High rates of virus-induced gene silencing by tobacco rattle virus in Populus.
Identifieur interne : 001D16 ( Main/Exploration ); précédent : 001D15; suivant : 001D17High rates of virus-induced gene silencing by tobacco rattle virus in Populus.
Auteurs : Zedan Shen [République populaire de Chine] ; Jian Sun [République populaire de Chine] ; Jun Yao [République populaire de Chine] ; Shaojie Wang [République populaire de Chine] ; Mingquan Ding [République populaire de Chine] ; Huilong Zhang [République populaire de Chine] ; Zeyong Qian [République populaire de Chine] ; Nan Zhao [République populaire de Chine] ; Gang Sa [République populaire de Chine] ; Rui Zhao [République populaire de Chine] ; Xin Shen [République populaire de Chine] ; Andrea Polle [Allemagne] ; Shaoliang Chen [République populaire de Chine]Source :
- Tree physiology [ 1758-4469 ] ; 2015.
Descripteurs français
- KwdFr :
- ARN messager (génétique), ARN messager (métabolisme), Extinction de l'expression des gènes (MeSH), Gènes de plante (MeSH), Maladies des plantes (virologie), Normes de référence (MeSH), Oxidoreductases (génétique), Photoblanchiment (MeSH), Phénotype (MeSH), Populus (génétique), Populus (virologie), Régulation de l'expression des gènes végétaux (MeSH), Vecteurs génétiques (métabolisme), Virus des plantes (physiologie).
- MESH :
- génétique : ARN messager, Oxidoreductases, Populus.
- métabolisme : ARN messager, Vecteurs génétiques.
- physiologie : Virus des plantes.
- virologie : Maladies des plantes, Populus.
- Extinction de l'expression des gènes, Gènes de plante, Normes de référence, Photoblanchiment, Phénotype, Régulation de l'expression des gènes végétaux.
English descriptors
- KwdEn :
- Gene Expression Regulation, Plant (MeSH), Gene Silencing (MeSH), Genes, Plant (MeSH), Genetic Vectors (metabolism), Oxidoreductases (genetics), Phenotype (MeSH), Photobleaching (MeSH), Plant Diseases (virology), Plant Viruses (physiology), Populus (genetics), Populus (virology), RNA, Messenger (genetics), RNA, Messenger (metabolism), Reference Standards (MeSH).
- MESH :
- chemical , genetics : Oxidoreductases, RNA, Messenger.
- genetics : Populus.
- metabolism : Genetic Vectors, RNA, Messenger.
- physiology : Plant Viruses.
- virology : Plant Diseases, Populus.
- Gene Expression Regulation, Plant, Gene Silencing, Genes, Plant, Phenotype, Photobleaching, Reference Standards.
Abstract
Virus-induced gene silencing (VIGS) has been shown to be an effective tool for investigating gene functions in herbaceous plant species, but has rarely been tested in trees. The establishment of a fast and reliable transformation system is especially important for woody plants, many of which are recalcitrant to transformation. In this study, we established a tobacco rattle virus (TRV)-based VIGS system for two Populus species, Populus euphratica and P. × canescens. Here, TRV constructs carrying a 266 bp or a 558 bp fragment of the phytoene desaturase (PDS) gene were Agrobacterium-infiltrated into leaves of the two poplar species. Agrobacterium-mediated delivery of the shorter insert, TRV2-PePDS266, into the host poplars resulted in expected photobleaching in both tree species, but not the longer insert, PePDS558. The efficiency of VIGS was temperature-dependent, increasing by raising the temperature from 18 to 28 °C. The optimized TRV-VIGS system at 28 °C resulted in a high silencing frequency and efficiency up to 65-73 and 83-94%, respectively, in the two tested poplars. Moreover, syringe inoculation of Agrobacterium in 100 mM acetosyringone induced a more efficient silencing in the two poplar species, compared with other agroinfiltration methods, e.g., direct injection, misting and agrodrench. There were plant species-related differences in the response to VIGS because the photobleaching symptoms were more severe in P. × canescens than in P. euphratica. Furthermore, VIGS-treated P. euphratica exhibited a higher recovery rate (50%) after several weeks of the virus infection, compared with TRV-infected P. × canescens plants (20%). Expression stability of reference genes was screened to assess the relative abundance of PePDS mRNA in VIGS-treated P. euphratica and P. × canescens. PeACT7 was stably expressed in P. euphratica and UBQ-L was selected as the most suitable reference gene for P. × canescens using three different statistical approaches, geNorm, NormFinder and BestKeeper. Quantitative real-time PCR showed significant reductions in PDS transcripts (55-64%) in the photobleached leaves of both VIGS-treated poplar species. Our results demonstrate that the TRV-based VIGS provides a practical tool for gene functional analysis in Populus sp., especially in those poplar species which are otherwise recalcitrant to transformation.
DOI: 10.1093/treephys/tpv064
PubMed: 26209619
Affiliations:
Links toward previous steps (curation, corpus...)
Le document en format XML
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<author><name sortKey="Sa, Gang" sort="Sa, Gang" uniqKey="Sa G" first="Gang" last="Sa">Gang Sa</name>
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<author><name sortKey="Zhao, Rui" sort="Zhao, Rui" uniqKey="Zhao R" first="Rui" last="Zhao">Rui Zhao</name>
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<term>Gene Silencing (MeSH)</term>
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<term>Oxidoreductases (genetics)</term>
<term>Phenotype (MeSH)</term>
<term>Photobleaching (MeSH)</term>
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<term>Populus (genetics)</term>
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<term>Maladies des plantes (virologie)</term>
<term>Normes de référence (MeSH)</term>
<term>Oxidoreductases (génétique)</term>
<term>Photoblanchiment (MeSH)</term>
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<term>Populus (virologie)</term>
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<term>Gènes de plante</term>
<term>Normes de référence</term>
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<front><div type="abstract" xml:lang="en">Virus-induced gene silencing (VIGS) has been shown to be an effective tool for investigating gene functions in herbaceous plant species, but has rarely been tested in trees. The establishment of a fast and reliable transformation system is especially important for woody plants, many of which are recalcitrant to transformation. In this study, we established a tobacco rattle virus (TRV)-based VIGS system for two Populus species, Populus euphratica and P. × canescens. Here, TRV constructs carrying a 266 bp or a 558 bp fragment of the phytoene desaturase (PDS) gene were Agrobacterium-infiltrated into leaves of the two poplar species. Agrobacterium-mediated delivery of the shorter insert, TRV2-PePDS266, into the host poplars resulted in expected photobleaching in both tree species, but not the longer insert, PePDS558. The efficiency of VIGS was temperature-dependent, increasing by raising the temperature from 18 to 28 °C. The optimized TRV-VIGS system at 28 °C resulted in a high silencing frequency and efficiency up to 65-73 and 83-94%, respectively, in the two tested poplars. Moreover, syringe inoculation of Agrobacterium in 100 mM acetosyringone induced a more efficient silencing in the two poplar species, compared with other agroinfiltration methods, e.g., direct injection, misting and agrodrench. There were plant species-related differences in the response to VIGS because the photobleaching symptoms were more severe in P. × canescens than in P. euphratica. Furthermore, VIGS-treated P. euphratica exhibited a higher recovery rate (50%) after several weeks of the virus infection, compared with TRV-infected P. × canescens plants (20%). Expression stability of reference genes was screened to assess the relative abundance of PePDS mRNA in VIGS-treated P. euphratica and P. × canescens. PeACT7 was stably expressed in P. euphratica and UBQ-L was selected as the most suitable reference gene for P. × canescens using three different statistical approaches, geNorm, NormFinder and BestKeeper. Quantitative real-time PCR showed significant reductions in PDS transcripts (55-64%) in the photobleached leaves of both VIGS-treated poplar species. Our results demonstrate that the TRV-based VIGS provides a practical tool for gene functional analysis in Populus sp., especially in those poplar species which are otherwise recalcitrant to transformation.</div>
</front>
</TEI>
<pubmed><MedlineCitation Status="MEDLINE" Owner="NLM"><PMID Version="1">26209619</PMID>
<DateCompleted><Year>2016</Year>
<Month>07</Month>
<Day>04</Day>
</DateCompleted>
<DateRevised><Year>2015</Year>
<Month>09</Month>
<Day>25</Day>
</DateRevised>
<Article PubModel="Print-Electronic"><Journal><ISSN IssnType="Electronic">1758-4469</ISSN>
<JournalIssue CitedMedium="Internet"><Volume>35</Volume>
<Issue>9</Issue>
<PubDate><Year>2015</Year>
<Month>Sep</Month>
</PubDate>
</JournalIssue>
<Title>Tree physiology</Title>
<ISOAbbreviation>Tree Physiol</ISOAbbreviation>
</Journal>
<ArticleTitle>High rates of virus-induced gene silencing by tobacco rattle virus in Populus.</ArticleTitle>
<Pagination><MedlinePgn>1016-29</MedlinePgn>
</Pagination>
<ELocationID EIdType="doi" ValidYN="Y">10.1093/treephys/tpv064</ELocationID>
<Abstract><AbstractText>Virus-induced gene silencing (VIGS) has been shown to be an effective tool for investigating gene functions in herbaceous plant species, but has rarely been tested in trees. The establishment of a fast and reliable transformation system is especially important for woody plants, many of which are recalcitrant to transformation. In this study, we established a tobacco rattle virus (TRV)-based VIGS system for two Populus species, Populus euphratica and P. × canescens. Here, TRV constructs carrying a 266 bp or a 558 bp fragment of the phytoene desaturase (PDS) gene were Agrobacterium-infiltrated into leaves of the two poplar species. Agrobacterium-mediated delivery of the shorter insert, TRV2-PePDS266, into the host poplars resulted in expected photobleaching in both tree species, but not the longer insert, PePDS558. The efficiency of VIGS was temperature-dependent, increasing by raising the temperature from 18 to 28 °C. The optimized TRV-VIGS system at 28 °C resulted in a high silencing frequency and efficiency up to 65-73 and 83-94%, respectively, in the two tested poplars. Moreover, syringe inoculation of Agrobacterium in 100 mM acetosyringone induced a more efficient silencing in the two poplar species, compared with other agroinfiltration methods, e.g., direct injection, misting and agrodrench. There were plant species-related differences in the response to VIGS because the photobleaching symptoms were more severe in P. × canescens than in P. euphratica. Furthermore, VIGS-treated P. euphratica exhibited a higher recovery rate (50%) after several weeks of the virus infection, compared with TRV-infected P. × canescens plants (20%). Expression stability of reference genes was screened to assess the relative abundance of PePDS mRNA in VIGS-treated P. euphratica and P. × canescens. PeACT7 was stably expressed in P. euphratica and UBQ-L was selected as the most suitable reference gene for P. × canescens using three different statistical approaches, geNorm, NormFinder and BestKeeper. Quantitative real-time PCR showed significant reductions in PDS transcripts (55-64%) in the photobleached leaves of both VIGS-treated poplar species. Our results demonstrate that the TRV-based VIGS provides a practical tool for gene functional analysis in Populus sp., especially in those poplar species which are otherwise recalcitrant to transformation.</AbstractText>
<CopyrightInformation>© The Author 2015. Published by Oxford University Press. All rights reserved. For Permissions, please email: journals.permissions@oup.com.</CopyrightInformation>
</Abstract>
<AuthorList CompleteYN="Y"><Author ValidYN="Y"><LastName>Shen</LastName>
<ForeName>Zedan</ForeName>
<Initials>Z</Initials>
<AffiliationInfo><Affiliation>College of Biological Sciences and Technology (Box 162), Beijing Forestry University, Beijing 100083, P.R. China.</Affiliation>
</AffiliationInfo>
</Author>
<Author ValidYN="Y"><LastName>Sun</LastName>
<ForeName>Jian</ForeName>
<Initials>J</Initials>
<AffiliationInfo><Affiliation>College of Life Science, Jiangsu Normal University, Xuzhou, Jiangsu Province 221116, P.R. China.</Affiliation>
</AffiliationInfo>
</Author>
<Author ValidYN="Y"><LastName>Yao</LastName>
<ForeName>Jun</ForeName>
<Initials>J</Initials>
<AffiliationInfo><Affiliation>College of Biological Sciences and Technology (Box 162), Beijing Forestry University, Beijing 100083, P.R. China.</Affiliation>
</AffiliationInfo>
</Author>
<Author ValidYN="Y"><LastName>Wang</LastName>
<ForeName>Shaojie</ForeName>
<Initials>S</Initials>
<AffiliationInfo><Affiliation>College of Biological Sciences and Technology (Box 162), Beijing Forestry University, Beijing 100083, P.R. China.</Affiliation>
</AffiliationInfo>
</Author>
<Author ValidYN="Y"><LastName>Ding</LastName>
<ForeName>Mingquan</ForeName>
<Initials>M</Initials>
<AffiliationInfo><Affiliation>College of Biological Sciences and Technology (Box 162), Beijing Forestry University, Beijing 100083, P.R. China.</Affiliation>
</AffiliationInfo>
</Author>
<Author ValidYN="Y"><LastName>Zhang</LastName>
<ForeName>Huilong</ForeName>
<Initials>H</Initials>
<AffiliationInfo><Affiliation>College of Biological Sciences and Technology (Box 162), Beijing Forestry University, Beijing 100083, P.R. China.</Affiliation>
</AffiliationInfo>
</Author>
<Author ValidYN="Y"><LastName>Qian</LastName>
<ForeName>Zeyong</ForeName>
<Initials>Z</Initials>
<AffiliationInfo><Affiliation>College of Biological Sciences and Technology (Box 162), Beijing Forestry University, Beijing 100083, P.R. China.</Affiliation>
</AffiliationInfo>
</Author>
<Author ValidYN="Y"><LastName>Zhao</LastName>
<ForeName>Nan</ForeName>
<Initials>N</Initials>
<AffiliationInfo><Affiliation>College of Biological Sciences and Technology (Box 162), Beijing Forestry University, Beijing 100083, P.R. China.</Affiliation>
</AffiliationInfo>
</Author>
<Author ValidYN="Y"><LastName>Sa</LastName>
<ForeName>Gang</ForeName>
<Initials>G</Initials>
<AffiliationInfo><Affiliation>College of Biological Sciences and Technology (Box 162), Beijing Forestry University, Beijing 100083, P.R. China.</Affiliation>
</AffiliationInfo>
</Author>
<Author ValidYN="Y"><LastName>Zhao</LastName>
<ForeName>Rui</ForeName>
<Initials>R</Initials>
<AffiliationInfo><Affiliation>College of Biological Sciences and Technology (Box 162), Beijing Forestry University, Beijing 100083, P.R. China.</Affiliation>
</AffiliationInfo>
</Author>
<Author ValidYN="Y"><LastName>Shen</LastName>
<ForeName>Xin</ForeName>
<Initials>X</Initials>
<AffiliationInfo><Affiliation>College of Biological Sciences and Technology (Box 162), Beijing Forestry University, Beijing 100083, P.R. China.</Affiliation>
</AffiliationInfo>
</Author>
<Author ValidYN="Y"><LastName>Polle</LastName>
<ForeName>Andrea</ForeName>
<Initials>A</Initials>
<AffiliationInfo><Affiliation>Forstbotanik und Baumphysiologie, Büsgen-Institut, Georg-August Universität Göttingen, Göttingen 37077, Germany.</Affiliation>
</AffiliationInfo>
</Author>
<Author ValidYN="Y"><LastName>Chen</LastName>
<ForeName>Shaoliang</ForeName>
<Initials>S</Initials>
<AffiliationInfo><Affiliation>College of Biological Sciences and Technology (Box 162), Beijing Forestry University, Beijing 100083, P.R. China lschen@bjfu.edu.cn.</Affiliation>
</AffiliationInfo>
</Author>
</AuthorList>
<Language>eng</Language>
<PublicationTypeList><PublicationType UI="D016428">Journal Article</PublicationType>
<PublicationType UI="D013485">Research Support, Non-U.S. Gov't</PublicationType>
</PublicationTypeList>
<ArticleDate DateType="Electronic"><Year>2015</Year>
<Month>07</Month>
<Day>23</Day>
</ArticleDate>
</Article>
<MedlineJournalInfo><Country>Canada</Country>
<MedlineTA>Tree Physiol</MedlineTA>
<NlmUniqueID>100955338</NlmUniqueID>
<ISSNLinking>0829-318X</ISSNLinking>
</MedlineJournalInfo>
<ChemicalList><Chemical><RegistryNumber>0</RegistryNumber>
<NameOfSubstance UI="D012333">RNA, Messenger</NameOfSubstance>
</Chemical>
<Chemical><RegistryNumber>EC 1.-</RegistryNumber>
<NameOfSubstance UI="D010088">Oxidoreductases</NameOfSubstance>
</Chemical>
<Chemical><RegistryNumber>EC 1.14.99.-</RegistryNumber>
<NameOfSubstance UI="C022398">phytoene dehydrogenase</NameOfSubstance>
</Chemical>
</ChemicalList>
<CitationSubset>IM</CitationSubset>
<MeshHeadingList><MeshHeading><DescriptorName UI="D018506" MajorTopicYN="N">Gene Expression Regulation, Plant</DescriptorName>
</MeshHeading>
<MeshHeading><DescriptorName UI="D020868" MajorTopicYN="Y">Gene Silencing</DescriptorName>
</MeshHeading>
<MeshHeading><DescriptorName UI="D017343" MajorTopicYN="N">Genes, Plant</DescriptorName>
</MeshHeading>
<MeshHeading><DescriptorName UI="D005822" MajorTopicYN="N">Genetic Vectors</DescriptorName>
<QualifierName UI="Q000378" MajorTopicYN="N">metabolism</QualifierName>
</MeshHeading>
<MeshHeading><DescriptorName UI="D010088" MajorTopicYN="N">Oxidoreductases</DescriptorName>
<QualifierName UI="Q000235" MajorTopicYN="N">genetics</QualifierName>
</MeshHeading>
<MeshHeading><DescriptorName UI="D010641" MajorTopicYN="N">Phenotype</DescriptorName>
</MeshHeading>
<MeshHeading><DescriptorName UI="D038761" MajorTopicYN="N">Photobleaching</DescriptorName>
</MeshHeading>
<MeshHeading><DescriptorName UI="D010935" MajorTopicYN="N">Plant Diseases</DescriptorName>
<QualifierName UI="Q000821" MajorTopicYN="N">virology</QualifierName>
</MeshHeading>
<MeshHeading><DescriptorName UI="D010942" MajorTopicYN="N">Plant Viruses</DescriptorName>
<QualifierName UI="Q000502" MajorTopicYN="Y">physiology</QualifierName>
</MeshHeading>
<MeshHeading><DescriptorName UI="D032107" MajorTopicYN="N">Populus</DescriptorName>
<QualifierName UI="Q000235" MajorTopicYN="N">genetics</QualifierName>
<QualifierName UI="Q000821" MajorTopicYN="Y">virology</QualifierName>
</MeshHeading>
<MeshHeading><DescriptorName UI="D012333" MajorTopicYN="N">RNA, Messenger</DescriptorName>
<QualifierName UI="Q000235" MajorTopicYN="N">genetics</QualifierName>
<QualifierName UI="Q000378" MajorTopicYN="N">metabolism</QualifierName>
</MeshHeading>
<MeshHeading><DescriptorName UI="D012015" MajorTopicYN="N">Reference Standards</DescriptorName>
</MeshHeading>
</MeshHeadingList>
<KeywordList Owner="NOTNLM"><Keyword MajorTopicYN="N">Agrobacterium infiltration</Keyword>
<Keyword MajorTopicYN="N">Populus euphratica</Keyword>
<Keyword MajorTopicYN="N">Populus × canescens</Keyword>
<Keyword MajorTopicYN="N">leaf photobleaching</Keyword>
<Keyword MajorTopicYN="N">phytoene desaturase</Keyword>
<Keyword MajorTopicYN="N">silencing efficiency</Keyword>
<Keyword MajorTopicYN="N">silencing frequency</Keyword>
<Keyword MajorTopicYN="N">tobacco rattle virus</Keyword>
</KeywordList>
</MedlineCitation>
<PubmedData><History><PubMedPubDate PubStatus="received"><Year>2014</Year>
<Month>12</Month>
<Day>16</Day>
</PubMedPubDate>
<PubMedPubDate PubStatus="accepted"><Year>2015</Year>
<Month>06</Month>
<Day>06</Day>
</PubMedPubDate>
<PubMedPubDate PubStatus="entrez"><Year>2015</Year>
<Month>7</Month>
<Day>26</Day>
<Hour>6</Hour>
<Minute>0</Minute>
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<Month>7</Month>
<Day>26</Day>
<Hour>6</Hour>
<Minute>0</Minute>
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<PubMedPubDate PubStatus="medline"><Year>2016</Year>
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<Hour>6</Hour>
<Minute>0</Minute>
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<ArticleId IdType="doi">10.1093/treephys/tpv064</ArticleId>
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<affiliations><list><country><li>Allemagne</li>
<li>République populaire de Chine</li>
</country>
<region><li>Basse-Saxe</li>
</region>
<settlement><li>Göttingen</li>
<li>Pékin</li>
</settlement>
<orgName><li>Université de Göttingen</li>
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</list>
<tree><country name="République populaire de Chine"><noRegion><name sortKey="Shen, Zedan" sort="Shen, Zedan" uniqKey="Shen Z" first="Zedan" last="Shen">Zedan Shen</name>
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<name sortKey="Chen, Shaoliang" sort="Chen, Shaoliang" uniqKey="Chen S" first="Shaoliang" last="Chen">Shaoliang Chen</name>
<name sortKey="Ding, Mingquan" sort="Ding, Mingquan" uniqKey="Ding M" first="Mingquan" last="Ding">Mingquan Ding</name>
<name sortKey="Qian, Zeyong" sort="Qian, Zeyong" uniqKey="Qian Z" first="Zeyong" last="Qian">Zeyong Qian</name>
<name sortKey="Sa, Gang" sort="Sa, Gang" uniqKey="Sa G" first="Gang" last="Sa">Gang Sa</name>
<name sortKey="Shen, Xin" sort="Shen, Xin" uniqKey="Shen X" first="Xin" last="Shen">Xin Shen</name>
<name sortKey="Sun, Jian" sort="Sun, Jian" uniqKey="Sun J" first="Jian" last="Sun">Jian Sun</name>
<name sortKey="Wang, Shaojie" sort="Wang, Shaojie" uniqKey="Wang S" first="Shaojie" last="Wang">Shaojie Wang</name>
<name sortKey="Yao, Jun" sort="Yao, Jun" uniqKey="Yao J" first="Jun" last="Yao">Jun Yao</name>
<name sortKey="Zhang, Huilong" sort="Zhang, Huilong" uniqKey="Zhang H" first="Huilong" last="Zhang">Huilong Zhang</name>
<name sortKey="Zhao, Nan" sort="Zhao, Nan" uniqKey="Zhao N" first="Nan" last="Zhao">Nan Zhao</name>
<name sortKey="Zhao, Rui" sort="Zhao, Rui" uniqKey="Zhao R" first="Rui" last="Zhao">Rui Zhao</name>
</country>
<country name="Allemagne"><region name="Basse-Saxe"><name sortKey="Polle, Andrea" sort="Polle, Andrea" uniqKey="Polle A" first="Andrea" last="Polle">Andrea Polle</name>
</region>
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