• The Plant Pathology Journal
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• v.37 no.5
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• pp.476-488
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• 2021

Xanthomonas campestris pv. campestris (Xcc) is the causal agent of black rot for cruciferous vegetables worldwide, especially for the cole crops such as cabbage and cauliflower. Due to the lack of resistant cabbage cultivars, black rot has brought about considerable yield losses in recent years in China. Understanding of the pathogen features is a key step for disease prevention, however, the pathogen diversity, population structure, and virulence are largely unknown. In this study, we studied 50 Xcc strains including 39 Xcc isolates collected from cabbage in 20 regions across China, using multilocus sequence genotyping (MLST), repetitive DNA sequence-based PCR (rep-PCR), and pathogenicity tests. For MLST analysis, a total of 12 allelic profiles (AP) were generated, among which the largest AP was AP1 containing 32 strains. Further cluster analysis of rep-PCR divided all strains into 14 DNA groups, with the largest group DNA I comprising of 34 strains, most of which also belonged to AP1. Inoculation tests showed that the representative Xcc strains collected from diverse regions performed differential virulence against three brassica hosts compared with races 1 and 4. Interestingly, these results indicated that AP1/DNA I was not only the main pathotype in China, but also a novel group that differed from the previously reported type races in both genotype and virulence. To our knowledge, this is the first extensive genetic diversity survey for Xcc strains in China, which provides evidence for cabbage resistance breeding and opens the gate for further cabbage-Xcc interaction studies.

• Microbiology and Biotechnology Letters
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• v.49 no.1
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• pp.120-129
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• 2021

Rotavirus A (RVA) is recognized as a major etiology responsible for the development of acute gastroenteritis in children worldwide. The purpose of the present study was to perform the molecular characterization of RVA. A total of 323 stool specimens collected from hospitalized children with acute gastroenteritis in Chiang Rai, Thailand, in 2016-2018 were identified for G- and P-genotypes through RT-PCR analysis. RVA was more prevalent in 2017-2018 (37.8%) than in 2016-2017 (23.2%). The seasonal peak of RVA occurred from March to April. G3P[8] was predominant in 2016-2017 (90.6%) and 2017-2018 (58.6%). Other genotypes including G1P[8], G8P[8], G9P[8], and mixed infections were also identified. G3P[8] strains clustered together in the same lineage with other novel human equine-like G3P[8] strains previously identified in multiple countries and presented a genotype 2 constellation (G3-P[8]-I2-R2-C2-M2-A2-N2-T2-E2-H2). Several amino acid differences were observed in the antigenic epitopes of the VP7 and VP8* capsid proteins of the equine-like G3P[8] compared with those of the RVA vaccine strains. The homology modeling of the VP7 and VP8* capsid proteins of the equine-like G3P[8] strains evidently exhibited that these residue differences were present on the surface-exposed area of the capsid structure. The emergence of the equine-like G3P[8] strains in Thailand indicates the rapid spread of strains with human and animal gene segments. Continuous surveillance for RVA is essential to monitor genotypes and genetic diversity, which will provide useful information for selecting rotavirus strains to develop a safe and effective RVA vaccine that is efficacious against multiple genotypes and variants.

• Journal of Microbiology and Biotechnology
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• v.33 no.11
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• pp.1448-1456
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• 2023

A Gram-positive, non-motile, and non-spore-forming lactic acid bacterium, designated as BK2^T, was isolated from kimchi, a Korean traditional fermented vegetable food, and the taxonomic characteristics of strain BK2^T, along with strain LMG 11983, were analyzed. Both strains optimally grew at 30℃, pH 7.0, and 1.0% NaCl. Cells of both strains were heterofermentative and facultatively anaerobic rods, demonstrating negative reactions for catalase and oxidase. Major fatty acids (>10%) identified in both strains were C_18:1 ω9c, C_16:0, and summed feature 7 (comprising C_19:1 ω6c and/or C_19:1 ω7c). The genomic DNA G+C contents of both strains were 44.7 mol%. The 16S rRNA gene sequence similarity (99.9%), average nucleotide identity (ANI; 99.9%), and digital DNA-DNA hybridization (dDDH; 99.7%) value between strains BK2^T and LMG 11983 indicated that they are different strains of the same species. Strain BK2^T was most closely related to Weissella confusa JCM 1093^T and Weissella cibaria LMG 17699^T, with 100% and 99.4% 16S rRNA gene sequence similarities, respectively. However, based on the ANI and dDDH values (92.3% and 48.1% with W. confusa, and 78.4% and 23.5% with W. cibaria), it was evident that strain BK2^T represents a distinct species separate from W. confusa and W. cibaria. Based on phylogenetic, phenotypic, and chemotaxonomic features, strains BK2^T and LMG 11983 represent a novel species of the genus Weissella, for which the name Weissella fermenti sp. nov. is proposed. The type of strain is BK2^T (=KACC 22833^T=JCM 35750^T).

• 한국생물공학회:학술대회논문집
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• 2001.11a
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• pp.625-628
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• 2001

To isolate novel strains that hydrolyzed the rac-ketoprofen ethyl ester to ketoprofen in the stereospecific manner, we screened broad ecological niches and soil samples in which the activity was expected to be found. From thousands of strains, we isolated a Pseudomonas sp. S34 producing a (S)-stereospecific esterase, and a thermostable esterase with (R)-form selectivity was also 。 btained from Bacillus stearothermophilus JYl44. To further analyse the gene structure and to induce a high level expression, two genes from each strain were cloned and sequenced. BLAST search results with the esterase gene from 534 revealed that both of gene (80-84 %) and amino acid sequences (89- 95 %) were highly conserved in the related esterases from Pseudomonas strains (fluorescens and aeruginosa). The thermostable esterase from JY144, however, revealed a relative low homology (45-52 %) to other esterase and/or lipase from related strains. Obviously, a complete conversion with pure enantiomer (R - or S) were readily achieved by recombinant clones expressing either (R)- or (S)- stereospecific esterase.

• Mycobiology
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• v.39 no.1
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• pp.33-39
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• 2011

Wild yeasts on the surface of various fruits including grapes were surveyed to obtain yeast strains suitable for fermenting a novel wine with higher alcohol content and supplemented with rice starch. We considered selected characteristics, such as tolerance to alcohol and osmotic pressure, capability of utilizing maltose, and starch hydrolysis. Among 637 putative yeast isolates, 115 strains exhibiting better growth in yeast-peptone-dextrose broth containing 30% dextrose, 7% alcohol, or 2% maltose were selected, as well as five ${\alpha}$-amylase producers. Nucleotide sequence analysis of the 26S rDNA gene classified the strains into 13 species belonging to five genera; Pichia anomala was the most prevalent (41.7%), followed by Wickerhamomyces anomalus (19.2%), P. guilliermondii (15%), Candida spp. (5.8%), Kodamaea ohmeri (2.5%), and Metschnikowia spp. (2.5%). All of the ${\alpha}$-amylase producers were Aureobasidium pullulans. Only one isolate (NK28) was identified as Saccharomyces cerevisiae. NK28 had all of the desired properties for the purpose of this study, except ${\alpha}$-amylase production, and fermented alcohol better than commercial wine yeasts.

• Journal of Microbiology and Biotechnology
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• v.27 no.5
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• pp.909-915
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• 2017

Bifidobacterium bifidum BGN4 (BGN4) has many proven beneficial effects, including antiallergy and anticancer properties. It has been commercialized and used in several probiotic products, and thus strain-specific identification of this strain is very valuable for further strain-dependent physiological study. For this purpose, we developed novel multiplex polymerase chain reaction (PCR) primer sets for strain-specific detection of BGN4 in commercial products and fecal samples of animal models. The primer set was tested on seven strains of B. bifidum and 75 strains of the other Bifidobacterium species. The BGN4-specific regions were derived using megaBLAST against genome sequences of various B. bifidum databases and four sets of primers were designed. As a result, only BGN4 produced four PCR products simultaneously whereas the other strains did not. The PCR detection limit using BGN4-specific primer sets was $2.8{\times}10^1CFU/ml$ of BGN4. Those primer sets also detected and identified BGN4 in the probiotic products containing BNG4 and fecal samples from a BGN4-fed animal model with high specificity. Our results indicate that the PCR assay from this study is an efficient tool for the simple, rapid, and reliable identification of BGN4, for which probiotic strains are known.

• Journal of Environmental Science International
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• v.21 no.2
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• pp.253-261
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• 2012

We isolated and characterized novel duck feather-degrading bacteria producing keratinase. Twelve strains were isolated from soil and faces at poultry farm, and decayed feathers. They were identified as Bacillus methylotrophicus, Pseudomonas geniculata, Pseudomonas hibiscicola, Exiquobacterium profundum, Bacillus pumilus, Bacillus amyloliquefaciens, Chryseobacterium indologenes, Bacillus thuringiensis, Thermomonas koreensis, respectively, by phenotypic characters and 16S rRNA gene analysis. Generally, the level of keratinase production was not proportional to feather degradation rate. The highest keratinolytic activity was observed in the culture inoculated with Chryseobacterium indologenes D27. Although all strains did not degrade human hair, strains tested effectively degraded chicken feather(53.8-91.4%), wool(40.4-93.0%) and human nail (51.0-82.9%). These results suggest that strains isolated could be not only used to improve the nutritional value of recalcitrant feather waste but also is a potential candidate for biotechnological processes of keratin hydrolysis.

• Journal of Microbiology and Biotechnology
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• v.26 no.11
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• pp.1983-1992
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• 2016

Pseudomonas strains isolated from the heavily contaminated Lubin copper mine and Zelazny Most post-flotation waste reservoir in Poland were screened for the presence of integrons. This analysis revealed that two strains carried homologous DNA regions composed of a gene encoding a DNA_BRE_C domain-containing tyrosine recombinase (with no significant sequence similarity to other integrases of integrons) plus a three-component array of putative integron gene cassettes. The predicted gene cassettes encode three putative polypeptides with homology to (i) transmembrane proteins, (ii) GCN5 family acetyltransferases, and (iii) hypothetical proteins of unknown function (homologous proteins are encoded by the gene cassettes of several class 1 integrons). Comparative sequence analyses identified three structural variants of these novel integron-like elements within the sequenced bacterial genomes. Analysis of their distribution revealed that they are found exclusively in strains of the genus Pseudomonas.

• Mycobiology
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• v.51 no.2
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• pp.79-86
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• 2023

In this study, fungal strains designated as KNUF-22-14A and KNUF-22-15A were isolated from soil samples in Korea. These two strains were identified based on cultural and morphological characteristics as well as phylogenetic analyses and were found to be morphologically and phylogenetically identical. Upon their morphological comparison with closely related species, such as Tolypocladium album, T. amazonense, T. endophyticum, T. pustulatum, and T. tropicale, a difference in the size of short phialides [0.6-2.4(-9.3)×0.8-1.4 ㎛] was observed. Meanwhile, these strains had larger conidia (1.2-3.0×1.2-3.0 ㎛) than T. album, T. amazonense, T. endophyticum, and T. tropicale and smaller conidia than T. pustulatum. Phylogenetic analyses using a multi-locus datasets based on ITS, LSU, and SSU showed that KNUF-22-14A and KNUF-22-15A formed a distinct cluster from previously identified Tolypocladium species. Thus, these fungal strains isolated from soil in Korea are proposed as a novel species according to their characteristics and are named Tolypocladium globosum sp. nov.

• ALGAE
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• v.39 no.2
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• pp.59-74
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• 2024

Terrestrial cyanobacteria are extremely diverse. In urban areas, they can be found as black stains on the surface of building walls, stone monuments, or man-made structures. Many of the terrestrial cyanobacteria are still understudied. To expand knowledge of terrestrial cyanobacterial diversity, a polyphasic characterization was performed to identify 12 strains isolated from campus of University of the Ryukyus, Okinawa, Japan. Multigene phylogenetic analyses based on 16S rRNA gene and 16S-23S rRNA internal transcribed spacer (ITS) region showed that the isolated strains formed two independent subclades within Nodosilinea, and were distantly related to all described Nodosilinea species. The 16S-23S rRNA ITS secondary structures showed variations for D1-D1' and Box B domain, while V3 domain was almost identical among entire species of Nodosilinea, including the studied strains. In addition, a unique morphological character, i.e. forming nodule or spiral shape, was also observed in certain studied strains. According to polyphasic characterization, Nodosilinea coculeatus sp. nov. and Nodosilinea terrestrialis sp. nov., were proposed as two new species of terrestrial cyanobacteria from Okinawa.