Kim, Yangseon;Goh, Jaeduk;Kang, Injeong;Shim, Hyeongkwon;Heu, Sunggi;Roh, Jaehwan
Proceedings of the Korean Society of Crop Science Conference
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2017.06a
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pp.112-112
/
2017
Rice blast caused by Magnaporthe oryzae is one of the most serious diseases that affect the quantity and quality of rice production. The use of resistant rice varieties would be the most effective way to control the rice blast. However R gene incorporation into the rice variety takes time and pathogen could overcome the R gene effects after for a while. For monitoring the rice blast resistance gene distribution in Korean varieties, the four major blast resistance genes against M. oryzae were screened in a number of Korean rice varieties using molecular markers. Of the 120 rice varieties tested, 40 were found to contain the Pi-5 gene, 25 for the Pi-9 gene, 79 for Pi-b and 40 for the Pi-ta gene. None of these rice varieties includes tested 4 R genes. 3 R genes combination, Pi-5/Pi-9/Pi-b, Pi-5, Pi-9.Pi-ta, or Pi-9/Pi-b/Pi-ta were found in 12 varieties, the rice blast disease severity were showed as resistant in the rice verities containing Pi-9/Pi-b/Pi-ta R genes combination, respectively. Also pathogenic diversity of M. oryzae isolates collected in the rice field from 2004 to 2015 in rice field in Korea were analyzed using rice blast monogenic lines, each harboring a single blast resistance gene. Compatibility of blast isolates against rice blast monogenic lines carrying the resistance genes Pi5, Pi9, Pib, and Piz showed dynamic changes by year. It indicates that pathogen has high evolutionary potential adapted host resistances to increase fitness and would lead to rice blast resistance bred into the cultivar becoming ineffective eventually.
Poly-ADP-ribosylation (PAR) is a post-translational modification of mainly chromosomal proteins. It is known to be strongly involved in several molecular events, including nucleosome-remodelling and carcinogenesis. In this investigation, it was attempted to evaluate PAR level as a reliable biomarker for early detection of cancer in blood lymphocyte histones. PAR of isolated histone proteins was monitored in normal and dimethylnitrosamine (DMN)-exposed mice tissues using a novel ELISA-based immuno-probe assay developed in our laboratory. An inverse relationship was found between the level of PAR and period of DMN exposure in various histone proteins of blood lymphocytes and spleen cells. With the increase in the DMN exposure period, there was reduction in the PAR level of individual histones in both cases. It was also observed that the decrease in the level of PAR of histones resulted in progressive relaxation of genomic DNA, perhaps triggering activation of genes that are involved in initiation of transformation. The observed effect of carcinogen on the PAR of blood lymphocyte histones provided us with a handy tool for monitoring biochemical or physiological status of individuals exposed to carcinogens without obtaining biopsies of cancerous tissues, which involves several medical and ethical issues. Obtaining blood from any patient and separating blood lymphocytes are routine medical practices involving virtually no medical intervention, post-procedure medical care or trauma to a patient. Moreover, the immuno-probe assay is very simple, sensitive, reliable and cost-effective. Therefore, combined with the ease of preparation of blood lymphocytes and the simplicity of the technique, immuno-probe assay of PAR has the potential to be applied for mass screening of cancer. It appears to be a promising step in the ultimate goal of making cancer detection simple, sensitive and reliable in the near future.
Bacteria have been regarded as a one of major etiologic factors in root canal infections. In endodontic treatment the effective removal of pathogenic microorganisms in the root canal is the key to successful outcome. Bacterial cell wall components may play an important role in the development of pulpal and periapical disease. The purpose of this study was to evaluate the effect of sonic extracts of Streptococcus spp. on cultured L929 cells and to characterize cell wall protein profiles of Streptococcus spp. Streptococcus spp. were isolated from infected root canals and identified with Vitek Systems(Biomeriux, USA). Five streptococci, namely S. sanguis, S. mitis, S uberis, S. mutans (ATCC 10449) and S. faecalis (ATCC 19433) weere enriched in brain heart infusion broth. Cell pellets were sonicated and cell wall extracts were dialyzed and membrane filtered. Prepared cell wall proteins were applied to cultured L929 cell. The cell reaction were evaluated by monitoring DNA synthesis, cell numbers and the change of cell morphology. The total cell wall protein profiles of microorganisms were characterized by sodium dodecyl sulfate polyacrylamide-gel eledruphoresis(SDS-PAGE). DNA synthesis of L929 cells were reduced by the increasing concentration of sonic extracts. DNA synthesis was significantly suppressed in more than $50{\mu}g$/ml of sonic extract conentration in five streptococci. S. nutans (ATCC 10449) showed stronger suppression on DNA synthesis than remaining four streptococci, which had the similar effect on DNA synthesis. Analysis of DNA synthesis measured by [$^3H$]-thymidine uptake was more sensitvie method than cell counting. Sonic extracts affected the microscopic findings of L929 cells. The protein profiles indicated that all five strains shared two major proteins with molecular masses of 70.8 and 57.5 kD respectively. S. uberis and S. mutans shared common minor proteins of which molecular weights were 147.9 and 112.2 kD respectively. However some minor proteins were unique for S. mitis, S. uberis and S. faecalis.
Proceedings of the Korea Society of Environmental Toocicology Conference
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2002.10a
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pp.156-156
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2002
The goal of this study is to develop a biomarker used in monitoring abnormal behaviors of Japanese medaka (Oryzias latipes) as a model organism caused by hazardous chemicals that are toxic and persistent in the ecosystem. A widely used insecticide, diazinon (O, O-diethyl O- (2-isopropyl-4-methyl-6-pyrimidinyl) phosphorothioate), is highly neurotoxic to fish, and it is also well known that it causes vertebral malformation and behavioral changes of fish at relatively low concentrations. The fish behaviors were observed on a real time basis using an image processing and automatic data acquisition system. The genes potentially involved in the abnormal behaviors were cloned using suppression subtractive hybridization (SSH) technique. The untreated individuals showed common behavioral characteristics. When the test fish was affected by diazinon at a concentration of 0.1 and 1 ppm, some specific patterns were observed in its behavioral activity and locomotive tracks. The typical patterns were enhanced surfacing activity, opercular movement, erratic movement, tremors and convulsions as reported previously. The number of genes up-regulated tty diazinon treatment were 97 which includes 27 of unknown genes. The number of down-regulated genes were 99 including 60 of unknown genes. These gene expression patterns will be analyzed by the artificial neural networks such as self organization map (SOM) and multilayer perceptron (MLP), revealing the role of genes responsible for the behaviors. These results may provide molecular biological and neurobehavioral bases of a biomonitoring system for diazinon using a model organism such as fish.
An aim of current study was to investigate the prevalence and the mechanism of quinolone-resistance in E. coli isolates obtained from chicken cecum in Korea. In addition, multilocus sequence typing (MLST) was also performed for the molecular characterization of E. coli isolates. In an antimicrobial susceptibility test by the disk diffusion method, the 63.5% (54/85) of E. coli isolates showed the resistance to quinolone group of antimicrobial agents. All of the 54 E. coli isolates showing resistant to quinolone group had sense mutations in gyrA gene and point mutations at the $57^{th}$, $80^{th}$, or $84^{th}$ residues in parC gene were detected in 90.7% of the isolates. Interestingly, E. coli ST was closely related to amino acid substitutions in parE gene. Our results indicated that the long-term use of antimicrobial agents in food-producing animals was strongly associated with a prevalence of antimicrobial resistance in commensal Enterobacteriaceae, suggesting the need for continuous surveillance and monitoring of antimicrobial resistant determinants in bacterial isolates from food animals.
Lee, Myungook;Ahn, Jong Il;Lee, Ah Ran;Ko, Dong Woo;Yang, Woo Sub;Lee, Gene;Ahn, Ji Yeon;Lim, Jeong Mook
Molecules and Cells
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v.40
no.8
/
pp.558-566
/
2017
Regular monitoring on experimental animal management found the fluctuation of ART outcome, which showed a necessity to explore whether superovulation treatment is responsible for such unexpected outcome. This study was subsequently conducted to examine whether superovulation treatment can preserve ultrastructural integrity and developmental competence of oocytes following oocyte activation and embryo culture. A randomized study using mouse model was designed and in vitro development (experiment 1), ultrastructural morphology (experiment 2) and functional integrity of the oocytes (experiment 3) retrieved after PMSG/hCG injection (superovulation group) or not (natural ovulation; control group) were evaluated. In experiment 1, more oocytes were retrieved following superovulation than following natural ovulation, but natural ovulation yielded higher (p < 0.0563) maturation rate than superovulation. The capacity of mature oocytes to form pronucleus and to develop into blastocysts in vitro was similar. In experiment 2, a notable (p < 0.0186) increase in mitochondrial deformity, characterized by the formation of vacuolated mitochondria, was detected in the superovulation group. Multivesicular body formation was also increased, whereas early endosome formation was significantly decreased. No obvious changes in other microorganelles, however, were detected, which included the formation and distribution of mitochondria, cortical granules, microvilli, and smooth and rough endoplasmic reticulum. In experiment 3, significant decreases in mitochondrial activity, ATP production and dextran uptake were detected in the superovulation group. In conclusion, superovulation treatment may change both maturational status and functional and ultrastuctural integrity of oocytes. Superovulation effect on preimplantation development can be discussed.
Lee, Kyung-Lak;Shin, Yuna;Lee, Jaean;Lee, Jae-Kwan;Kim, Han Soon
Korean Journal of Ecology and Environment
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v.49
no.4
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pp.393-399
/
2016
This study was conducted to investigate whether the presence of mcy gene and microcystin production are related to morphological characteristics of Korean Microcystis species. We isolated 6 different species of Microcystis (M. aeruginosa, M. ichthyoblabe, M. flos-aquae, M. novacekii, M. viridis and M. wesenbergii) from drinking water supply dams (Yeongchun, Ankei, Gachang), and used microscopic method for morphological identification, molecular method for amplifying a partial region of mcyB gene and ELISA method for microcystin analysis. In the present study, 80% of M. aeruginosa strains contained mcy gene, followed by 45% (10 strains) of M. icthyoblabe, 33% (1 strain) of M. wesenbergii, and 11% (4 strains) of M. flos-aquae. Each percentage of mcy gene in Microcystis morphospecies was similar to that of microcystin production in Microcystis morophospecies. In conclusion, the present study shows that molecular method using mcy gene primers can be used as an indirect indicator for the monitoring of toxic cyanobacteria (Microcystis).
Self-incompatibility (SI) prevents self-fertilization by inhibiting the pollen tube growth of self-pollen. Molecular analysis has revealed that the S locus comprises a number of genes, such as the S-locus glycoprotein (SLG), the S-locus receptor kinase (SRK), and SP11 (SCR). Although molecular markers related to those genes have been developed, a simple S-haplotype detecting method has not been reported due to the highly polymorphic and relatively small coding regions. In this study, the sequence characterized amplified region (SCAR) markers were used to establish an efficient radish genotyping method. We identified the S-haplotypes of 192 radish accessions using 19 different markers, which proved to be highly reliable. The accessions were assigned to 17 types of S-haplotypes, including 8 types of SRKs and 9 types of SLGs. Since the developed SCAR markers are based on their gene sequences, we could easily identify the S-haplotypes by a single specific band, with the highest frequencies detected for SLG 5, SRK 1, and SLG 1, in order. Among the tested markers, the SLG 1, SRK 1, and SRK 5 markers exhibited high reliability, compared to phenotypic results. Furthermore, we identified the seven types of unreported SLGs using SLG Class -I and -II specific markers. Although the developed SCAR markers still need to be improved for the genotyping of all S-haplotypes, these markers could be helpful for monitoring inbred lines, and for developing the MAS in radish breeding programs.
Human Sapovirus (HuSaV) is one of the major causes of acute gastroenteritis in humans, and it is used as a molecular diagnostic technique based on polymerase chain reaction (PCR) from humans, food, shellfish, and aquatic environments. In this study, the HuSaV diagnosis technique was used in an aquatic environment where a number of PCR inhibitors are included and pathogens, such as viruses, are estimated to exist at low concentration levels. HuSaV-specific primers are improved to detect 38 strains registered in the National Center for Biotechnology Information (NCBI). The established optimal condition and the composition, including the RT-nested PCR primers and SL® Non-specific reaction inhibitor, were found to have 100 times higher sensitivity based on HuSaV plasmid than the previously reported methods (100 ag based on HuSaV plasmid 1 ng/μL). Through an artificial infection test, the developed method was able to detect at least 1 fg/μL of HuSaV plasmid contaminated with total nucleic acid extracted from groundwater. In addition, RT-nested PCR primer sets for HuSaV detection can react, and a positive control is developed to verify false positives. This study is expected to be used as a HuSaV monitoring method in the future and applied to the safety response to HuSaV from water environments.
Jung, Ka-Young;Lee, Yun Kyung;Yoo, HaYoung;Nam, Gui-Sook;Hur, Jin
Journal of Korean Society on Water Environment
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v.37
no.6
/
pp.458-468
/
2021
In this study, we investigated the variations of dissolved organic matter (DOM) in an agricultural reservoir during the monsoon period (June to October, 2020) with respect to the organic carbon concentration (DOC), molecular weight distribution, and optical properties. The monsoon period was divided into three phases - beginning storm (BS), during storm (DS), and after storm (AS). Our results showed significant differences in the concentrations and characteristics of DOM during the summer monsoon. The DOC concentrations were decreased after the monsoon, probably due to a dilution effect. In contrast, increasing trends were observed in the specific UV absorbance (SUVA), and relative abundances of humic-like fluorescence and larger-sized compounds. These observations implied that the large-sized and humic-like organic components with terrestrial origins strongly affected the reservoir DOM after the summer monsoon. Meanwhile, biopolymer size fraction, which is associated with algal activity, became more abundant after the monsoon. These results suggest that DOM with autochthonous sources became dominant as a result of the inflow of nutrients into the reservoir after the storm. Spatial changes in DOM within the reservoir were not pronounced as much as the temporal variations. All taken, it can be concluded that the summer monsoon simply led to the decrease of DOM concentrations while the sources and the quality of DOM underwent substantial changes, which may enrich refractory organic matter in the reservoir. This study reveals the importance of in-depth DOM quality monitoring before and after summer monsoon for effective water quality management in agricultural reservoirs.
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