• Title/Summary/Keyword: species characterization

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Genome-wide identification and expression profiling of the pectin methylesterase gene family in Citrus sinensis (L.) Osbeck

  • Ho Bang Kim;Chang Jae Oh;Nam-Hoon Kim;Cheol Woo Choi;Minju Kim;Sukman Park;Seong Beom Jin;Su-Hyun Yun;Kwan Jeong Song
    • Journal of Plant Biotechnology
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    • v.49 no.4
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    • pp.271-291
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    • 2022
  • Pectin methylesterase (PME) plays an important role in vegetative and reproductive development and biotic/abiotic stress responses by regulating the degree of methyl-esterification of pectic polysaccharides in the plant cell wall. PMEs are encoded by a large multigene family in higher land plant genomes. In general, the expression of plant PME genes shows tissue- or cell-specific patterns and is induced by endogenous and exogenous stimuli. In this study, we identified PME multigene family members (CsPMEs) from the sweet orange genome and report detailed molecular characterization and expression profiling in different citrus tissues and two fruit developmental stages. We also discussed the possible functional roles of some CsPME genes by comparing them with the known functions of PMEs from other plant species. We identified 48 CsPME genes from the citrus genome. A phylogenetic tree analysis revealed that the identified CsPMEs were divided into two groups/types. Some CsPMEs showed very close phylogenetic relationships with the PMEs whose functions were formerly addressed in Arabidopsis, tomato, and maize. Expression profiling showed that some CsPME genes are highly or specifically expressed in the leaf, root, flower, or fruit. Based on the phylogenetic relationships and gene expression profiling results, we suggest that some CsPMEs could play functional roles in pollen development, pollen tube growth, cross incompatibility, root development, embryo/seed development, stomata movement, and biotic/abiotic stress responses. Our results shed light on the biological roles of individual CsPME isoforms and contribute to the search for genetic variations in citrus genetic resources.

Molecular Characterization of Hantavirus Isolates from Bandicota indica Captured in Indonesia and Thailand (인도네시아와 태국의 Bandicota indica 폐장조직에서 분리된 한타바이러스의 분자생물학적 특징)

  • Chu, Yong-Kyu;Cui, Longzhu;Song, Dae-Yong;Woo, Young-Dae;Praseno, Praseno;Leitmeyer, Katrin;Lee, Ho-Wang
    • The Journal of Korean Society of Virology
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    • v.30 no.3
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    • pp.203-210
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    • 2000
  • Hantaviruses are etiologic agents of hemorrhagic fever with renal syndrome (HFRS) and hantavirus pulmonary syndrome (HPS) in the world. Various hantaviruses were isolated from HFRS patients and several different rodent species in the world. Four hantavirus isolates from Indonesia and three isolates from Thailand among 89 Bandicotas captured in Yogyakarta, east region of Sumatra island, Indonesia and at Chiang Mai in Thailand during 1996 were made through several passages in Vero E6 cells. Viral genome M segment from two Indonesian isolates and three Thailand isolates were amplified using hantavirus generic primers of the M segment and cloned into pCRII vector. The genetic differences were analyzed by comparison of partial sequence of the M segment and antigenic differences were made by IFA. Nucleotide sequence homology of two isolates BC 8, BC 34 from Indonesia and two isolates thai 1322, thai 1330 to Seoul virus was 99% and 96%, respectively, but Thai 1164 was 80%Thai 1164 strain has shown 95% homology to Thai 749 virus. In conclusion it is indicated that two different serotype hantaviruses, Seoul and Thailand, are cocirculating among Bandicota in Thailand, in contrast Seoul serotype virus is circulating in Indonesia.

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Analysis and Characterization of Glutathione Peroxidases in an Environmental Microbiome and Isolated Bacterial Microorganisms

  • Yun-Juan Bao;Qi Zhou;Xuejing Yu;Xiaolan Yu;Francis J. Castellino
    • Journal of Microbiology and Biotechnology
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    • v.33 no.3
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    • pp.299-309
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    • 2023
  • Glutathione peroxidases (Gpx) are a group of antioxidant enzymes that protect cells or tissues against damage from reactive oxygen species (ROS). The Gpx proteins identified in mammals exhibit high catalytic activity toward glutathione (GSH). In contrast, a variety of non-mammalian Gpx proteins from diverse organisms, including fungi, plants, insects, and rodent parasites, show specificity for thioredoxin (TRX) rather than GSH and are designated as TRX-dependent peroxiredoxins. However, the study of the properties of Gpx in the environmental microbiome or isolated bacteria is limited. In this study, we analyzed the Gpx sequences, identified the characteristics of sequences and structures, and found that the environmental microbiome Gpx proteins should be classified as TRX-dependent, Gpx-like peroxiredoxins. This classification is based on the following three items of evidence: i) the conservation of the peroxidatic Cys residue; ii) the existence and conservation of the resolving Cys residue that forms the disulfide bond with the peroxidatic cysteine; and iii) the absence of dimeric and tetrameric interface domains. The conservation/divergence pattern of all known bacterial Gpx-like proteins in public databases shows that they share common characteristics with that from the environmental microbiome and are also TRX-dependent. Moreover, phylogenetic analysis shows that the bacterial Gpx-like proteins exhibit a star-like radiating phylogenetic structure forming a highly diverse genetic pool of TRX-dependent, Gpx-like peroxidases.

Crystal Structure and Biochemical Analysis of a Cytochrome P450 Steroid Hydroxylase (BaCYP106A6) from Bacillus Species

  • Ki-Hwa Kim;Hackwon Do;Chang Woo Lee;Pradeep Subedi;Mieyoung Choi;Yewon Nam;Jun Hyuck Lee;Tae-Jin Oh
    • Journal of Microbiology and Biotechnology
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    • v.33 no.3
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    • pp.387-397
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    • 2023
  • Cytochrome P450 (CYP) is a heme-containing enzyme that catalyzes hydroxylation reactions with various substrate molecules. Steroid hydroxylases are particularly useful for effectively introducing hydroxyl groups into a wide range of steroids in the pharmaceutical industry. This study reports a newly identified CYP steroid hydroxylase (BaCYP106A6) from the bacterium Bacillus sp. and characterizes it using an in vitro enzyme assay and structural investigation. Bioconversion assays indicated that BaCYP106A1 catalyzes the hydroxylation of progesterone and androstenedione, whereas no or low conversion was observed with 11β-hydroxysteroids such as cortisol, corticosterone, dexamethasone, and prednisolone. In addition, the crystal structure of BaCYP106A6 was determined at a resolution of 2.8 Å to investigate the configuration of the substrate-binding site and understand substrate preference. This structural characterization and comparison with other bacterial steroid hydroxylase CYPs allowed us to identify a unique Arg295 residue that may serve as the key residue for substrate specificity and regioselectivity in BaCYP106A6. This observation provides valuable background for further protein engineering to design commercially useful CYP steroid hydroxylases with different substrate specificities.

Isolation, Characterization, and Comparative Genomics of the Novel Potential Probiotics from Canine Feces

  • Ngamlak Foongsawat;Sirinthorn Sunthornthummas;Kwannan Nantavisai;Komwit Surachat;Achariya Rangsiruji;Siriruk Sarawaneeyaruk;Kedvadee Insian;Sirapan Sukontasing;Nuttika Suwannasai;Onanong Pringsulaka
    • Food Science of Animal Resources
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    • v.43 no.4
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    • pp.685-702
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    • 2023
  • Lactic acid bacteria (LAB) are commonly used as probiotics; however, not all LAB strains have the same beneficial effects. To successfully use LAB as probiotics in canines, LAB species should originate from the canine intestinal tract as they display host specificity. The objective of this study was to investigate the phenotypic and genomic traits of potential probiotic LAB isolated from canine fecal samples. Twenty LAB samples were evaluated for their potential probiotic characteristics including resistance to low pH, bile salts, hydrophobicity, auto-aggregation, co-aggregation, adhesion to epithelia or mucosa, and production of inhibitory compounds. Additionally, we evaluated their safety and other beneficial effects on canine health, such as DPPH free radical scavenging, and β-galactosidase. Four strains demonstrated potential probiotic characteristics and were selected: Enterococcus hirae Pom4, Limosilactobacillus fermentum Pom5, Pediococcus pentosaceus Chi8, and Ligilactobacillus animalis FB2. Safety evaluations showed that all strains lacked hemolytic activity, could not produce biogenic amines, and did not carry any pathogenic genes. In addition, L. fermentum Pom5 and P. pentosaceus Chi8 displayed susceptibility to all antibiotics and concordant with the absence of antibiotic resistance genes. Based on their phenotypic and genomic characteristics, L. fermentum Pom5 and P. pentosaceus Chi8 were identified as potential probiotic candidates for canines.

Genetic characterization of alloherpesvirus (cyprinid herpesvirus-2 and koi herpesvirus) and poxvirus (carp edema virus) identified from domestic and imported cyprinids in Korea

  • Ye Jin Jeong;Yu Gyeong Jeon;Hee Ju Choi;Eun Jin Baek;Guk Hyun Kim;Yun Jung Yang;Min Jae Kim;Joon Gyu Min;Kwang Il Kim
    • Fisheries and Aquatic Sciences
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    • v.26 no.7
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    • pp.437-446
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    • 2023
  • Cyprinids are popular species for aquaculture worldwide, with Asia being a significant contributor to their production. In Korea, common carp (Cyprinus carpio), koi carp (Cyprinus rubrofuscus), and goldfish (Carassius auratus) are cultivated domestically and imported for ornamental or human consumption purposes. Among the viruses that infect cyprinids, cyprinid herpesvirus-2 (CyHV-2), koi herpesvirus (KHV, also known as cyprinid herpesvirus-3), and carp edema virus (CEV) are of particular concern as they cause substantial economic losses to the aquaculture industry. In this study, we investigated these viruses in both of domestic and imported cyprinids. Our results revealed that CyHV-2 was only detected in imported goldfish from Thailand. To further investigate the genetic characteristics of them, the marker A region was analyzed. Despite belonging to the same cluster with isolates from China, France, Poland, and Israel, CyHV-2 detected in this study showed distinct differences in their repetitive sequence sizes. Furthermore, two different forms of KHV/CEV coinfection were identified from domestic koi carp, both of which exhibited typical symptoms. Phylogenetic analysis showed that one KHV isolate (ScKc-2105-K) was of the Asian type and closely related to isolates from Japan, Indonesia, Belgium, Taiwan, and China. Two CEV isolates (ScKc-2105-CE and GhKc-2207-CE) be- longed to the IIa type and showed high similarity with isolates from the USA, France, and Korea. Notably, koi carp injected with cultured KHV (ScKc-2105-K) showed 78.0% cumulative mortality within 14 days post-injection (dpi). Our findings support the importance of regular surveillance of viral diseases in cyprinids.

Nitric Oxide-Induced Downregulation of a NAD(P)-Binding Rossmann-Fold Superfamily Gene Negatively Impacts Growth and Defense in Arabidopsis thaliana

  • Tiba Nazar Ibrahim Al Azawi;Murtaza Khan;Bong-Gyu Mun;Song-Uk Lee;Da-sol Lee;Waqas Rahim;Anjali Pande;Nusrat Jahan Methela;Cho-Jun Ho;Byung-Wook Yun
    • Proceedings of the Korean Society of Crop Science Conference
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    • 2022.10a
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    • pp.143-143
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    • 2022
  • Plant defense systems against pathogens have been studied extensively and are currently a hot topic in plant science. Using a reverse genetics technique, this study looked into the involvement of the NO-downregulated NAD(P)-binding Rossmann-fold superfamily gene in plant growth and defense in Arabidopsis thaliana. For this purpose, the knockout and overexpressing plant of the candidate gene along with the relevant controls were exposed to control, oxidative and nitro-oxidative stresses. The results showed that candidate gene negatively regulates plants' root and shoot lengths. To investigate the role of the candidate gene in plant basal defense, R-gene-mediated resistance and systemic acquired resistance (SAR) plants were challenged with virulent or avirulent strains of Pseudomonas syringae pathovar tomato (Psf) DC3000. The results showed that the candidate gene negatively regulates plants' basal defense, R-gene-mediated resistance and SAR. Further characterization via GO analysis associated the candidate gene with metabolic and cellular processes and response to light stimulus, nucleotide binding and cellular location in the cytosol and nucleus. Protein structure analysis indicated the presence of a canonical Oxidoreductase family NAD (P)-binding Rossmann fold domain of 120 amino acids with a total of 121 plant homologs across 35 different plant species in the clad streptophyta. Arabidopsis eFP browser showed its expression in almost all the above-ground parts. Protein analysis indicated C225 and C359 as potential targets for S-Nitrosylation by NO. SMART analysis indicated possible interactions with mevalonate/galactokinase, galacturonic acid kinase, arabinose kinase, putative xylulose kinase, GroES-like zinc-binding alcohol dehydrogenase and various glyceraldehyde-3-phosphate dehydrogenases.

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Detection and characterization of potential virulence determinants in Staphylococcus pseudintermedius and S. schleiferi strains isolated from canine otitis externa in Korea

  • Gi Yong Lee;Soo In Lee;Ji Heon Park;Sun Do Kim;Geun-Bae Kim;Soo-Jin Yang
    • Journal of Veterinary Science
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    • v.24 no.6
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    • pp.85.1-85.13
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    • 2023
  • Background: A recent increase in the occurrence of canine skin and soft tissue infections, including otitis externa and pyoderma, caused by antimicrobial-resistant Staphylococcus pseudintermedius and S. schleiferi has become a significant public and veterinary health issues. Objective: We investigated the virulence potentials associated with the occurrence of canine otitis externa in S. pseudintermedius and S. schleiferi. Methods: In this study, the prevalence of genes encoding leukocidins, exfoliative toxins, and staphylococcal enterotoxins (SEs) was investigated using previously characterized S. pseudintermedius (n = 26) and S. schleiferi (n = 19) isolates derived from canine otitis externa. Susceptibility to cathelicidins (K9CATH and PMAP-36) and hydrogen peroxide (H2O2) was also examined in both staphylococcal species. Results: A high prevalence of genes encoding leukocidins (lukS/F-I, lukS1/F1-S, and lukS2/F2-S), exfoliative toxins (siet, expB, and sset), and SEs was identified in both S. pseudintermedius and S. schleiferi isolates. Notably, S. pseudintermedius isolates possessed higher number of SE genes, especially newer SE genes, than S. schleiferi isolates harboring egc clusters. Although no significant differences in susceptibility to K9CATH and H2O2 were observed between the two isolate groups, S. pseudintermedius isolates exhibited enhanced resistance to PMAP-36 compared to S. schleiferi isolates. Conclusions: These findings suggest that high a prevalence of various toxin genes together with enhanced resistance to cathelicidins may contribute to the pathogenicity of S. pseudintermedius and S. schleiferi in canine cutaneous infections.

Evaluation of Horticultural Characteristics on Water Dropwort (Oenanthe stolonifera DC.) Genetic Resources for Various Utilization

  • Eun Ji Kim;Sung Yong Jin;Hyun Soo Jung;Chi Seon Kim
    • Proceedings of the Plant Resources Society of Korea Conference
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    • 2022.09a
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    • pp.51-51
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    • 2022
  • Water dropwort (Oenanthe stolonifera DC.) is a perennial herbaceous plant that grows wild throughout Korea. As of 2020, 24,819 tons (51.7% of open field, 48.3% of facility) are produced in 1,278 ha (59.9% of open field, 40.1% of facility) nationwide. Water dropwort, which is rich in nutrients such as vitamins and iron, is mainly cultivated by vegetative propagation method using local traditional species, however, seed propagation and breeding of cultivars are insufficient so far. Since securing, propagating and continuous characterization of various genetic resources are required to breed new cultivars, this study was conducted to compare the main characteristics of domestic genetic resources and to improve their utilization. Growth characteristics such as plant height, fresh weight, plant type, and flowering date were investigated for the 89 varieties of genetic resources owned by Jeollabuk-do Agricultural Research and Extension Services after vegetative propagation in the individual pots. Also, the morphological image information of leaves and flowers was constructed. Genetic resources were collected nationwide and originated in 30 regions including Hwaseong, Siheung and Jeju. Their plant types could be classified into straight, intermediate and creeping types according to their morphological characteristics, and at this time, the number of intermediate types accounted for the largest proportion. Flowering was carried out under high-temperature and long-day conditions in summer. According to the flowering date, they could be classified into early, middle and late flowering varieties, and at this time, the middle flowering varieties occupied the largest proportion. As a result of the investigation of vegetative growth characteristics, varieties with long plant height, heavy fresh weight and thick stem thickness were identified. Along with this result, it is thought that classification and selection of genetic resources for various purposes will be possible through additional investigations such as analysis of components and antioxidant activity. Moreover, it is judged that such results can be used as basic data for breeding new water dropwort cultivars in the future.

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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.