• Title/Summary/Keyword: environmental microbiology

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Growth kinetics and chlorine resistance of heterotrophic bacteria isolated from young biofilms formed on a model drinking water distribution system (모델 상수관망에 형성된 초기 생물막에서 분리한 종속영양세균의 생장 동역학 및 염소 내성)

  • Park, Se-Keun;Kim, Yeong-Kwan;Oh, Young-Sook;Choi, Sung-Chan
    • Korean Journal of Microbiology
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    • v.51 no.4
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    • pp.355-363
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    • 2015
  • The present work quantified the growth of young biofilm in a model distribution system that was fed with chlorinated drinking water at a hydraulic retention time of 2 h. Bacterial biofilms grew on the surface of polyvinyl chloride (PVC) slides at a specific growth rate of $0.14{\pm}0.09day^{-1}$ for total bacteria and $0.16{\pm}0.08day^{-1}$ for heterotrophic bacteria, reaching $3.1{\times}10^4cells/cm^2$ and $6.6{\times}10^3CFU/cm^2$ after 10 days, respectively. The specific growth rates of biofilm-forming bacteria were found to be much higher than those of bulk-phase bacteria, suggesting that biofilm bacteria account for a major part of the bacterial production in this model system. Biofilm isolates exhibited characteristic kinetic properties, as determined by ${\mu}_{max}$ and $K_S$ values using the Monod model, in a defined growth medium containing various amounts of acetate. The lowest ${\mu}_{max}$ value was observed in bacterial species belonging to the genus Methylobacterium, and their slow growth seemed to confer high resistance to chlorine treatment (0.5 mg/L for 10 min). $K_S$ values (inversely related to substrate affinity) of Sphingomonas were two orders of magnitude lower for acetate carbon than those of other isolates. The Sphingomonas isolates may have obligate-oligotrophic characteristics, since the lower $K_S$ values allow them to thrive under nutrient-deficient conditions. These results provide a better understanding and control of multi-species bacterial biofilms that develop within days in a drinking water distribution system.

Efficient Remediation of Petroleum Hydrocarbon-Contaminated Soils through Sequential Fenton Oxidation and Biological Treatment Processes (펜톤산화 및 생물학적 연속처리를 통한 유류오염토양의 효율적 처리)

  • Bae, Jae-Sang;Kim, Jong-Hyang;Choi, Jung-Hye;Ekpeghere, Kalu I.;Kim, Soo-Gon;Koh, Sung-Cheol
    • Korean Journal of Microbiology
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    • v.47 no.4
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    • pp.356-363
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    • 2011
  • The accidental releases of total petroleum hydrocarbons (TPH) due to oil spills frequently ended up with soil and ground water pollution. TPH may be degraded through physicochemical and biological processes in the environment but with relatively slow rates. In this study an attempt has been made to develop an integrated chemical and biological treatment technology in order to establish an efficient and environment-friendly restoration technology for the TPH contaminated soils. A Fenton-like reaction was employed as a preceding chemical treatment process and a bioaugmentation process utilizing a diesel fuel degrader consortium was subsequently applied as a biological treatment process. An efficient chemical removal of TPH from soils occurred when the surfactant OP-10S (0.05%) and oxidants ($FeSO_4$ 4%, and $H_2O_2$ 5%) were used. Bioaugmentation of the degrader consortium into the soil slurry led to an increase in their population density at least two orders of magnitude, indicating a good survival of the degradative populations in the contaminated soils ($10^8-10^9$ CFU/g slurry). TPH removal efficiencies for the Fenton-treated soils increased by at least 57% when the soils were subjected to bioaugmentation of the degradative consortium. However, relatively lower TPH treatment efficiencies (79-83%) have been observed in the soils treated with Fenton and the degraders as opposed to the control (95%) that was left with no treatment. This appeared to be due to the presence of free radicals and other oxidative products generated during the Fenton treatment which might inhibit their degradation activity. The findings in this study will contribute to development of efficient bioremediation treatment technologies for TPH-contaminated soils and sediments in the environment.

Isolation and characterization of cellulolytic yeast belonging to Moesziomyces sp. from the gut of Grasshopper (메뚜기의 내장에서 분리한 Moesziomyces 속에 속하는 셀룰로오스 분해 효모의 분리 및 특성)

  • Kim, Ju-Young;Jung, Hee-Young;Park, Jong-Seok;Cho, Sung-Jin;Lee, Hoon Bok;Sung, Gi-Ho;Subramani, Gayathri;Kim, Myung Kyum
    • Korean Journal of Microbiology
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    • v.55 no.3
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    • pp.234-241
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    • 2019
  • An intensive interaction between yeasts and insects has highlighted their relevance for attraction to food and for the insect's development and behavior. Yeast associated in the gut of insects secretes cellulase which aided in the food digestion (cellulose degradation). Three strains of cellulose-degrading yeast were isolated from the gut of adult grasshoppers collected in Gyeonggi Province, South Korea. The strains $ON22^T$, $G10^T$, and $G15^T$, showed positive cellulolytic activity in the carboxymethyl cellulose (CMC)-plate assay. The phylogenetic tree based on sequence analysis of D1/D2 domains of the large subunit rRNA gene and the internal transcribed spacer (ITS) regions revealed that the strains $ON22^T$ (100 and 98.4% sequence similarities in D1/D2 domains and ITS) and $G10^T$ (99.8 and 99.5% in D1/D2 domain and ITS region) were most closely related to the species Moesziomyces aphidis JCM $10318^T$; $G15^T$ (100% in D1/D2 domains and ITS) belongs to the species Moesziomyces antarcticus JCM $10317^T$, respectively. Morphology and biochemical test results are provided in the species description. Cellulase with its massive applicability has been used in various industrial processes such as biofuels like bioethanol productions. Therefore, this is the first report of the cellulolytic yeast strains $ON22^T$, $G10^T$, and $G15^T$ related to the genus Moesziomyces in the family Ustilaginaceae (Ustilaginales), in Korea.

Sesquiterpenoids Bioconversion Analysis by Wood Rot Fungi

  • Lee, Su-Yeon;Ryu, Sun-Hwa;Choi, In-Gyu;Kim, Myungkil
    • 한국균학회소식:학술대회논문집
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    • 2016.05a
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    • pp.19-20
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    • 2016
  • Sesquiterpenoids are defined as $C_{15}$ compounds derived from farnesyl pyrophosphate (FPP), and their complex structures are found in the tissue of many diverse plants (Degenhardt et al. 2009). FPP's long chain length and additional double bond enables its conversion to a huge range of mono-, di-, and tri-cyclic structures. A number of cyclic sesquiterpenes with alcohol, aldehyde, and ketone derivatives have key biological and medicinal properties (Fraga 1999). Fungi, such as the wood-rotting Polyporus brumalis, are excellent sources of pharmaceutically interesting natural products such as sesquiterpenoids. In this study, we investigated the biosynthesis of P. brumalis sesquiterpenoids on modified medium. Fungal suspensions of 11 white rot species were inoculated in modified medium containing $C_6H_{12}O_6$, $C_4H_{12}N_2O_6$, $KH_2PO_4$, $MgSO_4$, and $CaCl_2$ for 20 days. Cultivation was stopped by solvent extraction via separation of the mycelium. The metabolites were identified as follows: propionic acid (1), mevalonic acid lactone (2), ${\beta}$-eudesmane (3), and ${\beta}$-eudesmol (4), respectively (Figure 1). The main peaks of ${\beta}$-eudesmane and ${\beta}$-eudesmol, which were indicative of sesquiterpene structures, were consistently detected for 5, 7, 12, and 15 days These results demonstrated the existence of terpene metabolism in the mycelium of P. brumalis. Polyporus spp. are known to generate flavor components such as methyl 2,4-dihydroxy-3,6-dimethyl benzoate; 2-hydroxy-4-methoxy-6-methyl benzoic acid; 3-hydroxy-5-methyl phenol; and 3-methoxy-2,5-dimethyl phenol in submerged cultures (Hoffmann and Esser 1978). Drimanes of sesquiterpenes were reported as metabolites from P. arcularius and shown to exhibit antimicrobial activity against Gram-positive bacteria such as Staphylococcus aureus (Fleck et al. 1996). The main metabolites of P. brumalis, ${\beta}$-Eudesmol and ${\beta}$-eudesmane, were categorized as eudesmane-type sesquiterpene structures. The eudesmane skeleton could be biosynthesized from FPP-derived IPP, and approximately 1,000 structures have been identified in plants as essential oils. The biosynthesis of eudesmol from P. brumalis may thus be an important tool for the production of useful natural compounds as presumed from its identified potent bioactivity in plants. Essential oils comprising eudesmane-type sesquiterpenoids have been previously and extensively researched (Wu et al. 2006). ${\beta}$-Eudesmol is a well-known and important eudesmane alcohol with an anticholinergic effect in the vascular endothelium (Tsuneki et al. 2005). Additionally, recent studies demonstrated that ${\beta}$-eudesmol acts as a channel blocker for nicotinic acetylcholine receptors at the neuromuscular junction, and it can inhibit angiogenesis in vitro and in vivo by blocking the mitogen-activated protein kinase (MAPK) signaling pathway (Seo et al. 2011). Variation of nutrients was conducted to determine an optimum condition for the biosynthesis of sesquiterpenes by P. brumalis. Genes encoding terpene synthases, which are crucial to the terpene synthesis pathway, generally respond to environmental factors such as pH, temperature, and available nutrients (Hoffmeister and Keller 2007, Yu and Keller 2005). Calvo et al. described the effect of major nutrients, carbon and nitrogen, on the synthesis of secondary metabolites (Calvo et al. 2002). P. brumalis did not prefer to synthesize sesquiterpenes under all growth conditions. Results of differences in metabolites observed in P. brumalis grown in PDB and modified medium highlighted the potential effect inorganic sources such as $C_4H_{12}N_2O_6$, $KH_2PO_4$, $MgSO_4$, and $CaCl_2$ on sesquiterpene synthesis. ${\beta}$-eudesmol was apparent during cultivation except for when P. brumalis was grown on $MgSO_4$-free medium. These results demonstrated that $MgSO_4$ can specifically control the biosynthesis of ${\beta}$-eudesmol. Magnesium has been reported as a cofactor that binds to sesquiterpene synthase (Agger et al. 2008). Specifically, the $Mg^{2+}$ ions bind to two conserved metal-binding motifs. These metal ions complex to the substrate pyrophosphate, thereby promoting the ionization of the leaving groups of FPP and resulting in the generation of a highly reactive allylic cation. Effect of magnesium source on the sesquiterpene biosynthesis was also identified via analysis of the concentration of total carbohydrates. Our current study offered further insight that fungal sesquiterpene biosynthesis can be controlled by nutrients. To profile the metabolites of P. brumalis, the cultures were extracted based on the growth curve. Despite metabolites produced during mycelia growth, there was difficulty in detecting significant changes in metabolite production, especially those at low concentrations. These compounds may be of interest in understanding their synthetic mechanisms in P. brumalis. The synthesis of terpene compounds began during the growth phase at day 9. Sesquiterpene synthesis occurred after growth was complete. At day 9, drimenol, farnesol, and mevalonic lactone (or mevalonic acid lactone) were identified. Mevalonic acid lactone is the precursor of the mevalonic pathway, and particularly, it is a precursor for a number of biologically important lipids, including cholesterol hormones (Buckley et al. 2002). Farnesol is the precursor of sesquiterpenoids. Drimenol compounds, bi-cyclic-sesquiterpene alcohols, can be synthesized from trans-trans farnesol via cyclization and rearrangement (Polovinka et al. 1994). They have also been identified in the basidiomycota Lentinus lepideus as secondary metabolites. After 12 days in the growth phase, ${\beta}$-elemene caryophyllene, ${\delta}$-cadiene, and eudesmane were detected with ${\beta}$-eudesmol. The data showed the synthesis of sesquiterpene hydrocarbons with bi-cyclic structures. These compounds can be synthesized from FPP by cyclization. Cyclic terpenoids are synthesized through the formation of a carbon skeleton from linear precursors by terpene cyclase, which is followed by chemical modification by oxidation, reduction, methylation, etc. Sesquiterpene cyclase is a key branch-point enzyme that catalyzes the complex intermolecular cyclization of the linear prenyl diphosphate into cyclic hydrocarbons (Toyomasu et al. 2007). After 20 days in stationary phase, the oxygenated structures eudesmol, elemol, and caryophyllene oxide were detected. Thus, after growth, sesquiterpenes were identified. Per these results, we showed that terpene metabolism in wood-rotting fungi occurs in the stationary phase. We also showed that such metabolism can be controlled by magnesium supplementation in the growth medium. In conclusion, we identified P. brumalis as a wood-rotting fungus that can produce sesquiterpenes. To mechanistically understand eudesmane-type sesquiterpene biosynthesis in P. brumalis, further research into the genes regulating the dynamics of such biosynthesis is warranted.

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Effects of Diet and Time on Feed on Fatty Acid Composition in Muscle of Charolais Steers (사료급원과 급여기간이 Charolais 거세우 근내 지방산 조성에 미치는 영향)

  • 최낙진;강수원;권응기;조원모;전병수;박병기
    • Journal of Animal Science and Technology
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    • v.48 no.6
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    • pp.847-860
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    • 2006
  • This study investigated the effects of feeding Charolais steers on diets rich in either n-6 or n-3 polyunsaturated fatty acids (PUFA) and time on feed (TOF) on muscle fatty acid composition and content. Twenty eight steers were fed on ad libitum forage and one of two concentrates varying in the source of fat; soya (high in C18:2 n-6) or whole linseed (high in C18:3 n-3) for either 60 or 90 days in IGER (Institute of Grassland and Environmental Research, UK). The concentrates were fed at approximately 0.73 of total DM intake. TOF influenced carcass weight, conformation and fatness scores, which were higher at 90 v. 60 days (P<0.05). Diet did not affect total fatty acid content of neutral lipid in m. longissimus thoracis but feeding linseed increased total phospholipid fatty acid by approx- imately 15%(P<0.05). Linseed increased the amount and proportion of C18:3 n-3 (P<0.001) and the proportion of CLA (cis-9, trans-11 conjugated linoleic acid), while soya increased the content (P<0.05) and proportion (P<0.001) of C18:2 n-6 in muscle neutral lipid. In muscle phospholipid, linseed significantly increased the amount of CLA, C18:3 n-3 and its longer chain derivatives as well as C14:0, C16:0, C18:0. C18:1 trans and C18:2 n-6. The amount and proportion of C18:2 n-6 and its longer chain C20 derivatives were higher on feeding soya. TOF (90 v. 60 day) increased the content of C14:0, C16:0, C16:1, CLA, C18:1 n-9, C18:2 n-6 and C18:3 n-3 in muscle neutral lipid. The P:S was not affected by diet or TOF. The ratio of C18:2 n-6 : C18:3 n-3 and sum of n-6 : n-3 fatty acids were higher in muscle from animals fed on linseed v. soya (P<0.001). The study indicates that the PUFA composition of beef muscle may be significantly modified by feeding contrasting dietary lipids, soya vs. linseed. Feeding linseed produced a better balance of muscle fatty acids, more in line with current nutritional recommendations with a lower C18:2 n-6:C18:3 n-3 ratio associated with higher muscle content of C18:3 n-3 and C20:5 n-3 and CLA and lower C20:4 n-6.

Effects of Environmental Factors on the Bacterial Community in Eutrophic Masan Reservoir (이화학적 수질인자가 부영양화된 마산저수지의 세균분포에 미치는 영향)

  • 남귀숙;손형식;차미선;조순자;이광식;이상준
    • Korean Journal of Microbiology
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    • v.39 no.2
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    • pp.95-101
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    • 2003
  • The total bacterial numbers, Eubacterial community structures and environmental factors which affect bacterial community were estimated monthly using DAPI and fluorescent in situ hybridization monthly, from June to November 2000 to evaluate the correlation between the bacterial community and environmental factors in eutrophic agricultural Masan reservoir in Asan. Average water temperatures varied from 12.3 to $27.5^{\circ}C$, pH 7.5 to 9.0, DO 7. I~12.8 mg/L, COD 6.4~13.0 mg/L, chlorophyll a 30.5~99.0 mg/㎥, SS 7.S~25.7 mg/L, TN 1.748~3.543 mg/L., and TP 0.104~0.581 mg/L, respectively. Total bacterial numbers showed high ranges from 0.4 to 9.6$\times$ $10^{6}$ cells/ml, and these indicated the mesotrophic or eutrophic state. The ratio of Eubacteria to total bacteria was 67.6-88.0%, which was higher than that in other reservoir. The relationships of total bacteria and Eubacteria community were more significant with organic nitrogen (Org-N), and organic phosphorus (Org-P) than with water temperature. Proteobacteria groups showed strongly significant relationships with Org-P and Org-N and significant relationships with water temperature, conductivity, COD, and inorganic nitrogen. C-F group was the most significant with Org-N, and HGC group with water temperature. However, relationships of Chl-a, pH, DO and SS showed no significance with any bacterial community. These results were different from other studies, because of the specific characteristics of Masan reservoir such as old, shallow and eutrophic states. The seasonal variation of bacterial community in Masan reservoir does not seem to depend on phytoplankton dynamics but on storm event and organic materials from watershed and the sediment of reservoir.

Evaluating the Impacts of Long-Term Use of Agricultural Chemicals on a Soil Ecosystem by Structural Analysis of Bacterial Community (세균군집의 구조분석을 통한 장기간 농약사용이 토양생태계에 미치는 영향 평가)

  • Yun, Byeong-Jun;Kim, Seong-Hyeon;Lee, Dong-Heon;O, Gye-Heon;Gang, Hyeong-Il
    • Korean Journal of Microbiology
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    • v.39 no.4
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    • pp.260-266
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    • 2003
  • In this study bacterial community was analyzed to evaluate the impacts of long-term use of agricultural chemicals on a soil ecosystem as well as to obtain fundamental data on the relationship. Sequences of 16S rRNA clones from a non-agricultural site and a tangerine orchard soil which has a history of long-term use of agricultural chemicals over 30 years were analyzed. This revealed that bacterial community containing 5 divisions and 18 genera was distributed in a tangerine orchard soil, while bacterial community containing 9 divisions and 44 genera was distributed. In a tangerine orchard soil site, the most abundant bacteria in subdivision level were placed into Proteobacteria γ group which occupied 56% of total clones. The other bacterial clones from the ocrhcard soil exposed to agricultural chemicals over 30 years were Acidobacteria group (25%), Fimicutes group (5%), Planctomycetes group (2%), Proteobacteria α (1%), δ group (1%), and Cyanobacteria group (1%). Whereas, the clones were from the non-agricultural site were distributed among the division or subdivision Acidobacteria group (14%), Planctomycetes group (13%), Proteobacteria α (10%), β (9%), δ (9%), Fimicutes group (8%), Verrucomicrobia group (8%), Actinobacteria group (6%), Proteobacteria γ group (3%), Bacteroidetes group (3%), Gemmatimonadetes group (3%), and Cyanobacteria group (1%). This finding suggests the possibility that long-term application of agricultural chemicals or fertilizers on a tangerine orchard might result in drastic reduction or alteration in the composition of the bacterial community in the contaminated soil site.

Degradation of Phenanthrene and Pyrene by Burkholderia sp. D5 (Burkholderia sp. D5에 의한 phenanthrene과 pyrene 분해)

  • Kim, Tae-Jeong;Jo, Gyeong-Suk;Ryu, Hui-Uk
    • Korean Journal of Microbiology
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    • v.39 no.4
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    • pp.267-271
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    • 2003
  • Burkholderia sp. D5, a polyaromatic hydrocarbons(PAHs)-degrading bacterium, was isolated from oil-contaminated soil. The bacterium could utilize phenanthrene (Phe) as a sole carbon source but could not use pyrene (Pyr). However, the strain could degrade Pyr when a cosubstrate such as yeast extract (YE) was supplemented. The PAH degradation rate of the bacterium was enhanced by the addition of other organic materials such as YE, peptone and glucose. YE was a particularly effective additive in stimulating cell growth as well as PAH degradation. When 1 g-YE/L was supplemented into the basal salt medium (BSM) with 215 mg-Phe/L, the specific growth rate (0.28 h-1) and Phe-degrading rate (29.30 μmol/L/h) were enhanced approximately ten and two times more than those obtained in the BSM with 215 mg-Phe/L, respectively. Through kinetic analysis, the maximum specific growth rate (μmax) and PAH degrading rate (Vmax) for Phe were obtained as 0.34/h and 289 ${\mu}mol$/L/h, respectively. Also, μmax and Vmax for Pyr were 0.27 h-1 and 50 ${\mu}mol$/L/h, respectively. The degradation rates for each Phe (2.20 μmol/L/h) and Pyr (2.18 μmol/L/h) were lower in mixture substrates than in a single substrate (29.30 ${\mu}mol$/L/h and 9.58 ${\mu}mol$/L/h, respectively). Burkholderia sp. D5 can degrade Phe and Pyr contained in soil, and the PAH degradation rates in soil were 20.03 ${\mu}mol$/L/h for Phe and 1.09 ${\mu}mol$/L/h for Pyr.

Analysis of Environmental Characteristics in Habitat of Amanita hemibapha (달걀버섯 발생지의 환경특성 분석)

  • Sou, Hong-Duck;Hur, Tae-Chul;Jung, Sung-Cheol;Joo, Sung-Hyun;Park, Hyun
    • The Korean Journal of Mycology
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    • v.39 no.3
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    • pp.164-170
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    • 2011
  • Amanita hemibapha is a kind of mycorrhizal mushroom which has a relation with host plants and environmental factors. For the purpose of studying the relationship between environmental factors and mushrooms, in this study we conducted to investigate the soil physicochemical properties, the distribution of plants and the diversity of mushrooms in Gwangyang, Chungdo and Daegu. Soil texture in habitate of Amanita hemibapha was Sandy Loam and Sandy Clay Loam which contains a high rate of sand. Soil pH was approximately 5.0 in all study sites. Total nitrogen contents, C/N ratio and available phosphate($P_2O_5$) were the highest at the Chungdo. However, Gwangyang and Chungdo represent similar Cation Exchange Capacity(C.E.C) and Daegu has much less than other areas. Exchangeable cations, $Ca^{2+}$ content was higher at Gwangyang and Chungdo than Daegu and $Na^+$ content was the highest at Chungdo among three sites. Daegu has much $Mg^{2+}$ contents followed by Chungdo and Gwangyang. Interestingly, $K^+$ content listed in reverse order of $Mg^{2+}$ at Gwangyang, Chungdo and Daegu. The main woody plants in study sites are confirmed as Carpinus laxiflora, Quercus mongolica, Q. serrata and Pinus densiflora. Quercus species are found as a common species in three study sites. Lastly, 8 family and 12 species of mushrooms are emerged in Gwangyang, also 8 family and 12 species and 5 family and 10 species are found in Chungdo and Daegu, respectively. Amanita pantherina, Boletus edulis, Tylopilus felleus and Marasmius maximus, which found in study sites, are also kind of mycorrhizal mushroom same as A. hemibaph. By using correspondence analysis, Q. mongolica, Q. serrata, Q. variabilis and C. laxiflora are expected to as a host plant of A. hemibapha.

Primer Evaluation for the Detection of Toxigenic Microcystis by PCR (독소 생성 Microcystis 검출을 위한 PCR primer의 평가)

  • 이현경;김준호;유순애;안태석;김치경;이동훈
    • Korean Journal of Microbiology
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    • v.39 no.3
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    • pp.166-174
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    • 2003
  • Microcystin produced by cyanobacteria in surface waters, such as eutrophic lake and river, is a kind of serious environmental problems due to its toxicity to human and wild animals. Microcystin is synthesized nonribosomally by the large modular multi-functional enzyme complex known as microcystin synthetase encoded by the mcy gene cluster. Amplification of mcy genes by PCR from cultures and environmental samples is a simple and efficient method to detect the toxigenic Microcystis. In order to evaluate primers designed to detect toxic microcystin-producing strains, 17 cyanobacterial strains and 20 environmental samples were examined by PCR with 7 pairs of primers. Some microcystin-producing cyanobacteria were not detected with FAA-RAA, TOX4F-TOX4R and FP-RP primers. The fragment of unexpected size was amplified with NSZW2-NSZW1 primers in Microcystis strains isolated from the lakes in Korea. TOX1P-TOX1F primers failed in amplification of toxin-producing strains. Only MSF-MSR and TOX2P- TOX2F primers amplified the fragments of mcy genes from 11 strains of microcystin-producing Microcystis. The water samples taken from 20 lakes in Korea were analyzed by PCR using each of the primers. In all the water samples, cyanobacteria capable of producing microcystin were detected by the PCR with TOX2P-TOX2F primers. These results indicate that TOX2P-TOX2F primers are better than the other primers for detection of microcystin-producing Microcystis strains in Korea. The nucleotide sequences of mcy gene in Microcystis aeruginosa NIER10010 suggest genetic diversity of Korean isolates.