Simultaneous detection of tomato resistance factors to Fusarium wilt in tomato by multiplex PCR

Abstract

The objective of this work was to develop and validate a protocol for simultaneous detection by multiplex polymerase chain reaction (multiplex PCR) of tomato (Solanum lycopersicum) genomic regions associated with factors of resistance to three physiological races of Fusarium oxysporum f. sp. lycopersici (FOL). The employed primer pairs were SSR‑67 (specific to the I‑1 gene), TFusrr (specific to the I‑2 gene), and SSRD (specific to the I‑3 gene). Genotyping results with molecular markers were compared with the phenotyping ones of a tomato germplasm collection, in bioassays of isolate inoculation of the three FOL races in seedlings by root dipping. The resistance or susceptibility was confirmed by PCR, through the visualization of specific amplicons corresponding to the target regions linked to the factors of resistance to distinct FOL races. The elaborated protocol for the joint use of the molecular markers, by multiplex PCR, allows of the selection of tomato accessions that are resistant to the races 1, 2, and 3 of F. oxysporum f. sp. lycopersici in a similar way to that done with each one separately. PCR multiplex is a viable tool to monitor the incorporation of these resistance factors into tomato inbred lines.