Identification of genotypes and in some cases species in Salvia L. is complicated due to the morphologic similarities. This study utilized a touch-down directed amplification of minisatellite DNA polymerase chain reaction (Td-DAMD-PCR) technique to develop species-specific Td-DAMD-PCR markers for Salvia L. A total of 22 minisatellite core sequences as primers, and individuals and bulked samples from five species of Salvia L. along with one Origanum L. and one Sideritis L. species were used. A total of 70 species-specific markers differentiating each species utilized were determined. Td-DAMD-PCR markers were obtained at higher annealing temperature and were reproducible. These markers could be used as diagnostic markers for identification of Salvia L. species. Utilization of Td-DAMD-PCR markers will be useful in germplasm characterization, plant genetic resources conservation and utilization inSalvia L. Furthermore integration of these DNA markers and analytical methods in Salvia L. improvement programs will lead to the development of a comprehensive system which can be conveniently applied at the industry level.
Key words: Directed amplification of minisatellite (DNA) markers, minisatellites, species identification, touch-down polymerase chain reaction (PCR).
AFLP, Amplified fragment length polymorphism; DAMD-PCR, directed amplification of minisatellite DNA region polymerase chain reaction; PCR, polymerase chain reactions; RAPD, random amplified polymorphic DNA; RFLP, restriction fragment length polymorphism; SSR, simple sequence repeat; TBE, TRIS-borate EDTA; Td-PCRs, touch-down polymerase chain reactions; BSA, bulked segregant analysis; TLC, thin layer chromatography; HPLC, high performance liquid chromatography.
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