• Journal of Genetic Medicine
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• v.12 no.2
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• pp.79-84
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• 2015

Purpose: We developed and validated a fetal trisomy detection method for use as a noninvasive prenatal test (NIPT) including a Clinical Laboratory Improvement Amendments (CLIA)-certified bioinformatics pipeline on a cloud-based computing system using both Illumina and Life Technology sequencing platforms for 221 Korean clinical samples. We determined the necessary proportions of the fetal fraction in the cell-free DNA (cfDNA) sample for NIPT of trisomies 13, 18, and 21 through a limit of quantification (LOQ) test. Materials and Methods: Next-generation sequencing libraries from 221 clinical samples and three positive controls were generated using Illumina and Life Technology chemistries. Sequencing results were uploaded to a cloud and mapped on the human reference genome (GRCh37/hg19) using bioinformatics tools. Based on Z-scores calculated by normalization of the mapped read counts, final aneuploidy reports were automatically generated for fetal aneuploidy determination. Results: We identified in total 29 aneuploid samples, and additional analytical methods performed to confirm the results showed that one of these was a false-positive. The LOQ test showed that the proportion of fetal fraction in the cfDNA sample would affect the interpretation of the aneuploidy results. Conclusion: Noninvasive chromosome examination (NICE), a CLIA-certified NIPT with a cloud-based bioinformatics platform, showed unambiguous success in fetus aneuploidy detection.

• Proceedings of the Korean Society of Crop Science Conference
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• 2017.06a
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• pp.139-139
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• 2017

The complete chloroplast genome sequence of Avena sterilis L., a dominant wild oat species in the family Poaceae, is first reported in this study. The complete cp genome sequence of A. sterilis is 135,887 bp in length with 38.5% overall GC content and exhibits a typical quadripartite structure comprising one pair of inverted repeats (21, 603 bp) separated by a small single-copy region (12,575 bp) and a large single-copy region (80,106). The A. sterilis cp genome encodes 111 unique genes, 76 of which are protein-coding genes, 4 rRNA genes, 30 tRNA genes and 18 duplicated genes in the inverted repeat region. Nine genes contain one or two introns. Pair-wise alignments of cp genome were performed for genome-wide comparison. This newly determined cp genome sequence of A. sterilis will provide valuable information for the future breeding programs of valuable cereal crops in the family Poaceae.

• Journal of Microbiology and Biotechnology
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• v.26 no.12
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• pp.2148-2158
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• 2016

To investigate the effects of Lactobacillus curvatus and Leuconostoc mesenteroides on suan cai (pickled Chinese cabbage) fermentation, L. curvatus and/or Ln. mesenteroides were inoculated into suan cai. Physicochemical indexes were measured, and the microbial dynamics during the fermentation were analyzed by Illumina MiSeq sequencing and quantitative polymerase chain reaction (qPCR). The results showed that inoculation with lactic acid bacteria (LAB) lowered the pH of the fermentation system more rapidly. The decrease in water-soluble carbohydrates in the inoculated treatments occurred more rapidly than in the control. The LAB counts in the control were lower than in other inoculated treatments during the first 12 days of fermentation. According to the Illumina MiSeq sequencing analyses, Firmicutes, Proteobacteria, Bacteroidetes, Actinobacteria, Cyanobacteria, Fusobacteria, and Verrucomicrobia were present in the fermentations, along with other unclassified bacteria. Generally, Firmicutes was predominant during the fermentation in all treatments. At the genus level, 16 genera were detected. The relative abundance of Lactobacillus in all inoculated treatments was higher than in the control. The relative abundance of Lactobacillus in the treatments containing L. curvatus was higher than in the Ln. mesenteroides-only treatment. The relative abundance of Leuconostoc in the Ln. mesenteroides-containing treatments increased continuously throughout the fermentation. Leuconostoc was highest in the Ln. mesenteroides-only treatment. According to the qPCR results, L. curvatus and/or Ln. mesenteroides inoculations could effectively inhabit the fermentation system. L. curvatus dominated the fermentation in the inoculated treatments.

• Journal of Microbiology and Biotechnology
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• v.28 no.7
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• pp.1168-1177
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• 2018

The aim of this study was to investigate the microbial community of three tannery wastewater treatment plants (WWTPs) involved in nitrification by Illumina MiSeq sequencing. The results showed that highly diverse communities were present in tannery wastewater. A total of six phyla, including Proteobacteria (37-41%), Bacteroidetes (6.04-16.80), Planctomycetes (3.65-16.55), Chloroflexi (2.51-11.48), Actinobacteria (1.91-9.21), and Acidobacteria (3.04-6.20), were identified as the main phyla, and Proteobacteria dominated in all the samples. Within Proteobacteria, Beta-proteobacteria was the most abundant class, with the sequence percentages ranging from 9.66% to 17.44%. Analysis of the community at the genus level suggested that Thauera, Gp4, Ignavibacterium, Phycisphaera, and Arenimonas were the core genera shared by at least two tannery WWTPs. A detailed analysis of the abundance of ammonia-oxidizing bacteria (AOB) and nitrite-oxidizing bacteria (NOB) indicated that Nitrosospira, Nitrosomonas, and Nitrospira were the main AOB and NOB in tannery wastewater, respectively, which exhibited relatively high abundance in all samples. In addition, real-time quantitative PCR was conducted to validate the results by quantifying the abundance of the AOB and total bacteria, and similar results were obtained. Overall, the results presented in this study may provide new insights into our understanding of key microorganisms and the entire community of tannery wastewater and contribute to improving the nitrogen removal efficiency.

• Food Science of Animal Resources
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• v.37 no.4
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• pp.535-541
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• 2017

This study investigated the psychrotrophic bacteria isolated from chicken meat to characterize their microbial composition during refrigerated storage. The bacterial community was identified by the Illumina MiSeq method based on bacterial DNA extracted from spoiled chicken meat. Molecular identification of the isolated psychrotrophic bacteria was carried out using 16S rDNA sequencing and their putrefactive potential was investigated by the growth at low temperature as well as their proteolytic activities in chicken meat. From the Illumina sequencing, a total of 187,671 reads were obtained from 12 chicken samples. Regardless of the type of chicken meat (i.e., whole meat and chicken breast) and storage temperatures ($4^{\circ}C$ and $10^{\circ}C$), Pseudomonas weihenstephanensis and Pseudomonas congelans were the most prominent bacterial species. Serratia spp. and Acinetobacter spp. were prominent in chicken breast and whole chicken meat, respectively. The 118 isolated strains of psychrotrophic bacteria comprised Pseudomonas spp. (58.48%), Serratia spp. (10.17%), and Morganella spp. (6.78%). All isolates grew well at $10^{\circ}C$ and they induced different proteolytic activities depending on the species and strains. Parallel analysis of the next generation sequencing and culture dependent approach provides in-depth information on the biodiversity of the spoilage microbiota in chicken meat. Further study is needed to develop better preservation methods against these spoilage bacteria.

• Genomics & Informatics
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• v.9 no.2
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• pp.69-73
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• 2011

The use of genomic information in genomic selection programs for dairy and beef cattle breeds has become a reality in recent years. In this investigation, we analyzed single-nucleotide polymorphisms (SNPs) for Hanwoo (n=50) and Holstein (n=50) breeds using the Illumina Bovine SNP50 BeadChip to facilitate genomic selection and utilization of the Hanwoo breed in Korea. Analysis of the entire genomes showed different spectra of SNP frequencies for Hanwoo and Holstein cattle. The study revealed a highly significant (p<0.001) difference between Hanwoo and Holstein cattle in minor allele frequency (MAF). The average MAFs were $0.19{\pm}0.16$ and $0.22{\pm}0.16$ for Hanwoo and Holstein, respectively. From the total of 52,337 SNPs that were successfully identified, about 72% and 79% were polymorphic in Hanwoos and Holsteins, respectively. Polymorphic and fixed SNPs were not distributed uniformly across the chromosomes within breeds or between the two breeds. The number of fixed SNPs on all chromosomes was higher in Hanwoo cattle, reflecting the genetic uniqueness of the Hanwoo breed. In general, the rate of polymorphisms detected in these two breeds suggests that the SNPs can be used for different applications, such as whole-genome association and comparative genetic studies, and are a helpful tool in developing breed identification genetic markers.

• 한국균학회소식:학술대회논문집
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• 2016.05a
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• pp.11-11
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• 2016

Penicillium species are commonly isolated from various outdoor and indoor environments, including marine environments such as sponges, algae and sand. Penicillium is especially important because numerous bioactive compounds have been isolated. Penicillium was the most common species in intertidal zone in Korea, however the diversity and ecological roles of Penicillium in intertidal zone are not clarified. We explored diversity and ecological roles of marine-derived Penicillium from tidal flat and sea sand in Korea. The diversity of marine-derived Penicillium from Korea was investigated using both culture-dependent and culture-independent approach by ${\beta}$-tubulin sequence. In addition, we evaluated optimal temperature, halo-tolerance, and enzyme activity of Penicillium strains, such as extracellular alginase, endoglucanase, ${\beta}$-glucosidase, and protease. For culture-dependent approach, a total of 182 strains of 62 Penicillium species were isolated, with 53 species being identified. The most common species was Penicillium oxalicum, followed by P. crustosum, P. brasilianum, P. koreense, and P. griseofulvum. Species richness and composition were not significantly different by season, substrates, and seaside. For culture-independent approach using Illumina sequencing, 73 OTUSs were detected. The most frequently observed species was P. antarcticum, followed by P. koreense, P. crustosum, and P. brevicompactum. Diversity of Penicillium was higher during winter season than during summer season and in western sea than in southern sea, respectively. Community structure was significantly different by season and sea side. 52 species were detected by both methods. Unique species were isolated from each of methods - 10 from culture methods and 21 from Illumina sequencing. Furthermore, salinity adaption of the Penicillium varied depending on species. Many Penicillium species showed endoglucanase, ${\beta}$-glucosidase, and protease activity. Some species including P. paneum and P. javanicum degraded the polycyclic aromatic hydrocarbons. Thus, our results demonstrate that intertidal zone in Korea harbors diverse Penicillium community and marine-derived Penicillium play important ecological roles as decomposers of organic material in marine environments.

• Korean Journal of Microbiology
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• v.53 no.2
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• pp.141-143
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• 2017

Caballeronia sordidicola strain PAMC 26577 was isolated from Cladonia sp., a lichen collected from Svalbard Archipelago in the Arctic Ocean. Draft genomic sequences of PAMC 26577 were determined using Illumina and 182 contigs were submitted to GenBank and N50 value was 159,226. The genome of PAMC 26577 was comprised of 8,334,211 base pairs and %G+C content was 59.4. The genome included 8 ribosomal RNA genes and 51 tRNA genes as non-coding sequences. Protein-coding genes were 8,065 in number and they included central metabolism genes as well as butanol/butyrate biosynthesis, polyhydroxybutyrate metabolism, serine cycle methylotrophy genes, and glycogen metabolism. Membrane transporters were more than two-hundreds in number, but sugar phosphotransferase system and TRAP transporters were lacking. PAMC 26577 lacked CRISPR-associated sequences and proteins. No transposable elements were observed and there were only limited number of phage remnant regions with 11 phage-related genes.

• Mycobiology
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• v.46 no.4
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• pp.421-428
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• 2018

The white button mushroom (Agaricus bisporus) is one of the most widely cultivated species of edible mushroom. Despite its economic importance, relatively little is known about the genetic diversity of this species. Illumina paired-end sequencing produced 43,871,558 clean reads and 69,174 contigs were generated from five offspring. These contigs were subsequently assembled into 57,594 unigenes. The unigenes were annotated with reference genome in which 6,559 unigenes were associated with clusters, indicating orthologous genes. Gene ontology classification assigned many unigenes. Based on genome data of the five offspring, 44 polymorphic simple sequence repeat (SSR) markers were developed. The major allele frequency ranged from 0.42 to 0.92. The number of genotypes and the number of alleles ranged from 1 to 4, and from 2 to 4, respectively. The observed heterozygosity and the expected heterozygosity ranged from 0.00 to 1.00, and from 0.15 to 0.64, respectively. The polymorphic information content value ranged from 0.14 to 0.57. The genetic distances and UPGMA clustering discriminated offspring strains. The SSR markers developed in this study can be applied in polymorphism analyses of button mushroom and for cultivar discrimination.

• Mycobiology
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• v.49 no.4
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• pp.385-395
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• 2021

Fungal endophytes have been recorded in various plant species with a richness of diversity, and their presence plays an essential role in host plant protection against biotic and abiotic stresses. This study applied the Illumina MiSeq sequencing platform based on the amplification of fungal ribosomal ITS2 region to analyze fungal endophytic communities of two oak species (Quercus mongolica and Q. serrata) with different oak wilt disease susceptibilities in Korea. The results showed a total of 230,768 sequencing reads were obtained and clustered at a 97% similarity threshold into 709 operational taxonomic units (OTUs). The OTUs of Q. serrata were higher than that of Q. mongolica with the number of 617 OTUs and 512 OTUs, respectively. Shannon index also showed that Q. serrata had a significantly higher level of fungal diversity than Q. mongolica. Total of OTUs were assigned into 5 fungal phyla, 17 classes, 60 orders, 133 families, 195 genera, and 280 species. Ascomycota was the dominant phylum with 75.11% relative abundance, followed by Basidiomycota with 5.28%. Leptosillia, Aureobasidium and Acanthostigma were the most abundant genera detected in Q. serrata with the average relative abundance of 2.85, 2.76, and 2.19%, respectively. On the other hand, Peltaster, Cladosporium and Monochaetia were the most common genera detected in Q. mongolica with the average relative abundance of 4.83, 3.03, and 2.87%, respectively. Our results indicated that fungal endophytic communities were significantly different between two oak species and these differences could influence responses of host trees to oak wilt disease caused by Raffaelea quercus-mongolicae.