TITLE:
The Use of Sequence-Based SSR Mining for the Development of a Vast Collection of Microsatellites in Aquilegia formosa
AUTHORS:
Brandon Schlautman, Vera Pfeiffer, Juan Zalapa, Johanne Brunet
KEYWORDS:
Aquilegia formosa, Microsatellites, SSR Mining
JOURNAL NAME:
American Journal of Plant Sciences,
Vol.5 No.15,
July
16,
2014
ABSTRACT: Numerous
microsatellite markers were developed forAquilegia
formosafrom sequences deposited within the Expressed Sequence Tag (EST),
Genomic Survey Sequence (GSS), and Nucleotide databases in NCBI. Microsatellites
(SSRs) were identified and primers were designed for 9 SSR containing sequences
in the Nucleotide database, 3803 sequences in the EST database, and 2226 sequences
in the GSS database. For validation purposes, 45 primer pairs were used to
amplify DNA from 16A. formosaindividuals
from the H. J. Andrews experimental forest in Oregon, a Long Term Ecological
Research (LTER) site. Genetic polymorphisms were identified at 30 of the 45
microsatellite loci with an average of 13.2 alleles per locus, and the observed
level of heterozygosity was greater than 0.8 for 21 of the 30 loci. The use of
these polymorphic loci was sufficient to individually separate the 16
individuals using a principal coordinate analysis. This comprehensive collection
of primers significantly increased the availability of microsatellite primers
forAquilegiaspp. and provided ample
material for future studies that required highly variable SSRs such as mapping
and association studies and investigation of plant mating system and gene flow.