First Report of Tomato chlorosis virus Infecting Sweet Pepper in Costa Rica

Abstract

In September 2008, a survey of whiteflies and whitefly-borne viruses was performed in 11 pepper-growing greenhouses in the province of Cartago, Costa Rica. During this survey, the vast majority of sweet pepper (Capsicum annuum cv. Nataly) plants showed interveinal chlorosis, enations, necrosis, and mild upward leaf curling. 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 with 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 ToCV in the samples: TocQ875F/TocQ998R primer set directed to a fragment of 123 bp of the HSP gene (3); ToCVp22RQF (5′-TGGATCTCACTGGTTGCTTG-3′)-ToCVp22RQR (5′-TAGTGTTTCAGCGCCAACAG-3′) primer pair that amplifies a 198-bp segment of the ToCV p22 gene (R. Hammond, E. Hernandez, J. Guevara, J. A. Vargas, A. Solorzano, R. Castro, N. Barboza, F. Mora, and P. Ramirez, unpublished) and the ToCVCPmRQF (5′-CATTGGTTGGGGATTACGTC-3′)-ToCVCPmRQR (5′-TCTCAGCCTTGACTTGAGCA-3′) primer pair designed to amplify a 170-bp portion of the ToCV CPm gene (R. Hammond, E. Hernandez, J. Guevara, J. A. Vargas, A. Solorzano, R. Castro, N. Barboza, F. Mora and P. Ramirez, unpublished). Fifteen symptomatic samples per greenhouse were tested for a total of 165 sweet pepper plants. From this total, seven samples from four different greenhouses produced amplification of PCR products with all three sets of primers. One of the seven samples showed mild chlorosis, but others were highly chlorotic with different levels of upward leaf curling. None of the other samples showed amplification with any of the primer sets; the symptoms on these plants could have been due to nutritional deficiencies or infection by viruses. Sequence analysis of the 460-bp HSP PCR products, produced using previously reported primers (3), and 150-bp fragment of the P22 revealed 100% sequence identity with a tomato isolate of ToCV from the United States (GenBank Accession No. AY903448). Because of the low number of samples infected with ToCV and the high incidence of symptoms, DNA extraction and a begomovirus PCR detection assay was performed using primer pair AV494/AC1048 (4). Negative results were obtained for all samples. To our knowledge, this is the first report of ToCV infecting sweet pepper plants in Costa Rica and the third one worldwide. ToCV has also been found to be infecting tomato in open field and greenhouses (1) and other weeds in greenhouses including Ruta chalepensis (Rutaceae), Phytolacca icosandra (Phytolaccaceae), Plantago major (Plantaginaceae), and Brassica sp. (Brassicaceae) (2) in the same region of Costa Rica, suggesting that it has adapted to the conditions of the area and poses an important threat to the vegetable production. References: (1) R. M. Castro et al. Plant Dis. 93:970, 2009. (2) A. Solorzano-Morales et al. Plant Dis. 95:497, 2011. (3) W. M. Wintermantel et al. Phytopathology 98:1340, 2008. (4) S. Wyatt and J. Brown. Phytopathology 86:1288, 1996.

Detección de crinivirus y begomovirus en plántulas de tomate y arvenses asociadas a semilleros

“…Castro et al, 2009;Nakhla et al, 1994;Barboza et al, 2014Solanum americanum ToCV -Vargas-Asencio et al, 2013 Solanum quitoense ToCV - Vargas-Asencio et al, 2013 Solanum nigrum ToCV - Vargas-Asencio et al, 2013 Capsicum annum ToCV MCLCuV-CR, PepGMV Lotrakul et al, 2000;Vargas et al, 2011 ToCV: virus de la clorosis del tomate, MCLCuV-CR: virus de la clorosis y del enrrollamiento de la hoja de melón-Costa Rica, SPCSV: virus del trastorno clorótico-enano del camote, SPLCV: virus del enrollamiento de la hoja del camote, BPYV: virus del amarillamiento falso de la remolacha, PepGMV: virus del mosaico dorado del chile, EuMV: virus del mosaico de la Euphorbia, BGMV: virus del mosaico dorado del frijol, BGYMV: virus del mosaico dorado amarillo del frijol, CalGMV: virus del mosaico dorado del Calopogonium; SiGMCRV: virus de Costa Rica del mosaico dorado de la Sida, TYMoV: virus del moteado amarillo del tomate, ToLCSinV: virus del enrrollamiento de la hoja de tomate de Sinaloa, TYLCV: virus del amarillamiento y enrollamiento de la hoja de tomate / ToCV: tomato chlorosis virus, MCLCuV-CR: melon chlorotic leaf curl virus-Costa Rica, SPCSV: sweet potato chlorotic stunt virus, SPLCV: sweet potato leaf curl virus, BPYV: beet pseudo-yellows virus, PepGMV: pepper golden mosaic virus, EuMV: Euphorbia mosaic virus, BGMV: bean golden mosaic virus, BGYMV: bean golden yellow mosaic virus, CalGMV: Calopogonium golden mosaic virus, SiGMCRV: sida golden mosaic Costa Rica virus, TYMoV: tomato yellow mottle virus, ToLCSinV: tomato leaf curl Sinaloa virus, TYLCV: tomato yellow leaf curl virus.…”

Advanced loop-mediated isothermal amplification method for sensitive and specific detection of Tomato chlorosis virus using a uracil DNA glycosylase to control carry-over contamination

“…ToCV has a bipartite genome of positive-sense single-stranded RNAs (RNA1 and RNA2) consisting of four and nine ORFs, respectively, that encode functional proteins (Lozano et al, 2006;Wintermantel and Wisler, 2006). In addition to tomato, many other cultivated crops such as sweet pepper (Capsicum annuum), potato (Solanum tuberosum), and zinnia (Zinnia elegans) have been identified as natural ToCV hosts (L. esculentum) (Lozano et al, 2004;Tsai et al, 2004;Barbosa et al, 2010;Vargas et al, 2011;Fortes and Navas-Castillo, 2012), and approximately 25 species of weeds are also ToCV hosts (Font et al, 2004;Segev et al, 2004;Morris et al, 2006;Wintermantel and Wisler, 2006;Alvarez-Ruiz et al, 2007;Trenado et al, 2007;Solórzano-Morales et al, 2011). ToCV has been reported in many countries throughout Europe, the Middle East, the Americas, and Africa since the first report was published by US researchers in 1998 (Accotto et al, 2001;Font et al, 2004;Segev et al, 2004;Dalmon et al, 2005;Alvarez-Ruiz et al, 2007;Barbosa et al, 2008;Fiallo-Olivé et al, 2011).…”

Identification of natural weed hosts of Tomato chlorosis virus in Korea by RT-PCR with root tissues

“…Approximately 36 species of plants have also been identified as ToCV hosts (Font et al 2004;Segev et al 2004;Lozano et al 2004;Tsai et al 2004;Morris et al 2006;Wintermantel and Wisler 2006;Alvarez-Ruiz et al 2007;Trenado et al 2007; Barbosa et al 2010;Solórzano-Morales et al 2011;Vargas et al 2011;Fortes and Navas-Castillo 2012). Most of the identified ToCV weed hosts are in the family Solanaceae, and Solanum nigrum, Physalis ixocarpa and P. wrightii were confirmed to be ToCV hosts in two independent studies (Font et al 2004;Segev et al 2004;Wintermantel and Wisler 2006;Trenado et al 2007).…”

“…ToCV was originally reported in tomato (S. lycopersicum), but the ToCV has been identified to infect a broad range of hosts as new hosts such as some cultivars of potato (Solanum tuberosum), sweet pepper (Capsicum annuum) and zinnia (Zinnia elegans) (Lozano et al 2004;Tsai et al 2004;Barbosa et al 2010;Vargas et al 2011;Fortes and Navas-Castillo 2012;Freitas et al 2012). Approximately 36 species of plants have also been identified as ToCV hosts (Font et al 2004;Segev et al 2004;Lozano et al 2004;Tsai et al 2004;Morris et al 2006;Wintermantel and Wisler 2006;Alvarez-Ruiz et al 2007;Trenado et al 2007; Barbosa et al 2010;Solórzano-Morales et al 2011;Vargas et al 2011;Fortes and Navas-Castillo 2012).…”

“…These positive results might reflect (Trenado et al 2007;Kataya et al 2008;Hirota et al 2010); however, our current observations suggest that shoot tissues are not good materials for the detection of viruses that are transmitted by insect vectors. According to previous studies on natural or experimental hosts of ToCV, 19 species (Capsicum annuum, Cyphomandra betacea, Datura stramonium, Nicotiana benthamiana, N. clevelandii, N. edwardsoni, N. glutinosa, N. megalosiphon, N. tabacum, Petunia hybrida, Physalis alkekengi, P. ixocarpa, P. peruviana, P. wrightii, Solanum acaule, S. lycopersicum, S. nigrescens, S. nigrum and S. tuberosum) out of 36 ToCV hosts belong to the Solanaceae family (Wisler et al 1998b;Wintermantel et al 2001;Font et al 2004;Lozano et al 2004;Segev et al 2004;Wintermantel and Wisler 2006;Alvarez-Ruiz et al 2007;Trenado et al 2007; Barbosa et al 2010;Vargas et al 2011). Eight Solanaceae species were previously confirmed as ToCV hosts in two or more references, but Datura stramonium showed different results in data from two research groups (Wintermantel and Wisler 2006;Alvarez-Ruiz et al 2007).…”