Development of tetra-primer amplicon refractory mutation system (T-ARMS) strategy for identification of the dark green mutant allele in tomato.

The high pigment (hp) tomato mutants increase the fruit pigment content in tomato. The dark green (dg) tomato mutant belonging to this category contains the hp-2 dg recessive allele, resulting from a single nucleotide polymorphism (SNP) in the DEETIOLATED1 (DET1) gene. Naturally, identification of this functional SNP is crucial for introgression of this important mutant allele in suitable tomato cultivar(s). Hence, we developed a functional co-dominant marker for detecting the hp-2 dg mutant allele using the tetra-primer amplicon refractory mutation system (T-ARMS) strategy. Although the other available techniques for identification of SNPs require specialized costly equipments and reagents, our method allows the reliable identification of this functional SNP in time-, labour- and cost-effective manner, using standard molecular biology equipments and reagents. To the best of our knowledge, this is the first report of T-ARMS strategy for the detection of the dg mutant allele in tomato, which will definitely help in identification and introgression of this allele by the breeders, working even under limited fund and resources.