Effects of Green Fluorescent Protein or β-Glucuronidase Tagging on the Accumulation and Pathogenicity of a Resistance-Breaking Lettuce mosaic virus Isolate in Susceptible and Resistant Lettuce Cultivars
The RNA genome of a resistance-breaking isolate of Lettuce mosaic virus (LMV-E) was engineered to express the jellyfish green fluorescent protein (GFP) or β-glucuronidase (GUS) fused to the helper-component proteinase (HC-Pro) to study LMV invasion and spread in susceptible and resistant lettuce cul...
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The American Phytopathological Society
2000-03-01
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Series: | Molecular Plant-Microbe Interactions |
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Online Access: | https://apsjournals.apsnet.org/doi/10.1094/MPMI.2000.13.3.316 |
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author | Sylvie German-Retana Thierry Candresse Emmanuel Alias René-Pierre Delbos Olivier Le Gall |
author_facet | Sylvie German-Retana Thierry Candresse Emmanuel Alias René-Pierre Delbos Olivier Le Gall |
author_sort | Sylvie German-Retana |
collection | DOAJ |
description | The RNA genome of a resistance-breaking isolate of Lettuce mosaic virus (LMV-E) was engineered to express the jellyfish green fluorescent protein (GFP) or β-glucuronidase (GUS) fused to the helper-component proteinase (HC-Pro) to study LMV invasion and spread in susceptible and resistant lettuce cultivars. Virus accumulation and movement were monitored by either histochemical GUS assays or detection of GFP fluorescence under UV light. The GFP- and GUS-tagged viruses spread systemically in the susceptible lettuce cultivars Trocadero and Vanguard, where they induced attenuated symptoms, compared with the wild-type virus. Accumulation of the GFP-tagged virus was reduced but less affected than in the case of the GUS-tagged virus. Systemic movement of both recombinant viruses was very severely affected in Vanguard 75, a lettuce cultivar nearly isogenic to Vanguard but carrying the resistance gene mo12 . Accumulation of the recombinant viruses in systemically infected leaves was either undetectable (GUS-tag) or erratic, strongly delayed, and inhibited by as much as 90% (GFP-tag). As a consequence, and contrary to the parental virus, the recombinant viruses were not able to overcome the protection afforded by the mo12 gene. Taken together, these results indicate that GUS or GFP tagging of the HC-Pro of LMV has significant negative effects on the biology of the virus, abolishing its resistance-breaking properties and reducing its pathogenicity in susceptible cultivars. |
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institution | Directory Open Access Journal |
issn | 0894-0282 1943-7706 |
language | English |
last_indexed | 2024-04-13T00:21:04Z |
publishDate | 2000-03-01 |
publisher | The American Phytopathological Society |
record_format | Article |
series | Molecular Plant-Microbe Interactions |
spelling | doaj.art-4c870722c8a54f76836c6f0a35704cda2022-12-22T03:10:45ZengThe American Phytopathological SocietyMolecular Plant-Microbe Interactions0894-02821943-77062000-03-0113331632410.1094/MPMI.2000.13.3.316Effects of Green Fluorescent Protein or β-Glucuronidase Tagging on the Accumulation and Pathogenicity of a Resistance-Breaking Lettuce mosaic virus Isolate in Susceptible and Resistant Lettuce CultivarsSylvie German-RetanaThierry CandresseEmmanuel AliasRené-Pierre DelbosOlivier Le GallThe RNA genome of a resistance-breaking isolate of Lettuce mosaic virus (LMV-E) was engineered to express the jellyfish green fluorescent protein (GFP) or β-glucuronidase (GUS) fused to the helper-component proteinase (HC-Pro) to study LMV invasion and spread in susceptible and resistant lettuce cultivars. Virus accumulation and movement were monitored by either histochemical GUS assays or detection of GFP fluorescence under UV light. The GFP- and GUS-tagged viruses spread systemically in the susceptible lettuce cultivars Trocadero and Vanguard, where they induced attenuated symptoms, compared with the wild-type virus. Accumulation of the GFP-tagged virus was reduced but less affected than in the case of the GUS-tagged virus. Systemic movement of both recombinant viruses was very severely affected in Vanguard 75, a lettuce cultivar nearly isogenic to Vanguard but carrying the resistance gene mo12 . Accumulation of the recombinant viruses in systemically infected leaves was either undetectable (GUS-tag) or erratic, strongly delayed, and inhibited by as much as 90% (GFP-tag). As a consequence, and contrary to the parental virus, the recombinant viruses were not able to overcome the protection afforded by the mo12 gene. Taken together, these results indicate that GUS or GFP tagging of the HC-Pro of LMV has significant negative effects on the biology of the virus, abolishing its resistance-breaking properties and reducing its pathogenicity in susceptible cultivars.https://apsjournals.apsnet.org/doi/10.1094/MPMI.2000.13.3.316infectious cDNAreporter-tagged potyvirusviral invasion |
spellingShingle | Sylvie German-Retana Thierry Candresse Emmanuel Alias René-Pierre Delbos Olivier Le Gall Effects of Green Fluorescent Protein or β-Glucuronidase Tagging on the Accumulation and Pathogenicity of a Resistance-Breaking Lettuce mosaic virus Isolate in Susceptible and Resistant Lettuce Cultivars Molecular Plant-Microbe Interactions infectious cDNA reporter-tagged potyvirus viral invasion |
title | Effects of Green Fluorescent Protein or β-Glucuronidase Tagging on the Accumulation and Pathogenicity of a Resistance-Breaking Lettuce mosaic virus Isolate in Susceptible and Resistant Lettuce Cultivars |
title_full | Effects of Green Fluorescent Protein or β-Glucuronidase Tagging on the Accumulation and Pathogenicity of a Resistance-Breaking Lettuce mosaic virus Isolate in Susceptible and Resistant Lettuce Cultivars |
title_fullStr | Effects of Green Fluorescent Protein or β-Glucuronidase Tagging on the Accumulation and Pathogenicity of a Resistance-Breaking Lettuce mosaic virus Isolate in Susceptible and Resistant Lettuce Cultivars |
title_full_unstemmed | Effects of Green Fluorescent Protein or β-Glucuronidase Tagging on the Accumulation and Pathogenicity of a Resistance-Breaking Lettuce mosaic virus Isolate in Susceptible and Resistant Lettuce Cultivars |
title_short | Effects of Green Fluorescent Protein or β-Glucuronidase Tagging on the Accumulation and Pathogenicity of a Resistance-Breaking Lettuce mosaic virus Isolate in Susceptible and Resistant Lettuce Cultivars |
title_sort | effects of green fluorescent protein or β glucuronidase tagging on the accumulation and pathogenicity of a resistance breaking lettuce mosaic virus isolate in susceptible and resistant lettuce cultivars |
topic | infectious cDNA reporter-tagged potyvirus viral invasion |
url | https://apsjournals.apsnet.org/doi/10.1094/MPMI.2000.13.3.316 |
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