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http://hdl.handle.net/10174/9134
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Title: | Molecular characterization of Olea europaea L. necrovirus transmission by fungal vectors |
Authors: | Varanda, Carla Marisa Reis |
Advisors: | Clara, Maria Ivone Esteves da Oliveira, Isabel Solange Martins de |
Keywords: | infecting necrovirus coat protein |
Issue Date: | 2011 |
Publisher: | Universidade Évora |
Abstract: | Abstract
OMMV is an olive infecting necrovirus, likely a recombinant between TNV-D, with which it
shares a high identity in its coat protein (CP), and OLV-1. Design of specific primers to use in
RT-PCR, allowed for the first time molecular discrimination of OMMV from TNV-D. RT-PCR
application to an olive orchard survey revealed OMMV predominance. Its transmission by
Olpidium brassicae was demonstrated here for the first time and further examined. OMMV
wild-type, a natural non-transmissible OMMVL11 mutant containing two mutations, a
construct OMMVWT/OMMVL11 and single mutants obtained by site-directed mutagenesis
were found similarly infectious. Sequencing, homology modelling and transmissibility assays
showed that the single mutation of Asn residue to Tyr in position 189 of the CP amino acid
sequence was located internally in the particle and abolished virus-zoospore adsorption and
transmissibility. This indicates that mutation altered CP conformation on zoospore recognition
sites. These findings may have important epidemiological implications in the olive crop.
Caracterização molecular da transmissão de Necrovirus (Tombusviridae) de Olea
europaea L. por fungos vectores
OMMV é um necrovirus que infecta a oliveira e que terá surgido a partir de recombinação
entre TNV-D, com o qual compartilha uma elevada identidade na cápside proteica (CP), e OLV-
1. O desenho de ‘primers’ específicos para utilização em RT-PCR permitiu, pela primeira vez, a
discriminação entre OMMV e TNV-D. A aplicação de RT-PCR a um olival revelou a
predominância de OMMV. A transmissão por Olpidium brassicae foi aqui pela primeira vez
demonstrada e examinada. OMMV ‘wild type’, um mutante natural OMMVL11 não
transmissível contendo duas mutações e mutantes obtidos por mutagénese dirigida
revelaram-se igualmente infecciosos. Sequenciação, modelagem por homologia e en |
URI: | http://hdl.handle.net/10174/9134 |
Type: | doctoralThesis |
Appears in Collections: | BIB - Formação Avançada - Teses de Doutoramento
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