Beet soil-borne mosaic virus (BSBMV) belongs to the Benyvirus genus together with Beet necrotic yellow vein virus (BNYVV). Both viruses share similar genomic organization and are vectored by Polymyxa betae. We recently demonstrated the ability of a BNYVV helper strain to replicate and encapsidate BSBMV RNA-3 suggesting a common and conserved viral RNA selection mechanism for both viruses. A 1,733 nts long BSBMV RNA-4 has been molecularly and functionally characterized by our group and full-length cDNA-derived infectious transcripts obtained. Similarly to BSBMV RNA-3, these RNA-4 transcripts are amplified in planta by BNYVV machinery. We demonstrated that BSBMV RNA-4 can substitute BNYVV RNA-4 for an efficient transmission through the vector P. betae in Beta vulgaris plants. Two putative ORFs have been identified and could encode for a 32 kDa (p32) and a 13 kDa (p13) protein. P32 sequence is close to BNYVV p31 protein. Using BNYVV helper strain, BSBMV RNA4’s role in the plant-virus-vector interaction investigation has been initiated using reverse genetics approaches. Mutated, deleted and Flag or GFP-tagged sequences of BSBMV RNA4 encoded proteins have been expressed in viral context in Chenopodium quinoa and Beta macrocarpa hosts. Necrotic local lesions phenotype has been associated specifically to the protein expression onto mechanically inoculated C. quinoa plants. Western blot analyses using FLAG-specific antibodies revealed a high molecular weight protein that suggest either a strong interaction of p32 protein with host protein(s) or post translational modifications that need to be better investigated. Infectious transcripts obtained from cDNA clones carrying deleted forms of BSBMV RNA-4 are being employed to identify the domains of the proteins involved in virus-vector interaction required for the efficient Benyviruses transmission.

Reverse genetics study of the Beet soil-borne mosaic virus RNA4 role in the plant-vector-virus interaction

D'ALONZO, MASSIMILIANO;RUBIES AUTONELL, CONCEPCION;RATTI, CLAUDIO
2010

Abstract

Beet soil-borne mosaic virus (BSBMV) belongs to the Benyvirus genus together with Beet necrotic yellow vein virus (BNYVV). Both viruses share similar genomic organization and are vectored by Polymyxa betae. We recently demonstrated the ability of a BNYVV helper strain to replicate and encapsidate BSBMV RNA-3 suggesting a common and conserved viral RNA selection mechanism for both viruses. A 1,733 nts long BSBMV RNA-4 has been molecularly and functionally characterized by our group and full-length cDNA-derived infectious transcripts obtained. Similarly to BSBMV RNA-3, these RNA-4 transcripts are amplified in planta by BNYVV machinery. We demonstrated that BSBMV RNA-4 can substitute BNYVV RNA-4 for an efficient transmission through the vector P. betae in Beta vulgaris plants. Two putative ORFs have been identified and could encode for a 32 kDa (p32) and a 13 kDa (p13) protein. P32 sequence is close to BNYVV p31 protein. Using BNYVV helper strain, BSBMV RNA4’s role in the plant-virus-vector interaction investigation has been initiated using reverse genetics approaches. Mutated, deleted and Flag or GFP-tagged sequences of BSBMV RNA4 encoded proteins have been expressed in viral context in Chenopodium quinoa and Beta macrocarpa hosts. Necrotic local lesions phenotype has been associated specifically to the protein expression onto mechanically inoculated C. quinoa plants. Western blot analyses using FLAG-specific antibodies revealed a high molecular weight protein that suggest either a strong interaction of p32 protein with host protein(s) or post translational modifications that need to be better investigated. Infectious transcripts obtained from cDNA clones carrying deleted forms of BSBMV RNA-4 are being employed to identify the domains of the proteins involved in virus-vector interaction required for the efficient Benyviruses transmission.
EMBO Workshop in Genomic approaches to interaction between plant viruses, their hosts and their vectors
P46
p46
M. D’Alonzo; D. Gilmer; C. Rubies Autonell; C. Ratti
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Utilizza questo identificativo per citare o creare un link a questo documento: http://hdl.handle.net/11585/99328
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