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Journal of Virology, March 2005, p. 3028-3037, Vol. 79, No. 5
0022-538X/05/$08.00+0     doi:10.1128/JVI.79.5.3028-3037.2005
Copyright © 2005, American Society for Microbiology. All Rights Reserved.

Identification and Genomic Characterization of a New Virus (Tymoviridae Family) Associated with Citrus Sudden Death Disease

Walter Maccheroni,1 Marcos C. Alegria,1 Christian C. Greggio,1 João Paulo Piazza,1 Rachel F. Kamla,1 Paula R. A. Zacharias,1 Moshe Bar-Joseph,1,{dagger} Elliot W. Kitajima,2 Laura C. Assumpção,1 Giovana Camarotte,1 Jussara Cardozo,1 Elaine C. Casagrande,1 Fernanda Ferrari,1 Sulamita F. Franco,1 Poliana F. Giachetto,1 Alessandra Girasol,1 Hamilton Jordão Jr.,1 Vitor H. A. Silva,1 Leonardo C. A. Souza,1 Carlos I. Aguilar-Vildoso,1 Almir S. Zanca,1 Paulo Arruda,1 João Paulo Kitajima,1 Fernando C. Reinach,1 Jesus A. Ferro,1 and Ana C. R. da Silva1*

Alellyx Applied Genomics, Campinas,1 Department of Entomology, Phytopathology, and Zoological Agriculture, ESALQ/USP, Piracicaba, São Paulo, Brazil2

Received 7 August 2004/ Accepted 21 October 2004

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.

* Corresponding author. Mailing address: Alellyx Applied Genomics, Techno Park (Rod. Anhanguera Km 104), Rua James Clerk Maxwell 320, 13069-380 Campinas, SP, Brazil. Phone: 55 19 3783 9406. Fax: 55 19 3783 9412. E-mail: ana.rasera{at}alellyx.com.br.

{dagger} Present address: The S. Tolkowsky Laboratory, Dept. of Virology, Agricultural Research Organization, the Volcani Center, Bet Dagan, Israel.

Journal of Virology, March 2005, p. 3028-3037, Vol. 79, No. 5
0022-538X/05/$08.00+0     doi:10.1128/JVI.79.5.3028-3037.2005
Copyright © 2005, American Society for Microbiology. All Rights Reserved.





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