Thirty-five-day-old tomato plants (cultivar Florasette) exhibited yellow leaf curling, stunting, and extremely reduced fruit set in spring 2001, in Guanica, Puerto Rico (PR). Twenty percent disease incidence was observed in this field and, 8 weeks later, 75% of the plants showed symptoms. These symptoms were distinct from those caused by other tomato-infecting begomoviruses reported previously from PR, namely Merremia mosaic virus, Tomato mottle virus (ToMoV), and Potato yellow mosaic virus (1). A colony of the B biotype of Bemisia tabaci (Genn.) was used to transmit the suspect virus from symptomatic plants collected in the field and established in the greenhouse in Rio Piedras, PR. The suspect virus was transmitted readily to tomato cultivar Roma (10 of 10 plants), and symptoms were like those observed in the field. Symptoms also were reminiscent of those described for several Old World begomoviruses, referred to as Tomato yellow leaf curl virus (TYLCV). Total nucleic acids were isolated from three symptomatic field samples and three greenhouse-inoculated tomato plants showing typical disease symptoms. Extracts were analyzed by polymerase chain reaction (PCR) with primers AV2466 and AC1145 to amplify a begomoviral fragment (approximately 1.1 bp) that contains a portion of the intergenic region and the viral coat protein gene (CP) (2). Amplicons were cloned, and their nucleotide sequences were determined. A comparison of CP with other well-studied begomoviral nucleotide sequences revealed that the CP sequences for field isolates 1 to 6 shared 99.7 to 100% identity with each other and 99.9 to 100% identity with TYLCV from Israel (TYLCV-IL; accession no. X76319) as well as TYLCV-IL isolates discovered in the Dominican Republic (DO; accession no. AF024715) and, subsequently, in Florida. TYLCV-specific PCR primers (forward) 5′-GAATTCCGCCTTTAA-TTTG-3′ and (reverse) 5′-GAATTCCCACTATCTTTCTC-3′ were used to amplify the complete viral genome form a PR field isolate. An expected-sized amplicon of approximately 2.8 kb was obtained, and the nucleotide sequence of two cloned amplicons was determined. Genome organization revealed a predicted precoat open reading frame of 351 bp, which is characteristic of other Old World begomoviruses, including TYLCV-IL. Nucleotide comparisons indicated that the PR isolate shared 99% nucleotide sequence identity with TYLCV-IL (first reported from Israel) and introduced TYLCV-IL isolates in DO and Florida, thereby confirming the introduction of TYLCV-IL into PR. TYLCV-IL was first identified several years ago in the Western Hemisphere, and the virus has been reported in five offshore locations and three continental U.S. states since its initial introduction into the DO in the early 1990s. Considering the extreme virulence of TYLCV-IL compared with most New World tomato-infecting begomoviruses, this introduction, which likely occurred from a nearby Caribbean country or Florida, has the potential to destroy the fresh-market tomato industry in PR, which supplies tomatoes to the continental United States during the winter months. There is compelling evidence for the routine movement of tomato seedlings from the continental United States to this location in PR throughout the last 10 years, including the previous introduction of ToMoV (1). These incidences and others indicate the need for those in infected areas to take precautions to avoid further spread of this highly damaging virus in and adjacent to the Caribbean region.
References: (1) A. M. Idris et al. Phytopathology 88:S42, 1998. (2) A. M. Idris and J. K. Brown, Phytopathology 88:648, 1998.
