Detection and Isolation of Microsatellites in Pistachio (Pistachia vera L.) for Evaluation of Iranian Varieties

Microsatellite repeats or simple sequence repeats ( SSRs) have become increasingly popular as a genetic marker in many plant species. the discovery and development of new microsatellite is a costly and technically demanding process. However, the major reason for the success of microsatellites as molecular tools is because they provide a higher incidence of detectable polymorphisms than other techniques such as RFLPs and RAPDs. The objective of the present study was to construct Pistachio microsatellite loci and evaluate their levels of polymorphisms in Pistachio genotypes. The recent development of new microsatellite enrichment techniques has shown ever increased efficiency of microsatellite characterization in species in which little or no previous sequence knowledge is needed. A genomic DNA library enriched for (AG) n , (GT) n and (ATT) n as a microsatellite motives has been developed from "Kale-Ghochi" Pistachio. The enrichment level for library was 45-60% and 35% of clones (5 out of 14) contained microsatellite sequences. Five primer pairs were designed based on flanking regions using the oligo software. Among the primer sets three pairs amplified cleanly and produced an easily interpretable PCR product in the pistachio genotypes and P. mutica (Baneh)

Keywords:

Pistachio , Microsatellite , Simple Sequence Repeat (SSR) , Enrichment