into 0.5 ml of dilution medium, which consisted of 0.25 M sucrose and 3 mg/ml BSA in dPBS. The thawed embryos were rehydrated in dilution medium for 10 min, washed 3 times with dPBS and then cultured in M16 medium at 37$^{\circ}C$, 5% CO$_2$ in air. Blastocysts that developed from frozen or refrozne biopsied embryos were transferred to recipients on Day 3 of pseudopregnancy, respectively. In vitro and in vivo developmental rates of the biopsied and intact 4~cell embryos after freezing and thawing were 78 (10l/130) and 25% (10/40), and 91 (114/125) and 30% (12/40), respectively. Although the rates of in vitro development of biopsied and intact embryos to blastocyst stage were significantly different after freezing and thawing (P

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The Flow-rate Measurements in a Multi-phase Flow Pipeline by Using a Clamp-on Sealed Radioisotope Cross Correlation Flowmeter (투과 감마선 계측신호의 Cross correlation 기법 적용에 의한 다중상 유체의 유량측정)

  • Kim, Jin-Seop;Kim, Jong-Bum;Kim, Jae-Ho;Lee, Na-Young;Jung, Sung-Hee
    • Journal of Radiation Protection and Research
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    • v.33 no.1
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    • pp.13-20
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    • 2008
  • The flow rate measurements in a multi-phase flow pipeline were evaluated quantitatively by means of a clamp-on sealed radioisotope based on a cross correlation signal processing technique. The flow rates were calculated by a determination of the transit time between two sealed gamma sources by using a cross correlation function following FFT filtering, then corrected with vapor fraction in the pipeline which was measured by the ${\gamma}$-ray attenuation method. The pipeline model was manufactured by acrylic resin(ID. 8 cm, L=3.5 m, t=10 mm), and the multi-phase flow patterns were realized by an injection of compressed $N_2$ gas. Two sealed gamma sources of $^{137}Cs$ (E=0.662 MeV, ${\Gamma}$ $factor=0.326\;R{\cdot}h^{-1}{\cdot}m^2{\cdot}Ci^{-1}$) of 20 mCi and 17 mCi, and radiation detectors of $2"{\times}2"$ NaI(Tl) scintillation counter (Eberline, SP-3) were used for this study. Under the given conditions(the distance between two sources: 4D(D; inner diameter), N/S ratio: $0.12{\sim}0.15$, sampling time ${\Delta}t$: 4msec), the measured flow rates showed the maximum. relative error of 1.7 % when compared to the real ones through the vapor content corrections($6.1\;%{\sim}9.2\;%$). From a subsequent experiment, it was proven that the closer the distance between the two sealed sources is, the more precise the measured flow rates are. Provided additional studies related to the selection of radioisotopes their activity, and an optimization of the experimental geometry are carried out, it is anticipated that a radioisotope application for flow rate measurements can be used as an important tool for monitoring multi-phase facilities belonging to petrochemical and refinery industries and contributes economically in the light of maintenance and control of them.

Analysis of Amperometric Response to Cholesterol according to Enzyme-Immobilization Methods (효소고정화 방법에 따른 콜레스테롤 검출용 바이오센서의 전류 감응도 분석)

  • Ji, Jung-Youn;Kim, Mee-Ra
    • Journal of the East Asian Society of Dietary Life
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    • v.21 no.5
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    • pp.731-738
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    • 2011
  • Cholesterol is the precursor of various steroid hormones, bile acid, and vitamin D with functions related to regulation of membrane permeability and fluidity. However, the presence of excess blood cholesterol may lead to arteriosclerosis and hypertension. Moreover, dietary cholesterol may affect blood cholesterol levels. Generally, cholesterol determination is performed by spectrophotometric or chromatographic methods, but these methods are very time consuming and costly, and require complicated pretreatment. Thus, the development of a rapid and simple analysis method for measuring cholesterol concentration in food is needed. Multi-walled carbon nanotube (MWCNT) was functionalized to MWCNT-$NH_2$ via MWCNT-COOH to have high sensitivity to $H_2O_2$. The fabricated MWCNT-$NH_2$ was attached to a glassy carbon electrode (GCE), after which Prussian blue (PB) was coated onto MWCNT-$NH_2$/GCE. MWCNT-$NH_2$/PB/GCE was used as a working electrode. An Ag/AgCl electrode and Pt wire were used as a reference electrode and counter electrode, respectively. The sensitivity of the modified working electrode was determined based on the amount of current according to the concentration of $H_2O_2$. The response increased with an increase of $H_2O_2$ concentration in the range of 0.5~500 ${\mu}M$ ($r^2$=0.96) with a detection limit of 0.1 ${\mu}M$. Cholesterol oxidase was immobilized to aminopropyl glass beads, CNBr-activated sepharose, Na-alginate, and toyopearl beads. The immobilized enzyme reactors with aminopropyl glass beads and CNBr-activated sepharose showed linearity in the range of 1~100 ${\mu}M$ cholesterol. Na-alginate and toyopearl beads showed linearity in the range of 5~50 and 1~50 ${\mu}M$ cholesterol, respectively. The detection limit of all immobilized enzyme reactors was 1 ${\mu}M$. These enzyme reactors showed high sensitivity; especially, the enzyme reactors with CNBr-activated sepharose and Na-alginate indicated high coupling efficiency and sensitivity. Therefore, both of the enzyme reactors are more suitable for a cholesterol biosensor system.

Association Between the Polymorphism on Intron 5 of the Lipoprotein Lipase Gene and Carcass Traits in Hanwoo (Korean cattle) (한우 Lipoprotein Lipase 유전자 Intron 5번의 Polymorphism과 경제 형질과의 관련성 분석)

  • Lee, H.J.;Lee, S.H.;Cho, Y.M.;Yoon, H.B.;Jeon, B. K.;Oh, S.J.;Kwon, M.S.;Yoon, D.H.
    • Journal of Animal Science and Technology
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    • v.46 no.6
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    • pp.947-956
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    • 2004
  • The primary role of lipoprotein lipase(LPL) is the hydrolysis of triglycerides(TG) from the core of triglyceride-rich lipoproteins such as chylomicrons and very low density lipoproteins in plasma. Fatty acids liberated by LPL on capillary endothelial surfaces are available for tissues as energy sources especially in muscles or for storage in the form of TG in adipose tissues. Therefore, as the candidate gene related to the carcass traits of the beef cattle, we have directly sequenced the exon 5${\sim}$exon 6 region in the bovine LPL gene for discovery of single nucleotide polymorphism(SNP) with 24 unrelated Hanwoo(Korean cattle). Novel eight sequence variants were detected: three loci on exon 5, three on intron 5 and two on exon 6. All SNPs identified were strongly linked each other, and one hundred twenty eight Hanwoo samples were genotyped one SNP on intron 5 using PCR-restriction fragment length polymorphism method by digestion with Hae III restriction enzyme. The allele frequency of the polymorphism was 0.76 and 0.24. The effects of this polymorphism on the breeding values of the carcass weight, loin muscle area, back fat thickness and marbling score were analyzed using least square methods of SAS GLM. The marbling score of BB genotype was significantly higher than those of AA and AB genotypes(P<0.05). This result indicates that this polymorphism may be associated with the variation of marbling score. Further study is warranted to investigate the phenotypic association in Hanwoo.

Effect of Plant Growth Regulator Treatments on the Growth and Lateral Root Formation in Soybean Sprouts - I. Effect of Plant Growth Regulator Treatments on the Growth in Soybean Sprouts (생장조절물질(生長調節物質) 처리(處理)가 콩나물의 생육(生育) 및 세근발생(細根發生)에 미치는 영향(影響) - I. 생장조절물질(生長調節物質)의 단용(單用) 및 혼용처리(混用處理)가 콩나물의 생육(生育)에 미치는 효과(效果))

  • Kang, C.K.;Lee, J.M.;Saka, H.
    • Korean Journal of Weed Science
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    • v.9 no.1
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    • pp.56-68
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    • 1989
  • aA series of experiments were conducted to investigate the effect of plant growth regulator treatments on the growth and lateral root formation in soybean sprouts in order to establish the effective method of producing root-less or short-rooted soybean sprouts with larger diameter in the hypocotyl. Major results can be summarized as follows. 1. Soybean sprouts showed fairly uniform elongation rate from 3 to g days after imbibition with daily increase of 3.8cm. The speed of elongation of hypocotyl was reduced whereas that of root accelerated 7 days after imbibition. Lateral roots began to emerge fairly evenly from 5 to 9 days after imbibition with a daily increase of 4.4. 2. Auxins(IAA, IBA, NAA, 2,4-D) inhibited hypocotyl elongation and formation of lateral roots and increased hypocotyl diameter without influencing root length and hook diameter at higher concentrations. The dry weight of cotyledon was increased significantly as compared to that of hypocotyl and root. Among the tested auxins, 2, 4-D was the most effective. 3. BA and 4PU-30 significantly reduced elongation of hypocotyl and root and resulted in the biggest diameter of hypocotyl when treated at higher concentrations. The lowest effective concentration of BA to prevent the formation of larval gal roots was 12.5ppm. The formation of lateral roots could be completely prevented by BA and 4PU-30 treatment but kinetin, zeatin, zeatin riboside resulted in many lateral roots and increased thickness of soybean sprouts with little influence. Cotyledon deformation was found in soybean sprouts treated by 4PU-30. 4. 2, 4-D was the most effective for increasing the hypocotyl diameter while 4PU-30 was the most effective for reducing no. of lateral roots. 5. It can be concluded that among the plant growth regulators tested, BA was effective in reducing root length and increasing hypocotyl diameter. BA 12.5 ppm or 15 ppm may thus be the more practical for production of soybean sprouts. 6. ABA showed no significant effect of growth parameter, however ABA 25 ppm inhibited only no of lateral roots with little influence on the growth of seedling. 7. Ethephon inhibited the elongation of hypocotyl and root and increased hypocotyl diameter at higher concentrations. 8. The combined effect of cytokinins and ethephon was very similar to result of BA treatment alone. As the ethephon concentration increased, hypocotyl diameter and dry weight of cotyledon tended to increase.

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