• Title/Summary/Keyword: ITS1 sequencing

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First Report of Gray Mold Disease of Sweet Basil (Ocimum basilicum) Caused by Botrytis cinerea in Korea

  • Aktaruzzaman, Md.;Kim, Joon-Young;Afroz, Tania;Hong, Sae-Jin;Kim, Byung-Sup
    • The Korean Journal of Mycology
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    • v.43 no.4
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    • pp.277-280
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    • 2015
  • In August 2015, we collected samples of gray mold from sweet basil growing in Sachunmeon, Gangneung, Gangwon Province, Korea. Symptoms included extensive growth of mycelia with gray conidia on young leaves, stems, and blossoms. The pathogen was isolated from infected leaves and blossoms and the fungus was cultured on potato dextrose agar. For identification of the fungus, morphology and rDNA sequencing analysis of the fungus were performed, which confirmed its pathogenicity according to Koch's postulates. The results of morphological examinations, pathogenicity tests, and the rDNA sequences of the internal transcribed spacer regions (ITS1 and ITS4) and the three nuclear protein-coding genes G3PDH, HSP60, and RPB2 showed that the causal agent was Botrytis cinerea. This is the first report of gray mold caused by Botrytis cinerea on sweet basil in Korea.

LITHOAUTOTROPHIC NITROGEN REMOVAL WITH ANAEROBIC GRANULAR SLUDGE AS SEED BIOMASS AND ITS MICROBIAL COMMUNITY

  • Ahn, Young-Ho;Lee, Jin-Woo;Kim, Hee-Chul;Kwon, Soo-Youl
    • Environmental Engineering Research
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    • v.11 no.4
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    • pp.173-180
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    • 2006
  • Autotrophic nitrogen removal and its microbial community from a laboratory scale upflow anaerobic sludge bed reactor were characterized with dynamic behavior of nitrogen removal and sequencing result of molecular technique (DNA extraction, PCR and amplification of 16S rDNA), respectively. In the experiment treating inorganic wastewater, the anaerobic granular sludge from a full-scale UASB reactor treating industrial wastewater was inoculated as seed biomass. The operating results revealed that an addition of hydroxylamine would result in lithoautotrophic ammonium oxidation to nitrite/nitrate, and also hydrazine would play an important role for the success of sustainable nitrogen removal process. Total N and ammonium removal of 48% and 92% was observed, corresponding to nitrogen conversion of 0.023 g N/L-d. The reddish brown-colored granular sludge with a diameter of $1{\sim}2\;mm$ was observed at the lower part of sludge bed. The microbial characterization suggests that an anoxic ammonium oxidizer and an anoxic denitrifying autotrophic nitrifier contribute mainly to the nitrogen removal in the reactor. The results revealed the feasibility on development of high performance lithoautotrophic nitrogen removal process with its microbial granulation.

Characterization of Newly Recorded Talaromyces veerkampii Isolated from Field Soil in Korea based on Morphology and Multigene Sequence Analysis

  • Mahesh Adhikari;Hyun Seung Kim;Hyo Bin Park;Ki Young Kim;In Kyu Lee;Eun Jeong Byeon;Ji Min Woo;Hyang Burm Lee;Youn Su Lee
    • The Korean Journal of Mycology
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    • v.50 no.4
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    • pp.347-355
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    • 2022
  • A fungal isolate belonging to the phylum Ascomycota was isolated and identified as Talaromyces veerkampii in 2017 during a survey of fungal diversity in field soils in Korea. This fungal isolate was identified and described based on macro- and micromorphological and molecular characterization. The identification was also based on partial 18S-ITS1-5.8S-ITS2-28S rDNA and calmodulin (CaM)-encoding gene sequencing data. Talaromyces veerkampii has not been previously reported in Korea. Thus, we report here a newly discovered species from soil in Korea along with its morphological and molecular characteristics.

CaGe: A Web-Based Cancer Gene Annotation System for Cancer Genomics

  • Park, Young-Kyu;Kang, Tae-Wook;Baek, Su-Jin;Kim, Kwon-Il;Kim, Seon-Young;Lee, Do-Heon;Kim, Yong-Sung
    • Genomics & Informatics
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    • v.10 no.1
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    • pp.33-39
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    • 2012
  • High-throughput genomic technologies (HGTs), including next-generation DNA sequencing (NGS), microarray, and serial analysis of gene expression (SAGE), have become effective experimental tools for cancer genomics to identify cancer-associated somatic genomic alterations and genes. The main hurdle in cancer genomics is to identify the real causative mutations or genes out of many candidates from an HGT-based cancer genomic analysis. One useful approach is to refer to known cancer genes and associated information. The list of known cancer genes can be used to determine candidates of cancer driver mutations, while cancer gene-related information, including gene expression, protein-protein interaction, and pathways, can be useful for scoring novel candidates. Some cancer gene or mutation databases exist for this purpose, but few specialized tools exist for an automated analysis of a long gene list from an HGT-based cancer genomic analysis. This report presents a new web-accessible bioinformatic tool, called CaGe, a cancer genome annotation system for the assessment of candidates of cancer genes from HGT-based cancer genomics. The tool provides users with information on cancer-related genes, mutations, pathways, and associated annotations through annotation and browsing functions. With this tool, researchers can classify their candidate genes from cancer genome studies into either previously reported or novel categories of cancer genes and gain insight into underlying carcinogenic mechanisms through a pathway analysis. We show the usefulness of CaGe by assessing its performance in annotating somatic mutations from a published small cell lung cancer study.

Estimation of Kinetic Coefficient and Assimilated Nutrients Mass in SBR Process (연속회분식 반응 공정에서 동역학적 계수 및 미생물합성에 사용된 영양물질 산정)

  • Ji, Dae-Hyun;Shin, Sang-Woo;Lee, Kwang-Ho;Lee, Jae-Kune
    • Journal of Korean Society on Water Environment
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    • v.23 no.5
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    • pp.607-612
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    • 2007
  • In this study, we investigated the variations of the kinetic coefficients and Chemical Oxygen Demand (COD), N and P mass used for assimilation of a sequencing batch reactor (SBR) system with the variation of SRTs; SRTs of 7.5, 10.0, 12.5, 15.0 and 20.0 days were tested in one cycle of SBR operation to determine the optimum conditions for the operation of the SBR and estimate its COD, nitrogen and phosphorus removal efficiencies. The SBR system was operated under the conditions as follows: an operation time of 6 hours per cycle, a hydraulic retention time (HRT) of 12 hours, an influent COD loading of $0.4kg/m^3/day$, and an influent nitrogen loading of $0.068kgT-N/m^3/day$. The yield coefficient (Y) and decay rate coefficient ($k_d$) were estimated to be 0.4198 kgMLVSS/kgCOD and $0.0107day^{-1}$ by calculating the removal rate of substrate according to the variation of SRT. Considering total nitrogen amount removed by sludge waste process, eliminated by denitrification, and in clarified water effluent with reference to 150 mg/cycle of influent nitrogen amount, the percentage of nitrogen mass balance from the ratio of the nitrogen amount in effluent (N output) to that in influent (N input) for Runs 1~5 were 95.5, 97.0, 95.5, 99.5, and 95.5%, respectively, which is well accounted for, with mass balances close to 100%.

Investigation of Ectomycorrhizal Fungal Colonization in Pinus thunbergii Seedlings at a Plantation Area in Gangneung, using Morphotyping and Sequencing the rDNA Internal Transcribed Spacer Region

  • Obase, Keisuke;Cha, Joo-Young;Lee, Jong-Kyu;Lee, Sang-Yong;Lee, Jin-Ho;Chun, Kun-Woo
    • Journal of Korean Society of Forest Science
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    • v.99 no.2
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    • pp.172-178
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    • 2010
  • The status of ectomycorrhizal (ECM) fungal colonization in Pinus thunbergii seedlings was investigated 2 years after planting in an eastern coastal area of Korea. We established three $10{\times}10$ m plots at a P. thunbergii plantation in Gangneung and sampled lateral roots from 10 seedlings in each plot. ECMs were classified into morphological groups and the number of root tips of each morphotype was counted. In total, 8 ECM morphotypes were observed and fungal species that form each morphotype were identified by sequencing of the internal transcribed spacer (ITS) region of the nuclear rDNA. Suillus granulatus was the most abundant species (44.1-65.7% of relative abundance) in all plots, followed by Tomentella ellisii (14.0-37.8%) and unidentified fungus belonged to Atheliaceae (10.6-20.1%). These 3 fungal species accounted for almost all of the ECM abundance in each plot (94.9-99.8%). The remaining 5 fungal species were uncommon and rare. There was no clear difference in ECM fungal communities among plots. Community structure of ECM fungi in the young P. thunbergii plantation was simple and composed of fungal species that were also observed in mature coastal pine forests.

Draft Genome of Toxocara canis, a Pathogen Responsible for Visceral Larva Migrans

  • Kong, Jinhwa;Won, Jungim;Yoon, Jeehee;Lee, UnJoo;Kim, Jong-Il;Huh, Sun
    • Parasites, Hosts and Diseases
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    • v.54 no.6
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    • pp.751-758
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    • 2016
  • This study aimed at constructing a draft genome of the adult female worm Toxocara canis using next-generation sequencing (NGS) and de novo assembly, as well as to find new genes after annotation using functional genomics tools. Using an NGS machine, we produced DNA read data of T. canis. The de novo assembly of the read data was performed using SOAPdenovo. RNA read data were assembled using Trinity. Structural annotation, homology search, functional annotation, classification of protein domains, and KEGG pathway analysis were carried out. Besides them, recently developed tools such as MAKER, PASA, Evidence Modeler, and Blast2GO were used. The scaffold DNA was obtained, the N50 was 108,950 bp, and the overall length was 341,776,187 bp. The N50 of the transcriptome was 940 bp, and its length was 53,046,952 bp. The GC content of the entire genome was 39.3%. The total number of genes was 20,178, and the total number of protein sequences was 22,358. Of the 22,358 protein sequences, 4,992 were newly observed in T. canis. Following proteins previously unknown were found: E3 ubiquitin-protein ligase cbl-b and antigen T-cell receptor, zeta chain for T-cell and B-cell regulation; endoprotease bli-4 for cuticle metabolism; mucin 12Ea and polymorphic mucin variant C6/1/40r2.1 for mucin production; tropomodulin-family protein and ryanodine receptor calcium release channels for muscle movement. We were able to find new hypothetical polypeptides sequences unique to T. canis, and the findings of this study are capable of serving as a basis for extending our biological understanding of T. canis.

Isolation and Identification of Rice Root Endophytic Antagonistic Serratia marcescens (벼 뿌리 내생 항균성 Serratia marcescens의 분리 및 동정)

  • Lee, Sook-Kyung;Song, Wan-Yeob;Kim, Hyung-Moo
    • Research in Plant Disease
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    • v.10 no.1
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    • pp.63-68
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    • 2004
  • Twenty-three strains of Serratia sp., isolated from surface-sterilized rice roots collected in Chonbuk and Chungnam province, were identified and characterized. They were Gram-negative, rod shaped and red pigmented typically and their endophytism was confirmed by inoculation and reisolation of the strains in planta. Their antifungal activity against 4 rice pathogenic fungi was compared and ranged from 62.4 to 85.2% against Rhizoctonia solani and 68.0 to 88.5% against Pyricularia grisea. Among the 23 strains tested, strain Rsm220 showed the strongest inhibition activity against 4 pathogenic fungi. The strain was, therefore, selected as a biocontrol candidate for both the pathogens and its bacteriological characteristics and 165 rDNA sequences were analyzed. Phenotypic and biochemical characteristics of the selected Rsm220 were highly related to the type strain of S. marcescens and 165 rDNA sequencing of Rsm220 showed a homology of 98.2% to the type strain of S. marcescens. The strain Rsm220 was identified as S. marcescens and the inhibition result of this endophytic strain indicates that it is a potential biocontrol agent for R. solani and R grisea.

Whole-transcriptome analyses of the Sapsaree, a Korean natural monument, before and after exercise-induced stress

  • Kim, Ji-Eun;Choe, Junkyung;Lee, Jeong Hee;Kim, Woong Bom;Cho, Whan;Ha, Ji Hong;Kwon, Ki Jin;Han, Kook Il;Jo, Sung-Hwan
    • Journal of Animal Science and Technology
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    • v.58 no.4
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    • pp.17.1-17.7
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    • 2016
  • Background: The Sapsaree (Canis familiaris) is a Korean native dog that is very friendly, protective, and loyal to its owner, and is registered as a natural monument in Korea (number: 368). To investigate large-scale gene expression profiles and identify the genes related to exercise-induced stress in the Sapsaree, we performed whole-transcriptome RNA sequencing and analyzed gene expression patterns before and after exercise performance. Results: We identified 525 differentially expressed genes in ten dogs before and after exercise. Gene Ontology classification and KEGG pathway analysis revealed that the genes were mainly involved in metabolic processes, such as programmed cell death, protein metabolic process, phosphatidylinositol signaling system, and cation binding in cytoplasm. The ten Sapsarees could be divided into two groups based on the gene expression patterns before and after exercise. The two groups were significantly different in terms of their basic body type ($p{\leq}0.05$). Seven representative genes with significantly different expression patterns before and after exercise between the two groups were chosen and characterized. Conclusions: Body type had a significant effect on the patterns of differential gene expression induced by exercise. Whole-transcriptome sequencing is a useful method for investigating the biological characteristics of the Sapsaree and the large-scale genomic differences of canines in general.

Application of Molecular Methods for the Identification of Acetic Acid Bacteria Isolated from Blueberries and Citrus Fruits

  • Gerard, Liliana Mabel;Davies, Cristina Veronica;Solda, Carina Alejandra;Corrado, Maria Belen;Fernandez, Maria Veronica
    • Microbiology and Biotechnology Letters
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    • v.48 no.2
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    • pp.193-204
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    • 2020
  • Sixteen acetic acid bacteria (AAB) were isolated from blueberries and citric fruits of the Salto Grande region (Concordia, Entre Rios, Argentina) using enrichment techniques and plate isolation. Enrichment broths containing ethanol and acetic acid enabled maximum AAB recovery, since these components promote their growth. Biochemical tests allowed classification of the bacteria at genus level. PCR-RFLP of the 16S rRNA and PCR-RFLP of the 16S-23S rRNA intergenic spacer allowed further classification at the species level; this required treatment of the amplified products of 16S and 16S-23S ITS ribosomal genes with the following restriction enzymes: AluI, RsaI, HaeIII, MspI, TaqI, CfoI, and Tru9I. C7, C8, A80, A160, and A180 isolates were identified as Gluconobacter frateurii; C1, C2, C3, C4, C5, C6, A70, and A210 isolates as Acetobacter pasteurianus; A50 and A140 isolates as Acetobacter tropicalis; and C9 isolate as Acetobacter syzygii. The bacteria identified by 16S rRNA PCR-RFLP were validated by 16S-23S PCR-RFLP; however, the C1 isolate showed different restriction patterns during identification and validation. Partial sequencing of the 16S gene resolved the discrepancy.