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This study focused on the development of EST-simple sequence repeats markers and the detection of their transferability and their utility for evaluating wheat leaf rust pathogen diversity. A total of 44,407 publicly available EST sequences derived from Puccinia triticina were computationally mined. Di-nucleotide repeat density covered the vast majority of assembled ESTs (45%). The tri-repeat motif (TCT) and pentarepeat motif (TTCTT) were the most repeated motif. A set of 103 Class I type sequences containing simple sequence repeats were further analyzed by BLASTX similarity. Nineteen primer pairs flanking regions of EST-SSRs were designed. Of the 19 primer pairs tested, 10 successfully amplified fragments. Their polymorphism was evaluated with 8 Puccinia triticina (Pt) single-uredinal isolates collected from the different regions of Turkey. These newly developed EST-SSR primer pairs can be implicated as stable markers for pathogen diversity analysis. It was also shown that some leaf rust EST-SSR markers were capable of cross-amplification in P. graminis f. sp. tritici. [ABSTRACT FROM AUTHOR]

Abstract: In the present study, mitochondrial DNA polymerase chain reaction-restriction-fragment length polymorphism (PCR-RFLP) and nuclear DNA inter-simple sequence repeat (ISSR) assays were used to assess the phylogenetic and phylogeographic relationships among Garra rufa samples from Anatolia. The complete mtDNA NADH 3/4 dehydrogenase (ND-3/4) gene amplified by PCR was digested with eight restriction enzymes. These enzymes produced 20 composite haplotypes for G. rufa populations. All the mtDNA haplotypes detected were highly diverged from each other and each lineage had a unique genetic profile. The evaluation of mtDNA PCR-RFLP data coupled with geological history of Anatolia indicated a deep genetic divergence among the mtDNA haplotypes of G. rufa populations from drainages of the Mediterranean and Persian Gulf, suggesting an early isolation of Tigris-Euphrates with Orontes river and other rivers draining into the Mediterranean Sea. In general, data from both mtDNA and nDNA were congruent. [Copyright &y& Elsevier]

Abstract: In this study, the aim is to determine allele frequencies of D1S1677, D2S441, D4S2364 miniSTR loci and to provide these 3 loci for using routinely in Turkish Criminal Laboratories. Allele frequencies and forensic parameters for the three miniSTR loci were investigated in a sample of 200 unrelated healthy Turkish individuals. We use Powerstats Excel workbook, in order to use a simple, quick and reliable method to analyze the race-based population statistics of potentially useful loci. This population data suggests that these three miniSTR loci will serve as useful complements to the CODIS loci to aid in the forensic analysis of degraded DNA, as well as missing persons work and parentage testing with limited next-of-kin reference samples, with a validated method in Turkish Criminal Laboratories. In Turkish population, powers of discrimination for these three loci are more than 0.837. Information content of polymorphism results are 0.75, 0.83 and 0.63 respectively. [Copyright &y& Elsevier]

Abstract: In this study; validation and optimization study of 6 new miniSTR was made to use these loci on forensic laboratory in Turkey. Coble and Butler''s method was based in this study but several modifications were made on this PCR method. Primer dye set should be FAM-TET-HEX rather than FAM-VIC-NED. Primer concentrations were increased. Top Taq™ Master Mix Kit (Qiagen) was used for PCR rather than use of single by single PCR solution. 6 loci were studied as two triple multiplex sets. After the method was optimized with these conditions, validation studies were carried out by working the sensitivity and reproducibility of this procedure and analyzing mixed samples. [Copyright &y& Elsevier]