Development of Genetic Modification Tools for Hanseniaspora uvarum
Apiculate yeasts belonging to the genus are commonly isolated from viticultural settings and often dominate the initial stages of grape must fermentations. Although considered spoilage yeasts, they are now increasingly becoming the focus of research, with several whole-genome sequencing studies publ...
Gespeichert in:
| Veröffentlicht in: | International journal of molecular sciences 2021-02, Vol.22 (4) |
|---|---|
| Hauptverfasser: | , , , , , , |
| Format: | Artikel |
| Sprache: | eng |
| Schlagworte: | |
| Online-Zugang: | Volltext |
| Tags: |
Tag hinzufügen
Keine Tags, Fügen Sie den ersten Tag hinzu!
|
| container_end_page | |
|---|---|
| container_issue | 4 |
| container_start_page | |
| container_title | International journal of molecular sciences |
| container_volume | 22 |
| creator | Badura, Jennifer van Wyk, Niël Brezina, Silvia Pretorius, Isak S Rauhut, Doris Wendland, Jürgen von Wallbrunn, Christian |
| description | Apiculate yeasts belonging to the genus
are commonly isolated from viticultural settings and often dominate the initial stages of grape must fermentations. Although considered spoilage yeasts, they are now increasingly becoming the focus of research, with several whole-genome sequencing studies published in recent years. However, tools for their molecular genetic manipulation are still lacking. Here, we report the development of a tool for the genetic modification of
. This was employed for the disruption of the
gene, which encodes a putative alcohol acetyltransferase involved in acetate ester formation. We generated a synthetic marker gene consisting of the
promoter controlling a hygromycin resistance open reading frame (ORF). This new marker gene was used in disruption cassettes containing long-flanking (1000 bp) homology regions to the target locus. By increasing the antibiotic concentration, transformants were obtained in which both alleles of the putative
gene were deleted in a diploid
.
strain. Phenotypic characterisation including fermentation in Müller-Thurgau must showed that the null mutant produced significantly less acetate ester, particularly ethyl acetate. This study marks the first steps in the development of gene modification tools and paves the road for functional gene analyses of this yeast. |
| doi_str_mv | 10.3390/ijms22041943 |
| format | Article |
| fullrecord | <record><control><sourceid>pubmed</sourceid><recordid>TN_cdi_pubmedcentral_primary_oai_pubmedcentral_nih_gov_7920042</recordid><sourceformat>XML</sourceformat><sourcesystem>PC</sourcesystem><sourcerecordid>33669299</sourcerecordid><originalsourceid>FETCH-LOGICAL-p233t-2364af92ffbce88a2bd59d5b4bb72ad7ebb83b05bcd0f1bd1237fe6382be867d3</originalsourceid><addsrcrecordid>eNpVkE1LxDAURYMgzji6cy35A9X0vTZpNoKOOiOMuBnXJWkSzdA2JWkH_PcO-IGu7uJwD5dLyEXOrhAlu_a7LgGwIpcFHpF5XgBkjHExI6cp7RgDhFKekBki5xKknJO7e7u3bRg62480OLqyvR19Q5-D8c43avShp9sQ2kRdiHSt-mR7r9IQoqLTXsWpOyPHTrXJnn_ngrw-PmyX62zzsnpa3m6yARDHDJAXyklwTje2qhRoU0pT6kJrAcoIq3WFmpW6Mczl2uSAwlmOFWhbcWFwQW6-vMOkO2uaw-Ko2nqIvlPxow7K1_9J79_rt7CvhQTGCjgILv8Kfps_b-AnGh1izQ</addsrcrecordid><sourcetype>Open Access Repository</sourcetype><iscdi>true</iscdi><recordtype>article</recordtype></control><display><type>article</type><title>Development of Genetic Modification Tools for Hanseniaspora uvarum</title><source>MEDLINE</source><source>Elektronische Zeitschriftenbibliothek - Frei zugängliche E-Journals</source><source>MDPI - Multidisciplinary Digital Publishing Institute</source><source>PubMed Central</source><creator>Badura, Jennifer ; van Wyk, Niël ; Brezina, Silvia ; Pretorius, Isak S ; Rauhut, Doris ; Wendland, Jürgen ; von Wallbrunn, Christian</creator><creatorcontrib>Badura, Jennifer ; van Wyk, Niël ; Brezina, Silvia ; Pretorius, Isak S ; Rauhut, Doris ; Wendland, Jürgen ; von Wallbrunn, Christian</creatorcontrib><description>Apiculate yeasts belonging to the genus
are commonly isolated from viticultural settings and often dominate the initial stages of grape must fermentations. Although considered spoilage yeasts, they are now increasingly becoming the focus of research, with several whole-genome sequencing studies published in recent years. However, tools for their molecular genetic manipulation are still lacking. Here, we report the development of a tool for the genetic modification of
. This was employed for the disruption of the
gene, which encodes a putative alcohol acetyltransferase involved in acetate ester formation. We generated a synthetic marker gene consisting of the
promoter controlling a hygromycin resistance open reading frame (ORF). This new marker gene was used in disruption cassettes containing long-flanking (1000 bp) homology regions to the target locus. By increasing the antibiotic concentration, transformants were obtained in which both alleles of the putative
gene were deleted in a diploid
.
strain. Phenotypic characterisation including fermentation in Müller-Thurgau must showed that the null mutant produced significantly less acetate ester, particularly ethyl acetate. This study marks the first steps in the development of gene modification tools and paves the road for functional gene analyses of this yeast.</description><identifier>EISSN: 1422-0067</identifier><identifier>DOI: 10.3390/ijms22041943</identifier><identifier>PMID: 33669299</identifier><language>eng</language><publisher>Switzerland: MDPI</publisher><subject>Acetates - metabolism ; Alleles ; Fermentation - genetics ; Gene Deletion ; Genes, Fungal ; Genetic Engineering - methods ; Hanseniaspora - enzymology ; Hanseniaspora - genetics ; Microorganisms, Genetically-Modified - genetics ; Open Reading Frames ; Phenotype ; Promoter Regions, Genetic ; Proteins - genetics ; Saccharomyces cerevisiae - genetics ; Saccharomyces cerevisiae Proteins - genetics ; Vitis - metabolism ; Wine</subject><ispartof>International journal of molecular sciences, 2021-02, Vol.22 (4)</ispartof><rights>2021 by the authors. 2021</rights><lds50>peer_reviewed</lds50><oa>free_for_read</oa><woscitedreferencessubscribed>false</woscitedreferencessubscribed><orcidid>0000-0003-3031-8821 ; 0000-0001-9127-3175 ; 0000-0003-0615-9250 ; 0000-0001-8350-253X</orcidid></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktopdf>$$Uhttps://www.ncbi.nlm.nih.gov/pmc/articles/PMC7920042/pdf/$$EPDF$$P50$$Gpubmedcentral$$Hfree_for_read</linktopdf><linktohtml>$$Uhttps://www.ncbi.nlm.nih.gov/pmc/articles/PMC7920042/$$EHTML$$P50$$Gpubmedcentral$$Hfree_for_read</linktohtml><link.rule.ids>230,314,727,780,784,885,27923,27924,53790,53792</link.rule.ids><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/33669299$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Badura, Jennifer</creatorcontrib><creatorcontrib>van Wyk, Niël</creatorcontrib><creatorcontrib>Brezina, Silvia</creatorcontrib><creatorcontrib>Pretorius, Isak S</creatorcontrib><creatorcontrib>Rauhut, Doris</creatorcontrib><creatorcontrib>Wendland, Jürgen</creatorcontrib><creatorcontrib>von Wallbrunn, Christian</creatorcontrib><title>Development of Genetic Modification Tools for Hanseniaspora uvarum</title><title>International journal of molecular sciences</title><addtitle>Int J Mol Sci</addtitle><description>Apiculate yeasts belonging to the genus
are commonly isolated from viticultural settings and often dominate the initial stages of grape must fermentations. Although considered spoilage yeasts, they are now increasingly becoming the focus of research, with several whole-genome sequencing studies published in recent years. However, tools for their molecular genetic manipulation are still lacking. Here, we report the development of a tool for the genetic modification of
. This was employed for the disruption of the
gene, which encodes a putative alcohol acetyltransferase involved in acetate ester formation. We generated a synthetic marker gene consisting of the
promoter controlling a hygromycin resistance open reading frame (ORF). This new marker gene was used in disruption cassettes containing long-flanking (1000 bp) homology regions to the target locus. By increasing the antibiotic concentration, transformants were obtained in which both alleles of the putative
gene were deleted in a diploid
.
strain. Phenotypic characterisation including fermentation in Müller-Thurgau must showed that the null mutant produced significantly less acetate ester, particularly ethyl acetate. This study marks the first steps in the development of gene modification tools and paves the road for functional gene analyses of this yeast.</description><subject>Acetates - metabolism</subject><subject>Alleles</subject><subject>Fermentation - genetics</subject><subject>Gene Deletion</subject><subject>Genes, Fungal</subject><subject>Genetic Engineering - methods</subject><subject>Hanseniaspora - enzymology</subject><subject>Hanseniaspora - genetics</subject><subject>Microorganisms, Genetically-Modified - genetics</subject><subject>Open Reading Frames</subject><subject>Phenotype</subject><subject>Promoter Regions, Genetic</subject><subject>Proteins - genetics</subject><subject>Saccharomyces cerevisiae - genetics</subject><subject>Saccharomyces cerevisiae Proteins - genetics</subject><subject>Vitis - metabolism</subject><subject>Wine</subject><issn>1422-0067</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2021</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNpVkE1LxDAURYMgzji6cy35A9X0vTZpNoKOOiOMuBnXJWkSzdA2JWkH_PcO-IGu7uJwD5dLyEXOrhAlu_a7LgGwIpcFHpF5XgBkjHExI6cp7RgDhFKekBki5xKknJO7e7u3bRg62480OLqyvR19Q5-D8c43avShp9sQ2kRdiHSt-mR7r9IQoqLTXsWpOyPHTrXJnn_ngrw-PmyX62zzsnpa3m6yARDHDJAXyklwTje2qhRoU0pT6kJrAcoIq3WFmpW6Mczl2uSAwlmOFWhbcWFwQW6-vMOkO2uaw-Ko2nqIvlPxow7K1_9J79_rt7CvhQTGCjgILv8Kfps_b-AnGh1izQ</recordid><startdate>20210216</startdate><enddate>20210216</enddate><creator>Badura, Jennifer</creator><creator>van Wyk, Niël</creator><creator>Brezina, Silvia</creator><creator>Pretorius, Isak S</creator><creator>Rauhut, Doris</creator><creator>Wendland, Jürgen</creator><creator>von Wallbrunn, Christian</creator><general>MDPI</general><scope>CGR</scope><scope>CUY</scope><scope>CVF</scope><scope>ECM</scope><scope>EIF</scope><scope>NPM</scope><scope>5PM</scope><orcidid>https://orcid.org/0000-0003-3031-8821</orcidid><orcidid>https://orcid.org/0000-0001-9127-3175</orcidid><orcidid>https://orcid.org/0000-0003-0615-9250</orcidid><orcidid>https://orcid.org/0000-0001-8350-253X</orcidid></search><sort><creationdate>20210216</creationdate><title>Development of Genetic Modification Tools for Hanseniaspora uvarum</title><author>Badura, Jennifer ; van Wyk, Niël ; Brezina, Silvia ; Pretorius, Isak S ; Rauhut, Doris ; Wendland, Jürgen ; von Wallbrunn, Christian</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-p233t-2364af92ffbce88a2bd59d5b4bb72ad7ebb83b05bcd0f1bd1237fe6382be867d3</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2021</creationdate><topic>Acetates - metabolism</topic><topic>Alleles</topic><topic>Fermentation - genetics</topic><topic>Gene Deletion</topic><topic>Genes, Fungal</topic><topic>Genetic Engineering - methods</topic><topic>Hanseniaspora - enzymology</topic><topic>Hanseniaspora - genetics</topic><topic>Microorganisms, Genetically-Modified - genetics</topic><topic>Open Reading Frames</topic><topic>Phenotype</topic><topic>Promoter Regions, Genetic</topic><topic>Proteins - genetics</topic><topic>Saccharomyces cerevisiae - genetics</topic><topic>Saccharomyces cerevisiae Proteins - genetics</topic><topic>Vitis - metabolism</topic><topic>Wine</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Badura, Jennifer</creatorcontrib><creatorcontrib>van Wyk, Niël</creatorcontrib><creatorcontrib>Brezina, Silvia</creatorcontrib><creatorcontrib>Pretorius, Isak S</creatorcontrib><creatorcontrib>Rauhut, Doris</creatorcontrib><creatorcontrib>Wendland, Jürgen</creatorcontrib><creatorcontrib>von Wallbrunn, Christian</creatorcontrib><collection>Medline</collection><collection>MEDLINE</collection><collection>MEDLINE (Ovid)</collection><collection>MEDLINE</collection><collection>MEDLINE</collection><collection>PubMed</collection><collection>PubMed Central (Full Participant titles)</collection><jtitle>International journal of molecular sciences</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Badura, Jennifer</au><au>van Wyk, Niël</au><au>Brezina, Silvia</au><au>Pretorius, Isak S</au><au>Rauhut, Doris</au><au>Wendland, Jürgen</au><au>von Wallbrunn, Christian</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Development of Genetic Modification Tools for Hanseniaspora uvarum</atitle><jtitle>International journal of molecular sciences</jtitle><addtitle>Int J Mol Sci</addtitle><date>2021-02-16</date><risdate>2021</risdate><volume>22</volume><issue>4</issue><eissn>1422-0067</eissn><abstract>Apiculate yeasts belonging to the genus
are commonly isolated from viticultural settings and often dominate the initial stages of grape must fermentations. Although considered spoilage yeasts, they are now increasingly becoming the focus of research, with several whole-genome sequencing studies published in recent years. However, tools for their molecular genetic manipulation are still lacking. Here, we report the development of a tool for the genetic modification of
. This was employed for the disruption of the
gene, which encodes a putative alcohol acetyltransferase involved in acetate ester formation. We generated a synthetic marker gene consisting of the
promoter controlling a hygromycin resistance open reading frame (ORF). This new marker gene was used in disruption cassettes containing long-flanking (1000 bp) homology regions to the target locus. By increasing the antibiotic concentration, transformants were obtained in which both alleles of the putative
gene were deleted in a diploid
.
strain. Phenotypic characterisation including fermentation in Müller-Thurgau must showed that the null mutant produced significantly less acetate ester, particularly ethyl acetate. This study marks the first steps in the development of gene modification tools and paves the road for functional gene analyses of this yeast.</abstract><cop>Switzerland</cop><pub>MDPI</pub><pmid>33669299</pmid><doi>10.3390/ijms22041943</doi><orcidid>https://orcid.org/0000-0003-3031-8821</orcidid><orcidid>https://orcid.org/0000-0001-9127-3175</orcidid><orcidid>https://orcid.org/0000-0003-0615-9250</orcidid><orcidid>https://orcid.org/0000-0001-8350-253X</orcidid><oa>free_for_read</oa></addata></record> |
| fulltext | fulltext |
| identifier | EISSN: 1422-0067 |
| ispartof | International journal of molecular sciences, 2021-02, Vol.22 (4) |
| issn | 1422-0067 |
| language | eng |
| recordid | cdi_pubmedcentral_primary_oai_pubmedcentral_nih_gov_7920042 |
| source | MEDLINE; Elektronische Zeitschriftenbibliothek - Frei zugängliche E-Journals; MDPI - Multidisciplinary Digital Publishing Institute; PubMed Central |
| subjects | Acetates - metabolism Alleles Fermentation - genetics Gene Deletion Genes, Fungal Genetic Engineering - methods Hanseniaspora - enzymology Hanseniaspora - genetics Microorganisms, Genetically-Modified - genetics Open Reading Frames Phenotype Promoter Regions, Genetic Proteins - genetics Saccharomyces cerevisiae - genetics Saccharomyces cerevisiae Proteins - genetics Vitis - metabolism Wine |
| title | Development of Genetic Modification Tools for Hanseniaspora uvarum |
| url | https://sfx.bib-bvb.de/sfx_tum?ctx_ver=Z39.88-2004&ctx_enc=info:ofi/enc:UTF-8&ctx_tim=2025-01-08T07%3A26%3A27IST&url_ver=Z39.88-2004&url_ctx_fmt=infofi/fmt:kev:mtx:ctx&rfr_id=info:sid/primo.exlibrisgroup.com:primo3-Article-pubmed&rft_val_fmt=info:ofi/fmt:kev:mtx:journal&rft.genre=article&rft.atitle=Development%20of%20Genetic%20Modification%20Tools%20for%20Hanseniaspora%20uvarum&rft.jtitle=International%20journal%20of%20molecular%20sciences&rft.au=Badura,%20Jennifer&rft.date=2021-02-16&rft.volume=22&rft.issue=4&rft.eissn=1422-0067&rft_id=info:doi/10.3390/ijms22041943&rft_dat=%3Cpubmed%3E33669299%3C/pubmed%3E%3Curl%3E%3C/url%3E&disable_directlink=true&sfx.directlink=off&sfx.report_link=0&rft_id=info:oai/&rft_id=info:pmid/33669299&rfr_iscdi=true |