• Title/Summary/Keyword: axenic

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Effects of Temperature and Irradiance on Growth Rate of Skeletonema marinoi-dohrnii Complex Isolated from Gyeonggi Bay, Korea (경기만 해역에서 분리된 Skeletonema marinoi-dohrnii complex의 생장률에 대한 수온과 광도의 영향)

  • Song, Tae Yoon;Yoo, Man Ho;Lee, Youngju;Choi, Joong Ki
    • Korean Journal of Environmental Biology
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    • v.32 no.2
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    • pp.118-128
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    • 2014
  • The growth rate of Skeletonema marinoi-dohrnii complex isolated from Gyeonggi Bay was studied in on axenic batch cultures, under non-nutrient limited condition to determine the growth rate over a wide range of light intensities ($5{\sim}500{\mu}mol\;m^{-2}s^{-1}$) and temperature ($2{\sim}35^{\circ}C$). This species exhibited its maximum specific growth rate of $2.48d^{-1}$ at a combination of $26.1^{\circ}C$ and light intensity of $197{\mu}mol\;m^{-2}s^{-1}$ as associated to optimal conditions of light and temperature. The results supported that S. marinoi-dohrnii complex are more likely to occur in late winter-spring blooms in the western and southern Korea as well as Gyeonggi Bay due to relatively high growth rates ($0.79{\sim}1.61d^{-1}$), considering the effect of temperature on nutrient competition among Skeletonema species. This study might be helpful to improve the precision and reality of a coastal ecosystem model.

Electron-microscopic studies on fine structure and enzyme activity in the axenic and conventional strains of Entamoeba histolytica (이질아메바(Entamoeba histolytica)의 미세구조 및 효소활성에 관한 전자현미경적 연구)

  • Yong, Tae-Sun;Jeong, Pyeong-Rim;Lee, Geun-Tae
    • Parasites, Hosts and Diseases
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    • v.23 no.2
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    • pp.269-284
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    • 1985
  • The metabolism of Entamoeba histolytica would be affected by various environmental factors, and alteration of the environment was known to afEect the fine structure of 5. histolytica. The present study was designed electronmicroscopically to investigate the ultrastructure and enzyme activities in the aEonic and conventional strains of 5. histolytica. The trophozoites of axenically cultivated HK-9 strain and conventional YS-27 and YS-49 strains of 5. histolytica were collected and liKed with 4% paraformaldehyde/0.1M cacodylate buffier(pH 74), After washing them by centrifugation, 1% warm agar was added in the sediment. Solidified agar with the trophozoites was cut into $lmm^3$ cubes, and incubated in the various substrates to observe enzyme activities. Then, the specimen was post-fixed with 3% glutaraldehyde/0.1M cacodylate buffer (PH 7.4) and 1% osmium tetroBide/0.1M cacodylate buffier (pH 7.4) , dehydrated in ascending ethanol series and embedded in epoxy resin. These were sectioned on an ultramicrotome and observed with a transmission electronmicroscope. The procedures for the observation of the fine structure were same as the above, except for the incubation in the substrate. The sections were stained with uranyl acetate and lead citrate. For the observation of the surface of the amoebae, scanning-electronmicroscopy was carried out. The results obtained in the present study are summarized as follows: 1. The fuzzy coat around double-layered plasma membrane of 5. histolytica was more irregularly and densely distributed in the conventional strains (YS-27, YS-49 strains) than in the axonic strain (HK-9 strain). 2. The endosomes, button bodies and chromatin material were surrounded by a double-layered nuclear membrane having scattered nuclear fores. The paranuclear body, mono- or double-layered vacuoles, vacuolar membrane whorls, rosette-like cylindrical bodies, aggregation of cylindrical bodies and helical bodies were found in the cytoplasm of the amoebae. Helical bodies and glycogen granules were generally abundant, while a few smooth endoplasmic reticula were observed in the cytoplasm. 3. Alkaline phosphatase activity was mainly demonstrated in the plasma membrane, limiting membranes of vacuoles and smooth endoplasmic reticula. ATPase activity was observed in the nucleus, limiting membranes of vacuoles and vacuolar membrane whorls. 4. Acid phosphatase activity was commonly demonstrated in the limiting membranes an contents of vacuoles, Iysosome-like organelles, plasma membrane and the button bodies in the nucleus. The activity was more weakly demonstrated in the HK-9 strain than in the other conventional strains of 5. histolytica. No peroBidase activity was observed in the amoeba strains employed in the present study. 5. With a scanning electron-microscope, no distinct structural differences were observed between the amoeba strains. All the trophozoite forms of the amoebae showed crater-like depressions and rugged features on the outer surface.

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Somatic embryogenesis and plant regeneration of Hovenia dulcis Thunb (헛개나무의 체세포배발생 및 식물체 재분화)

  • Eom, Seung-Hee;Shin, Dong-Yong;Lee, Hyeon-Yong;Kim, Myong-Jo;Kim, Jong-Dai;Choi, Won-Cheol;Heo, Kwon;Yu, Chang-Yeon
    • Korean Journal of Medicinal Crop Science
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    • v.10 no.1
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    • pp.41-45
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    • 2002
  • An efficient and reproducible procedure for the large scale propagation of Hovenia dulcis Thunb. is described. Shoot primodia emerging from the leaf surface was induced from MS medium supplemented with NAA. Stem cuttings were suitable explants for multiple shoot proliferation. They produced axillary shoots which branched repeatedly, yielding an average of 7 shoots per explants after 4 weeks in culture, when cultured on a woody plant medium (WPM) containing 0.1mg/l BA and 0.1mg/l NAA. Stem, leaf and root segments from axenic seedlings were used as explant source to induce somatic embryogenesis. A high frequency of somatic embryos were induced directly from leaf in MS medium with NAA, 2,4-D and in medium containing NAA, 2,4-D with BA. Somatic embryos were germinated in MS medium supplemented with 1mg/ l $GA_3$. Somatic embryos proliferated secondary somatic embryos rapidly after transfer to MS medium supplemented with 1mg/ l kinetin, 1mg/ l $GA_3$ and 2% dextrose.

Mycelial Growth and in vitro Ectomycorrhizal Synthesis on Pinus densiflora Seedlings of Tricholoma bakamatsutake in Korea (한국산 가송이(가칭)의 균사생장 특성과 소나무에 외생균근 형성)

  • Jeon, Sung-Min;Ka, Kang-Hyeon;Hong, Ki-Sung
    • The Korean Journal of Mycology
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    • v.42 no.4
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    • pp.312-321
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    • 2014
  • Tricholoma bakamatsutake is one of the edible ectomycorrhizal mushrooms as an allied species of Tricholoma matsutake. This is the first report on physical characteristics of T. bakamatsutake strains collected from Quercus mongolica forests in Korea. The pure cultures of these strains were isolated from the tissues of fruit bodies and the culture characteristics were investigated under different conditions (media, temperatures, nitrogen sources). Most strains showed the highest mycelial growth on potato dextrose agar (PDA) at 20 or $25^{\circ}C$. Two strains of T. bakamatsutake preferred the ammonium-form rather than the nitrate-form as an inorganic nitrogen source. T. bakamatsutake showed significantly slower mycelial growth when compared with T. matsutake from a Korean forest, although the optimum culture conditions for the two allied species were similar. We also tested the ability to form mycorrhizae as well as cellulase activity of T. bakamatsutake. All strains showed cellulase activity on a carboxymethylcellulose (CMC) agar plate. The mycorrhizae on axenic Pinus densiflora seedlings were formed by two strains of T. bakamatsutake after 3 or 8 months of inoculation. P. densiflora seedlings inoculated with T. bakamatsutake had a much higher biomass than un-inoculated seedlings.

Properties of Dissolved Organic Carbon (DOC) released by Three Species of Blue- green Algae (남조류에 의해 배출된 용존유기탄소의 특성)

  • Choi, Kwang-Soon;Imai, Akio;Kim, Bom-Chul;Matsushige, Kazuo
    • Korean Journal of Ecology and Environment
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    • v.34 no.1 s.93
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    • pp.20-29
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    • 2001
  • The amount, chemical composition and optical property of extracellular dissolved organic carbon (EOC) by phytoplankton were examined using axenic cultures of Microcystis aeruginosa, Anabaena flos-aquae, and Oscillatoria agardhii. The extracellular organic matter was categorized into five fractions (hydrophobic acids; AHSs, hydrophobic neutrals; HoNs, hydrophilic acids; HiAs, hydrophilic bases; HiBs, and hydrophilic neutrals; HiNs) using three adsorbent resins(XAD-8, cation, and anion). The release pattern and chemical composition of EOC varied with algal species and their growth phases. Percentage of extracellular release increased with age in all cultures. HiAs were the dominant component of EOC in all cultures, whereas the proportion of HiAs decreased with age in all cultures. In contrast, the proportions of HiBs and HiNs increased as cultures aged. In particular, the HiN fraction increased from 0% to 44% of EOC in M. aeruginosa and from 3.0% to 28% in A. flos-aquae, respectively. The proportion of AHSs was higher in the cultures of A. flos-aquae(7.5${\sim}$16%) and O. agardhii (8.7${\sim}$16%) than M. aeruginosa(0.2${\sim}$2.5%). The proportions of AHSs increased with culture age in M. aeruginosa and O. agardhii, but decreased in A. flos-aquae. The specific UV absorbance also varied among species; 1.9 Abs${\cdot}$cm$^{-1}$/mgC${\cdot}$L$^{-1}$ for M. aeruginosa, 3.7 Abs${\cdot}$cm$^{-1}$/mgC${\cdot}$L$^{-1}$ for A. flos-aquae, and 13.0 Abs${\cdot}$cm$^{-1}$/mgC${\cdot}$L^{-1}$ for O. agardhii. The results of this study indicates that DOC excreted by three blue-green algae differed with species and the growth phase.

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Physico-Chemical Factors on the Growth of Cochlodinium polykrikoides and Nutrient Utilization (Cochlodinium polykrikoides의 성장에 미치는 물리$\cdot$화학적 요인과 영양염 이용)

  • KIM Hyung Chul;LEE Chang Ku;LEE Sam Geun;KIM Hak Gyoon;PARK Chung Kil
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.34 no.5
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    • pp.445-456
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    • 2001
  • In the 1990s, Cochlodinium polykikoides red tide has been annually occurred in the southern coast of Korea and caused the mass damage to the fisheries with a huge amount of economic loss. The present study was done to establish the biological foundation for the elucidation of the mechanism of C. polykikoiaes red tide. The growth response of C. polykikoides to physico-chemical factors such as temperature, salinity, pH, and light intensity were examined using axenic cultures to evaluate the relative importance of these factors on the dynamics of natural populations, It was found that the highest growth conditions were $25^{\circ}C,\;40\%_{\circ}$, pH 7.5, and 7,500 lux, respectively. The tolerable salinity range of growth was relatively wide at an optimum temperature and was reduced to a much narrower range at a sub-optimum temperature. These findings indicate that C. polykikoides is an eurythermal and euryhaline organism. The organism demanded higher light intensity and oceanic pH narrow in its growth. C. polykikoides utilize inorganic nutrients, such as nitrate and ammonium as N, and phosphate as P. The nutritional requirements of C. polykikoides were $40{\mu}M$ for nitrate, $50{\mu}M$ for ammonium, and $5{\mu}M$ for phosphate. The half saturation constant (Ks) for growth was $2.10{\mu}M$ for nitrate, $1.03{\mu}M$ for ammonium, and $0.57{\mu}M$ for phosphate. These values were comparatively smaller than those of other dinoflagellates reported previously. We confirmed that the organism is characterized as an eutrophic species. However, ammonium Ks value is smaller than that of other eutrophic species, This result indicates that C. polykikoides red tide may outbreak in the waters which eutrophication is in progress rather than eutrophicated waters. C. polykikoides preferred ammonium better than nitrate as a nitrogen source when in a growth stage, Therefore, our results indicate that ammonium is more important nutrient on the growth of the organism in comparison with other inorganic nutrients and C. polykikoides red tide is related with the increased ammonium concentration in the coastal waters.

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Photoalteration in Biodegradability and Chemical Compositions of Algae- derived Dissolved Organic Matter (자외선에 의한 조류기원 용존유기물의 생분해도 및 화학조성변환.)

  • Imai, Akio;Matsushige, Kazuo;Nagai, Takashi;Kim, Yong-Hwan;Kim, Bom-Chul;Choi, Kwang-Soon
    • Korean Journal of Ecology and Environment
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    • v.36 no.3 s.104
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    • pp.235-241
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    • 2003
  • The effect of ultraviolet (UV) radiation on the characteristics of algae-derived dissolved organic matter (DOM) was examined by comparing the biodegradability and DOM fraction distribution of algal DOM before and after UV exposure. Algal DOM from two axenic cultures of Microcystis aeruginosa and Oscillatoria agardhii were irradiated for 24 h at a UV intensity of 42 W/$m^2$. A complete degradation of algal DOM during the UV exposure did not occur, remaining at constant concentrations of dissolved organic carbon(DOC). After UV exposure, however, microbial degradations were reduced by 17% in M. aeruginosa and 53% in O. agardhii, respectively, and decomposition rates also were two times lower in UV exposed algal DOM. In addition, the chemical compositions of algal DOM altered substantially after UV radiation exposure. The proportions of hydrophilic bases (HiB; protein-like DOM) decreased considerably in both algal DOM sources after UV exposure (16.8% and 20.0% of DOM, respectively), whereas those of hydrophilic acids (HiA; carboxylic acids-like DOM) increased as much as the decrease of the HiB fraction. Capillary ion electrophoresis (CE) analysis showed that several carboxylic acids increased significantly after UV exposure, further confirming an increase in HiA fractions. The results of this study clearly indicate that algal DOM can be changed in its chemical composition as well as biodegradability without complete degradation by UV radiation.

Changes of Bacterial Diversity Depend on the Spoilage of Fresh Vegetables (신선 채소류의 부패에 따른 세균의 다양성 변화 및 세균에 의한 채소 부패 조사)

  • Lee, Dong-Hwan;Ryu, Jung-El;Park, So-Yeon;Roh, Eun-Jung;Oh, Chang-Sik;Jung, Kyu-Suk;Yoon, Jong-Chul;Heu, Sung-Gi
    • Research in Plant Disease
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    • v.17 no.1
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    • pp.38-43
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    • 2011
  • Almost 10~30% of vegetables were discarded by the spoilage from farms to tables. After harvest, vegetables are often spoiled by a wide variety of microorganisms including many bacterial and fungal species. This investigation was conducted to extent the knowledge of relationship the spoilage of vegetables and the diversity of microbes. The total aerobic bacterial numbers in fresh lettuce, perilla leaf, and chicory were $2.6{\sim}2.7{\times}10^6$, $4.6{\times}10^5$, $1.2{\times}10^6\;CFU/g$ of fresh weight, respectively. The most common bacterial species were Pseudomonas spp., Alysiella spp., and Burkholderia spp., and other 18 more genera were involved in. After one week of incubation of those vegetables at $28^{\circ}C$, the microbial diversity had been changed. The total aerobic bacterial numbers increased to $1.1{\sim}4.6{\times}10^8$, $4.9{\times}10^7$, and $7.6{\times}10^8\;CFU/g$ of fresh weight for lettuce, perilla leaf, and chicory that is about $10^2$ times increased bacterial numbers than that before spoilage. However, the diversity of microbes isolated had been simplified and fewer bacterial species had been isolated. The most bacterial population (~48%) was taken up by Pseudomonas spp., and followed by Arthrobacter spp. and Bacillus spp. The spoilage activity of individual bacterial isolates had been tested using axenic lettuce plants. Among tested isolates, Pseudomonas fluorescence and Pantoea agglomerans caused severe spoilage on lettuce.

Axenic Culture Production and Growth of a Dinoflagellate, Cochlodinium polykrikoides (적조 와편모조류, Cochlodinium polykrikoides의 순수분리 및 성장)

  • SEO Pil-Soo;LEE Sang-Jun;Kim Yoon;LEE Jeong-Ho;KIM Hak-Gyoon;LEE Jae-Dong
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.31 no.1
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    • pp.71-76
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    • 1998
  • To know the antibiotic specificity of a Dinoflagellate, Cochlodinium polykrikoides, we investigated the survival time of C. polykrikoides against several concentrations of antibiotics and judged the selective specificity of antibiotics based on the $LT_50$ ($50\%$ of lethal time). The result showed that C. polykrikoides was sensitive to tetracycline and chloramphenicol, and resistant to polymixin-B, ampicillin, penicillin-G, dihydrostreptomycin, and neomycin. In the case of sensitive antibiotics to C. polykrikoides, tetracycline and chloramphenicol, the safety concentrations of both antibiotics were determined and the antibiotic specificity based or the plotted survival curve was analyzed. Before antibiotic treatment, we tested the antibiotic susceptibility of the contaminated bacterial population in tile culture of C. polykrikoides, and decided the proper kinds of antibiotics and concentrations before percoll-centrifugation. By percoll-centrifugation, we reduced bacteria, removed fungi, collected the algal pellet, and made axonic culture by antibiotic cascade procedure based on the result of antibiotic susceptibility test. We observed that axonic C. polykrikoides culture entered the logarthmic phase of growth when cell density was over 740 cells/ml and propagated to 5,800 cells/ml maximally. Divisions per day, k value of C. polykrikoides represented a good index for growth at the low density of cells. There was a highest k value shift before reaching to the logarithmic phase. We suggested that the preceeding highest k value shift stage is a good indicator for accurate broadcasting for red. tide blooming in the field, and the stage is also a good time for controlling red tide blooming in the filed, either.

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