In silico search, characterization and validation of new EST-SSR markers in the genus Prunus

BMC Res Notes. 2016 Jul 7:9:336. doi: 10.1186/s13104-016-2143-y.

Abstract

Background: Simple sequence repeats (SSRs) are defined as sequence repeat units between 1 and 6 bp that occur in both coding and non-coding regions abundant in eukaryotic genomes, which may affect the expression of genes. In this study, expressed sequence tags (ESTs) of eight Prunus species were analyzed for in silico mining of EST-SSRs, protein annotation, and open reading frames (ORFs), and the identification of codon repetitions.

Results: A total of 316 SSRs were identified using MISA software. Dinucleotide SSR motifs (26.31 %) were found to be the most abundant type of repeats, followed by tri- (14.58 %), tetra- (0.53 %), and penta- (0.27 %) nucleotide motifs. An attempt was made to design primer pairs for 316 identified SSRs but these were successful for only 175 SSR sequences. The positions of SSRs with respect to ORFs were detected, and annotation of sequences containing SSRs was performed to assign function to each sequence. SSRs were also characterized (in terms of position in the reference genome and associated gene) using the two available Prunus reference genomes (mei and peach). Finally, 38 SSR markers were validated across peach, almond, plum, and apricot genotypes. This validation showed a higher transferability level of EST-SSR developed in P. mume (mei) in comparison with the rest of species analyzed.

Conclusions: Findings will aid analysis of functionally important molecular markers and facilitate the analysis of genetic diversity.

Keywords: Breeding; EST-SSR; Functional domain marker; In silico analysis; Prunus; SSR mining.

Publication types

  • Validation Study

MeSH terms

  • Computer Simulation
  • DNA, Plant / genetics*
  • Expressed Sequence Tags*
  • Genetic Markers*
  • Genome, Plant*
  • Genotype
  • Microsatellite Repeats*
  • Models, Genetic
  • Molecular Sequence Annotation
  • Nucleotide Motifs
  • Open Reading Frames
  • Polymorphism, Genetic
  • Prunus / classification
  • Prunus / genetics*
  • Software

Substances

  • DNA, Plant
  • Genetic Markers