• 제목/요약/키워드: restriction fragment length polymorphism(RFLP)

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Molecular Characterization of Burkholderia cepacia Complex Isolates Causing Bacterial Fruit Rot of Apricot

  • Li, Bin;Fang, Yuan;Zhang, Guoqing;Yu, Rongrong;Lou, Miaomiao;Xie, Guanlin;Wang, Yanli;Sun, Guochang
    • The Plant Pathology Journal
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    • 제26권3호
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    • pp.223-230
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    • 2010
  • The Burkholderia cepacia complex isolates causing bacterial fruit rot of apricot were characterized by speciesspecific PCR tests, recA-HaeIII restriction fragment length polymorphism (RFLP) assays, rep-PCR genomic fingerprinting, recA gene sequencing, and multilocus sequence typing (MLST) analysis. Results indicated that the isolates Bca 0901 and Bca 0902 gave positive amplifications with primers specific for B. vietnamiensis while the two bacterial isolates showed different recA-RFLP and rep-PCR profiles from those of B. vietnamiensis strains. In addition, the two bacterial isolates had a higher proteolytic activity compared with that of the non-pathogenic B. vietnamiensis strains while no cblA and esmR marker genes were detected for the two bacterial isolates and B. vietnamiensis strains. The two bacterial isolates were identified as Burkholderia seminalis based on recA gene sequence analysis and MLST analysis. Overall, this is the first characterization of B. seminalis that cause bacterial fruit rot of apricot.

MUTYH Association with Esophageal Adenocarcinoma in a Han Chinese Population

  • Kong, Feng;Han, Xue-Ying;Luan, Yun;Qi, Tong-Gang;Sun, Chao;Wang, Jue;Hou, Hua-Ying;Jiang, Yu-Hua;Zhao, Jing-Jie;Cheng, Guang-Hui
    • Asian Pacific Journal of Cancer Prevention
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    • 제14권11호
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    • pp.6411-6413
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    • 2013
  • Adenocarcinoma of esophagus (AE) is a complex disease, affected by a variety of genetic and environmental factors. Much evidence has shown that the MutY glycosylase homologue (MUTYH) plays a key role in the pathogenesis of many cancers. However, there have been no reports on influence on AE in the Han Chinese population. The objective of this study was to investigate this issue. A gene-based association study was conducted using three single nucleotide polymorphisms(SNPs) reported in previous studies. The three SNPs (rs3219463, rs3219472, rs3219489) were genotyped in 207 unrelated AE patients and 249 healthy controls in a case-control study using polymerase chain reaction and restriction fragment length polymorphism (PCR-RFLP). The results revealed that the genotype distribution of rs3219472 differed between the case and control groups (OR=1.66,95%CI=1.11-2.48, P=0.012), indicating that an association may exist between MUTYH and AE. These findings support a signifcant role for MUTYH in AE pathogenesis in the Han Chinese population.

Rice genotype, parental lineage and physiological tolerance to soil salinity shapes the community structure of rice seed bacterial endophytes

  • Walitang, Denver I.;Kim, Kiyoon;Chatterjee, Poulami;Kang, Yeongyeong;Sa, Tongmin
    • 한국작물학회:학술대회논문집
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    • 한국작물학회 2017년도 9th Asian Crop Science Association conference
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    • pp.342-342
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    • 2017
  • Rice seeds are a home to endophytic bacterial communities which serve as a source of the plant's endophytes. As rice undergo physiological and adaptive modifications through cross breeding in the process of attaining salinity tolerance, this may also lead to changes in the endophytic bacterial community especially those residing in the seeds. This study explores the community structure of seed bacterial endophytes as influenced by rice parental lineage, genotype and physiological adaptation to salinity stress. Endophytic bacterial diversity was studied through culture dependent technique, cloning and Terminal-Restriction Fragment Length Polymorphism (T-RFLP) analysis. Results revealed considerably diverse communities of bacterial endophytes in the interior of rice seeds. The richness of ribotypes ranges from 5-14 T-RFs corresponding to major groups of bacterial endophytes in the seeds. Endophytic bacterial diversity of the salt-sensitive IR29 is significantly more diverse compared to those of salt-tolerant cultivars. Proteobacteria followed by Actinobacteria and Firmicutes dominated the overall endophytic bacterial communities of the indica rice seeds based on 16S rDNA analysis of clones and isolates. Community profiles show common ribotypes found in all cultivars of the indica subspecies representing potential core microbiota belonging to Curtobacterium, Flavobacterium, Enterobacter, Xanthomonas, Herbaspirillum, Microbacterium and Stenotrophomonas. Multivariate analysis showed that the bacterial endophytic community and diversity of rice seeds are mainly influenced by their host's genotype, physiological adaptation to salt stress and parental lineage.

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Soil salinity shifts the community structure and diversity of seed bacterial endophytes of salt-sensitive and tolerant rice cultivars

  • Walitang, Denver I.;Ahmed, Shamim;Jeon, Sunyoung;Pyo, Chaeeun;Sa, Tongmin
    • 한국작물학회:학술대회논문집
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    • 한국작물학회 2017년도 9th Asian Crop Science Association conference
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    • pp.244-244
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    • 2017
  • Soil salinity due to accumulation of salts particularly sodium chloride affects agricultural lands and their vegetation. Generally, rice is a moderately sensitive plant with some cultivars with varying tolerance to salinity. Though there are physiological differences between salt-sensitive and salt-tolerant rice cultivars, both are still affected especially during high salinity and prolonged exposure. This also ultimately affects their indigenous bacterial endophytes particularly those that inhabit the rice seed endosphere. This study investigates the dynamic structure of seed bacterial endophytes of salt-sensitive and tolerant rice cultivars grown in different levels of soil salinity. Endophytic bacterial diversity was studied Terminal-Restriction Fragment Length Polymorphism (T-RFLP) analysis. Results revealed a very interesting pattern of diversity and shifts in community structure of bacterial endophytes in the rice seeds. There is a general decrease in diversity for the salt-sensitive rice cultivar, IR29 as soil salinity increases. For the salt-tolerant cultivars, IC32 and IC37, diversity interestingly increased at moderate salinity then decreased at high soil salinity. The patterns of community structure is also strikingly different for the salt-sensitive and salt-tolerant rice cultivars. IR29 has a more even distribution of abundance, but under soil salinity, the community shifted where Curtobacterium, Pantoea, Flavobacterium and Microbacterium become the more dominant bacterial communities. For IC32 and IC37, the dominant bacterial groups under normal stress conditions were also the dominant bacterial groups during salt stress conditions. Their seed bacterial community is dominated by endophytes belonging to Microbacterium, Flavobacterium, Pantoea, Kosakonia and Enterobacter. Stenotrophomonas and Xanthomonas have not changed in terms of abundance under different salinity stress level in the salt-sensitive and salt-tolerant rice cultivars. This study showed that soil salinity greatly influenced the seed bacterial communities of rice seeds irrespective of their physiological tolerance to salinity.

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Genetic identification of anisakid nematodes isolated from largehead hairtail (Trichiurus japonicus) in Korea

  • Kim, Jeong-Ho;Nam, Woo-Hwa;Jeon, Chan-Hyeok
    • Fisheries and Aquatic Sciences
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    • 제19권5호
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    • pp.26.1-26.8
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    • 2016
  • Background: The nematode species belonging to genus Anisakis occur at their third larval stage in numerous marine teleost fish species worldwide and known to cause accidental human infection through the ingestion of raw or undercooked fish or squids. They may also draw the attention of consumers because of the visual impact of both alive and dead worms. Therefore, the information on their geographical distribution and clear species identification is important for epidemiological survey and further prevention of human infection. Results: For identification of anisakid nematodes species isolated from largehead hairtail (Trichiurus japonicus), polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) analysis of internal transcribed spacers of ribosomal DNA were conducted. Mitochondrial cytochrome c oxidase subunit 2 gene was also sequenced, and phylogenetic analysis was conducted. From the largehead hairtail (n = 9), 1259 nematodes were isolated in total. Most of the nematodes were found encapsulated throughout the viscera (56.2 %, 708/1259) or moving freely in the body cavity (41.5 %, 523/1259), and only 0.3 % (4/1259) was found in the muscles. By PCR-RFLP, three different nematode species were identified. Anisakis pegreffii was the most dominantly found (98.7 %, 1243/1259) from the largehead hairtail, occupying 98.7 % (699/708) of the nematodes in the mesenteries and 98.1 % (513/523) in the body cavity. Hybrid genotype (Anisakis simplex ${\times}$ A. pegreffii) occupied 0.5 %, and Hysterothylacium sp. occupied 0.2 % of the nematodes isolated in this study. Conclusions: The largehead hairtail may not significantly contribute accidental human infection of anisakid nematode third stage larvae because most of the nematodes were found from the viscera or body cavity, which are not consumed raw. But, a high prevalence of anisakid nematode larvae in the largehead hairtail is still in concern because they may raise food safety problems to consumers. Immediate evisceration or freezing of fish after catch will be necessary before consumption.

Stem Rot on Adzuki Bean (Vigna angularis) Caused by Rhizoctonia solani AG 4 HGI in China

  • Sun, Suli;Xia, Changjian;Zhang, Jiqing;Duan, Canxing;Wang, Xiaoming;Wu, Xiaofei;Lee, Suk-Ha;Zhu, Zhendong
    • The Plant Pathology Journal
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    • 제31권1호
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    • pp.67-71
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    • 2015
  • During late August and early September 2011, stem rot symptoms were observed on adzuki bean plants (Vigna angularis) growing in fields located in Beijing and Hebei Province, China, respectively. In this study, four isolates were obtained from infected stems of adzuki bean plants. Based on their morphology, and sequence and polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) analyses of the ribosomal DNA internal transcribed spacers (rDNA-ITS) region, the four isolates were identified as Rhizoctonia solani in anastomosis group (AG) 4 HGI. Pathogenicity tests showed that all isolates were strongly pathogenic to adzuki bean and resulted in serious wilt symptoms which was similar to observations in the fields. Additionally, the isolates infected several other crops and induced related rot on the roots and basal stems. To our knowledge, this is the first report of Rhizoctonia solani AG 4 HGI causing stem rot on adzuki bean.

Polychlorobiphenyl (PCB) 토양오염복원: PCB 제거 토양미생물들의 군집과 기능을 효과적으로 분석하는 신 genomics 방법개발에 관한 연구

  • 박준홍
    • 한국지하수토양환경학회:학술대회논문집
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    • 한국지하수토양환경학회 2005년도 총회 및 춘계학술발표회
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    • pp.28-30
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    • 2005
  • Because of high population diversity in soil microbial communities, it is difficult to accurately assess the capability of biodegradation of toxicant by microbes in soil and sediment. Identifying biodegradative microorganisms is an important step in designing and analyzing soil bioremediation. To remove non-important noise information, it is necessary to selectively enrich genomes of biodegradative microorganisms fromnon-biodegradative populations. For this purpose, a stable isotope probing (SIP) technique was applied in selectively harvesting the genomes of biphenyl-utilizing bacteria from soil microbial communities. Since many biphenyl-using microorganisms are responsible for aerobic PCB degradation In soil and sediments, biphenyl-utilizing bacteria were chosen as the target organisms. In soil microcosms, 13C-biphenyl was added as a selective carbon source for biphenyl users, According to $13C-CO_2$ analysis by GC-MS, 13C-biphenyl mineralization was detected after a 7-day of incubation. The heavy portion of DNA(13C-DNA) was separated from the light portion of DNA (12C-DNA) using equilibrium density gradient ultracentrifuge. Bacterial community structure in the 13C-DNAsample was analyzed by t-RFLP (terminal restriction fragment length polymorphism) method. The t-RFLP result demonstates that the use of SIP efficiently and selectively enriched the genomes of biphenyl degrading bacteria from non-degradative microbes. Furthermore, the bacterial diversity of biphenyl degrading populations was small enough for environmental genomes tools (metagenomics and DNA microarrays) to be used to detect functional (biphenyl degradation) genes from soil microbial communities, which may provide a significant progress in assessing microbial capability of PCB bioremediation in soil and groundwater.

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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
    • 한국수정란이식학회지
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    • 제31권1호
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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.

Genetic Diversity of Sweet potato feathery mottle virus from Sweet Potatoes in Korea

  • Kwak, Hae-Ryun;Kim, Mi-Kyeong;Jung, Mi-Nam;Lee, Su-Heon;Park, Jin-Woo;Kim, Kook-Hyung;Ko, Sug-Ju;Choi, Hong-Soo
    • The Plant Pathology Journal
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    • 제23권1호
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    • pp.13-21
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    • 2007
  • Sweet potato feathery mottle virus(SPFMV) is one of the most prevalent viruses infecting sweet potatoes and occurs widely in sweet potato cultivating areas in Korea. To assess their genetic variation, a total of 28 samples infected with SPFMV were subjected to restriction fragment length polymorphism(RFLP) analysis using DNAs amplified by RT-PCR with specific primer sets corresponding to the coat protein(CP) region of the virus. The similarity matrix by UPGMA procedure indicated that 28 samples infected with SPFMV were classified into three groups based on the number and size of DNA fragments by digestion of CP-encoding regions with 7 enzymes including SalI, AluI, EcoRI, HindIII, FokI, Sau3AI, and DraI bands. Four primer combinations out of 5 designed sets were able to differentiate SPFMV and sweet potato virus G infection, suggesting that these specific primers could be used to differentiate inter-groups of SPFMV. Sequence analysis of the CP genes of 17 SPFMV samples were 97-99% and 91-93% identical at the intra-group and inter-groups of SPFMV, respectively. The N-terminal region of the CP is highly variable and examination of the multiple alignments of amino acid sequences revealed two residues(residues 31 and 32) that were consistently different between SPFMV-O and SPFMV-RC.

일천궁과 토천궁에서 발생하는 총채벌레류의 분자동정 (Molecular Identification of Thrips in Two Medicinal Crops, Cnidium officinale Makino and Ligusticum chuanxiong Hort)

  • 정충렬;정대희;박홍우;김현준;전권석;윤정범
    • 한국약용작물학회지
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    • 제27권1호
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    • pp.17-23
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    • 2019
  • Background: Cnidium officinale Makino and Ligusticum chuanxiong Hort. are important medicinal crops in Korea. However, there is insufficient information on the identification of thrips, which attack these plants. Until now, one species of thrips has been recorded as a main pest. Methods and Results: To identify the thrips emerging in C. officinale Makino and L. chuanxiong Hort., these plants were independently cultivated in two local areas. Thirty individuals of each plant species were selected randomly and surveyed for the presence of thrips. After confirming the existence of thrips, 100 thrips individuals were collected from each crop using the beating method. To identify thrips species, we performed PCR-restriction fragment length polymorphism (RFLP)-based analysis using ITS2 primer sets. Six thrips species were identified: western flower thrips (Frankliniella occidentalis), flower thrips (F. intonsa), onion thrips (Thrips tabaci), chrysanthemum thrips (T. nigropilosus), chilli thrips (Scirtothrips dorsalis), and grass thrips (Anaphothrips obscurus). The proportion of these species differed between the host plant species. Conclusions: Six thrips species were major pests of two medicinal crops. Integrated pest management is required to control these thrips species, and will enhance the yield and quality of C. officinale and L. chuanxiong.