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Development of a locus-specific,co-dominant SCAR marker for assisted-selection of the Sw-5 (Tospovirus resistance) gene cluster in a wide range of tomato accessions
Authors:Érico C. Dianese  Maria Esther N. de Fonseca  Rob Goldbach  Richard Kormelink  Alice K. Inoue-Nagata  Renato O. Resende  Leonardo S. Boiteux
Affiliation:1.National Center for Vegetable Crops Research (CNPH),Embrapa Vegetable Crops,Brasília,Brazil;2.Phytopathology Department,University of Brasilia (UnB),Brasília,Brazil;3.Laboratory of Virology,Wageningen University,Wageningen,The Netherlands
Abstract:The best levels of broad-spectrum Tospovirus resistance reported in tomatoes thus far are conferred by the Sw-5 locus. This locus contains at least five paralogues (denoted Sw-5a through Sw-5e), of which Sw-5b represents the actual resistance gene. Here we evaluated a panel of seven PCR primer pairs matching different sequences within a genomic region spanning the Sw-5a and Sw-5b gene cluster. Primer efficiency evaluation was done employing tomato isolines with and without the Sw-5 locus. One primer pair produced a single and co-dominant polymorphism between susceptible and resistant isolines. Sequence analysis of these amplicons indicated that they were specific for the Sw-5 locus and their differences were due to insertions/deletions. The polymorphic SCAR amplicon encompass a conserved sequence of the promoter region of the functional Sw-5b gene, being located in the position −31 from its open reading frame. This primer pair was also evaluated in field assays and with a collection of accessions known to be either susceptible or resistant to tospoviruses. An almost complete correlation was found between resistance under greenhouse/field conditions and the presence of the marker. Therefore, this primer pair is a very useful tool in marker-assisted selection systems in a large range of tomato accessions.
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