|Freitas-Astua, J. - CENTRO APTA CITROS, BR|
|Locali, E. - CENTRO APTA CITROS, BR|
|Antonioli-Luizon, R. - CENTRO APTA CITROS, BR|
|Kitajima, E. W. - NAP/MEPA/ESALQ/USP, BR|
|Machado, M. A. - CENTRO APTA CITROS, BR|
Submitted to: International Organization of Citrus Virologists Proceedings
Publication Type: Proceedings
Publication Acceptance Date: November 15, 2004
Publication Date: November 30, 2004
Citation: Freitas-Astua, J., Locali, E., Antonioli-Luizon, R., Kitajima, E., Hilf, M.E., Gottwald, T.R., Machado, M. 2004. Nationwide detection of citrus leprosis virus in brazil using rt-pcr. International Organization of Citrus Virologists Proceedings. Interpretive Summary: Citrus leprosis virus (CiLV) causes a widespread and destructive disease of citrus in Brazil called leprosis. This paper describes the development of methods to detect the virus that causes leprosis. The ability to detect CiLV will substantially decrease the risk that leprosis disease will become established in the citrus areas of the United States.
Technical Abstract: Citrus leprosis virus (CiLV) is transmitted by the Tenuipalpidae mite Brevipalpus phoenicis and causes estimated yearly losses of US $60-100 million in Brazil. Two forms of CiLV have been recognized based on particle morphology and cytopathology: the prevalent cytoplasmic type (CiLV-C) and the rare, nuclear type (CiLV-N). Using RT-PCR with primers that specifically amplify a region within the putative movement protein gene of the CiLV-C, we detected the virus in a limited number of citrus samples from several municipalities within Sao Paulo State, Brazil. To expand upon these initial findings, we analyzed 152 symptomatic samples of five species and 21 varieties of citrus from 57 municipalities in nine states representing the main citrus producing areas of the country. Citrus leaves, but also fruits and stems were assessed using both symptomatic and asymptomatic samples. RT-PCR analysis of nucleic acid extracts from all samples with typical leprosis symptoms yielded bands of the expected size as assessed in agarose gels. Using the same conditions, RT-PCR did not amplify any bands from extracts from asymptomatic samples. The results demonstrate the usefulness of RT-PCR as a tool for the detection of CiLV isolates from different citrus varieties and geographic regions.