Microsatellite variability and heterozygote deficiency in the arctic–alpine Alaskan wheatgrass (Elymus alaskanus) complex

Genetic variation in the allotetraploid grass Elymus alaskanus complex was assessed using microsatellites in seven populations from Canada, Greenland, and the U.S.A. Microsatellite variation was compared with allozyme and RAPD variation. Our results indicated that E. alaskanus was highly homozygous but also highly variable. The polymorphic loci ranged from 50 to 100% with a mean of 78.6%, and the mean number of allele per locus was 3.14. Average expected heterozygosity value (HE, gene diversity) varied across populations and ranged from 0.244 to 0.651 with mean of 0.414. The mean value of HE across Canadian populations (0.517) was significantly higher than that across populations in Greenland (0.367). The correlation between allozyme and microsatellite gene diversity value (HE) showed a high positive correlation (r = 0.68), but between RAPD and microsatellite showed a low positive correlation (r = 0.08). Populations were highly differentiated, with 38% of variation among populations. Interpopulation genetic distance showed no association with geographic distance between the population sites of origin. A Hardy-Weinberg exact test for all loci and all populations reveals a significant heterozygote deficiency. Possible explanations for heterozygote deficiency are discussed.Key words: Elymus alaskanus, microsatellites, heterozygote deficiency, genetic differentiation, variability.

Characterization of microsatellite loci from Elymus alaskanus and length polymorphism in several Elymus species (Triticeae: Poaceae)

A size-selected genomic library from Elymus alaskanus was screened for the presence of (GA)n, (GT)n, (CAC)n, and (TCT)n microsatellite sequences. A total of 28 positive clones were found for the two dinucleotide repeats, whereas no positive clones were found for the trinucleotide repeats. Positive clones were sequenced to validate the presence of microsatellites and to generate polymerase chain reaction (PCR) primers, based on the sequences flanking the microsatellite. Primer pairs were designed and evaluated for 18 selected microsatellites. The resulting loci were analysed by PCR for their usefulness as molecular markers in an array of 18 accessions representing E. alaskanus and 10 other Elymus species. PCR amplification revealed alleles for the 18 loci, which varied in having 1-10 alleles in these accessions. In the 18 accessions tested, 7 of the 18 loci were polymorphic, with gene diversity values ranging from 0.54 to 0.80 among all species. Within E. alaskanus, gene diversity values ranged from 0.20 to 0.72, with a mean of 0.48. Polymorphism was also detected within accessions. No clear relationship between total repeat length and the degree of polymorphism was observed in this study. Primer pairs designed to amplify microsatellites in E. alaskanus can be used to generate polymorphism products in other species within the genus. Hence, microsatellites are useful markers for studying both inter- and intra-specific genetic variability within Elymus.Key words: Elymus alaskanus, Triticeae, microsatellites, simple sequence repeats, SSRs, polymorphism.