• Title/Summary/Keyword: Whole-Genome Sequencing

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Safety Evaluation of Bifidobacterium breve IDCC4401 Isolated from Infant Feces for Use as a Commercial Probiotic

  • Choi, In Young;Kim, Jinhee;Kim, Su-Hyeon;Ban, O-Hyun;Yang, Jungwoo;Park, Mi-Kyung
    • Journal of Microbiology and Biotechnology
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    • v.31 no.7
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    • pp.949-955
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    • 2021
  • Previously, our research group isolated Bifidobacterium breve IDCC4401 from infant feces as a potential probiotic. For this study, we evaluated the safety of B. breve IDCC4401 using genomic and phenotypic analyses. Whole genome sequencing was performed to identify genomic characteristics and investigate the potential presence of genes encoding virulence, antibiotic resistance, and mobile genetic elements. Phenotypic analyses including antibiotic susceptibility, enzyme activity, production of biogenic amines (BAs), and proportion of D-/L-lactate were evaluated using E-test, API ZYM test, high-performance liquid chromatography (HPLC), and D-/L-lactic acid assay respectively. The genome of B. breve IDCC4401 consists of 2,426,499 bp with a GC content of 58.70% and 2,016 coding regions. Confirmation of the genome as B. breve was provided by its 98.93% similarity with B. breve DSM20213. Furthermore, B. breve IDCC4401 genes encoding virulence and antibiotic resistance were not identified. Although B. breve IDCC4401 showed antibiotic resistance against vancomycin, we confirmed that this was an intrinsic feature since the antibiotic resistance gene was not present. B. breve IDCC4401 showed leucine arylamidase, cystine arylamidase, α-galactosidase, β-galactosidase, and α-glucosidase activities, whereas it did not show production of harmful enzymes such as β-glucosidase and β-glucuronidase. In addition, B. breve IDCC4401 did not produce any tyramine, histamine, putrescine, cadaverine, or 2-phenethylamine, which are frequently detected BAs during fermentation. B. breve IDCC4401 produced 95.08% of L-lactate and 4.92% of D-lactate. Therefore, our findings demonstrate the safety of B. breve IDCC 4401 as a potential probiotic for use in the food industry.

Novel Qnr Families as Conserved and Intrinsic Quinolone Resistance Determinants in Aeromonas spp.

  • Sang-Gyu Kim;Bo-Eun Kim;Jung Hun Lee;Dae-Wi Kim
    • Journal of Microbiology and Biotechnology
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    • v.34 no.6
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    • pp.1276-1286
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    • 2024
  • The environment has been identified as an origin, reservoir, and transmission route of antibiotic resistance genes (ARGs). Among diverse environments, freshwater environments have been recognized as pivotal in the transmission of ARGs between opportunistic pathogens and autochthonous bacteria such as Aeromonas spp. In this study, five environmental strains of Aeromonas spp. exhibiting multidrug resistance (MDR) were selected for whole-genome sequencing to ascertain their taxonomic assignment at the species-level and to delineate their ARG repertoires. Analyses of their genomes revealed the presence of one protein almost identical to AhQnr (A. hydrophila Qnr protein) and four novel proteins similar to AhQnr. To scrutinize the classification and taxonomic distribution of these proteins, all Aeromonas genomes deposited in the NCBI RefSeq genome database (1,222 genomes) were investigated. This revealed that these Aeromonas Qnr (AQnr) proteins are conserved intrinsic resistance determinants of the genus, exhibiting species-specific diversity. Additionally, structure prediction and analysis of contribution to quinolone resistance by AQnr proteins of the isolates, confirmed their functionality as quinolone resistance determinants. Given the origin of mobile qnr genes from aquatic bacteria and the crucial role of Aeromonas spp. in ARG dissemination in aquatic environments, a thorough understanding and strict surveillance of AQnr families prior to the clinical emergence are imperative. In this study, using comparative genome analyses and functional characterization of AQnr proteins in the genus Aeromonas, novel Aeromonas ARGs requiring surveillance has suggested.

Current status and prospects of citrus genomics (감귤 유전체 연구 동향 및 전망)

  • Kim, Ho Bang;Lim, Sanghyun;Kim, Jae Joon;Park, Young Cheol;Yun, Su-Hyun;Song, Kwan Jeong
    • Journal of Plant Biotechnology
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    • v.42 no.4
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    • pp.326-335
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    • 2015
  • Citrus is an economically important fruit tree with the largest amount of fruit production in the world. It provides important nutrition such as vitamin C and other health-promoting compounds including its unique flavonoids for human health. However, it is classified into the most difficult crops to develop new cultivars through conventional breeding approaches due to its long juvenility and some unique reproductive biological features such as gamete sterility, nucellar embryony, and high level of heterozygosity. Due to global warming and changes in consumer trends, establishing a systematic and efficient breeding programs is highly required for sustainable production of high quality fruits and diversification of cultivars. Recently, reference genome sequences of sweet orange and clementine mandarin have been released. Based on the reference whole-genome sequences, comparative genomics, reference-guided resequencing, and genotyping-by-sequencing for various citrus cultivars and crosses could be performed for the advance of functional genomics and development of traits-related molecular markers. In addition, a full understanding of gene function and gene co-expression networks can be provided through combined analysis of various transcriptome data. Analytic information on whole-genome and transcriptome will provide massive data on polymorphic molecular markers such as SNP, INDEL, and SSR, suggesting that it is possible to construct integrated maps and high-density genetic maps as well as physical maps. In the near future, integrated maps will be useful for map-based precise cloning of genes that are specific to citrus with major agronomic traits to facilitate rapid and efficient marker-assisted selection.

Prediction of Genomic Relationship Matrices using Single Nucleotide Polymorphisms in Hanwoo (한우의 유전체 표지인자 활용 개체 혈연관계 추정)

  • Lee, Deuk-Hwan;Cho, Chung-Il;Kim, Nae-Soo
    • Journal of Animal Science and Technology
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    • v.52 no.5
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    • pp.357-366
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    • 2010
  • The emergence of next-generation sequencing technologies has lead to application of new computational and statistical methodologies that allow incorporating genetic information from entire genomes of many individuals composing the population. For example, using single-nucleotide polymorphisms (SNP) obtained from whole genome amplification platforms such as the Ilummina BovineSNP50 chip, many researchers are actively engaged in the genetic evaluation of cattle livestock using whole genome relationship analyses. In this study, we estimated the genomic relationship matrix (GRM) and compared it with one computed using a pedigree relationship matrix (PRM) using a population of Hanwoo. This project is a preliminary study that will eventually include future work on genomic selection and prediction. Data used in this study were obtained from 187 blood samples consisting of the progeny of 20 young bulls collected after parentage testing from the Hanwoo improvement center, National Agriculture Cooperative Federation as well as 103 blood samples from the progeny of 12 proven bulls collected from farms around the Kyong-buk area in South Korea. The data set was divided into two cases for analysis. In the first case missing genotypes were included. In the second case missing genotypes were excluded. The effect of missing genotypes on the accuracy of genomic relationship estimation was investigated. Estimation of relationships using genomic information was also carried out chromosome by chromosome for whole genomic SNP markers based on the regression method using allele frequencies across loci. The average correlation coefficient and standard deviation between relationships using pedigree information and chromosomal genomic information using data which was verified using a parentage test andeliminated missing genotypes was $0.81{\pm}0.04$ and their correlation coefficient when using whole genomic information was 0.98, which was higher. Variation in relationships between non-inbred half sibs was $0.22{\pm}0.17$ on chromosomal and $0.22{\pm}0.04$ on whole genomic SNP markers. The variations were larger and unusual values were observed when non-parentage test data were included. So, relationship matrix by genomic information can be useful for genetic evaluation of animal breeding.

Genomics and Molecular Markers for Major Cucurbitaceae Crops (주요 박과작물의 유전체 및 분자마커 연구 현황)

  • Park, Girim;Kim, Nahui;Park, Younghoon
    • Journal of Life Science
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    • v.25 no.9
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    • pp.1059-1071
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    • 2015
  • Watermelon and melon are economically important Cucurbitaceae crops. Recently, the development of molecular markers based on the construction of genetic linkage maps and detection of DNA sequence variants through next generation sequencing are essential as molecular breeding strategies for crop improvement that uses marker-assisted selection and backcrossing. In this paper, we intended to provide useful information for molecular breeding of watermelon and melon by analyzing the current status of international and domestic research efforts on genomics and molecular markers. Due to diverse genetic maps constructed and the reference genome sequencing completed in the past, DNA markers that are useful for selecting important traits including yield, fruit quality, and disease resistances have been reported and publicly available. To date, more than 16 genetic maps and loci and linked markers for more than 40 traits have reported for each watermelon and melon. Furthermore, the functional genes that are responsible for those traits are being continuously discovered by high-density genetic map and map-based cloning. In addition, whole genome resequencing of various germplasm is under progress based on the reference genome. Not only by the efforts for developing novel molecular markers, but application of public marker information currently available will greatly facilitate breeding process through genomics-assisted breeding.

Endless debates on the extant basal-most angiosperm (현생 기저 피자식물에 대한 끝나지 않는 논쟁)

  • Kim, Sangtae
    • Korean Journal of Plant Taxonomy
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    • v.40 no.1
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    • pp.1-15
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    • 2010
  • Recognizing a basal group in a taxon is one of the most important factors involved in understanding the evolutionary history of that group of life. Many botanists have suggested a sister to all other angiosperms to understand the origin and rapid diversification of angiosperms based on morphological and fossil evidence. Recent technical advances in molecular biology and the accumulation of molecular phylogenetic data have provided evidence of the extant basal-most angiosperm which is a sister to all other angiosperms. Although it is still arguable, most plant taxonomists agree that Amborella trichopoda Baill., a species (monotypic genus and monotypic family) distributed in New Caledonia, is a sister to all other extant angiosperms based on evidence from the following molecular approaches: 1) classical phylogenetic analyses based on multiple genes (or DNA regions), 2) analyses of a tree network of duplicated gene families, and 3) gene-structural evidence. As an alternative hypothesis with relatively minor evidence, some researchers have also suggested that Amborella and Nymphaeaceae form a clade that is a sister to all other angiosperms. Debate regarding the basal-most angiosperms is still ongoing and is currently one of the hot issues in plant evolutionary biology. We expect that sequencing of the whole genome of Amborella as an evolutionary model plant and subsequent studies based on this genome sequence will provide information regarding the origin and rapid diversification of angiosperms, which is Darwin's so called abominable mystery.

Historical Record of Mushroom Research and Industry in Korea

  • Yoo, Young Bok;Oh, Youn Lee;Shin, Pyung Gyun;Jang, Kab Yeul;Sung, Gi-Ho;Kong, Won-Sik
    • 한국균학회소식:학술대회논문집
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    • 2014.05a
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    • pp.13-13
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    • 2014
  • Two kinds of mushrooms, Gumji (金芝; Ganoderma) and Soji, were described in old book of Samguksagi (History of the three kingdoms; B.C 57~A.D 668; written by Bu Sik Kim in 1145) in Korea-dynasty. Many kinds of mushrooms were also described in more than 17 kinds of old books during Chosun-dynasty (1392~1910) in Korea. Nowadays, mushroom cultivation has been increased through out the world last decade years. Production of mushrooms has also been increased 10-20% and many varieties have been cultivated. Similar trends were also observed in Korea. Approximately two hundred commercial strains of 37 species in mushrooms were developed and distributed to cultivators. Somatic hybrid variety of oyster mushroom 'Wonhyeong-neutari' were developed by protoplast fusion, and distributed to grower in 1989. The fruiting body yield index of somatic hybrids of Pleurotus ranged between 27 and 155 compared to parental values of 100 and 138. In addition, more diverse mushroom varieties such as Phellinus baumi, Auricularia spp., Pleurotus ferulae, Hericium erinaceus, Hypsizigus marmoreus, Grifola frondosa, Agrocybe aegerita and Pleurotus cornucopiae have been attempted to cultivate in small scale cultivation. Production of mushrooms as food was 190,111 metric tons valued at 800 billion Korean Won (one trillion won if include mushroom factory products; 1dollar = 1,040 Won) in 2011. Major cultivated species are Pleurotus ostreatus, Pleurotus eryngii, Flammulina velutipes, Lentinula edodes, Agaricus bisporus, and Ganoderma lucidum, which cover 90% of total production. Since mushroom export was initiated from 1960 to 1980, the export and import of mushrooms have been increased in Korea. Technology developed for liquid spawn production and automatic cultivation systems lead to the reduction of the production cost resulting in the increasement of mushroom export. However some species were imported because of high production cost for these mushrooms requiring the effective cultivation methods. Developing of effective post-harvest system will be also directly related to mushroom export. In academic area, RDA scientists have been conducting mushroom genome projects. One of the main results is the whole genome sequencing of Flammulina velutipes for molecular breeding. An electrophoretic karyotype of of F. velutipes was obtained using CHEF with 7 chromosomes, with a total genome size of approximately 26.7 Mb. The mususcript of the genome of F. velutipes was published in PLOS ONE this year. For medicinal mushrooms, we have been conducting the genome research on Cordyceps and its related species for developing functional foods using this mushroom. In 2013, Korea Food and Drug Administraion (KFDA) approved Cordyceps mushroom for its value as an immune booster.

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Chromosomal Assembly of Tegillarca granosa Genome using Third-generation DNA Sequencing and Hi-C Technology (3세대 DNA 염기서열 분석과 Hi-C기술을 이용한 꼬막 게놈의 유전체 연구)

  • Kim, Jinmu;Lee, Seung Jae;Jo, Euna;Choi, Eunkyung;Cho, Minjoo;Shin, So Ryung;Lee, Jung Sick;Park, Hyun
    • Journal of Marine Life Science
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    • v.6 no.2
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    • pp.97-105
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    • 2021
  • Tegillarca granosa, is one of the most important fishery resources throughout Asia. However, due to industrialization factories, marine environmental pollution, and global warming, the marine fishery production has drop sharply. In order to understand the genetic factors of the blood clam, which is a major fishery resource on the southern coast of Korea, the whole genome of blood clam was studied. The assembled genome of T. granosa was 915.4 Mb, and 19 chromosomes were identified. 25,134 genes were identified, and 22,745 genes were functionally annotated. As a result of performing gene gain and loss analysis between the blood clam genome and eight other types of shellfish, it was confirmed that 725 gene groups were expanded, and 479 gene groups were contracted. The homeobox gene cluster of blood clam showed a well-preserved genetic structure within lophotrochozoan ancestor. T. granosa genome showed high similarity between three hemoglobin genes with Scarpharca broughtonii. The blood clam genome will provide information for the genetic and physiological characteristics of blood clam adaptation, evolution, and the development of aquaculture industry.

Elucidation of the Biosynthetic Pathway of Vitamin B Groups and Potential Secondary Metabolite Gene Clusters Via Genome Analysis of a Marine Bacterium Pseudoruegeria sp. M32A2M

  • Cho, Sang-Hyeok;Lee, Eunju;Ko, So-Ra;Jin, Sangrak;Song, Yoseb;Ahn, Chi-Yong;Oh, Hee-Mock;Cho, Byung-Kwan;Cho, Suhyung
    • Journal of Microbiology and Biotechnology
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    • v.30 no.4
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    • pp.505-514
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    • 2020
  • The symbiotic nature of the relationship between algae and marine bacteria is well-studied among the complex microbial interactions. The mutual profit between algae and bacteria occurs via nutrient and vitamin exchange. It is necessary to analyze the genome sequence of a bacterium to predict its symbiotic relationships. In this study, the genome of a marine bacterium, Pseudoruegeria sp. M32A2M, isolated from the south-eastern isles (GeoJe-Do) of South Korea, was sequenced and analyzed. A draft genome (91 scaffolds) of 5.5 Mb with a DNA G+C content of 62.4% was obtained. In total, 5,101 features were identified from gene annotation, and 4,927 genes were assigned to functional proteins. We also identified transcription core proteins, RNA polymerase subunits, and sigma factors. In addition, full flagella-related gene clusters involving the flagellar body, motor, regulator, and other accessory compartments were detected even though the genus Pseudoruegeria is known to comprise non-motile bacteria. Examination of annotated KEGG pathways revealed that Pseudoruegeria sp. M32A2M has the metabolic pathways for all seven vitamin Bs, including thiamin (vitamin B1), biotin (vitamin B7), and cobalamin (vitamin B12), which are necessary for symbiosis with vitamin B auxotroph algae. We also identified gene clusters for seven secondary metabolites including ectoine, homoserine lactone, beta-lactone, terpene, lasso peptide, bacteriocin, and non-ribosomal proteins.

Species Transferability of Klebsiella pneumoniae Carbapenemase-2 Isolated from a High-Risk Clone of Escherichia coli ST410

  • Lee, Miyoung;Choi, Tae-Jin
    • Journal of Microbiology and Biotechnology
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    • v.30 no.7
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    • pp.974-981
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    • 2020
  • Sequence type 410 (ST410) of Escherichia coli is an extraintestinal pathogen associated with multi drug resistance. In this study, we aimed to investigate the horizontal propagation pathway of a high-risk clone of E. coli ST410 that produces Klebsiella pneumoniae carbapenemase (KPC). blaKPC-encoding E. coli and K. pneumoniae isolates were evaluated, and complete sequencing and comparative analysis of blaKPC-encoding plasmids from E. coli and K. pneumoniae, antimicrobial susceptibility tests, polymerase chain reaction, multilocus sequence typing, and conjugal transfer of plasmids were performed. Whole-genome sequencing was performed for plasmids mediating KPC-2 production in E. coli and K. pneumoniae clinical isolates. Strains E. coli CPEc171209 and K. pneumoniae CPKp171210 were identified as ST410 and ST307, respectively. CPEc171209 harbored five plasmids belonging to serotype O8:H21, which is in the antimicrobial-resistant clade C4/H24. The CPKp171210 isolate harbored three plasmids. Both strains harbored various additional antimicrobial resistance genes. The IncX3 plasmid pECBHS_9_5 harbored blaKPC-2 within a truncated Tn4401a transposon, which also contains blaSHV-182 with duplicated conjugative elements. This plasmid displayed 100% identity with the IncX3 plasmid pKPBHS_10_3 from the K. pneumoniae CPKp171210 ST307 strain. The genes responsible for the conjugal transfer of the IncX3 plasmid included tra/trb clusters and pil genes coding the type IV pilus. ST410 can be transmitted between patients, posing an elevated risk in clinical settings. The emergence of a KPC-producing E. coli strain (ST410) is concerning because the blaKPC-2-bearing plasmids may carry treatment resistance across species barriers. Transgenic translocation occurs among carbapenem-resistant bacteria, which may spread rapidly via horizontal migration.