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Identification and Characterization of Pseudomonas syringae pv. syringae, a Causative Bacterium of Apple Canker in Korea

  • Seunghee, Lee;Wonsu, Cheon;Hyeok Tae, Kwon;Younmi, Lee;Jungyeon, Kim;Kotnala, Balaraju;Yongho, Jeon
    • The Plant Pathology Journal
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    • v.39 no.1
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    • pp.88-107
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    • 2023
  • In the present investigation, bacterial isolates from infected apple trees causing apple canker during winter were studied in the northern Gyeongbuk Province, Korea. The pathogen was identified as Pseudomonas syringae pv. syringae (Pss) through various physiological and biochemical characterization assays such as BIOLOG, gas chromatography of fatty acid methyl esters, and 16S rRNA. Bioassays for the production of phytotoxins were positive for syringopeptin and syringomycin against Bacillus megaterium and Geotrichum candidum, respectively. The polymerase chain reaction (PCR) method enabled the detection of toxin-producing genes, syrB1, and sypB in Pss. The differentiation of strains was performed using LOPAT and GATTa tests. Pss further exhibited ice nucleation activity (INA) at a temperature of -0.7℃, indicating an INA+ bacterium. The ice-nucleating temperature was -4.7℃ for a non-treated control (sterilized distilled water), whereas it was -9.6℃ for an INA- bacterium Escherichia coli TOP10. These methods detected pathogenic strains from apple orchards. Pss might exist in an apple tree during ice injury, and it secretes a toxin that makes leaves yellow and cause canker symptoms. Until now, Korea has not developed antibiotics targeting Pss. Therefore, it is necessary to develop effective disease control to combat Pss in apple orchards. Pathogenicity test on apple leaves and stems showed canker symptoms. The pathogenic bacterium was re-isolated from symptomatic plant tissue and confirmed as original isolates by 16S rRNA. Repetitive element sequence-based PCR and enterobacterial repetitive intergenic consensus PCR primers revealed different genetic profiles within P. syringae pathovars. High antibiotic susceptibility results showed the misreading of mRNA caused by streptomycin and oxytetracycline.

Genomic Analysis of the Carrot Bacterial Blight Pathogen Xanthomonas hortorum pv. carotae in Korea

  • Mi-Hyun Lee;Sung-Jun Hong;Dong Suk Park;Hyeonheui Ham;Hyun Gi Kong
    • The Plant Pathology Journal
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    • v.39 no.4
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    • pp.409-416
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    • 2023
  • Bacterial leaf blight of carrots caused by Xanthomonas hortorum pv. carotae (Xhc) is an important worldwide seed-borne disease. In 2012 and 2013, symptoms similar to bacterial leaf blight were found in carrot farms in Jeju Island, Korea. The phenotypic characteristics of the Korean isolation strains were similar to the type strain of Xhc. Pathogenicity showed symptoms on the 14th day after inoculation on carrot plants. Identification by genetic method was multi-position sequencing of the isolated strain JJ2001 was performed using four genes (danK, gyrB, fyuA, and rpoD). The isolated strain was confirmed to be most similar to Xhc M081. Furthermore, in order to analyze the genetic characteristics of the isolated strain, whole genome analysis was performed through the next-generation sequencing method. The draft genome size of JJ2001 is 5,443,372 bp, which contains 63.57% of G + C and has 4,547 open reading frames. Specifically, the classification of pathovar can be confirmed to be similar to that of the host lineage. Plant pathogenic factors and determinants of the majority of the secretion system are conserved in strain JJ2001. This genetic information enables detailed comparative analysis in the pathovar stage of pathogenic bacteria. Furthermore, these findings provide basic data for the distribution and diagnosis of Xanthomonas hortorum pv. carotae, a major plant pathogen that infects carrots in Korea.

Genetic Stability Analysis of in vitro Regenerated Wolly Grass (Imperata cylindrica 'Rubra') Based on Inter-Simple Sequence Repeats (ISSR) Markers (ISSR 표지에 의한 기내재생 홍띠(Imperata cylindrica 'Rubra')의 유전적 안정성 분석)

  • Ye-Jin Lee;In-jin Kang;Chang-Hyu Bae
    • Proceedings of the Plant Resources Society of Korea Conference
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    • 2020.12a
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    • pp.54-54
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    • 2020
  • 지구온난화에 따라 농업부문 신재생에너지의 중요성이 증대되고 있으며, 화본과 식물은 바이오에너지작물의 중요한 소재를 제공하고 있다. 화본과 식물의 기내대량증식연구의 일환으로 홍띠식물의 기내 재생 식물체의 유전적 안정성에 대한 기초자료를 제공할 목적으로 기내배양으로 재분화시킨 홍띠(Imperata cylindrica 'Rubra') 재분화 식물체 중 녹색체 재생식물체를 대상으로 ISSR 표지를 사용하여 유전적 안정성을 조사하였다. 재분화식물체는 MS (Murashige and Skoog, 1962)배지에 생장조절제를 첨가한 배지에서 배양하였다. 생장점 부위를 적출하여 캘러스를 유도하고(0.1 mg/L 2,4-D와 2 mg/L BA), 캘러스 증식(0.1 mg/L 2,4-D와 0.05 mg/L BA), 신초 재분화( 0.01 mg/L NAA와 2 mg/L BA) 후 MS배지에서 식물체를 양성하고 순화시켰다. 배양은 26±2℃, 25 µmol/m2/s, 14h/10h (day/night) 광조건 하에서 실시하였다. 재분화식물체는 홍띠 및 녹색 재분화식물체 2 종류로 나타났는데, 이는 생장점에서는 홍띠가 분화되었음에도 불구하고 생장점 주변조직에서 유래한 녹새체가 분화된 후 우세하게 자라서 녹색재생체가 우점하는 것으로 추정된다. ISSR 분석은 대조구로 모식물체 홍띠를(8개체), 재분화식물체는 녹색체 중, 1년간 노지포장에서 재배중인 녹색체(10개체)와 실험실내 화분에서 재배중인 시료를(10개체) 사용하였다. ISSR 밴드패턴을 비교한 결과, 재분화체는 실내포트 재배식물체 10.3%, 노지1년 재배식물체 8.3%로 대조구의 4.1%보다 유전적 다형성 비율이 2배 이상 높게 나타났다. 또한 재분화식물체들의 유전적 유사도를 평가하고 군집분석을 실시하였다.

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Clinical and molecular detection of fowl pox in domestic pigeons in Basrah Southern of Iraq

  • Isam Azeez Khaleefah;Hassan M. Al-Tameemi;Qayssar Ali Kraidi;Harith Abdulla Najem;Jihad Abdulameer Ahmed;Haider Rasheed Alrafas
    • Korean Journal of Veterinary Research
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    • v.64 no.1
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    • pp.7.1-7.6
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    • 2024
  • Bird species, particularly poultry and other bird types, including domestic pigeons, are susceptible to fowl pox, a contagious viral disease. The main goal of this study was to validate clinical avipoxvirus diagnoses using molecular analytical methods. The essential components of the investigation were the clinical signs, visible abnormalities, histological changes, and polymerase chain reaction analysis. Twenty out of 120 pigeons had clinical symptoms, which included yellowish crust or nodules near the feet, eyes, and beak. An erosive epidermal lesion and an epidermal acanthotic papular lesion with basal vacuolation were maculopapular evidence associated with significant epidermal hyperkeratosis, as confirmed by histological analysis. In addition, the results showed keratinocyte necrosis beneath the hyperkeratotic epidermal layer, together with superficial and deep dermal perivascular lymphocytic infiltration. In addition, the P4b core protein gene underwent phylogenetic analysis. The sequence analysis results indicated a high degree of similarity across the local strains, with just minor variations observed. Five sample sequences were selected and submitted to the NCBI database. These sequences were identified as OR187728, OR187729, OR187730, OR187731, and OR187732. All the various strains in this research may be classified under clade A of the chicken pox virus phylogenetic classification. This study presents the first description and characterization of pox virus infections in domestic pigeons inside the Basrah governorate.

Comparison study between recovered carbon black and commercial carbon black filled epoxy conductive materials

  • Huai M. Ooi;Pei L. Teh;Cheow K. Yeoh;Wee C. Wong;Chong H. Yew;Xue Y. Lim;Kai K. Yeoh;Nor A. Abdul Rahim;Chun H. Voon
    • Advances in materials Research
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    • v.13 no.3
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    • pp.221-232
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    • 2024
  • Waste tire management and recycling have grown to be significant issues because they bring up a global environmental concern. Thus, turning recycled waste tires into useful products may help tackle the environmental issue. This research aims to study and compare the effect of recycled carbon black (rCB) and commercial carbon black (CB) at certain 15 vol. % of filler loading on the mechanical, thermal, morphology and electrical properties of epoxy/CB composites. For this project, epoxy resin, diethyltoluenediamine (DETDA), recovered carbon black (rCB) and commercial carbon black (CB) graded N330, N550, N660 and N774 were mixed and compared accordingly to the formulation determined. The CB content was dispersed in the epoxy matrix using the mechanical mixing technique. The distribution and dispersion of CB in the epoxy matrix affect the characteristics of the conductive composites. rCB content at 15 vol% was selected at fixed content for comparison purposes due to the optimum value in electrical conductivity results. The flexural strength results followed the sequence of rCB>N774>N660>N550>N330. As for electrical conductivity results, epoxy/N330 exhibited the highest conductivity value, while the others achieved a magnitude of X10-3 due to the highest external surface area of N330. In terms of thermal stability, epoxy/N330 and epoxy/N774 were slightly more stable than epoxy/rCB.

Molecular Characterization and Ontogenetic Expression Patterns of Recombination Activating Genes (RAG1/2) in Marine Medaka Oryzias dancena (바다송사리(Oryzias dancena)의 재조합활성화 유전자 RAG1/2의 분자 특성 및 개체발생학적 발현 패턴)

  • Tae-Su Kim;Juhwan Park;Yoon Kwon Nam;Chan-Hee Kim
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.57 no.3
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    • pp.239-252
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    • 2024
  • Recombination activating genes (RAGs) play a crucial role in initiating V(D)J recombination, which is essential for developing adaptive immunity in vertebrates. In this study, we cloned and characterized RAG1/2 cDNA from the marine medaka Oryzias dancena (OdRAG1/2) and investigated their mRNA expression patterns during ontogenetic developmental stages. The OdRAG1 and OdRAG2 cDNA contained open reading frames (ORFs) encoding proteins containing 1,078 and 531 amino acids, respectively. Multiple sequence alignment and phylogenetic analysis revealed that OdRAG1 and OdRAG2 are highly conserved with their corresponding orthologs, featuring distinct core and non-core regions. Notably, expression analysis showed that, in contrast to other fish RAGs studied, OdRAG1/2 expression peaked at 0 days post-hatching (DPH). Additionally, for the expression of T and B cell differentiation markers, CD3γ and CD20, also peaked at 0 DPH. Collectively, adaptive immunity in O. dancena potentially begins during embryonic development, which is critical for V(D)J recombination and essential immune component development, suggesting the early ontogenetic stage interactions between innate and adaptive immunity.

Genetic Characteristics of Extended-Spectrum Beta-Lactamase-Producing Salmonella Isolated from Retail Meats in South Korea

  • Haiseong Kang;Hansol Kim;Hyochin Kim;Ji Hye Jeon;Seokhwan Kim;Yongchjun Park;Soon Han Kim
    • Journal of Microbiology and Biotechnology
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    • v.34 no.5
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    • pp.1101-1108
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    • 2024
  • Earlier studies have validated the isolation of extended-spectrum beta-lactamase-producing Salmonella (ESBL-Sal) strains from food. While poultry is recognized as a reservoir for Salmonella contamination, pertinent data regarding ESBL-Sal remains limited. Consequently, the Ministry of Food and Drug Safety has isolated Salmonella spp. from retail meat and evaluated their antibiotic susceptibility and genetic characteristics via whole-genome sequencing. To further elucidate these aspects, this study investigates the prevalence, antibiotic resistance profiles, genomic characteristics, and homology of ESBL-Sal spp. obtained from livestock-derived products in South Korean retail outlets. A total of 653 Salmonella spp. were isolated from 1,876 meat samples, including 509 beef, 503 pork, 555 chicken, and 309 duck samples. The prevalence rates of Salmonella were 0.0%, 1.4%, 17.5%, and 28.2% in the beef, pork, chicken, and duck samples, respectively. ESBL-Sal was exclusively identified in poultry meat, with a prevalence of 1.4% in the chicken samples (8/555) and 0.3% in the duck samples (1/309). All ESBL-Sal strains carried the blaCTX-M-1 gene and exhibited resistance to ampicillin, ceftiofur, ceftazidime, nalidixic acid, and tetracycline. Eight ESBL-Sal isolates were identified as S. Enteritidis with sequence type (ST) 11. The major plasmid replicons of the Enteritidis-ST11 strains were IncFIB(S) and IncFII(S), carrying antimicrobial resistance genes (β-lactam, tetracycline, and aminoglycoside) and 166 virulence factor genes. The results of this study provide valuable insights for the surveillance and monitoring of ESBL-Sal in South Korean food chain.

Epidemiological, and molecular investigation of Canine parvovirus-2 infection in Egypt

  • Eman Farag Ammar;Yamen Mohammed Hegazy;Magdy Al-gaabary;Samah M. Mosad;Mohamed Salem;Mohamed Marzok;Fadhel Housawi;Mohamed Al-ali;Abdulrahman Alhaider;Amin Tahoun
    • Journal of Veterinary Science
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    • v.25 no.4
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    • pp.56.1-56.13
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    • 2024
  • Importance: Canine parvovirus enteritis (CPE) is a contagious viral disease of dogs caused by the canine parvovirus-2 (CPV-2) associated with high morbidity and mortality rates. CPV-2 has a high global evolutionary rate. Molecular characterization of CPV-2 and understanding its epidemiology are essential for controlling CPV-2 infections. Objective: This study examined the risk factors and survival outcomes of dogs infected with CPV-2. Molecular characterization of CPV-2 genotypes circulating in Egypt was performed to determine the evolution of CPV-2 nationally and globally. Methods: An age-matched case-control study was conducted on 47 control and 47 CPV-infected dogs. Conditional logistic regression analysis examined the association between the potential risk factors and CPE in dogs. Survival analysis was performed to determine the survival pattern of the infected dogs. Thirteen fecal samples from infected dogs were collected to confirm the CPV genotype by CPV-2 VP2 gene sequencing, assembly of nucleotide sequences, and phylogenic analysis. Results: Unvaccinated and roamer dogs had eight and 2.3 times higher risks of CPV infection than vaccinated dogs and non-roamer dogs, respectively. The risk of death from CPE was high among dogs without routine visits to veterinary clinics and among non-roamer dogs. Molecular characterization of CPV-2 confirmed its genotype identity and relationship with the CPV-2 c and b clade types. Conclusions and Relevance: This study highlights the potential factors for CPE control, especially vaccination and preventing dogs from roaming freely outside houses. Isolated CPV genotypes are closely related to southern Asian genotypes, suggesting a substantial opportunity for global transmission.

Exploring the molecular characteristics, detoxification functions, and immune responses of two glutathione S-transferases in redlip mullet (Liza haematochelia)

  • Jeongeun Kim;Welivitiye Kankanamge Malithi Omeka;Qiang Wan;Jehee Lee
    • Fisheries and Aquatic Sciences
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    • v.27 no.5
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    • pp.314-328
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    • 2024
  • The mechanism for the elimination of xenobiotics undergoes three different phases of reactions in organisms. Among these, glutathione S-transferases (GSTs) are classified as phase II detoxification enzymes, catalyzing the conjugation of electrophilic substrates to glutathione or reduced hydroperoxides. This study aimed to investigate the molecular characteristics, detoxification functions, and immune responses of GST omega (LhGSTO1) and kappa (LhGSTK1) in redlip mullet. The open reading frames of LhGSTO1 (720 bp) and LhGSTK1 (687 bp) encoded proteins of 239 and 228 amino acids, respectively. Sequence analysis revealed that LhGSTO1 and LhGSTK1 possessed GSH-binding sites in their N-terminal domains. Substrate-binding sites in the C-terminal domain were exclusively identified in LhGSTO1. In the tissue-specific transcription profile analysis, both LhGSTO1 and LhGSTK1 were ubiquitously expressed in all tissues of healthy mullets. Temporal expression analysis of LhGSTO1 and LhGSTK1 in the blood showed that their expression was significantly modulated by polyinosinic:polycytidylic (poly I:C), lipopolysaccharide (LPS), and Lactococcus garvieae. Different chemical and cellular assays were performed to assess the detoxification and cellular protective abilities of the two proteins. A substrate specificity test using the recombinant proteins revealed that both proteins possessed specific activity toward 1-chloro-2,4-dinitrobenzene (CDNB). In the disk diffusion assay, the smallest clearance zones were observed for LhGSTO1 and LGSTK1 against CdCl2. In the cell protection assay, both LhGSTO1 and LhGSTK1 showed significant Cd detoxification ability compared to the control. Collectively, these results demonstrate that GST omega and kappa are involved in host defense against immune stimulants and xenobiotics in redlip mullet.

Molecular cloning, identification, transcriptional analysis, and silencing of enolase on the life cycle of Haemaphysalis longicornis (Acari, Ixodidae) tick

  • Md. Samiul Haque;Md. Khalesur Rahman;Mohammad Saiful Islam;Myung-Jo You
    • Parasites, Hosts and Diseases
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    • v.62 no.2
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    • pp.226-237
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    • 2024
  • Ticks, blood-sucking ectoparasites, spread diseases to humans and animals. Haemaphysalis longicornis is a significant vector for tick-borne diseases in medical and veterinary contexts. Identifying protective antigens in H. longicornis for an anti-tick vaccine is a key tick control strategy. Enolase, a multifunctional protein, significantly converts D-2-phosphoglycerate and phosphoenolpyruvate in glycolysis and gluconeogenesis in cell cytoplasm. This study cloned a complete open reading frame (ORF) of enolase from the H. longicornis tick and characterized its transcriptional and silencing effect. We amplified the full-length cDNA of the enolase gene using rapid amplification of cDNA ends. The complete cDNA, with an ORF of 1,297 nucleotides, encoded a 432-amino acid polypeptide. Enolase of the Jeju strain H. longicornis exhibited the highest sequence similarity with H. flava (98%), followed by Dermacentor silvarum (82%). The enolase motifs identified included N-terminal and C-terminal regions, magnesium binding sites, and several phosphorylation sites. Reverse transcription-polymerase chain reaction (RT-PCR) analysis indicated that enolase mRNA transcripts were expressed across all developmental stages of ticks and organs such as salivary gland and midgut. RT-PCR showed higher transcript levels in syn-ganglia, suggesting that synganglion nerves influence enolase's role in tick salivary glands. We injected enolase double-stranded RNA into adult unfed female ticks, after which they were subsequently fed with normal unfed males until they spontaneously dropped off. RNA interference significantly (P<0.05) reduced feeding and reproduction, along with abnormalities in eggs (no embryos) and hatching. These findings suggest enolase is a promising target for future tick control strategies.