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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Bibliographic Details
Published in:Viruses 2020-06, Vol.12 (6), p.688
Main Authors: Kato, Maiko, Harding, Robert, Dale, James, Dugdale, Benjamin
Format: Article
Language:English
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)
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Summary: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.
ISSN:1999-4915
1999-4915
DOI:10.3390/v12060688