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Title: First report of Tomato chlorosis virus infecting sweet pepper in Costa Rica

Author
item VARGAS, JOSE - Universidad De Costa Rica
item Hammond, Rosemarie
item HERNANDEZ, EDUARDO - Universidad De Costa Rica
item BARBOZA, NATALIA - Universidad De Costa Rica
item MORA, FLORI-BETH - Universidad De Costa Rica
item RAMIREZ, PILAR - Universidad De Costa Rica

Submitted to: Plant Disease
Publication Type: Peer Reviewed Journal
Publication Acceptance Date: 7/11/2011
Publication Date: 11/1/2011
Citation: Vargas, J.A., Hammond, R., Hernandez, E., Barboza, N., Mora, F., Ramirez, P. 2011. First report of Tomato chlorosis virus infecting sweet pepper in Costa Rica. Plant Disease. 95:1482.

Interpretive Summary: Tomato chlorosis virus (ToCV) is a worldwide emerging whitefly-transmitted virus. In 2008, a survey of whiteflies and whitefly-borne viruses was performed in greenhouses in the province of Cartago, Costa Rica. During this survey, sweet pepper plants showing viral symptoms were observed and large populations of whiteflies were present. Molecular analyses conducted by an ARS scientist in Beltsville, Maryland and collaborators in Costa Rica determined that the plants were infected with ToCV. This is the first report of ToCV infecting sweet pepper in Costa Rica. This information has been communicated to growers in Costa Rica and is being used to assess the incidence and economic impact of the virus in sweet pepper production.. The results impact U.S. agriculture as ToCV is present in the U.S. and is a serious threat to vegetable production in North America.

Technical Abstract: In September 2008, a survey of whiteflies and whitefly-borne viruses was performed in greenhouses in the province of Cartago, Costa Rica. During this survey, sweet pepper (Capsicum annuum cv. Nataly) plants showing interveinal chlorosis, enations, necrosis, and mild upward leaf curling were observed. Large populations of whiteflies were present and they were found to be composed only of Trialeurodes vaporariorum. Total RNA from frozen plant samples was extracted using TRI Reagent (Molecular Research Inc., Cincinnati, OH). RevertAid H Minus reverse transcriptase kit (Fermentas, Hanover, MD) was used for reverse transcription of the total RNA extract, with cDNA synthesis directed using random primers. A Real Time PCR assay was performed to detect Tomato chlorosis virus (ToCV) (genus Crinivirus; family Closteroviridae) using the SYBR Green PCR Master Mix (Applied Biosystems, Carlsbad, CA). Three sets of primers were used to confirm the presence of Tomato chlorosis virus (ToCV) in the samples, the TocQ875F-TocQ998R primer set directed to the HSP gene, the ToCVp22RQF (5'-TGGATCTCACTGGTTGCTTG-3')-ToCVp22RQR(5'- TAGTGTTTCAGCGCCAACAG- 3') primer pair which amplifies a segment of the ToCV p22 gene (Hammond et al. unpublished) and the ToCVCPmRQF (5'- CATTGGTTGGGGATTACGTC- 3') –ToCVCPmRQR (5'- TCTCAGCCTTGACTTGAGCA-3') primer pair designed to amplify a portion of the of ToCV CPm gene (Hammond et al. unpublished). A total of 165 sweet pepper samples were tested. From this total, seven produced amplification of PCR products with all three sets of primers. Except for one of the seven samples, which only showed mild chlorosis, they all showed severe chlorosis and different levels of upward leaf curling. None of the other samples showed amplification with any of the primer sets. Sequence analysis of the HSP PCR products produced using previously reported primers revealed 100% sequence identity with a tomato isolate of ToCV from the United States (GenBank Accession No. AY903448). ToCV was first reported infecting sweet pepper plants in Spain in 2004, follow by a report in Brazil in 2010 and now this is the first report of ToCV infecting sweet pepper plants in Costa Rica. If ToCV has now adapted to infect sweet pepper plants it is anticipated that many other reports will follow in the future. This virus has also been found infecting tomato and other weeds associated with greenhouses in the same region of Costa Rica, suggesting that is has adapted to the conditions of the area and represents an important threat to the vegetable production.