• Title/Summary/Keyword: codA gene

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Enhanced drought and oxidative stress tolerance in transgenic sweetpotato expressing a codA gene (CodA 고발현 형질전환 고구마의 산화 및 건조 스트레스 내성 증가)

  • Park, Sung-Chul;Kim, Myoung Duck;Kim, Sun Ha;Kim, Yun-Hee;Jeong, Jae Cheol;Lee, Haeng-Soon;Kwak, Sang-Soo
    • Journal of Plant Biotechnology
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    • v.42 no.1
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    • pp.19-24
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    • 2015
  • Glycine betaine (GB) is one of the compatible solutes that accumulate in the chloroplasts of certain halotolerant plants under salt or cold stress. The codA gene for choline oxidase, the enzyme that converts choline into GB, has been cloned from a soil bacterium Arthrobacter globiformis. We generated transgenic sweetpotato plants [Ipomoea batatas (L.) Lam] expressing codA gene in chloroplasts under the control of the SWPA2 promoter (referred to as SC plants) and evaluated SC plants under oxidative and drought stresses. SC plants showed enhanced tolerance to methyl viologen (MV)-mediated oxidative stress and drought stress due to induced expression of codA. At $5{\mu}M$ of MV treatment, all SC plants showed enhanced tolerance to MV-mediated oxidative stress through maintaining low ion leakage and increased GB levels compared to wild type plants. When plants were subjected to drought conditions, SC plants showed enhanced tolerance to drought stress through maintaining high relative water contents and increased codA expression compared to wild type plants. These results suggest that the SC plants generated in this study will be useful for enhanced biomass production on global marginal lands.

Comparative Assessment of Specific Genes of Bacteria and Enzyme over Water Quality Parameters by Quantitative PCR in Uncontrolled Landfill (정량 PCR을 이용한 비위생 매립지의 특정 세균 및 효소 유전자와 수질인자와의 상관관계 평가)

  • Han, Ji-Sun;Sung, Eun-Hae;Park, Hun-Ju;Kim, Chang-Gyun
    • Journal of Korean Society of Environmental Engineers
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    • v.29 no.8
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    • pp.895-903
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    • 2007
  • As for the increasing demanding on the development of direct-ecological landfill monitoring methods, it is needed for critically defining the condition of landfills and their influence on the environment, quantifying the amount of enzymes and bacteria mainly concerned with biochemical reaction in the landfills. This study was thus conducted to understand the fates of contaminants in association with groundwater quality parameters. For the study, groundwater was seasonally sampled from four closed unsanitary landfills(i.e. Cheonan(C), Wonju(W), Nonsan(N), Pyeongtaek(P) sites) in which microbial diversity was simultaneously obtained by 16S rDNA methods. Subsequently, a number of primer sets were prepared for quantifying the specific gene of representative bacteria and the gene of encoding enzymes dominantly found in the landfills. The relationship between water quality parameters and gene quantification were compared based on correlation factors. Correlation between DSR(Sulfate reduction bacteria) gene and BOD(Biochemical Oxygen Demand) was greater than 0.8 while NSR(Nitrification bacteria-Nitrospira sp.) gene and nitrate were related more than 0.9. A stabilization indicator(BOD/COD) and MTOT(Methane Oxidation bacteria), MCR(Methyl coenzyme M reductase), Dde(Dechloromonas denitrificans) genes were correlated over 0.8, but ferric iron and Fli(Ferribacterium limineticm) gene were at the lowest of 0.7. For MTOT, it was at the highest related at 100% over BOD/COD. In addition, anaerobic genes(i.e., nirS-Nitrite reductase, MCR. Dde, DSR) and DO were also related more than 0.8, which showing anaerobic reactions generally dependant upon DO. As demonstrated in the study, molecular biological investigation and water quality parameters are highly co-linked, so that quantitative real-time PCR could be cooperatively used for assessing landfill stabilization in association with the conventional monitoring parameters.

The Pulation Structure of the Pacific Cod (Gadus macrocephalus Tilesius) Based on Mitochondrial DNA Sequences (미토콘드리아 유전자 염기서열 분석에 의한 대구 계군 분석)

  • Seo, Young-Il;Kim, Joo-Il;Oh, Taeg-Yun;Lee, Sun-Kil;Park, Jong-Hwa;Kim, Hee-Yong;Cho, Eun-Seob
    • Journal of Life Science
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    • v.20 no.3
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    • pp.336-344
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    • 2010
  • To assess population structure and genetic diversity among the Pacific cod (Gadus macrocephalus), we investigated mtDNA COI gene sequences of 7 populations. Samples were obtained from Sokcho, Wolsung, Geojedo, Yeosu, Geomundo and Westsouth in 2008 and 2009 (n=28). The sequence analysis of 28 individual samples showed 8 haplotypes, ranging in sequence divergence by pairwise comparisons from 0.2 to 2.2% (1 bp-11 bp). The Gal haplotype was found in Wolsung, Geojedo, Yeosu, Geomundo and Westsouth, and was regarded as the main haplotype of Korean Pacific cod. Ga2, Ga3, Ga6 and Ga7 haplotypes were found only in Sokcho. In the PHYLIP analysis, 8 haplotypes formed two independent groups: cladeA consisted of Ga2, Ga3, Ga6 and Ga7 haplotypes, whereas cladeB contained Gal, Ga4, Ga5 and Ga8 haplotypes. The genetic relationship between the two groups was weakly supported by bootstrap analysis(<50%). In pairwise comparisons between 6 populations other than that from Sokcho, a very high per generation migration ratio ($N_m$=infinite) and a very low level of geographic distance ($F_{sr}=-0.0123-(-0.0423)$) were observed. The estimates of genetic distance between Sokcho and the other localities were all statistically significant (p<0.05, p<0.01, p<0.001), indicating a limited mtDNA-based gene flow between Sokcho and other regions. The finding of the lowest genetic diversity in the Sokcho population (nucleotide diversity=0.00589) may be a result of relatively small population size and interrupted gene flow to other localities. Consequently, the overall considerable migration of Pacific cod population in Korea caused a genetically homogeneous structure to form, although a distinct population was found in this study.

Identification of Red Tide-causing Organism and Characteristics of Red Tide Occurrence in the Oncheon Down Stream, Busan (온천천 하류 적조 원인생물의 동정 및 발생 특성)

  • Kim, Mi-hee;Ji, Hwa-seong;Cho, Jeong-goo;Cho, Sunja
    • Journal of Korean Society on Water Environment
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    • v.34 no.3
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    • pp.285-292
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    • 2018
  • This study was performed in order to identify the red tide-causing organism and to understand the characteristics of the water quality during the winter of 2015 and 2016 in the Oncheon stream, a tidal river in Busan, where red tide often occurs in the wintertime. Two sites were selected on the stream and the surface water was sampled a total of 28 times during the experimental period. Twelve water quality characteristics, including water temperature, pH, DO, COD, total-N (T-N), total-P (T-P), and salinity were analyzed in order to test water quality. The cell numbers of cryptomonads were counted directly by microscopic observation. The nucleotide sequences of the partial 28S rRNA gene and psbA gene from metagenomic DNA, derived from each sampling site, were analyzed. According to the results, the alga most responsible for the bloom was identified as Teleaulax OC1 sp., which belongs to the cryptomonads. Three items of chl-a, pH, and DO were positively correlated with the cell numbers of the cryptomonads counted at the upper stream of the tidal area (St 1) while eight items of chl-a, TOC, BOD, total-N, COD, SS, pH, and DO were positively correlated with the cells located at the junction between the stream and Su-young river (St 2) in the order.

New nirS-Harboring Denitrifying Bacteria Isolated from Activated Sludge and Their Denitrifying Functions in Various Cultures

  • LEE, SOO-YOUN;LEE, SANG-HYON;PARK, YONG-KEUN
    • Journal of Microbiology and Biotechnology
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    • v.15 no.1
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    • pp.14-21
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    • 2005
  • By using PCR with nirS gene primers, three nirSharboring denitrifying bacteria (strain N6, strain N23, and strain R13) were newly isolated from activated sludge of a weak municipal wastewater treatment plant. Small-subunit rRNA gene-based analysis indicated that strain N6, strain N23, and strain R13 were closely related to Arthrobacter sp.,Staphylococcus sp., and Bacillus sp., respectively. In an attempt to identify their roles in biological nitrate and nitrite removal from sewage, we investigated their specific denitrification rates (SDNRs) for $NO_-^3$ - and $NO_-^2$ - in various cultures. All purecultures of each isolated nirS-harboring bacterial strain could remove $NO_-^3$ - and $NO_-^2$ - simultaneously in high efficiency, and the carbon requirements for $NO_-^3$ - removal of strain N6 and strain R13 were effectively low at 3.1 and 4.1 g COD/g $NO_3N$, respectively. In the case of mix-cultures of the strains (N6+N23, N6+R13, N23+R13, and N6+N23+R13), their SDNRs for $NO_-^3$ - were also effective, and their carbon requirements for $NO_-^3$ - removal were also effective at 3.0- 3.8 g COD/g NO3N. However, all tested mix-cultures accumulated $NO_-^2$ - in their culture media. On the other hand, the continuous culture of activated sludge mixed with strain N6 showed no significant increase of $NO_-^3$ - removal in comparison with strain N6's pure culture. These results suggest that nitrate and nitrite removal in biological wastewater treatment might be dependent on complicated bacterial interactions, including several effective denitrifying bacteria isolated in this study, rather than the specific bacterial types present and the number of bacterial types in activated sludge.

Municipal Wastewater Treatment and Microbial Diversity Analysis of Microalgal Mini Raceway Open Pond (미세조류 옥외 배양시스템을 이용한 도시하수 정화 및 미생물 군집다양성 분석)

  • Kang, Zion;Kim, Byung-Hyuk;Shin, Sang-Yoon;Oh, Hee-Mock;Kim, Hee-Sik
    • Korean Journal of Microbiology
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    • v.48 no.3
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    • pp.192-199
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    • 2012
  • Microalgal biotechnology has gained prominence because of the ability of microalgae to produce value-added products including biodiesel through photosynthesis. However, carbon and nutrient source is often a limiting factor for microalgal growth leading to higher input costs for sufficient biomass production. Use of municipal wastewater as a low cost alternative to grow microalgae as well as to treat the same has been demonstrated in this study using mini raceway open ponds. Municipal wastewater was collected after primary treatment and microalgae indigenous in the wastewater were encouraged to grow in open raceways under optimum conditions. The mean removal efficiencies of TN, TP, COD-$_{Mn}$, $NH_3$-N after 6 days of retention time was 80.18%, 63.56%, 76.34%, and 96.74% respectively. The 18S rRNA gene analysis of the community revealed the presence of Chlorella vulgaris and Scenedesmus obliquus as the dominant microalgae. In addition, 16S rRNA gene analysis demonstrated that Rhodobacter, Luteimonas, Porphyrobacter, Agrobacterium, and Thauera were present along with the microalgae. From these results, it is concluded that microalgae could be used to effectively treat municipal wastewater without aerobic treatment, which incurs additional energy costs. In addition, municipal wastewater shall also serve as an excellent carbon and nitrogen source for microalgal growth. Moreover, the microalgal biomass shall be utilized for commercial purposes.

Microbial Community Analysis in the Wastewater Treatment of Hypersaline-Wastewater (고농도 염분폐수의 정화능이 우수한 기능성 미생물 커뮤니티의 군집 분석)

  • Lee, Jae-Won;Kim, Byung-Hyuk;Park, Yong-Seok;Song, Young-Chae;Koh, Sung-Cheol
    • Microbiology and Biotechnology Letters
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    • v.42 no.4
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    • pp.377-385
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    • 2014
  • In this study, a wastewater treatment system for hypersaline wastewater utilizing the Hypersaline Wastewater Treatment Community (HWTC) has been developed. The hypersaline wastewater treatment efficiency and microbial community of the HWTC were investigated. The average removal efficiencies of chemical oxygen demand were 84% in an HRT of 2.5 days. Microbial community analysis, by denaturing gradient gel electrophoresis (DGGE) of PCR-amplified 16S rRNA gene fragments and 16S rRNA gene clone library, revealed community diversity. The 16S rRNA gene analysis of dominant microbial bacteria in 4% hypersaline wastewater confirmed the presence of Halomonas sp. and Paenibacillus sp. Phylogenetic analysis suggested that the taxonomic affiliation of the dominant species in the HWTC was ${\gamma}$-proteobacteria and firmicutes. These results indicate the possibility that an appropriate hypersaline wastewater treatment system can be designed using acclimated sludge with a halophilic community.

Mutagenic effects of industrial wastewaters by using umu-test (umu-test에 의한 일부 배출시설별 폐수의 변이원성 조사연구)

  • 김영환;손종렬;문영환;배은상
    • Journal of environmental and Sanitary engineering
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    • v.11 no.2
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    • pp.9-20
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    • 1996
  • Genotoxicity/mutagenicity of organic chemicals in industrial wastewater was investigated using umu-test with a Salmonella typhimurium TA1535 strain. The tester strain was derived by introducing plasmid pSK 1002, which carried a umu C - lac Z fusion gene into S typhimurium TA1535, and tester strain in the presence microsomal activation proved to be the more sensitive maker of genotoxicity. Genotoxic responses were observed in concentrated with a blue-rayon column, from 14 plants tested. The results were as follow; 1. Genotoxic responses were observed in concentrated from nine plants(64.3%) tested. 2. The results show that genotoxic activity was particulary high in the untreated wastewaters and decreased in the treated wastewaters(35.7%) 3. No significant correlation was found between genotoxicity and water ollution indicators, such as COD and BOD.

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Molecular cloning and expression analysis of an interferon stimulated gene 15 from rock bream Oplegnathus fasciatus

  • Kim, Ju-Won;Kwon, Mun-Gyeong;Park, Myoung-Ae;Hwang, Jee-Youn;Park, Hyung-Jun;Baeck, Gun-Wook;Kim, Mu-Chan;Park, Chan-Il
    • Journal of fish pathology
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    • v.23 no.2
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    • pp.177-187
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    • 2010
  • The Interferon stimulated gene 15 (ISG15) is strongly induced in many cell types by IFNs, viral infections, and double-stranded RNA (poly I:C). The ISG15 homologue cDNA was isolated from the rock bream LPS stimulated leukocyte cDNA library. The rock bream ISG15 homologue was found to consist of 833 bp encoding 157 amino acid residues. Compared with other known ISG15 peptide sequences, the most conserved regions of the rock bream ISG15 peptide were found to be the tandem ubiquitin-like domains and a C-terminal LRLRGG conjugating motif, characteristic of mammalian and non-mammalian ISG15 proteins. Phylogenetic analysis based on the deduced amino acid sequence revealed a homologous relationship between the ISG15 sequence of rock bream and that of Atlantic salmon, Atlantic cod, northern snake head, black rockfish and olive flounder. The expression of the rock bream ISG15 molecule was induced in the peripheral blood leukocytes (PBLs) from 1 to 24 h following poly I:C stimulation, with a peak at 3 h post-stimulation. The rock bream ISG15 gene was predominantly expressed in the PBLs, spleen and gill.

Molecular Sexing and Species Identification of the Processed Meat and Sausages of Horse, Cattle and Pig

  • Kim, Yoo-Kyung;Kang, Yong-Jun;Kang, Geun-Ho;Seong, Pil-Nam;Kim, Jin-Hyoung;Park, Beom-Young;Cho, Sang-Rae;Jeong, Dong Kee;Oh, Hong-Shik;Cho, In-Cheol;Han, Sang-Hyun
    • Journal of Embryo Transfer
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    • v.31 no.1
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    • pp.61-64
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    • 2016
  • We developed a polymerase chain reaction (PCR)-based molecular method for sexing and identification using sexual dimorphism between the Zinc Finger-X and -Y (ZFX-ZFY) gene and polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) for mitochondrial DNA (mtDNA) cytochrome B (CYTB) gene in meat pieces and commercial sausages from animals of different origins. Sexual dimorphism based on the presence or absence of SINE-like sequence between ZFX and ZFY genes showed distinguishable band patterns between male and female DNA samples and were easily detected by PCR analyses. Male DNA had two PCR products appearing as distinct two bands (ZFX and ZFY), and female DNA had a single band (ZFX). Molecular identification was carried out using PCR-RFLP of CYTB gene, and showed clear species classification results. The results yielded identical information on the sexes and the species of the meat samples collected from providers without any records. The analyses for DNA isolated from commercial sausage showed that pig was the major source but several sausages originated from chicken and Atlantic cod. Applying this PCR-based molecular method was useful and yielded clear sex information and identified the species of various tissue samples originating from livestock.