Olachea C.A, Agua Viva, Cabudare, Estado Lara, ZIP 3023, Venezuela. K. Zambrano. Universidad Centro-Occidental Lisandro Alvarado, Estado Lara, ZIP 3023, ...
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Identification of Tomato spotted wilt virus associated with fruit damage during a recent virus outbreak in pepper in Venezuela Y. Pérez-Colmenares, A. Mejías, E. Rodríguez-Román, D. Avilán, J.C Gómez, and E. Marys
Plant Disease "First Look" paper • http://dx.doi.org/10.1094/PDIS-11-14-1202-PDN • posted 01/21/2015 This paper has been peer reviewed and accepted for publication but has not yet been copyedited or proofread. The final published version may differ.
Instituto Venezolano de Investigaciones Científicas (IVIC), Centro de Microbiología y Biología Celular, ZIP 20632, Caracas 1020-A, Venezuela. J. E. Olachea Olachea C.A, Agua Viva, Cabudare, Estado Lara, ZIP 3023, Venezuela K. Zambrano Universidad Centro-Occidental Lisandro Alvarado, Estado Lara, ZIP 3023, Venezuela In August 2014, several diseased pepper (Capsicum annuum L.) plants showing severe stunting, necrotic spots and leaf deformation were noticed in commercial pepper fields located in Tintorero, Lara State, Venezuela. Affected plants produced deformed unmarketable fruits with striking chlorotic and necrotic ringspots and irregular coloring. On the basis of these symptoms, infection with Tospovirus was suspected. To confirm this, RT-PCR using universal tospovirus primers L1 (5’AATTGCCTTGCAACCAATTC-3’) and L2 (5'-ATCAGTCGAAATGGTCGGCA-3’) which amplify part of the L RNA within the open reading frame of the RNA polymerase gene (Chatzivassiliou et al. 2000) was performed on total RNA extracted from symptomatic leaf tissue with the TRIZOL Reagent (Invitrogen, Carlsbad, CA). The reaction from 22 diseased plants yielded amplicons of the expected size (~276). Two amplicons were directly sequenced in both orientations and the sequences were deposited in Genbank (Accession Nos. KP006412 and KP006413). BLAST analysis (National Center for Biotechnology Information database) showed that the fragment sequences shared best nucleotide sequence identity (99%) to Tomato spotted wilt virus (TSWV) isolates deposited in GenBank from different hosts from Russia (JX452818.1), Italy (KJ575619.1), South Korea (KC261947.1), Venezuela (KC202213.1) and Iran (KC767959.1). Further TSWV identity was confirmed in symptomatic plants by RT-PCR assays with primers CP1-TSWV (TTAACTTACAGCTGCTTT) and CP2-TSWV (CAAAGCATATAAGAACTT) for the amplification of the TSWV coat protein. A single DNA product of ~823 bp was amplified from all samples. A search of GenBank revealed a sequence identity of 95 to 99% with the coat protein gene of known TSWV isolates. Pepper plants were inoculated mechanically with sap from symptomatic pepper from field. Foliage developed mild bronzing and leaf deformation, and the presence of TSWV was confirmed using RT-PCR. To our knowledge, this is the first report of TSWV infecting pepper in Venezuela. Considering its broad range, this emerging virus could pose a significant threat to the Venezuelan vegetable industry. References: Chatzivassiliou et al. Plant. Dis. 137:127, 2000.
Plant Disease "First Look" paper • http://dx.doi.org/10.1094/PDIS-11-14-1202-PDN • posted 01/21/2015 This paper has been peer reviewed and accepted for publication but has not yet been copyedited or proofread. The final published version may differ.
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Ringspots on pepper fruit associated with TSWV during virus outbreak in Venezuela 863x1151mm (72 x 72 DPI)
