Molecular Characterization of a New Begomovirus Associated with Tomato Yellow Leaf Curl and Eggplant Yellow Mosaic Diseases in Thailand

Abstract

Leaf curl and yellowing symptoms on tomato, and yellow mosaic symptoms on eggplant, are frequently observed in Kanchanaburi Province, Thailand. DNA was extracted from leaves of 13 symptomatic tomato and six symptomatic eggplant samples by the method of Gilbertson et al (1). Viral DNA was amplified by polymerase chain reaction (PCR) using the begomovirus-specific degenerate primer pair PAL1v1978/PAR1c715 (3), which amplified a 1.4-kb fragment of DNA-A. All samples, except one eggplant sample, yielded the expected product. The 1.4-kb PCR products of one tomato and one eggplant sample were cloned and sequenced. Both begomoviruses from tomato and eggplant contained a DNA-B component, amplified using two degenerate primer pairs, DNABLC1/DNABLV2 and DNABLC2/DNABLV2 (2). Based on sequences of the DNA products amplified by the degenerate primer pairs, specific primers were designed to complete the DNA-A and DNA-B sequences. Computer-assisted sequence comparisons were performed with geminivirus sequences available in the laboratory at the Asian Vegetable Research and Development Center, Taiwan and in the GenBank sequence database. Both tomato (GenBank Accession Nos. AF511529 and AF511528) and eggplant (GenBank Accession Nos. AF511530 and AF511527) virus isolates contain the conserved geminivirus sequence-TAATATTAC on the DNA-A and B. Based on the high sequence identities of 99% DNA-A and 98% DNA-B, these two virus isolates are considered to be the same species. Although the genome organization of these two isolates was the same as Tomato yellow leaf curl virus Thailand (TYLCTHV; GenBank Accession Nos. X63015 and X63016), including six open reading frames (ORFs) on the DNA-A (AV1, AV2, AC1, AC2, AC3, and AC4) and two ORFs on the DNA-B (BV1 and BC1), sequence comparisons showed highest DNA-A sequence identity (73%) with Ageratum yellow vein virus from Singapore (GenBank Accession No. X74516) and highest DNA-B identity (77%) with the TYLCTHV (X63016). To our knowledge, these tomato- and eggplant-infecting viruses from Thailand constitute a distinctly novel bipartite Begomovirus species. References: (1) R. L. Gilbertson et al. J. Gen. Virol. 72:2843, 1991. (2) S. K. Green et al. Plant Dis. 85:1286, 2001. (3) M. R. Rojas et al. Plant Dis. 77:340, 1993.