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De novo assembly and comparative transcriptome analysis of Euglena gracilis in response to anaerobic conditions

BACKGROUND: The phytoflagellated protozoan, Euglena gracilis, has been proposed as an attractive feedstock for the accumulation of valuable compounds such as β-1,3-glucan, also known as paramylon, and wax esters. The production of wax esters proceeds under anaerobic conditions, designated as wax est...

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Autores principales: Yoshida, Yuta, Tomiyama, Takuya, Maruta, Takanori, Tomita, Masaru, Ishikawa, Takahiro, Arakawa, Kazuharu
Formato: Online Artículo Texto
Lenguaje:English
Publicado: BioMed Central 2016
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4778363/
https://www.ncbi.nlm.nih.gov/pubmed/26939900
http://dx.doi.org/10.1186/s12864-016-2540-6
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author Yoshida, Yuta
Tomiyama, Takuya
Maruta, Takanori
Tomita, Masaru
Ishikawa, Takahiro
Arakawa, Kazuharu
author_facet Yoshida, Yuta
Tomiyama, Takuya
Maruta, Takanori
Tomita, Masaru
Ishikawa, Takahiro
Arakawa, Kazuharu
author_sort Yoshida, Yuta
collection PubMed
description BACKGROUND: The phytoflagellated protozoan, Euglena gracilis, has been proposed as an attractive feedstock for the accumulation of valuable compounds such as β-1,3-glucan, also known as paramylon, and wax esters. The production of wax esters proceeds under anaerobic conditions, designated as wax ester fermentation. In spite of the importance and usefulness of Euglena, the genome and transcriptome data are currently unavailable, though another research group has recently published E.gracilis transcriptome study during our submission. We herein performed an RNA-Seq analysis to provide a comprehensive sequence resource and some insights into the regulation of genes including wax ester metabolism by comparative transcriptome analysis of E.gracilis under aerobic and anaerobic conditions. RESULTS: The E.gracilis transcriptome analysis was performed using the Illumina platform and yielded 90.3 million reads after the filtering steps. A total of 49,826 components were assembled and identified as a reference sequence of E.gracilis, of which 26,479 sequences were considered to be potentially expressed (having FPKM value of greater than 1). Approximately half of all components were estimated to be regulated in a trans-splicing manner, with the addition of protruding spliced leader sequences. Nearly 40 % of 26,479 sequences were annotated by similarity to Swiss-Prot database using the BLASTX program. A total of 2080 transcripts were identified as differentially expressed genes (DEGs) in response to anaerobic treatment for 24 h. A comprehensive pathway enrichment analysis using the KEGG pathway revealed that the majority of DEGs were involved in photosynthesis, nucleotide metabolism, oxidative phosphorylation, fatty acid metabolism. We successfully identified a candidate gene set of paramylon and wax esters, including novel β-1,3-glucan and wax ester synthases. A comparative expression analysis of aerobic- and anaerobic-treated E.gracilis cells indicated that gene expression changes in these components were not extensive or dynamic during the anaerobic treatment. CONCLUSION: The RNA-Seq analysis provided comprehensive transcriptome information on E.gracilis for the first time, and this information will advance our understanding of this unique organism. The comprehensive analysis indicated that paramylon and wax ester metabolic pathways are regulated at post-transcriptional rather than the transcriptional level in response to anaerobic conditions. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (doi:10.1186/s12864-016-2540-6) contains supplementary material, which is available to authorized users.
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spelling pubmed-47783632016-03-05 De novo assembly and comparative transcriptome analysis of Euglena gracilis in response to anaerobic conditions Yoshida, Yuta Tomiyama, Takuya Maruta, Takanori Tomita, Masaru Ishikawa, Takahiro Arakawa, Kazuharu BMC Genomics Research Article BACKGROUND: The phytoflagellated protozoan, Euglena gracilis, has been proposed as an attractive feedstock for the accumulation of valuable compounds such as β-1,3-glucan, also known as paramylon, and wax esters. The production of wax esters proceeds under anaerobic conditions, designated as wax ester fermentation. In spite of the importance and usefulness of Euglena, the genome and transcriptome data are currently unavailable, though another research group has recently published E.gracilis transcriptome study during our submission. We herein performed an RNA-Seq analysis to provide a comprehensive sequence resource and some insights into the regulation of genes including wax ester metabolism by comparative transcriptome analysis of E.gracilis under aerobic and anaerobic conditions. RESULTS: The E.gracilis transcriptome analysis was performed using the Illumina platform and yielded 90.3 million reads after the filtering steps. A total of 49,826 components were assembled and identified as a reference sequence of E.gracilis, of which 26,479 sequences were considered to be potentially expressed (having FPKM value of greater than 1). Approximately half of all components were estimated to be regulated in a trans-splicing manner, with the addition of protruding spliced leader sequences. Nearly 40 % of 26,479 sequences were annotated by similarity to Swiss-Prot database using the BLASTX program. A total of 2080 transcripts were identified as differentially expressed genes (DEGs) in response to anaerobic treatment for 24 h. A comprehensive pathway enrichment analysis using the KEGG pathway revealed that the majority of DEGs were involved in photosynthesis, nucleotide metabolism, oxidative phosphorylation, fatty acid metabolism. We successfully identified a candidate gene set of paramylon and wax esters, including novel β-1,3-glucan and wax ester synthases. A comparative expression analysis of aerobic- and anaerobic-treated E.gracilis cells indicated that gene expression changes in these components were not extensive or dynamic during the anaerobic treatment. CONCLUSION: The RNA-Seq analysis provided comprehensive transcriptome information on E.gracilis for the first time, and this information will advance our understanding of this unique organism. The comprehensive analysis indicated that paramylon and wax ester metabolic pathways are regulated at post-transcriptional rather than the transcriptional level in response to anaerobic conditions. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (doi:10.1186/s12864-016-2540-6) contains supplementary material, which is available to authorized users. BioMed Central 2016-03-03 /pmc/articles/PMC4778363/ /pubmed/26939900 http://dx.doi.org/10.1186/s12864-016-2540-6 Text en © Yoshida et al. 2016 Open AccessThis article is distributed under the terms of the Creative Commons Attribution 4.0 International License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution, and reproduction in any medium, provided you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated.
spellingShingle Research Article
Yoshida, Yuta
Tomiyama, Takuya
Maruta, Takanori
Tomita, Masaru
Ishikawa, Takahiro
Arakawa, Kazuharu
De novo assembly and comparative transcriptome analysis of Euglena gracilis in response to anaerobic conditions
title De novo assembly and comparative transcriptome analysis of Euglena gracilis in response to anaerobic conditions
title_full De novo assembly and comparative transcriptome analysis of Euglena gracilis in response to anaerobic conditions
title_fullStr De novo assembly and comparative transcriptome analysis of Euglena gracilis in response to anaerobic conditions
title_full_unstemmed De novo assembly and comparative transcriptome analysis of Euglena gracilis in response to anaerobic conditions
title_short De novo assembly and comparative transcriptome analysis of Euglena gracilis in response to anaerobic conditions
title_sort de novo assembly and comparative transcriptome analysis of euglena gracilis in response to anaerobic conditions
topic Research Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4778363/
https://www.ncbi.nlm.nih.gov/pubmed/26939900
http://dx.doi.org/10.1186/s12864-016-2540-6
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