Development Of SSR Markers Research Articles

Development and characterization of microsatellite markers for genetic analysis of a Brazilian endangered tree species Caryocar brasiliense

In this work we report the development and characterization of 10 microsatellite loci for the endangered tree species Caryocar brasiliense. Using genomic library enrichment, the efficiency of SSR marker development was 14.4% from sequencing data to operationally useful loci. Primer sequences for this set of 10 loci are made available together with their estimates of expected heterozygosity, probability of paternity exclusion and probability of identity. Mendelian inheritance and segregation was confirmed for all 10 loci in open-pollinated half-sib families as well as the absolute transferability of these 10 loci to five other species of the same genus. Number of alleles per locus ranged from 10 to 22 with a mean value of 16 and expected heterozygosity varying from 0.84 to 0.94. The combined probability of genetic identity was on the order of 10–17 clearly demonstrating that SSR multilocus genotypes are likely to be unique and capable of readily discriminating individuals of C. brasiliense. The very high combined probability of paternity exclusion (0.99999995) also indicates that these markers will permit detailed parentage studies in natural populations even in situations where both maternity and paternity are unknown. The battery of microsatellite markers developed and characterized in this study opens a new perspective for the generation of fundamental population genetic data for devising sound collection and conservation procedures for C. brasiliense and related species of the genus.

Occurrence of microsatellites in spinach sequences from computer databases and development of polymorphic SSR markers

AbstractMicrosatellites are valuable tools as molecular markers in plant breeding. To establish genetic linkage maps or for population studies, information about the occurrence and usability of microsatellite markers in different species is necessary. Sequences of spinach Spinacia oleracea from computer databases were therefore searched for the presence of microsatellites. Sixty simple sequence repeats were found in 237 spinach sequences with a total of 349.4 kb DNA. After removing duplicated sequences, 50 different microsatellites with various motifs remained. Differences between nuclear and chloroplast DNA were not in the number of microsatellites but in their type and length. Chloroplast sequences from spinach contain only short strings of A and AT repeats, whereas nuclear sequences show a wider variety of motifs.Flanking primers for polymerase chain reaction (PCR) analysis were designed for 13 of these microsatellites and tested with two different varieties of spinach. Twelve primer pairs gave amplification products and seven of these showed polymorphisms in the variety ‘Wiremona’ but only one in the variety ‘Monatol’. These markers may be used for linkage analysis or population studies in spinach.