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Content archived on 2024-04-30

Risk assessment with genetically engineered woody plants expressing virus coat protein gene

Objective



Coat protein mediated protection is a very promising way to control detrimental virus diseases. The objective of the project is to assess possible risks associated with the use of woody perennial crops (Prunus and grapevine) expressing viral coat protein (CP). The main risks to be evaluated are possible transencapsidation and RNA recombination leading to a change in the host range of the viruses and /or emergence of new viruses. Experiments will be conducted both in confined environment and small scale field conditions. Transgenic Prunus armeniaca and P. domestica expressing the CP gene of plum pox polyviruses and grapevines transformed with the CP gene of grapevine fan leaf and arabis mosaic nepoviruses have been already obtained. Grapevines expressing the CP gene of grapevine trichoviruses A and B will be produced under this project. In addition herbaceous Nicotiana plant models have also been transformed with the same constructs. In this project we want to compare the transencapsidation phenomenon of transgenic Prunus expressing the native form of PPV CP and those transformed with a CP gene deleted of the DAG amino acid triplet involved in the virus vector recognition. Due to the lack of data about nepoviruses transencaspidation will be studied by simultaneously inoculating Nicotiana benthamiana with infectious transcripts derived from GFLV RNAl and ArMV RNA2. Further transgenic grapevines and/or N. benthamiana expressing GFLV CP (resp. GVA CP) will be inoculated with ArMV (resp. GVB) and conversely. For the evaluation of RNA recombination Prunus expressing PPV CP mutant lacking the DAG triplet will be inoculated with PPV variants. For GFLV, available mutants will be inoculated to N. benthamiana. The one is a tripartite mutant which has an RNA2 split into 2 RNA species, one of them bearing the CP gene, the second are CP defective mutants which infect protoplats but do not spread to whole plants. For GVA and GVB, infectious transcripts will be prepared within the project and will be use as sources of chimeric viruses by exchanging the CP of GVA and GVB.
A small scale field will be planned with transgenic Prunus to evaluate the impact of possible transencapsidation and RNA recombination under natural conditions especially on aphid transmissibility. Similar experiments will also be set up with transgenic grapevine rootstocks expressing GFLV CP to monitor viral populations and transgene expression in the vineyard. Results of these experiments will bring scientific informations on the frequency of heteroencapsidation and RNA recombination with the concerned viruses. The project aims also the assessment of transgene expression on 2 woody perennials in field conditions and to design alternative constructs before their large distribution in the environment

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Coordinator

Institut National de la Recherche Agronomique
EU contribution
No data
Address
71,Avenue Edouard Bourleaux
33883 Villenave-d'Ornon
France

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Total cost

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Participants (7)

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