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Genome-scale analyses and characteristics of putative pathogenicity genes of Stagonosporopsis cucurbitacearum, a pumpkin gummy stem blight fungus
Outbreaks of gummy stem blight (GSB), an emerging seed pumpkin disease, have increased in number and have become more widespread in recent years. Previously we reported that Stagonosporopsis cucurbitacearum ( Sc. ) is the dominant fungal cause of pumpkin seedling GSB in Northeast China, where it has...
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Published in: | Scientific reports 2020-10, Vol.10 (1), p.18065-18065, Article 18065 |
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description | Outbreaks of gummy stem blight (GSB), an emerging seed pumpkin disease, have increased in number and have become more widespread in recent years. Previously we reported that
Stagonosporopsis cucurbitacearum
(
Sc.
) is the dominant fungal cause of pumpkin seedling GSB in Northeast China, where it has greatly reduced crop yields in that region. Here, high-throughput whole-genome sequencing and assembly of the
Sc.
genome were conducted toward revealing pathogenic molecular regulatory mechanisms involved in fungal growth and development
.
Zq-1 as representative
Sc.
strain, DNA of Zq-1was prepared for genomic sequencing, we obtained 5.24 Gb of high-quality genomic sequence data via PacBio RS II sequencing. After sequence data was processed to filter out low quality reads, a hierarchical genome-assembly process was employed that generated a genome sequence of 35.28 Mb in size. A total of 9844 genes were predicted, including 237 non-coding RNAs, 1024 genes encoding proteins with signal peptides, 2066 transmembrane proteins and 756 secretory proteins.Transcriptional identification revealed 54 differentially expressed secretory proteins. Concurrently, 605, 130 and 2869 proteins were matched in the proprietary databases Carbohydrate-Active EnZymes database (CAZyme), Transporter Classification Database (TCDB) and Pathogen–Host Interactions database (PHI), respectively. And 96 and 36 DEGs were identified form PHI database and CAZyme database, respectively. In addition, contig00011.93 was an up-regulated DEG involving ATP-binding cassette metabolism in the procession of infection. In order to test relevance of gene predictions to GSB, DEGs with potential pathogenic relevance were revealed through transcriptome data analysis of
Sc.
strains pre- and post-infection of pumpkin. Interestingly,
Sc.
and
Leptosphaeria maculans
(
Lm.
) exhibited relatively similar with genome lengths, numbers of protein-coding genes and other characteristics. This work provides a foundation for future exploration of additional
Sc.
gene functions toward the development of more effective GSB control strategies. |
doi_str_mv | 10.1038/s41598-020-75235-x |
format | article |
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Stagonosporopsis cucurbitacearum
(
Sc.
) is the dominant fungal cause of pumpkin seedling GSB in Northeast China, where it has greatly reduced crop yields in that region. Here, high-throughput whole-genome sequencing and assembly of the
Sc.
genome were conducted toward revealing pathogenic molecular regulatory mechanisms involved in fungal growth and development
.
Zq-1 as representative
Sc.
strain, DNA of Zq-1was prepared for genomic sequencing, we obtained 5.24 Gb of high-quality genomic sequence data via PacBio RS II sequencing. After sequence data was processed to filter out low quality reads, a hierarchical genome-assembly process was employed that generated a genome sequence of 35.28 Mb in size. A total of 9844 genes were predicted, including 237 non-coding RNAs, 1024 genes encoding proteins with signal peptides, 2066 transmembrane proteins and 756 secretory proteins.Transcriptional identification revealed 54 differentially expressed secretory proteins. Concurrently, 605, 130 and 2869 proteins were matched in the proprietary databases Carbohydrate-Active EnZymes database (CAZyme), Transporter Classification Database (TCDB) and Pathogen–Host Interactions database (PHI), respectively. And 96 and 36 DEGs were identified form PHI database and CAZyme database, respectively. In addition, contig00011.93 was an up-regulated DEG involving ATP-binding cassette metabolism in the procession of infection. In order to test relevance of gene predictions to GSB, DEGs with potential pathogenic relevance were revealed through transcriptome data analysis of
Sc.
strains pre- and post-infection of pumpkin. Interestingly,
Sc.
and
Leptosphaeria maculans
(
Lm.
) exhibited relatively similar with genome lengths, numbers of protein-coding genes and other characteristics. This work provides a foundation for future exploration of additional
Sc.
gene functions toward the development of more effective GSB control strategies.</description><identifier>ISSN: 2045-2322</identifier><identifier>EISSN: 2045-2322</identifier><identifier>DOI: 10.1038/s41598-020-75235-x</identifier><identifier>PMID: 33093634</identifier><language>eng</language><publisher>London: Nature Publishing Group UK</publisher><subject>631/326/193/2541 ; 631/326/421 ; Ascomycota - genetics ; Ascomycota - growth & development ; Ascomycota - pathogenicity ; China ; Crop Production ; Crop yield ; Cucurbita - microbiology ; DNA sequencing ; Fungi ; Gene expression ; Genes ; Genome, Fungal - genetics ; Genomes ; Host-Pathogen Interactions - genetics ; Humanities and Social Sciences ; Membrane proteins ; multidisciplinary ; Non-coding RNA ; Nucleotide sequence ; Pathogenicity ; Pathogens ; Peptides ; Plant Diseases - microbiology ; Proteins ; Science ; Science (multidisciplinary) ; Seedlings ; Signal peptides ; Stagonosporopsis cucurbitacearum ; Stem blight ; Transcription ; Whole Genome Sequencing</subject><ispartof>Scientific reports, 2020-10, Vol.10 (1), p.18065-18065, Article 18065</ispartof><rights>The Author(s) 2020</rights><rights>The Author(s) 2020. This work is published under http://creativecommons.org/licenses/by/4.0/ (the “License”). Notwithstanding the ProQuest Terms and Conditions, you may use this content in accordance with the terms of the License.</rights><lds50>peer_reviewed</lds50><oa>free_for_read</oa><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c577t-b9d8b95f72142be7cfe0a6ebdc841bc6e94eda5352e006f985dc4811635884cb3</citedby><cites>FETCH-LOGICAL-c577t-b9d8b95f72142be7cfe0a6ebdc841bc6e94eda5352e006f985dc4811635884cb3</cites></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktopdf>$$Uhttps://www.proquest.com/docview/2471524068/fulltextPDF?pq-origsite=primo$$EPDF$$P50$$Gproquest$$Hfree_for_read</linktopdf><linktohtml>$$Uhttps://www.proquest.com/docview/2471524068?pq-origsite=primo$$EHTML$$P50$$Gproquest$$Hfree_for_read</linktohtml><link.rule.ids>230,314,723,776,780,881,25731,27901,27902,36989,36990,44566,53766,53768,74869</link.rule.ids><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/33093634$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Zhao, Qian</creatorcontrib><creatorcontrib>Wu, Jianzhong</creatorcontrib><creatorcontrib>Zhang, Liyan</creatorcontrib><creatorcontrib>Yan, Chao</creatorcontrib><creatorcontrib>Jiang, Shukun</creatorcontrib><creatorcontrib>Li, Zhugang</creatorcontrib><creatorcontrib>Sun, Dequan</creatorcontrib><creatorcontrib>Lai, Yongcai</creatorcontrib><creatorcontrib>Gong, Zhenping</creatorcontrib><title>Genome-scale analyses and characteristics of putative pathogenicity genes of Stagonosporopsis cucurbitacearum, a pumpkin gummy stem blight fungus</title><title>Scientific reports</title><addtitle>Sci Rep</addtitle><addtitle>Sci Rep</addtitle><description>Outbreaks of gummy stem blight (GSB), an emerging seed pumpkin disease, have increased in number and have become more widespread in recent years. Previously we reported that
Stagonosporopsis cucurbitacearum
(
Sc.
) is the dominant fungal cause of pumpkin seedling GSB in Northeast China, where it has greatly reduced crop yields in that region. Here, high-throughput whole-genome sequencing and assembly of the
Sc.
genome were conducted toward revealing pathogenic molecular regulatory mechanisms involved in fungal growth and development
.
Zq-1 as representative
Sc.
strain, DNA of Zq-1was prepared for genomic sequencing, we obtained 5.24 Gb of high-quality genomic sequence data via PacBio RS II sequencing. After sequence data was processed to filter out low quality reads, a hierarchical genome-assembly process was employed that generated a genome sequence of 35.28 Mb in size. A total of 9844 genes were predicted, including 237 non-coding RNAs, 1024 genes encoding proteins with signal peptides, 2066 transmembrane proteins and 756 secretory proteins.Transcriptional identification revealed 54 differentially expressed secretory proteins. Concurrently, 605, 130 and 2869 proteins were matched in the proprietary databases Carbohydrate-Active EnZymes database (CAZyme), Transporter Classification Database (TCDB) and Pathogen–Host Interactions database (PHI), respectively. And 96 and 36 DEGs were identified form PHI database and CAZyme database, respectively. In addition, contig00011.93 was an up-regulated DEG involving ATP-binding cassette metabolism in the procession of infection. In order to test relevance of gene predictions to GSB, DEGs with potential pathogenic relevance were revealed through transcriptome data analysis of
Sc.
strains pre- and post-infection of pumpkin. Interestingly,
Sc.
and
Leptosphaeria maculans
(
Lm.
) exhibited relatively similar with genome lengths, numbers of protein-coding genes and other characteristics. This work provides a foundation for future exploration of additional
Sc.
gene functions toward the development of more effective GSB control strategies.</description><subject>631/326/193/2541</subject><subject>631/326/421</subject><subject>Ascomycota - genetics</subject><subject>Ascomycota - growth & development</subject><subject>Ascomycota - pathogenicity</subject><subject>China</subject><subject>Crop Production</subject><subject>Crop yield</subject><subject>Cucurbita - microbiology</subject><subject>DNA sequencing</subject><subject>Fungi</subject><subject>Gene expression</subject><subject>Genes</subject><subject>Genome, Fungal - genetics</subject><subject>Genomes</subject><subject>Host-Pathogen Interactions - genetics</subject><subject>Humanities and Social Sciences</subject><subject>Membrane proteins</subject><subject>multidisciplinary</subject><subject>Non-coding RNA</subject><subject>Nucleotide sequence</subject><subject>Pathogenicity</subject><subject>Pathogens</subject><subject>Peptides</subject><subject>Plant Diseases - microbiology</subject><subject>Proteins</subject><subject>Science</subject><subject>Science (multidisciplinary)</subject><subject>Seedlings</subject><subject>Signal peptides</subject><subject>Stagonosporopsis cucurbitacearum</subject><subject>Stem blight</subject><subject>Transcription</subject><subject>Whole Genome Sequencing</subject><issn>2045-2322</issn><issn>2045-2322</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2020</creationdate><recordtype>article</recordtype><sourceid>PIMPY</sourceid><sourceid>DOA</sourceid><recordid>eNp9ks1u1DAUhSMEolXpC7BAltiwIODfxNkgoaqUSpVYAGvLdm4yLkkc_FN1HoM3xjNTSssCb3zlc-5nX_lU1UuC3xHM5PvIiehkjSmuW0GZqG-fVMcUc1FTRunTB_VRdRrjNS5L0I6T7nl1xBjuWMP4cfXrAhY_Qx2tngDpRU_bCLEUPbIbHbRNEFxMzkbkB7TmpJO7AbTqtPEjLM66tEWlgL3-NenRLz6uPvg1uohstjkYl7QFHfL8FunCmNcfbkFjnuctiglmZCY3bhIa8jLm-KJ6NugpwundflJ9_3T-7exzffXl4vLs41VtRdum2nS9NJ0YWko4NdDaAbBuwPRWcmJsAx2HXgsmKGDcDJ0UveWSkIYJKbk17KS6PHB7r6_VGtysw1Z57dT-wIdR6VAGn0DZjjc9t-0AtueaC8PsMBgpWiYE4R0trA8H1prNDL2FJQU9PYI-Vha3UaO_Ua2QpKW4AN7cAYL_mSEmNbtoYZr0Aj5HRbngBBPBZLG-_sd67XMoH7dztURQjpudix5cNvgYAwz3jyFY7QKkDgFSJUBqHyB1W5pePRzjvuVPXIqBHQyxSMsI4e_d_8H-BmFD1jg</recordid><startdate>20201022</startdate><enddate>20201022</enddate><creator>Zhao, Qian</creator><creator>Wu, Jianzhong</creator><creator>Zhang, Liyan</creator><creator>Yan, Chao</creator><creator>Jiang, Shukun</creator><creator>Li, Zhugang</creator><creator>Sun, Dequan</creator><creator>Lai, Yongcai</creator><creator>Gong, Zhenping</creator><general>Nature Publishing Group UK</general><general>Nature Publishing Group</general><general>Nature Portfolio</general><scope>C6C</scope><scope>CGR</scope><scope>CUY</scope><scope>CVF</scope><scope>ECM</scope><scope>EIF</scope><scope>NPM</scope><scope>AAYXX</scope><scope>CITATION</scope><scope>3V.</scope><scope>7X7</scope><scope>7XB</scope><scope>88A</scope><scope>88E</scope><scope>88I</scope><scope>8FE</scope><scope>8FH</scope><scope>8FI</scope><scope>8FJ</scope><scope>8FK</scope><scope>ABUWG</scope><scope>AEUYN</scope><scope>AFKRA</scope><scope>AZQEC</scope><scope>BBNVY</scope><scope>BENPR</scope><scope>BHPHI</scope><scope>CCPQU</scope><scope>DWQXO</scope><scope>FYUFA</scope><scope>GHDGH</scope><scope>GNUQQ</scope><scope>HCIFZ</scope><scope>K9.</scope><scope>LK8</scope><scope>M0S</scope><scope>M1P</scope><scope>M2P</scope><scope>M7P</scope><scope>PIMPY</scope><scope>PQEST</scope><scope>PQQKQ</scope><scope>PQUKI</scope><scope>Q9U</scope><scope>7X8</scope><scope>5PM</scope><scope>DOA</scope></search><sort><creationdate>20201022</creationdate><title>Genome-scale analyses and characteristics of putative pathogenicity genes of Stagonosporopsis cucurbitacearum, a pumpkin gummy stem blight fungus</title><author>Zhao, Qian ; Wu, Jianzhong ; Zhang, Liyan ; Yan, Chao ; Jiang, Shukun ; Li, Zhugang ; Sun, Dequan ; Lai, Yongcai ; Gong, Zhenping</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c577t-b9d8b95f72142be7cfe0a6ebdc841bc6e94eda5352e006f985dc4811635884cb3</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2020</creationdate><topic>631/326/193/2541</topic><topic>631/326/421</topic><topic>Ascomycota - genetics</topic><topic>Ascomycota - growth & development</topic><topic>Ascomycota - pathogenicity</topic><topic>China</topic><topic>Crop Production</topic><topic>Crop yield</topic><topic>Cucurbita - microbiology</topic><topic>DNA sequencing</topic><topic>Fungi</topic><topic>Gene expression</topic><topic>Genes</topic><topic>Genome, Fungal - genetics</topic><topic>Genomes</topic><topic>Host-Pathogen Interactions - genetics</topic><topic>Humanities and Social Sciences</topic><topic>Membrane proteins</topic><topic>multidisciplinary</topic><topic>Non-coding RNA</topic><topic>Nucleotide sequence</topic><topic>Pathogenicity</topic><topic>Pathogens</topic><topic>Peptides</topic><topic>Plant Diseases - microbiology</topic><topic>Proteins</topic><topic>Science</topic><topic>Science (multidisciplinary)</topic><topic>Seedlings</topic><topic>Signal peptides</topic><topic>Stagonosporopsis cucurbitacearum</topic><topic>Stem blight</topic><topic>Transcription</topic><topic>Whole Genome Sequencing</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Zhao, Qian</creatorcontrib><creatorcontrib>Wu, Jianzhong</creatorcontrib><creatorcontrib>Zhang, Liyan</creatorcontrib><creatorcontrib>Yan, Chao</creatorcontrib><creatorcontrib>Jiang, Shukun</creatorcontrib><creatorcontrib>Li, Zhugang</creatorcontrib><creatorcontrib>Sun, Dequan</creatorcontrib><creatorcontrib>Lai, Yongcai</creatorcontrib><creatorcontrib>Gong, Zhenping</creatorcontrib><collection>SpringerOpen</collection><collection>Medline</collection><collection>MEDLINE</collection><collection>MEDLINE (Ovid)</collection><collection>MEDLINE</collection><collection>MEDLINE</collection><collection>PubMed</collection><collection>CrossRef</collection><collection>ProQuest Central (Corporate)</collection><collection>Health & Medical Collection</collection><collection>ProQuest Central (purchase pre-March 2016)</collection><collection>Biology Database (Alumni Edition)</collection><collection>Medical Database (Alumni Edition)</collection><collection>Science Database (Alumni Edition)</collection><collection>ProQuest SciTech Collection</collection><collection>ProQuest Natural Science Collection</collection><collection>Hospital Premium Collection</collection><collection>Hospital Premium Collection (Alumni Edition)</collection><collection>ProQuest Central (Alumni) (purchase pre-March 2016)</collection><collection>ProQuest Central (Alumni)</collection><collection>ProQuest One Sustainability</collection><collection>ProQuest Central</collection><collection>ProQuest Central Essentials</collection><collection>Biological Science Collection</collection><collection>AUTh Library subscriptions: ProQuest Central</collection><collection>Natural Science Collection</collection><collection>ProQuest One Community College</collection><collection>ProQuest Central</collection><collection>Health Research Premium Collection</collection><collection>Health Research Premium Collection (Alumni)</collection><collection>ProQuest Central Student</collection><collection>SciTech Premium Collection</collection><collection>ProQuest Health & Medical Complete (Alumni)</collection><collection>Biological Sciences</collection><collection>Health & Medical Collection (Alumni Edition)</collection><collection>Medical Database</collection><collection>Science Database</collection><collection>Biological Science Database</collection><collection>Publicly Available Content Database</collection><collection>ProQuest One Academic Eastern Edition (DO NOT USE)</collection><collection>ProQuest One Academic</collection><collection>ProQuest One Academic UKI Edition</collection><collection>ProQuest Central Basic</collection><collection>MEDLINE - Academic</collection><collection>PubMed Central (Full Participant titles)</collection><collection>Open Access: DOAJ - Directory of Open Access Journals</collection><jtitle>Scientific reports</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Zhao, Qian</au><au>Wu, Jianzhong</au><au>Zhang, Liyan</au><au>Yan, Chao</au><au>Jiang, Shukun</au><au>Li, Zhugang</au><au>Sun, Dequan</au><au>Lai, Yongcai</au><au>Gong, Zhenping</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Genome-scale analyses and characteristics of putative pathogenicity genes of Stagonosporopsis cucurbitacearum, a pumpkin gummy stem blight fungus</atitle><jtitle>Scientific reports</jtitle><stitle>Sci Rep</stitle><addtitle>Sci Rep</addtitle><date>2020-10-22</date><risdate>2020</risdate><volume>10</volume><issue>1</issue><spage>18065</spage><epage>18065</epage><pages>18065-18065</pages><artnum>18065</artnum><issn>2045-2322</issn><eissn>2045-2322</eissn><abstract>Outbreaks of gummy stem blight (GSB), an emerging seed pumpkin disease, have increased in number and have become more widespread in recent years. Previously we reported that
Stagonosporopsis cucurbitacearum
(
Sc.
) is the dominant fungal cause of pumpkin seedling GSB in Northeast China, where it has greatly reduced crop yields in that region. Here, high-throughput whole-genome sequencing and assembly of the
Sc.
genome were conducted toward revealing pathogenic molecular regulatory mechanisms involved in fungal growth and development
.
Zq-1 as representative
Sc.
strain, DNA of Zq-1was prepared for genomic sequencing, we obtained 5.24 Gb of high-quality genomic sequence data via PacBio RS II sequencing. After sequence data was processed to filter out low quality reads, a hierarchical genome-assembly process was employed that generated a genome sequence of 35.28 Mb in size. A total of 9844 genes were predicted, including 237 non-coding RNAs, 1024 genes encoding proteins with signal peptides, 2066 transmembrane proteins and 756 secretory proteins.Transcriptional identification revealed 54 differentially expressed secretory proteins. Concurrently, 605, 130 and 2869 proteins were matched in the proprietary databases Carbohydrate-Active EnZymes database (CAZyme), Transporter Classification Database (TCDB) and Pathogen–Host Interactions database (PHI), respectively. And 96 and 36 DEGs were identified form PHI database and CAZyme database, respectively. In addition, contig00011.93 was an up-regulated DEG involving ATP-binding cassette metabolism in the procession of infection. In order to test relevance of gene predictions to GSB, DEGs with potential pathogenic relevance were revealed through transcriptome data analysis of
Sc.
strains pre- and post-infection of pumpkin. Interestingly,
Sc.
and
Leptosphaeria maculans
(
Lm.
) exhibited relatively similar with genome lengths, numbers of protein-coding genes and other characteristics. This work provides a foundation for future exploration of additional
Sc.
gene functions toward the development of more effective GSB control strategies.</abstract><cop>London</cop><pub>Nature Publishing Group UK</pub><pmid>33093634</pmid><doi>10.1038/s41598-020-75235-x</doi><tpages>1</tpages><oa>free_for_read</oa></addata></record> |
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source | Open Access: PubMed Central; Publicly Available Content Database; Full-Text Journals in Chemistry (Open access); Springer Nature - nature.com Journals - Fully Open Access |
subjects | 631/326/193/2541 631/326/421 Ascomycota - genetics Ascomycota - growth & development Ascomycota - pathogenicity China Crop Production Crop yield Cucurbita - microbiology DNA sequencing Fungi Gene expression Genes Genome, Fungal - genetics Genomes Host-Pathogen Interactions - genetics Humanities and Social Sciences Membrane proteins multidisciplinary Non-coding RNA Nucleotide sequence Pathogenicity Pathogens Peptides Plant Diseases - microbiology Proteins Science Science (multidisciplinary) Seedlings Signal peptides Stagonosporopsis cucurbitacearum Stem blight Transcription Whole Genome Sequencing |
title | Genome-scale analyses and characteristics of putative pathogenicity genes of Stagonosporopsis cucurbitacearum, a pumpkin gummy stem blight fungus |
url | http://sfxeu10.hosted.exlibrisgroup.com/loughborough?ctx_ver=Z39.88-2004&ctx_enc=info:ofi/enc:UTF-8&ctx_tim=2025-02-03T14%3A38%3A29IST&url_ver=Z39.88-2004&url_ctx_fmt=infofi/fmt:kev:mtx:ctx&rfr_id=info:sid/primo.exlibrisgroup.com:primo3-Article-proquest_doaj_&rft_val_fmt=info:ofi/fmt:kev:mtx:journal&rft.genre=article&rft.atitle=Genome-scale%20analyses%20and%20characteristics%20of%20putative%20pathogenicity%20genes%20of%20Stagonosporopsis%20cucurbitacearum,%20a%20pumpkin%20gummy%20stem%20blight%20fungus&rft.jtitle=Scientific%20reports&rft.au=Zhao,%20Qian&rft.date=2020-10-22&rft.volume=10&rft.issue=1&rft.spage=18065&rft.epage=18065&rft.pages=18065-18065&rft.artnum=18065&rft.issn=2045-2322&rft.eissn=2045-2322&rft_id=info:doi/10.1038/s41598-020-75235-x&rft_dat=%3Cproquest_doaj_%3E2454101538%3C/proquest_doaj_%3E%3Cgrp_id%3Ecdi_FETCH-LOGICAL-c577t-b9d8b95f72142be7cfe0a6ebdc841bc6e94eda5352e006f985dc4811635884cb3%3C/grp_id%3E%3Coa%3E%3C/oa%3E%3Curl%3E%3C/url%3E&rft_id=info:oai/&rft_pqid=2471524068&rft_id=info:pmid/33093634&rfr_iscdi=true |