• Title/Summary/Keyword: environmental DNA

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Comparison of Radical Scavenging, Anticytotoxic, and Anti-Inflammatory Effects of Euphorbia Maculata and E. supina (큰땅빈대와 애기땅빈대의 라디칼 소거, 세포독성 억제 및 항염증 활성 비교)

  • Rhim, Tae-Jin
    • Journal of Environmental Science International
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    • v.25 no.8
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    • pp.1131-1142
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    • 2016
  • This study was conducted to compare the antioxidant, anticytotoxic, and anti-inflammatory properties of Euphorbia maculata ethanol extract with those of E. supina ethanol extract. 2,2-Diphenyl-1-picrylhydrazyl (DPPH) radical and superoxide scavenging activities of E. maculata at $50{\mu}g/mL$ were $38.3{\pm}3.7$ and $21.5{\pm}1.2%$, respectively, whereas those of E. supina at the same concentration were $109.4{\pm}0.9$ and $59.5{\pm}4.8%$, respectively. Oxygen radical absorbance capacities of E. maculata and E. supina at $10{\mu}g/mL$ were $14.70{\pm}0.63$ and $26.17{\pm}1.36nmol/mL$ Trolox, respectively. Cupric reducing antioxidant capacities of E. maculata and E. supina at $10{\mu}g/mL$ were $10.22{\pm}0.97$ and $62.99{\pm}5.28nmol/mL$ Trolox, respectively. Total phenolic contents of E. maculata and E. supina at $50{\mu}g/mL$ were $29.03{\pm}0.14$ and $87.89{\pm}0.20nmol/mL$ gallic acid, respectively. E. maculata and E. supina were reported to prevent supercoiled DNA breakage induced by peroxyl and hydroxyl radicals in a concentration-dependent manner, where protection against the supercoiled DNA breakage provided by E. supina was greater than that provided by E. maculata. E. maculata and E. supina at $100{\mu}g/mL$ inhibited tert-butyl hydroperoxide-induced cytotoxicity in HepG2 cells by $49.4{\pm}4.3$ and $87.3{\pm}4.5%$, respectively. E. maculata and E. supina at $500{\mu}g/mL$ inhibited lipopolysaccharide-induced nitric oxide production in RAW 264.7 cells by $63.1{\pm}7.0$ and $85.2{\pm}1.6%$, respectively. The antioxidant capacities including DPPH radical scavenging, superoxide scavenging, oxygen radical absorbance, and cupric reducing antioxidant activity were found to be highly correlated with total phenolic content (0.896 < r < 0.983, p < 0.01) and anticytotoxic activities (0.915 < r < 0.960, p < 0.01). However, the superoxide scavenging activity was not significantly correlated (r = 0.604, p > 0.05) with the anti-inflammatory activity. Thus, these findings demonstrated that the radical scavenging, anticytotoxic, and anti-inflammatory capacities of E. supina were more potent than those of E. maculata. Further studies are needed to elucidate the properties of polyphenolic constituents in E. supina responsible for these effects and the underlying mechanisms.

First Report of Fusarium Wilt Caused by Fusarium oxysporum on Kohlrabi in Korea (Fusarium oxysporum에 의한 콜라비 시들음병)

  • Choi, In-Young;Kim, Ju;Ju, Ho-Jong;Park, Ji-Hyun;Shin, Hyeon-Dong
    • Research in Plant Disease
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    • v.21 no.1
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    • pp.27-31
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    • 2015
  • In April 2014, Fusarium wilt was found on kohlrabi seedlings in Iksan, Korea. Symptoms included wilting of foliage, drying and withering of older leaves, and stunting of the plants. The infected plants eventually died during growth. Colonies on potato dextrose agar were pinkish white, and felted with cottony and aerial mycelium. Macroconidia were falcate to almost straight, thin walled and usually 3-septate. Microconidia were usually formed abundantly in false-heads on short monophialides on the hyphae and were hyaline, smooth, oval to ellipsoidal, aseptate or medianly 1-septate, very occasionally 2-septate, slightly constricted at the septa, $4-11{\times}2.5-5{\mu}m$. On the basis of the morphological characteristics and phylogenetic analyses of molecular markers (internal transcribed spacer rDNA and translation elongation factor $1{\alpha}$), the fungus was identified as Fusarium oxysporum. Pathogenicity of a representative isolate was proved by artificial inoculation, fulfilling Koch's postulates. To our knowledge, this is the first report on the occurrence of Fusarium oxysporum on kohlrabi in Korea.

Use of 16S-23S rRNA Intergenic Spacer Region for Rapid Detection of Vibrio fluvialis (16S-23S rRNA Intergenic Spacer Region을 이용한 Vibrio fluvialis의 검출)

  • 강현실;허문수;이제희
    • Korean Journal of Environmental Biology
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    • v.21 no.1
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    • pp.77-85
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    • 2003
  • We have examined the 16S-23S rRNA intergenic spacer regions (ISR) of Vibrio fluvialis. ISRs were PCR amplified, cloned into a plasmid vector and then sequenced. As results of ISR nucleotide sequence analysis, total of 6 clones were isolated depending on the size. The clones were different in both the number and the composition of the tRNA genes, and were designated ISR-A, ISR-E, ISR-El, ISR-lA, ISR-EKV, ISR-EKAV. ISR-A contains $tRNA^{Ala}$; ISR-lA, $tRNA^{Ile}$-$tRNA^{Ala}$; ISR-EKV, $tRNA^{GIu}$-$tRNA^{Lys}$-$tRNA^{Val}$;ISE-EKAV, $tRNA^{GIu}$-$tRNA^{Lys}$-$tRNA^{Ala}$-$tRNA^{Val}$; ISR -E and E1, $tRNA^{GIu}$ clusters. ISR-EKV was shown to be a minor type out of the six ISR types and showed a very limited homology between ISR-EKV from V, fluvialis and ISRa from other Vibrio species. Therefore ISR-EKV sequence was used to design species-specific primers to detect V, fiuvialis from other Vibrio species by PCR reaction. The specificity of the primers was examined using genomic DNA of other Vibrios as templates for PCR reaction. The result showed that PCR can be a useful method to detect V. fluvialis among Vibrio species in a single PCR reaction.

Development of a PCR Assay for Detection of the Protozoan Parasite Perkinsus (PCR 기법을 이용한 바지락포자충 Perkinsus 진단 기술개발)

  • 박경일;박영미;이제희;최광식
    • Korean Journal of Environmental Biology
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    • v.20 no.2
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    • pp.109-117
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    • 2002
  • Detection of protozoan parasites Perkinsus sp. and P. atlanticus was developed in this study using a specific polymerase chain reaction (PCR) to diagnose the presence of those organisms that causes extensive mortalities of marine shellfishes. The PCR was conducted together with fluid thioglycollate medium (FTM) method and 2 M NaOH lysis method. For the test, Manila clams, Ruditapes philippinarum, were collected from four coastal locations in Korea including Wando Island, Gimnyeong, Sungsan and Sogwipo in Jeju. In addition, trophozites of Perkinsus sp. cultivated in vitro and the granular ark clam, Tegillarca granosa, taken from Gangjin on the south coast of Korea, were used as positive and negative controls, respectively. Expected DNA bands were detected in the samples from Wando Island, Sungsan and the in vitro cultured Perkinsus sp. when the probes specific for the genus Perkinsus and P. atlanticus were used. The samples were also positively diagnosed by the FTM and 2 M NaOH methods. In contrast, the Manila clams from Gimnyeong and Sogwipo, and the granular arks clams from Gangjin showed no detectable signs of infection with the PCR, the FTM method and the 2 M NaOH lysis method. On the other hand, being amplified by p. atlanticus specific primer, it is suggested that the protozoan parasite Perkinsus sp. found in the Korean Manila clam is P. atlanticus. Finally the PCR- based assay developed in the present study can be used in detection of Perkinsus infection and discrimination of Peykinsus species in quarantine stations or laboratories due to the high sensitivity and specificity as well as its rapid detection.

Microbial Structure and Community of RBC Biofilm Removing Nitrate and Phosphorus from Domestic Wastewater

  • Lee, Han-Woong;Choi, Eui-So;Yun, Zu-Whan;Park, Yong-Keun
    • Journal of Microbiology and Biotechnology
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    • v.18 no.8
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    • pp.1459-1469
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    • 2008
  • Using a rotating biological contactor modified with a sequencing bath reactor system (SBRBC) designed and operated to remove phosphate and nitrogen [58], the microbial community structure of the biofilm from the SBRBC system was characterized based on the extracellular polymeric substance (EPS) constituents, electron microscopy, and molecular techniques. Protein and carbohydrate were identified as the major EPS constituents at three different biofilm thicknesses, where the amount of EPS and bacterial cell number were highest in the initial thickness of 0-100${\mu}m$. However, the percent of carbohydrate in the total amount of EPS decreased by about 11.23%, whereas the percent of protein increased by about 11.15% as the biofilm grew. Thus, an abundant quantity of EPS and cell mass, as well as a specific quality of EPS were apparently needed to attach to the substratum in the first step of the biofilm growth. A FISH analysis revealed that the dominant phylogenetic group was $\beta$- and $\gamma$-Proteobacteria, where a significant subclass of Proteobacteria for removing phosphate and/or nitrate was found within a biofilm thickness of 0-250${\mu}m$. In addition, 16S rDNA clone libraries revealed that Klebsiella sp. and Citrobacter sp. were most dominant within the initial biofilm thickness of 0-250${\mu}m$, whereas sulfur-oxidizing bacteria, such as Beggiatoa sp. and Thiothrix sp., were detected in a biofilm thickness over 250${\mu}m$. The results of the bacterial community structure analysis using molecular techniques agreed with the results of the morphological structure based on scanning electron microscopy. Therefore, the overall results indicated that coliform bacteria participated in the nitrate and phosphorus removal when using the SBRBC system. Moreover, the structure of the biofilm was also found to be related to the EPS constituents, as well as the nitrogen and phosphate removal efficiency. Consequently, since this is the first identification of the bacterial community and structure of the biofilm from an RBC simultaneously removing nitrogen and phosphate from domestic wastewater, and it is hoped that the present results may provide a foundation for understanding nitrate and phosphate removal by an RBC system.

Increased Genotoxicity of N'-methyl-N'-nitroguanidine by Oxidative Stress (산화적 스트레스에 의한 N'-methyl-N'-nitroguanidine의 유전독성증가)

  • Kang, Jin-Seok;Jung, Ki-Kyung;Suh, Soo-Kyung;Kim, Joo-Hwan;Lee, Hwa-Ok;Jung, Hai-Kwan;Kim, Seung-Hee;Park, Sue-Nie
    • Environmental Analysis Health and Toxicology
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    • v.22 no.4
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    • pp.357-366
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    • 2007
  • To investigate the possible enhancement of genotoxicity in stress environment, we examined the of effect of genotoxic material in oxidative stress-induced condition using human tell line. Human lymphoblast cell line, TK6 was treated with hydrogen peroxide ($H_2O_2$) for induction of oxidative stress, and treated with N'-methyl-N'-nitroguanidine (MNNG), af a genetoxic material. We carried out MTS assay to set treatment doses. TK6 was treated with $H_2O_2$ at 6.75 (low dote) or $13.5\;{\mu}M$ (high dose) for 2 h, and treated with MNNG af 0.117 (low dose), 0.234 (middle dose), $0.468\;{\mu}M$ (high dose) for 2 h. As results, a treatment of MNNG induced DNA dam age as dose dependently. And TK6 treated with $H_2O_2$ at low as well as high dose followed by MNNG treatment showed higher DNA damage compared to MNNG alone treated groups. Malondialdehyde, as a marker of lipid peroxidation was increased in $H_2O_2$ and MNNG treated groups. Real-time RT-PCR analyses for expression of several antioxidative enzymes showed that catalase mRNA and glutathione peroxidase 1 mRNA expression were decreased in $H_2O_2$ and MNNG treated groups. Taken together, we conclude that genotoxicity induced by MNNG is enhanced in a condition of oxidative stress induced by $H_2O_2$ and it suggests that it should be associated with induction of lipid peroxidation and decrease of antioxidant enzymes.

PLCE1 Gene in Esophageal Cancer and Interaction with Environmental Factors

  • Guo, Li-Yan;Zhang, Shen;Suo, Zhen;Yang, Chang-Shuang;Zhao, Xia;Zhang, Guo-An;Hu, Die;Ji, Xing-Zhao;Zhai, Min
    • Asian Pacific Journal of Cancer Prevention
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    • v.16 no.7
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    • pp.2745-2749
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    • 2015
  • Objective: To study the PLCE1 gene rs2274223 polymorphism with regard to esophageal cancer and its interaction with diet, lifestyle, psychological and environmental factors in Southwest Shandong province. Materials and Methods: A case series study (case-case) was conducted. Questionnaire data were collected and 3 ml-5ml venous blood was drawn for DNA extraction among the qualified research subjects. PLCE1 gene polymorphism was detected after PCR amplification of DNA. SPSS 13.0 software was used for statistical analysis of the data. Results: The three genotypes A/A, A/G and G/G PLCE1 gene rs2274223 was 31, 16 and 4 cases, accounting for 60.8%, 31.4%, 0.08% respectively. The difference of three genotypes (AA/GA/GG) proportion between negative and positive family history of patients was statistically significant, ${\chi}^2=6.213$, p=0.045. There was no statistically significant relationship between PLCE1 gene rs2274223 polymorphism and smoking, drinking, ${\chi}^2=0.119$, p=0.998, and ${\chi}^2=1.727$, p=0.786. There was no linkage of the three rs2274223 PLCE1 gene genotypes (AA/GA/GG) proportion with eating fried, pickled, hot, mildew, overnight, smoked, excitant food, eat speed, salt taste or not (p>0.05). or with living environment pollution and nine risk factors of occupational exposure (p>0.05). There was no statistically significant difference in TS scores between different genotype of rs2274223 PLCE1 gene. Conclusions: The PLCE1 rs2274223 polymorphism has a relationship with family history of esophageal cancer, but does not have any significant association with age, gender, smoking, alcohol drinking, food hygiene, eating habits, living around the environment and occupation in cases.

Molecular Analysis of Microbial Community in Soils Cultivating Bt Chinese Cabbage (분자생물학적 분석을 통한 Bt 배추의 토양미생물상 영향 비교평가)

  • Sohn, Soo-In;Oh, Young-Ju;Oh, Sung-Dug;Kim, Min-Kyung;Ryu, Tae-Hoon;Lee, Ki-Jong;Suh, Seok-Choel;Baek, Hyeong-Jin;Park, Jong-Sug
    • Korean Journal of Environmental Agriculture
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    • v.29 no.3
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    • pp.293-299
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    • 2010
  • The aim of this study was to investigate the possible impact of Bt Chinese cabbage on the soil microbial community. Microbial communities were isolated from the rhizosphere of one Bt Chinese cabbage variety and four varieties of conventional ones and were subjected to be analyzed using both culture-dependent and molecular methods. The total counts of bacteria, fungi, and actinomycetes in the rhizosphere of transgenic and conventional Chinese cabbages were observed to have an insignificant difference. Denaturing gradient gel electrophoresis (DGGE) analysis of PCR-amplified 16S rRNA genes revealed that the bacterial community structures were very similar to each other and this genetic stability of microbial communities was maintained throughout the culture periods. Analysis of dominant isolates in the rhizosphere of transgenic and conventional Chinese cabbages showed that the dominant isolates from the soil of transgenic Chinese cabbage belonged to the Bacilli and Alphaproteobacteria, while the dominant isolates from the soil of conventional cabbage belonged to the Holophagae and Planctomycetacia, respectively. These results indicate that the Bt transgenic cabbage has no significant impact on the soil microbial communities.

Bacterial Productivity and Degradability of Organic Compounds in the Coastal Area of East Sea (동해 연안역의 박테리아 생산력과 유기물질의 분해능)

  • 이기성;고동규;김근호;이영근;최청일;최영길
    • Korean Journal of Environmental Biology
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    • v.17 no.1
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    • pp.59-69
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    • 1999
  • Bacterial population, signal compounds of pollution, bacterial productivity, heterotrophic activity and the activities of extracellular enzymes were analyzed four times from July 1994 to April 1995 at Daejin, Galnam, Pohang, and Ulsan in the coastal area of Bast Sea in order to investigate the state of eutrophication. The population of heavy metal resistant bacteria in Pohang was the highest among all stations. The bacterial populations which could utilize C -P compounds (phosphonate) showed the highest at Pohang that was expected for the industrial and agricultural effluents to inpour. It could be useful as an index of expectation about the influx of industrial sewages and agricultural effluents. The activities of extracellular enzymes for the degradation of organic matter were the highest in the Ulsan Chuyongam, but low in Daejin. These results showed positive relationship with the population of heterotrophic bacteria and bacterial productivity The ratio of poly-P and DNA in Pohang was the highest, and that in Ulsan, Galnam, and Daejin consecutively, corresponding with the distribution of heterotrophic bacteria. In case of the heterotrophic activities, each of the stations gives difference of seasonal variation in heterotrophic activities. Bacterial productivity seemed to be dependent on seasons; the highest in summer and the lowest in fall and winter. These seasonal variations of bacterial productivity seems to be similar with those of heterotrophic bacteria. It is suggested that the state of eutrophication could be evaluated by the criteria of the population sizes and the productivities of microorganisms, the amount of poly-P, heterotrophic activity, the activities of extracellular enzymes, etc.

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Characterization of the Lsi1 Homologs in Cucurbita moschata and C. ficifolia for Breeding of Stock Cultivars Used for Bloomless Cucumber Production

  • Jung, Jaemin;Kim, Joonyup;Jin, Bingkui;Choi, Youngmi;Hong, Chang Oh;Lee, Hyun Ho;Choi, Youngwhan;Kang, Jumsoon;Park, Younghoon
    • Horticultural Science & Technology
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    • v.35 no.3
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    • pp.333-343
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    • 2017
  • Bloomless cucumber fruits are commercially produced by grafting onto the pumpkin stocks (Cucurbita moschata) to restricted silicon ($SiO_2$) absorption. Inhibition of silicon absorption in bloomless stocks is conferred by a mutant allele of the CmLsi1 homologous to Lsi1 in rice. In this study, we characterized the Lsi1 homologs in pumpkin (C. moschata) and its cold-tolerant wild relative C. ficifolia ('Heukjong') in order to develop a DNA marker for selecting a bloomless trait and to establish the molecular basis for breeding bloomless stock cultivars of C. ficifolia. A Cleaved amplified polymorphic sequence (CAPS) marker (CM1-CAPS) was designed based on a non-sysnonymous single nucleotide polymorphism (SNP, C>T) of the CmLsi1 mutant-type allele, and its applicability for Marker-assisted selection (MAS) was confirmed by evaluating three bloom and five bloomless pumpkin stock cultivars. Quantitative RT-PCR of the CmLsi1 for these stock cultivers implied that expression level of the CmLsi1 gene does not appear to be associated with the bloom/bloomless trait and may differ depending on plant species and tissues. A full length cDNA of the Lsi1 homolog [named CfLsi1($B^+$)] of 'Heukjong' (C. ficifolia), was cloned and sequence comparison between CmLsi1($B^+$) and CfLsi1($B^+$) revealed that there exists total 24 SNPs, of which three were non-synonymous. Phylogenetic analysis of CfLsi1($B^+$) and Lsi1 homologs further revealed that CfLsi1($B^+$) is closesly related to Nodulin 26-like intrinsic proteins (NIPs) and most similar to CpNIP1 of C. pepo than C. moschata.