Conservation of microsatellite flanking sequences in different taxa of Leguminosae

Abstract

To test if locus-specific microsatellite markers designed for one genus are informative when used with related genera, the conservation of microsatellite-flanking intergeneric primer binding sites was tested in the closely related tribes Vicieae and Cicereae, from the subfamily Papilionoideae of the Leguminosae family. A total of 123 sequence-tagged microsatellite sites (STMS) markers derived from chickpea were used to amplify loci in lentil (Lens) and dry pea (Pisum). The percentage of chickpea primer binding sites conserved between the three genera was 54.4%. Hybridisation of 63 selected amplified loci to the digoxigenin-labelled oligonucleotide probe (TAA)5 showed that 69.8% of loci from dry pea and 66.6% of loci from lentil hybridised to the probe. Sequencing of amplified products from chickpea with the primer Ta176 demonstrated that one amplicon contained a microsatellite, whereas another amplicon amplified with the same particular STMS primer pair did not. Amplicons produced from lentil and pea with this primer pairs did not contain microsatellite sequences. Results obtained with Tr7, which amplified a PCR product in lentil and chickpea but not in pea, showed that microsatellite sequences were present in chickpea and absent in lentil. Similar results were obtained with Ts35, which produces amplicons in pea and chickpea; but, again, microsatellite sequences were only present in chickpea. We therefore conclude that STMS derived from chickpea could be used to detect variability between other Leguminosae genera, but it is necessary to verify whether homologous loci are revealed.