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Localization of Tobacco Yellow Dwarf Virus Replication Using the In Plant Activation (INPACT) Expression Platform
Geminiviruses and their diseases are a considerable economic threat to a vast number of crops worldwide. Investigating how and where these viruses replicate and accumulate in their hosts may lead to novel molecular resistance strategies. In this study, we used the Rep-inducible In Plant Activation (...
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Published in: | Viruses 2020-06, Vol.12 (6), p.688 |
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description | Geminiviruses and their diseases are a considerable economic threat to a vast number of crops worldwide. Investigating how and where these viruses replicate and accumulate in their hosts may lead to novel molecular resistance strategies. In this study, we used the Rep-inducible In Plant Activation (INPACT) expression platform, based on the genome of tobacco yellow dwarf virus (TYDV), to determine where this model mastrevirus replicates in its host tobacco. By developing an infectious clone of TYDV and optimizing its delivery by agroinfiltration, we first established an efficient artificial infection process. When delivered into transgenic tobacco plants containing a TYDV-based INPACT cassette encoding the β-glucuronidase (GUS) reporter, we showed the virus activates GUS expression. Histology revealed that reporter gene expression was limited to phloem-associated cell types suggesting TYDV replication has a restricted tissue tropism. |
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Investigating how and where these viruses replicate and accumulate in their hosts may lead to novel molecular resistance strategies. In this study, we used the Rep-inducible In Plant Activation (INPACT) expression platform, based on the genome of tobacco yellow dwarf virus (TYDV), to determine where this model mastrevirus replicates in its host tobacco. By developing an infectious clone of TYDV and optimizing its delivery by agroinfiltration, we first established an efficient artificial infection process. When delivered into transgenic tobacco plants containing a TYDV-based INPACT cassette encoding the β-glucuronidase (GUS) reporter, we showed the virus activates GUS expression. Histology revealed that reporter gene expression was limited to phloem-associated cell types suggesting TYDV replication has a restricted tissue tropism.</description><identifier>ISSN: 1999-4915</identifier><identifier>EISSN: 1999-4915</identifier><identifier>DOI: 10.3390/v12060688</identifier><identifier>PMID: 32604765</identifier><language>eng</language><publisher>Switzerland: MDPI AG</publisher><subject>Cloning ; Communication ; Deoxyribonucleic acid ; DNA ; Enzymes ; Ethanol ; Geminiviridae - genetics ; Geminiviridae - growth & development ; geminivirus ; Gene expression ; Gene Expression Regulation, Plant - genetics ; Genome, Viral - genetics ; Genomes ; Glucuronidase - genetics ; Host Microbial Interactions - genetics ; Host Microbial Interactions - physiology ; In Plant Activation (INPACT) ; Infections ; Localization ; Nicotiana - virology ; Pathogenesis ; phloem ; Plant Diseases - virology ; Plants, Genetically Modified - virology ; Proteins ; Replication ; Reporter gene ; Tobacco ; Transgenic plants ; Tropism ; Viral Tropism - genetics ; Virus Replication - genetics ; Virus Replication - physiology ; Viruses ; β-glucuronidase (GUS)</subject><ispartof>Viruses, 2020-06, Vol.12 (6), p.688</ispartof><rights>2020. 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Investigating how and where these viruses replicate and accumulate in their hosts may lead to novel molecular resistance strategies. In this study, we used the Rep-inducible In Plant Activation (INPACT) expression platform, based on the genome of tobacco yellow dwarf virus (TYDV), to determine where this model mastrevirus replicates in its host tobacco. By developing an infectious clone of TYDV and optimizing its delivery by agroinfiltration, we first established an efficient artificial infection process. When delivered into transgenic tobacco plants containing a TYDV-based INPACT cassette encoding the β-glucuronidase (GUS) reporter, we showed the virus activates GUS expression. Histology revealed that reporter gene expression was limited to phloem-associated cell types suggesting TYDV replication has a restricted tissue tropism.</description><subject>Cloning</subject><subject>Communication</subject><subject>Deoxyribonucleic acid</subject><subject>DNA</subject><subject>Enzymes</subject><subject>Ethanol</subject><subject>Geminiviridae - genetics</subject><subject>Geminiviridae - growth & development</subject><subject>geminivirus</subject><subject>Gene expression</subject><subject>Gene Expression Regulation, Plant - genetics</subject><subject>Genome, Viral - genetics</subject><subject>Genomes</subject><subject>Glucuronidase - genetics</subject><subject>Host Microbial Interactions - genetics</subject><subject>Host Microbial Interactions - physiology</subject><subject>In Plant Activation (INPACT)</subject><subject>Infections</subject><subject>Localization</subject><subject>Nicotiana - virology</subject><subject>Pathogenesis</subject><subject>phloem</subject><subject>Plant Diseases - virology</subject><subject>Plants, Genetically Modified - virology</subject><subject>Proteins</subject><subject>Replication</subject><subject>Reporter gene</subject><subject>Tobacco</subject><subject>Transgenic plants</subject><subject>Tropism</subject><subject>Viral Tropism - genetics</subject><subject>Virus Replication - genetics</subject><subject>Virus Replication - physiology</subject><subject>Viruses</subject><subject>β-glucuronidase (GUS)</subject><issn>1999-4915</issn><issn>1999-4915</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2020</creationdate><recordtype>article</recordtype><sourceid>PIMPY</sourceid><sourceid>DOA</sourceid><recordid>eNpdkk1vEzEQhi0EoqVw4A8gS1zaQ8Bef6x9QYrSFiJFUKEUiZPltb2po806tb0p5dfjsCVqOXk08-jxq9EA8BajD4RI9HGHK8QRF-IZOMZSygmVmD1_VB-BVymtEeJcovolOCIVR7Tm7BjcLoLRnf-tsw89DC1chkYbE-BP13XhDp7f6djCHz4OCX53286bkbxOvl_BfOPgvIdXne4znJrsd-P0dP71ajpbnsGLX9voUtr3CpTbEDevwYtWd8m9eXhPwPXlxXL2ZbL49nk-my4mhnKZS-wSUnPZUsYFs0zUwlBiqK4rLQjiBAnTNtJibjgRxDrOROOQkFI4VxtLTsB89Nqg12ob_UbHexW0V38bIa6UjtmbzqmmqbhkjBJky3cVFbYhrEYaV0JibnlxfRpd26HZOGtcn6PunkifTnp_o1Zhp2rCKOWkCE4fBDHcDi5ltfHJlBXr3oUhqYpiSTHBDBf0_X_oOgyxL6vaU0JSzsieOhspE0NK0bWHMBip_VGow1EU9t3j9Afy3xWQP99xsCM</recordid><startdate>20200626</startdate><enddate>20200626</enddate><creator>Kato, Maiko</creator><creator>Harding, Robert</creator><creator>Dale, James</creator><creator>Dugdale, Benjamin</creator><general>MDPI AG</general><general>MDPI</general><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>7U9</scope><scope>7X7</scope><scope>7XB</scope><scope>88E</scope><scope>8FE</scope><scope>8FH</scope><scope>8FI</scope><scope>8FJ</scope><scope>8FK</scope><scope>ABUWG</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>H94</scope><scope>HCIFZ</scope><scope>K9.</scope><scope>LK8</scope><scope>M0S</scope><scope>M1P</scope><scope>M7P</scope><scope>PIMPY</scope><scope>PQEST</scope><scope>PQQKQ</scope><scope>PQUKI</scope><scope>PRINS</scope><scope>7X8</scope><scope>5PM</scope><scope>DOA</scope><orcidid>https://orcid.org/0000-0003-2281-4809</orcidid><orcidid>https://orcid.org/0000-0002-9600-7287</orcidid></search><sort><creationdate>20200626</creationdate><title>Localization of Tobacco Yellow Dwarf Virus Replication Using the In Plant Activation (INPACT) Expression Platform</title><author>Kato, Maiko ; 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subjects | Cloning Communication Deoxyribonucleic acid DNA Enzymes Ethanol Geminiviridae - genetics Geminiviridae - growth & development geminivirus Gene expression Gene Expression Regulation, Plant - genetics Genome, Viral - genetics Genomes Glucuronidase - genetics Host Microbial Interactions - genetics Host Microbial Interactions - physiology In Plant Activation (INPACT) Infections Localization Nicotiana - virology Pathogenesis phloem Plant Diseases - virology Plants, Genetically Modified - virology Proteins Replication Reporter gene Tobacco Transgenic plants Tropism Viral Tropism - genetics Virus Replication - genetics Virus Replication - physiology Viruses β-glucuronidase (GUS) |
title | Localization of Tobacco Yellow Dwarf Virus Replication Using the In Plant Activation (INPACT) Expression Platform |
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