EvaluatingWheat Microsatellite Markers for the Use in Genetic Analysis of inopyrum , Dasypyrum , and Pseudoroegneria Species

A set of 42 SSRs of wheat were evaluated for their cross-ampli�cation on the DNA of inopyrum ponticum, inopyrum intermedium,inopyrum elongatum,inopyrum bessarabicum, Pseudoroegneria stipifolia, andDasypyrum villosum.e number of thewheat SSRmarkers that ampli�edDNA fragmentswith determined size for.ponticumwas 33 (78.6%); for. intermedium, 28 (66.7%); for . elongatum, 24 (57.1%); for . bessarabicum, 24 (57.1%); for P. stipifolia, 26 (69.1%); and for D. villosum, 29 (69.0%). �wenty-four primer pairs of wheat SSR markers were successfully ampli�ed from all investigated species. e dataset can be used for phylogenetic studies of wild relatives of wheat, for the estimation of their diversity, and for the introgression of agronomically valuable genes into wheat genome.


Introduction
Wheat is a major food source for most world population.However, its cultivation is strictly limited by such challenges as pests, diseases, droughts, extreme temperatures, and others.In order to manage sustainable wheat production, it is necessary to develop new cultivars of wheat resistant and tolerant to adverse environmental factors.e genetic diversity of wheat is restricted due to the bottleneck effect which occurred during domestication and, on the other hand, due to the limited diversity of wheat germplasm used in the breeding programs [1,2].e genetic variability of cultivated wheat can be improved by wide hybridization involving wild relative species.One of the important gene pools is different species of inopyrum, Dasypyrum, and Pseudoroegneria genera [3,4].ese genetic resources can be efficiently used in wheat breeding by application of molecular microsatellite markers.Microsatellites also known as Simple Sequence Repeats (SSRs) are tandem repeats of two to six nucleotide DNA motifs randomly distributed all over genome.ey have been successfully used as DNA markers to estimate genetic diversity, genome mapping, and marker-assisted selection for agronomically important traits [5].SSR markers are a powerful tool for the investigation of cereal genomes because of the high level of polymorphism they reveal [6].In addition, SSRs can be used in wide hybridization of wheat to monitor and map desirable alien genes in segregating populations as they are locus speci�c and inherited in codominant manner [7].
A large number of the primer sequences of wheat SSRs are publicly available.However, the development of new SSR markers de novo for wild relatives of wheat is a time-and costconsuming process.e �anking regions of microsatellites show sufficient homology between closely related species.us, the primers of wheat SSR markers can be used in the analysis of other Poaceae species.is is called "transferability" and it has been successfully used for cultivated (barley, oats, and rye) [8] and wild (Elymus, Aegilops) species [9,10].e goal of our study was to test a set of 42 microsatellite markers of wheat for their ability to cross-amplify DNA from the accessions of . intermedium, . elongatum, . ponticum, . bessarabicum, P. stipifolia, and D. villosum.
Polymerase chain reaction (PCR) and fragment analysis were performed as follows.PCR was performed in �nal volume of 25 L in thermal cycler using the conditions recommended at GrainGenes 2.0 [11] according to the protocol by Schuelke [12].e �rst round of PCR was conducted using reverse (0.16 pmol/L) and forward primer with M13 tail (0.04 pmol/L).During the second round, the �uorescent-labeled M13 primer was incorporated into the PCR product at annealing temperature 53 ∘ C. Fragment analysis of PCR product was carried out using ABI 3130xl Genetic Analyzer (Applied Biosystems, Foster City, CA, USA) with GeneScan 500 LIZ size standard according to the manufacturer's recommendations.e sizes of fragments were analyzed using Peak Scanner Soware v1.0 (Applied Biosystems, Foster City, CA, USA).

Dataset Description
e dataset associated with this Dataset Paper consists of one item which is described as follows.).Size of the microsatellite fragments (base pair (bp)) ampli�ed on DNA of . intermedium, . elongatum, P. stipifolia, . bessarabicum, . ponticum, and D. villosum.A total of 42 SSR markers of wheat were evaluated on their cross-ampli�cation on DNA of . intermedium, . elongatum, . ponticum, . bessarabicum, P. stipifolia, and D. villosum.For each accession, two plants were analyzed.e sizes of the peaks are obtained by means of the fragment analysis.e number of the wheat SSR markers that ampli�ed DNA fragments with determined size for . ponticum was 33 (78.6%); for . intermedium, 28 (66.7%); for . elongatum, 24 (57.1%); for . bessarabicum, 24 (57.1%); for P. stipifolia, 26 (69.1%); and for D. villosum, 29 (69.0%).us, the transferability of the SSR markers to the genomes of the investigated species is comparable to that of oat, rye [8], Aegilops [9,13], Elymus, and Pseudoroegneria [10].Twentyfour primer pairs of wheat SSR markers were successfully ampli�ed from all investigated species.e absence of ampli-�cation may result from either null allele in a locus or signi�cant differences between wheat and studied species in the �anking regions where primers should anneal [14].e transferred SSRs can be linked to useful genes or QTLs as among selected markers there are some previously used for the mapping and detection of resistance genes of . intermedium (Xwmc221, Xwmc121, and Xcfd68) [15] and . elongatum (Xgwm325, Xgwm179, and Xgwm335) [16,17] and others are linked to QTLs of valuable traits on different wheat chromosomes [18].e size shown in the table is the sum of the real size of a fragment and 25 bp of M13 tail of forward primer.e localization on wheat chromosomes is shown according to Graingenes 2.0 database [10].If no peaks are detected, they are shown as "0" size.Where the variants between plant genotypes are observed, they are shown through slash "/".If a group of peaks is found for one plant and it differs from the group of another plant, the sizes of peaks are shown in brackets.Cases when it is difficult to determine major peaks among multiple peaks are designated "ND" (not determined).

Concluding Remarks
In conclusion, it has been shown that most of 42 SSRs are applicable for the investigation of genomes of . ponticum, . intermedium, . elongatum, . bessarabicum, P. stipifolia, and D. villosum.e polymorphism in SSR fragment size between different analyzed species has been shown.e transferred SSR markers can be used in assaying the genetic variability of these wild species and establishment of phylogenetic relationships between them.In addition, they can be used in wide hybridization of wheat to monitor and map alien introgressions.