Development of PCR-based markers from the tomato glutamate oxaloacetate transaminase isozyme gene family as a means of revitalising old isozyme markers and recruiting new ones

Abstract

Sequences annotated as aspartate aminotransferases (synonymous with glutamate oxaloacetate transaminases) in the SOL Genomics Network unigene database were used to design 10 pairs of PCR primers for genetic marker development. These primer pairs generated nine CAPS markers, two SCAR markers and one SSR marker, which were bin-mapped using a set of tomato introgression lines (IL) derived from Lycopersicon esculentum cv. M82 and Lycopersicon pennellii LA716. Based on their bin locations, these markers are largely dispersed throughout the tomato genome and appear to have tagged all four of the glutamate oxaloacetate transaminase (Got) isozyme marker genes placed on the classical genetic map of tomato. Orthologous relationships with Arabidopsis aspartate aminotransferase (Asp) genes suggest the existence of at least two additional functional Got genes in tomato that have also been tagged by these markers and likewise an additional functional Asp gene in Arabidopsis. The Got-2 isozyme marker has often been used for the marker-assisted breeding of the I-3 gene for Fusarium wilt resistance introgressed from L. pennellii LA716. The Got-2 CAPS marker that we have developed offers a facile PCR-based alternative to the isozyme marker for the marker-assisted breeding of I-3. However, all of the PCR-based markers we have developed have the potential to assist the breeding of linked traits introgressed from wild relatives of tomato.