Seyed Mostafa Pirseyedi

Isolation and Characterization of Novel Microsatellite Markers in Pomegranate (Punica granatum L.)

Pomegranate (Punica granatum L.) has been cultivated from ancient times for its economic, ornamental and medicinal properties globally. Here, we report the isolation and characterization of 12 polymorphic microsatellite markers from a repeat-enriched genomic library of Punica granatum L. The genetic diversity of these loci was assessed in 60 genotypes of Punica granatum L. All loci were variable: the number of polymorphic alleles per locus ranged from two to five (average 2.9). The observed and expected heterozygosities ranged from 0.15 to 0.87 and 0.29 to 0.65, respectively. The polymorphic information content ranged from 0.26 to 0.61 (average: 0.43). To the best of our knowledge, this is the first time that polymorphic microsatellite markers have been reported for P. granatum L. These new markers should allow studies of the population structure and genetic diversity of pomegranate to be performed in the future.

Comparison of genetic variation among accessions of Aegilops tauschii using AFLP and SSR markers

Genetic diversity of 54 accessions of Aegilops tauschii from five countries was assessed using sequence-tagged microsatellites (or simple sequence repeats, SSRs) and amplified fragment length polymorphisms (AFLPs). In the case of AFLP analysis, a total of 256 amplification products obtained, 234 of them were polymorphic across all the 54 accessions. A total of 224 fragments were obtained from the 24 SSR primers and 219 of fragments were polymorphic across all the genotypes screened. Based on both AFLP and SSR markers, the highest percentage of polymorphisms were obtained in Iranian and accessions of unknown origin. The highest polymorphic information content (PIC) value was observed for SSRs (0.82) while the highest marker index (MI) value was for AFLPs (8.5) reflecting the hyper-variability of the first and the distinctive nature of the second system. Principal co-ordinate analysis (PCO) revealed congruent patterns of genetic relationships for both data sets, but did not group accessions strictly according to their geographical origins. Poor correlation was found between AFLP and SSR marker loci. This low association may be due to low number of AFLP and SSR markers. These results show that molecular markers can help to organize the genetic variability and expose useful diversity for breeding purposes.

Genetic diversity and relationships among Pistacia species and cultivars

Iran is one of the two major centres of Pistacia diversity and the main producer of pistachios in the world. About 282 Iranian pistachio genotypes (Pistacia spp.), together with 22 foreign cultivars (P. vera), were genotyped using 10 simple sequence repeat (SSR) markers to analyse the genetic diversity and relationships among Pistacia species and cultivars. The results revealed that the genetic diversity within P. atlantica subsp. kurdica was considerably lower than in P. vera or P. khinjuk. Principal coordinate analysis revealed a clear separation between the different Pistacia spices, as well as between the Iranian and foreign cultivars. AMOVA analysis showed that the variation between the species, between different populations, and within populations accounted for 41, 9, and 50% of the total variation, respectively. The results demonstrated that the study of genetic diversity and relationships among Pistacia species and cultivars using SSR markers provides important information for the collection and conservation of pistachio germplasm. In addition, the Iranian cultivars had a broader genetic background than that of the foreign cultivars. Thus, they are very important for genetic conservation and the planning of future breeding programmes. We also determined the different levels of genetic diversity that exist between and within the species and populations and showed that gene flow occurs between the Iranian cultivars and wild-type P. vera populations. The study provides practical information that policy-makers and scientists can apply to the conservation and sustainable use of all the species studied.

Transposable element junctions in marker development and genomic characterization of barley

Barley is a model plant in genomic studies of Triticeae species. However, barleys large genome size and high repetitive sequence content complicate the whole‐genome sequencing. The majority of the barley genome is composed of transposable elements (TEs). In this study, TE repeat junctions (RJs) were used to develop a large‐scale molecular marker platform, as a prerequisite to genome assembly. A total of 10.22 Gb of barley nonassembled 454 sequencing data were screened with RJPrimers pipeline. In total, 9,881,561 TE junctions were identified. From detected RJs, 400,538 polymerase chain reaction (PCR)‐based RJ markers (RJMs) were designed across the genome, with an average of 39 markers/Mb. The utility of designed markers was tested using a random subset of RJMs. Over 94% of the markers successfully amplified amplicons, among which ∼90% were genome specific. In addition to marker design, identified RJs were utilized to detect 1190,885 TEs across the genome. In gene‐poor regions of the genome Gypsy elements comprised the majority of TEs (∼65%), while in gene‐rich regions Gypsy, Copia, and Mariner were the main transposons, each representing an average ∼23% of total TEs. The numerous RJ primer pairs developed in this study will be a valuable resource for barley genomic studies including genomic selection, fine mapping, and genome assembly. In addition, the results of this study show that characterizing RJs provides insight into TE composition of species without a sequenced genome but for which short‐read sequence data is available.

Radiation hybrid map of barley chromosome 3H

Assembly of the barley (Hordeum vulgare L.) genome is complicated by its large size (5.1 Gb) and proportion of repetitive elements (84%). This process is facilitated by high resolution maps for aligning bacterial artificial chromosome (BAC) contigs along chromosomes. Available genetic maps, however, do not provide accurate information on the physical position of a large portion of the genome located in recombination‐poor regions. Radiation hybrid (RH) mapping is an alternative approach, which is based on radiation‐induced deletions along the length of chromosomes. In this study, the first RH map for barley chromosome 3H was developed. In total, 373 in vivo RH lines were generated by irradiating wheat (Triticum aestivum L.)–barley chromosome 3H addition lines and crossing them to a normal wheat cultivar. Each RH informative line (containing deletions) had, on average, three deletions. The induced deletion size varied from 36.58 Kb to 576.00 Mb, with an average length of 52.42 Mb. This initial chromosome 3H radiation hybrid (3H‐RH) map had a 9.53× higher resolution than an analogous genetic map, reaching a maximum of >262.40× resolution in regions around the centromere. The final RH map was 3066.1 cR in length, with a 0.76 Mb resolution. It was estimated that the map resolution can be improved to an average of 30.34 Kb by saturating the 3H‐RH map with molecular markers. The generated RH panel enabled alignment of BAC and sequenced contigs as small as 1.50 Kb in size. The high resolution and the coverage of poor‐recombination regions make RH maps an ideal resource for barley genome assembly, as well as other genetic studies.

Novel nuclear-cytoplasmic interaction in wheat (Triticum aestivum) induces vigorous plants

Interspecific hybridization can be considered an accelerator of evolution, otherwise a slow process, solely dependent on mutation and recombination. Upon interspecific hybridization, several novel interactions between nuclear and cytoplasmic genomes emerge which provide additional sources of diversity. The magnitude and essence of intergenomic interactions between nuclear and cytoplasmic genomes remain unknown due to the direction of many crosses. This study was conducted to address the role of nuclear-cytoplasmic interactions as a source of variation upon hybridization. Wheat (Triticum aestivum) alloplasmic lines carrying the cytoplasm of Aegilops mutica along with an integrated approach utilizing comparative quantitative trait locus (QTL) and epigenome analysis were used to dissect this interaction. The results indicate that cytoplasmic genomes can modify the magnitude of QTL controlling certain physiological traits such as dry matter weight. Furthermore, methylation profiling analysis detected eight polymorphic regions affected by the cytoplasm type. In general, these results indicate that novel nuclear-cytoplasmic interactions can potentially trigger an epigenetic modification cascade in nuclear genes which eventually change the genetic network controlling physiological traits. These modified genetic networks can serve as new sources of variation to accelerate the evolutionary process. Furthermore, this variation can synthetically be produced by breeders in their programs to develop epigenomic-segregating lines.

Identification and Validation of a New Source of Low Grain Cadmium Accumulation in Durum Wheat

Cadmium (Cd) is a heavy metal that has no known biological function and is toxic for many living organisms. The maximum level of Cd concentration allowed in the international market for wheat grain is 0.2 mg kg−1. Because phenotyping for Cd uptake is expensive and time consuming, molecular markers associated with genes conferring low Cd uptake would expedite selection and lead to the development of durum cultivars with reduced Cd concentrations. Here, we identified single nucleotide polymorphisms (SNPs) associated with a novel low Cd uptake locus in the durum experimental line D041735, which has hexaploid common wheat in its pedigree. Genetic analysis revealed a single major QTL for Cd uptake on chromosome arm 5BL within a 0.3 cM interval flanked by SNP markers. Analysis of the intervening sequence revealed a gene with homology to an aluminum-induced protein as a candidate gene. Validation and allelism tests revealed that the low Cd uptake gene identified in this study is different from the closely linked Cdu1-B gene, which also resides on 5BL. This study therefore showed that the durum experimental line D041735 contains a novel low Cd uptake gene that was likely acquired from hexaploid wheat.

Isolation and characterization of novel microsatellite loci in Vaccinium arctostaphylos L

For the first time, this study has described the characterization of 25 novel polymorphic microsatellite loci from a repeat-enriched genomic library of V. arctostaphylos using a modified FIASCO method. Polymorphism of each locus was assessed in 64 individuals of the Vaccinium. The average allele number of the microsatellites, observed (HO) and expected (HE) heterozygosities were 5.88, 0.85 and 0.67 per locus, respectively. The polymorphic information content (PIC) value and inbreeding coefficient (F) ranged from 0.26 to 0.87 and −0.72 to 0.03, respectively. Ten of the twenty-five loci showed significant departures from Hardy–Weinberg equilibrium. These microsatellite loci could be useful to study genetic diversity and population structure of V.arctostaphylos.