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Identification and genomic characterization of a new virus (Tymoviridae family) associated with citrus sudden death disease.

ABSTRACT: Citrus sudden death (CSD) is a new disease that has killed approximately 1 million orange trees in Brazil. Here we report the identification of a new virus associated with the disease. RNAs isolated from CSD-affected and nonaffected trees were used to construct cDNA libraries. A set of viral sequences present exclusively in libraries of CSD-affected trees was used to obtain the complete genome sequence of the new virus. Phylogenetic analysis revealed that this virus is a new member of the genus Marafivirus. Antibodies raised against the putative viral coat proteins allowed detection of viral antigens of expected sizes in affected plants. Electron microscopy of purified virus confirmed the presence of typical isometric Marafivirus particles. The screening of 773 affected and nonaffected citrus trees for the presence of the virus showed a 99.7% correlation between disease symptoms and the presence of the virus. We also detected the virus in aphids feeding on affected trees. These results suggest that this virus is likely to be the causative agent of CSD. The virus was named Citrus sudden death-associated virus.

SUBMITTER: Maccheroni W

PROVIDER: S-EPMC548464 | biostudies-literature | 2005 Mar

REPOSITORIES: biostudies-literature

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Identification and genomic characterization of a new virus (Tymoviridae family) associated with citrus sudden death disease.

Maccheroni Walter W Alegria Marcos C MC Greggio Christian C CC Piazza João Paulo JP Kamla Rachel F RF Zacharias Paula R A PR Bar-Joseph Moshe M Kitajima Elliot W EW Assumpção Laura C LC Camarotte Giovana G Cardozo Jussara J Casagrande Elaine C EC Ferrari Fernanda F Franco Sulamita F SF Giachetto Poliana F PF Girasol Alessandra A Jordão Hamilton H Silva Vitor H A VH Souza Leonardo C A LC Aguilar-Vildoso Carlos I CI Zanca Almir S AS Arruda Paulo P Kitajima João Paulo JP Reinach Fernando C FC Ferro Jesus A JA da Silva Ana C R AC

Journal of virology 20050301 5

Citrus sudden death (CSD) is a new disease that has killed approximately 1 million orange trees in Brazil. Here we report the identification of a new virus associated with the disease. RNAs isolated from CSD-affected and nonaffected trees were used to construct cDNA libraries. A set of viral sequences present exclusively in libraries of CSD-affected trees was used to obtain the complete genome sequence of the new virus. Phylogenetic analysis revealed that this virus is a new member of the genus ...[more]

PMID: 15709023

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Project description:BACKGROUND: The xylem-inhabiting bacterium Xylella fastidiosa (Xf) is the causal agent of Pierce's disease (PD) in vineyards and citrus variegated chlorosis (CVC) in orange trees. Both of these economically-devastating diseases are caused by distinct strains of this complex group of microorganisms, which has motivated researchers to conduct extensive genomic sequencing projects with Xf strains. This sequence information, along with other molecular tools, have been used to estimate the evolutionary history of the group and provide clues to understand the capacity of Xf to infect different hosts, causing a variety of symptoms. Nonetheless, although significant amounts of information have been generated from Xf strains, a large proportion of these efforts has concentrated on the study of North American strains, limiting our understanding about the genomic composition of South American strains - which is particularly important for CVC-associated strains. RESULTS: This paper describes the first genome-wide comparison among South American Xf strains, involving 6 distinct citrus-associated bacteria. Comparative analyses performed through a microarray-based approach allowed identification and characterization of large mobile genetic elements that seem to be exclusive to South American strains. Moreover, a large-scale sequencing effort, based on Suppressive Subtraction Hybridization (SSH), identified 290 new ORFs, distributed in 135 Groups of Orthologous Elements, throughout the genomes of these bacteria. CONCLUSION: Results from microarray-based comparisons provide further evidence concerning activity of horizontally transferred elements, reinforcing their importance as major mediators in the evolution of Xf. Moreover, the microarray-based genomic profiles showed similarity between Xf strains 9a5c and Fb7, which is unexpected, given the geographical and chronological differences associated with the isolation of these microorganisms. The newly identified ORFs, obtained by SSH, represent an approximately 10% increase in our current knowledge of the South American Xf gene pool and include new putative virulence factors, as well as novel potential markers for strain identification. Surprisingly, this list of novel elements include sequences previously believed to be unique to North American strains, pointing to the necessity of revising the list of specific markers that may be used for identification of distinct Xf strains.

Dataset Information

Identification and genomic characterization of a new virus (Tymoviridae family) associated with citrus sudden death disease.

Publications

Identification and genomic characterization of a new virus (Tymoviridae family) associated with citrus sudden death disease.

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