Tomato-infecting begomoviruses have been reported throughout Brazil since the introduction of the B biotype of Bemisia tabaci. Here, we report a large scale survey on the distribution and genetic diversity of tomato-infecting begomoviruses. Tomato samples with typical begomovirus symptoms were collected in seven different states, comprising the major tomato growing areas of the country. Viruses were detected by polymerase chain reaction (PCR) using universal primers for the genus Begomovirus. PCR-amplified fragments were cloned and sequenced. Based on sequence comparisons and phylogenetic analyses, at least seven previously undescribed species of begomoviruses were found. Four of the new viruses were found exclusively in the Southeastern states, two exclusively in the Northeastern states, and one was found in both regions. Sequence comparisons reveal strong evidence of recombination among the Brazilian begomoviruses. Together, the results indicate the existence of a high degree of pre-existing genetic diversity among tomato-infecting begomoviruses in Brazil and suggest that these viruses have emerged after being transferred from natural hosts to tomatoes, due to the introduction into Brazil of a novel polyfagous biotype of the whitefly vector.

A viral complex causing golden mosaic and leaf distortion (rugosity) in tomato plants was obtained from viruliferous whiteflies, and named TGV-Ub1. This complex was sap-transmitted from tomato to Nicotiana benthamiana . PCR amplification using universal begomovirus primers yielded two distinct fragments for DNA-A, suggesting that the TGV-Ub1 complex comprised at least two distinct viruses. Clones corresponding to full-length viral genomes were obtained from tomato plants infected with TGV-Ub1. Comparisons of the complete sequences of clones pUb1-49 (DNA-A), pUb1-62 and pUb1-81 (both DNA-B) indicated that they constitute novel western hemisphere begomoviruses. Clones pUb1-49 and pUB1-81 have identical common regions, thus representing the cognate DNA-A and -B of a novel begomovirus, named Tomato rugose mosaic virus (ToRMV). Clone pUb1-62 has a distinct common region from ToRMV and all other geminiviruses. A cognate DNA-A for pUb1-62 was not found. Clones containing 1·8 copies of the genomic components were constructed. Infectivity assays of these clones in tomato and N. benthamiana demonstrated that the clones corresponding to ToRMV systemically infected both hosts. Symptoms were analogous to those observed when using the pure isolates obtained in this study. The combination of pUb1-49 and -62 did not result in systemic infection, indicating that these components do not form a viable virus. ToRMV was sap-transmitted from N. benthamiana to N. benthamiana , and by grafting to Solanum tuberosum and Datura stramonium . ToRMV-A and ToRMV-B were detected in plants of Nicandra physaloides and Phaseolus vulgaris , respectively, growing in nearby tomato fields, in association with distinct DNA components.

Although tomato golden mosaic virus (TGMV) was reported in Brazil more than 20 years ago (3), tomato-infecting geminiviruses have not been of economic significance in the country until recently. However, a sharp increase in the incidence of geminivirus-like symptoms in tomatoes has been reported in several areas of Brazil since 1994. This has coincided with the appearance of the B biotype of Bemisia tabaci, which, as opposed to the A biotype, readily colonizes solanaceous plants (2). We have isolated geminiviruses from symptomatic tomato plants in the Federal District, in two different areas of the state of Minas Gerais, and in the state of Pernambuco. Tomato plants in these areas showed a variety of symptoms, including yellow mosaic, severe leaf distortion, down-cupping, and epinasty. Whitefly infestation was high in all fields sampled, and in some fields, particularly in Pernambuco, incidence of virus-like symptoms was close to 100%, and no tomatoes of commercial value were harvested (1). Using primer pairs PAL1v1978/PAR1c496 and PCRc1/PBL1v2040 (4), DNA-A and -B fragments were polymerase chain reaction (PCR)-amplified from total DNA extracted from diseased plants, cloned, and sequenced. Sequence comparisons of the PCR fragments indicated the existence of at least six different geminiviruses. The nucleotide sequence homologies for DNA-A fragments ranged from 67 to 80% for the 5′ end of the cp gene, and from 44 to 80% for the 5′ end of the rep gene. Data base comparisons indicated the viruses are most closely related to TGMV, bean golden mosaic virus from Brazil (BGMV-Br), and tomato yellow vein streak virus (ToYVSV), although homologies were less than 80% for the fragments compared. A similar lack of a close relationship with each other and other geminiviruses was obtained with two DNA-B component PCR products compared, corresponding to the 5′ end of the BC1 open reading frame. Infectious, full-length genomic clones from the tomato viruses are being generated for biological and molecular characterization. References: (1) I. C. Bezerra et al. Fitopatol. Bras. 22:331, 1997. (2) F. H. França et al., Ann. Soc. Entomol. Bras. 25:369, 1996. (3) J. C. Matyis et al. Summa Phytopathol. 1:267, 1975. (4) M. R. Rojas et al. Plant Dis. 77:340, 1993.

A mancha-angular (Phaeoisariopsis griseola) e a ferrugem (Uromyces appendiculatus) são algumas das mais prejudiciais doenças do feijoeiro na região do Triângulo Mineiro, Minas Gerais. O objetivo deste estudo foi determinar a influência das semeaduras sucessivas na severidade da mancha-angular e da ferrugem, bem como as perdas na produção em decorrência dessas doenças. Os experimentos foram conduzidos em condições de campo, nas safras outono-inverno (1994) e das águas (1994/95), cada uma com três semeaduras sucessivas de feijão. A severidade da mancha-angular e da ferrugem aumentou significativamente da primeira para a terceira semeadura, atingindo nível mais elevado na última. As semeaduras anteriores foram a fonte de inóculo para os cultivos subseqüentes. Aplicações de fungicida foram eficientes no controle das doenças, permitindo um aumento significativo da produção de grãos, quando comparado com o controle. A mancha-angular apresentou maior incidência no outono-inverno e nas águas, e a ferrugem, apenas no outono-inverno. Nessas safras as condições climáticas (temperatura e umidade relativa) favoreceram a evolução das doenças, resultando num aumento da severidade. Houve uma correlação negativa e altamente significativa entre a severidade da mancha-angular e da ferrugem com a produção de grãos nas duas safras.