• Parasites, Hosts and Diseases
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• 제56권6호
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• pp.559-565
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• 2018

The identification and characterization of pathogenic and zoonotic tick-borne diseases like granulocytic anaplasmosis are essential for developing effective control programs. The differential diagnosis of pathogenic Anaplasma phagocytophilum and non-pathogenic A. phagocytophilum-like Anaplasma spp. is important for implementing effective treatment from control programs. The objective of the present study was to investigate the prevalence of Anaplasma spp. in horses in Korea by nucleotide sequencing and restriction enzyme fragment length polymorphism assay. Of the 627 horses included in the study, only 1 (0.2%) was infected with A. phagocytophilum. Co-infection with A. phagocytophilumlike Anaplasma spp. was not detected in the study. The 16S rRNA sequence of A. phagocytophilum was similar (99.5-100%) to A. phagocytophilum 16S rRNA isolated from horses in other countries. PCR adapted to amplify A. phagocytophilum groEL and msp2 genes failed to generate amplicons, suggesting genetic diversity in these genes. This study is the first molecular detection of A. phagocytophilum in horses in Korea. Human granulocytic anaplasmosis and animal infection of A. phagocytophilum have been reported in Korea recently. Because of vector tick distribution, global warming, and the increase of the horse industry, horses should be considered as a potential reservoir for A. phagocytophilum, and cross infectivity should be evaluated even though a low prevalence of infection was detected in this study. Furthermore, continuous surveillance and effective control measures for A. phagocytophilum should be established to prevent disease distribution and possible transmission to humans.

• The Korean Journal of Mycology
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• 제47권2호
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• pp.165-172
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• 2019

To investigate the diversity of aquatic fungi, we collected deposits of soil, plants, and plant litter from ponds and streams. NNIBRFG1552 was isolated from soil deposits and NNIBRFG3013 from plant deposits in Namsaengi-mot in Jeju, Korea in 2016; NNIBRFG5472 was isolated from plant litter in Bocheong-cheon, Boeun, Korea in 2018. Based on morphological characteristics and phylogenetic analysis using rDNA sequences of the internal transcribed spacer (ITS), NNIBRFG1552, NNIBRFG3013, and NNIBRFG5472 were identified as Filoporella exilis (100% similarity with KC834046), F. fistucella (99.8% with KC834047), and F. cf. annelidica (100% with KC834044), respectively. Furthermore, cultural and morphological characteristics were analyzed to confirm the molecular identification. No species of the genus Filosporella has yet been reported in Korea.

• Microbiology and Biotechnology Letters
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• 제47권3호
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• pp.401-411
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• 2019

All Idiomarina species are isolated from saline environments; microorganisms in such extreme habitats develop metabolic adaptations and can produce compounds such as proteases with an industrial potential. ARDRA and 16S rRNA gene sequencing are established methods for performing phylogenetic analysis and taxonomic identification. However, 16S-23S ITS is more variable than the 16S rRNA gene within a genus, and is therefore, used as a marker to achieve a more precise identification. In this study, ten protease producing Idiomarina strains isolated from the Peruvian salterns were characterized using biochemical and molecular methods to determine their bacterial diversity and industrial potential. In addition, comparison between the length and nucleotide sequences of a 16S-23S ITS region allowed us to assess the inter and intraspecies variability. Based on the 16S rRNA gene, two species of Idiomarina were identified (I. zobellii and I. fontislapidosi). However, biochemical tests revealed that there were differences between the strains of the same species. Moreover, it was found that the ITS contains two tRNA genes, $tRNA^{Ile(GAT)}$ and $tRNA^{Ala(TGC)}$, which are separated by an ISR of a variable size between strains of I. zobellii. In one strain of I. zobellii (PM21), we found nonconserved nucleotides that were previously not reported in the $tRNA^{Ala}$ gene sequences of Idiomarina spp. Thus, based on the biochemical and molecular characteristics, we can conclude that protease producing Idiomarina strains have industrial potential; only two I. zobellii strains (PM48 and PM72) exhibited the same properties. The differences between the other strains could be explained by the presence of subspecies.

• Journal of Species Research
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• 제8권2호
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• pp.176-190
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• 2019

A total of 38 bacterial strains within the classes Bacilli and Deinococci were isolated from various sources in Korea. Samples were collected from animal intestine, urine, soil, tidal flat mud, and kimchi. In the sequence comparison and phylogenetic analysis of 16S rRNA sequences, the 38 isolates were assigned to the classes Bacilli and Deinococci with sequence similarities more than 98.7%. Twenty-four strains and 13 strains were classified the order Bacillales and Lactobacillales in the class Bacilli, respectively. In the order Bacillales, there were nine species in the genus Bacillus, seven species in the genus Paenibacillus, and the remaining eight species in the genera Domibacillus, Halobacillus, Virgibacillus, Lysinibacillus, Paenisporosarcina, Planococcus, Savagea, and Staphylococcus. In the order Lactobacillales, there were four species in the genus Lactobacillus, three species in the genus Leuconostoc, three species in the genus Lactococcus, and the remaining three species in the genera Aerococcus, Enterococcus, and Streptococcus. One species was related to the genus Deinococcus of the order Deinococcales. Most of the isolated strains were Gram-stain-positive, but some were Gram-stain-variable or Gram-stain-negative. Cells were rod or cocci-shaped. Based on the results of 16S rRNA analysis, we report 38 strains as previously unrecorded species to Korea, and the basic characteristics of strains are described herein.

• Mycobiology
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• 제48권6호
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• pp.476-483
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• 2020

The genus Pholiota (Strophariaceae, Basidiomycota) is made up of wood-rotting saprotrophic mushrooms characterized by a yellow or brown pileus with scales and/or slimy, and by a brownish smooth spore with a germ pore. However, these features are not enough to distinguish its species, or separate the genus Pholiota from other brown-spored wood-rotting genera such as Hypholoma and Stropharia. Although internal transcribed spacer (ITS) sequencebased identification has improved identification accuracy for species of Pholiota, most Pholiota species in Korea are reported based on morphological features. To evaluate the taxonomy of Pholiota species, we investigated 62 specimens collected from 1999 to 2019 in Korea using ITS sequence analysis and morphological observation. Twelve of the 16 recorded Pholiota species in Korea were identified. While eight species were clearly separated, the ITS analysis did not distinguish three in the Pholiota adiposa complex. Therefore, further investigation is required to distinguish these three species. ITS sequences deposited in GenBank confirm that P. highlandensis exists in Korea. The presence of the other four Pholiota species could not be confirmed through specimens or sequence information in GenBank. A taxonomic key and the ITS sequence data for Korean Pholiota species are included and can be good baselines for further research on Pholiota taxonomy and diversity.

• Animal Systematics, Evolution and Diversity
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• 제36권4호
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• pp.336-346
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• 2020

Mitochondrial genome is an important molecule for systematic and evolutionary studies in metazoans. The development of next-generation sequencing (NGS) technique has rapidly increased the number of mitogenome sequences. The process of generating mitochondrial genome based on NGS includes different steps, from DNA preparation, sequencing, assembly, and annotation. Despite the effort to improve sequencing, assembly, and annotation methods of mitogenome, the low quality and/or quantity sequence in the final map can still be generated through the work. Therefore, it is necessary to check and curate mitochondrial genome sequence after annotation for proofreading and feedback. In this study, we introduce the pipeline for sequencing and curation for mitogenome based on NGS. For this purpose, two mitogenome sequences of Dermatobranchus otome were sequenced by Illumina Miseq system with different amount of raw read data. Generated reads were targeted for assembly and annotation with commonly used programs. As abnormal repeat regions present in the mitogenomes after annotation, primers covering these regions were designed and conventional PCR followed by Sanger sequencing were performed to curate the mitogenome sequences. The obtained sequences were used to replace the abnormal region. Following the replacement, each mitochondrial genome was compared with the other as well as the sequences of close species available on the Genbank for confirmation. After curation, two mitogenomes of D. otome showed a typically circular molecule with 14,559 bp in size and contained 13 protein-coding genes, 22 tRNA genes, two rRNA genes. The phylogenetic tree revealed a close relationship between D. otome and Tritonia diomea. The finding of this study indicated the importance of caution and curation for the generation of mitogenome from NGS.

• Microbiology and Biotechnology Letters
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• 제50권1호
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• pp.126-134
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• 2022

Norovirus (NoV) is an important pathogen causing acute gastroenteritis worldwide. The purpose of the present study was the molecular characterization of NoV. A total of 408 stool specimens collected from hospitalized children associated with acute gastroenteritis in Chiang Rai, Thailand, 2015-2017 were investigated for the presence of NoVs by RT-PCR. NoV GII was detected in 32 samples (7.8%). Five distinct genotypes were identified, including GII.4 (13/32, 40.6%), GII.3 (11/32, 34.3%), GII.17 (4/32, 12.5%), GII.2 (2/32, 6.3%), and GII.14 (2/32, 6.3%). NoV infection occurred mostly in young children under 3 years of age (31/32, 96.9%) and showed the main peak in summer months from March to April (18/32, 56.3%). Phylogenetic analysis revealed that all 13 GII.4 strains clustered with GII.4 Sydney 2012 variant. Representative GII.3 strains were analyzed as a recombinant GII.3P[12] strain. Several amino acid differences were found in the antigenic epitopes and antibody binding sites of the VP1 capsid of the GII.3P[12]. Homology modeling of the P domain of the GII.3P[12] strain demonstrated that 10/13 amino acid differences were predicted to be located on the surface-exposed area of the capsid structure. These amino acid changes might affect the infectivity and the antigenicity of the recombinant GII.3P[12]. The prevalence of GII.4 Sydney 2012 and recombinant GII.3P[12] strains indicates the genetic diversity of circulating NoVs in Thailand, emphazing the importance of continuous surveillance to mornitor newly emerging NoV strains in the future.

• Journal of Plant Biotechnology
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• 제49권1호
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• pp.46-60
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• 2022

The genetic variability and population structure of apple mosaic virus (ApMV) have been studied; however, synonymous codon usage patterns influencing the survival rates and fitness of ApMV have not been reported. Based on phylogenetic analyses of 52 ApMV coat protein (CP) sequences obtained from apple, pear, and hazelnut, ApMV isolates were clustered into two groups. High molecular diversity in GII may indicate their recent expansion. A constant and conserved genomic composition of the CP sequences was inferred from the low codon usage bias. Nucleotide composition and relative synonymous codon usage (RSCU) analysis indicated that the ApMV CP gene is AU-rich, but G- and U-ending codons are favored while coding amino acids. This unequal use of nucleotides together with parity rule 2 and the effective number of codon (ENC) plots indicate that mutation pressure together with natural selection drives codon usage patterns in the CP gene. However, in this combination, selection pressure plays a more crucial role. Based on principal component analysis plots, ApMV seems to have originated from apple trees in Europe. However, according to the relative codon deoptimization index and codon adaptation index (CAI) analyses, ApMV exhibited the greatest fitness to hazelnut. As inferred from the results of the similarity index analysis, hazelnut has a major role in shaping ApMV RSCU patterns, which is consistent with the CAI analysis results. This study contributes to the understanding of plant virus evolution, reveals novel information about ApMV evolutionary fitness, and helps find better ApMV management strategies.

• Journal of Species Research
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• 제11권4호
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• pp.266-277
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• 2022

The current surveys of Ulva in the subtidal area around Jeju Island give a chance to discover unrecorded green algal species of the Korean macroalgal flora. As a result of this investigation, we found Ulva adhaerens Matusmoto & Shimada, inhabiting the subtidal regions, up to 15 m deep, and conducted the DNA barcoding on plastid rbcL-3P and tufA regions with describing the morphological characteristics. Our specimens of U. adhaerens forms a monophyletic clade with the Japanese type specimen and U. piritoka Ngāti Kuri, Heesch & W.A. Nelson from New Zealand exhibiting each 0.3% sequence divergences, respectively, in the plastid rbcL-3P. The genetic variation of U. adhaerens clade is 1.0-3.9% in rbcL-3P and 4.8-9.8% in tufA to each Ulva species, including the generic type, U. lactuca Linneaus. The morphology of Korean U. adhaerens specimens is identical to the type specimens of U. adhaerens from Japan having the development of rhizoidal filaments from both of the cell layers of the distromatic blade and the extension of rhizoidal clumps with adhesive trait between blades by extended rhizoidal clumps at the basal blades. The thallus attachment to substrate is by numerous minute discoidal plates made up of rhizoids originating from the inner part of distromatic blades in basal. Although there are still some problems to resolve the relationship between U. adhaerens and U. piritoka in the rbcL dataset and the phylogenetic pattern of the Group II intron of rbcL, we propose the new record of U. adhaerens in Korean macroalgal flora based on the morphological characteristics of Korean specimens. Continued study of the genus Ulva by morphological and molecular assessment will delimit the species of Ulva, elucidate the relationships between them, and uncover the species diversity.

• Microbiology and Biotechnology Letters
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• 제50권4호
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• pp.563-573
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• 2022

Xanthomonas arboricola pv. juglandis (Xaj) is a globally important bacterial pathogen of walnut trees that causes substantial economic losses in commercial walnut production. Although prophages are common in bacterial plant pathogens and play important roles in bacterial diversity and pathogenicity, there has been limited investigation into the distribution and function of prophages in Xaj. In this study, we identified and characterized 13 predicted prophages from the genomes of 12 Xaj isolates from around the globe. These prophages ranged in length from 11.8 kb to 51.9 kb, with between 11-75 genes and 57.82-64.15% GC content. The closest relatives of these prophages belong to the Myoviridae and Siphoviridae families of the Caudovirales order. The phylogenetic analysis allowed the classification of the prophages into five groups. The gene constitution of these predicted prophages was revealed via Roary analysis. Amongst 126 total protein groups, the most prevalent group was only present in nine prophages, and 22 protein groups were present in only one prophage (singletons). Also, bioinformatic analysis of the 13 identified prophages revealed the presence of 431 genes with an average length of 389.7 bp. Prokka annotation of these prophages identified 466 hypothetical proteins, 24 proteins with known function, and six tRNA genes. The proteins with known function mainly comprised prophage integrase IntA, replicative DNA helicase, tyrosine recombinase XerC, and IS3 family transposase. There was no detectable insertion site specificity for these prophages in the Xaj genomes. The identified Xaj prophage genes, particularly those of unknown function, merit future investigation.