Fine-tuning the performance of ddRAD-seq in the peach genome

Sci Rep. 2021 Mar 18;11(1):6298. doi: 10.1038/s41598-021-85815-0.

Abstract

The advance of Next Generation Sequencing (NGS) technologies allows high-throughput genotyping at a reasonable cost, although, in the case of peach, this technology has been scarcely developed. To date, only a standard Genotyping by Sequencing approach (GBS), based on a single restriction with ApeKI to reduce genome complexity, has been applied in peach. In this work, we assessed the performance of the double-digest RADseq approach (ddRADseq), by testing 6 double restrictions with the restriction profile generated with ApeKI. The enzyme pair PstI/MboI retained the highest number of loci in concordance with the in silico analysis. Under this condition, the analysis of a diverse germplasm collection (191 peach genotypes) yielded 200,759,000 paired-end (2 × 250 bp) reads that allowed the identification of 113,411 SNP, 13,661 InDel and 2133 SSR. We take advantage of a wide sample set to describe technical scope of the platform. The novel platform presented here represents a useful tool for genomic-based breeding for peach.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Computational Biology / methods
  • DNA, Plant / genetics
  • DNA, Plant / isolation & purification
  • Deoxyribonucleases, Type II Site-Specific / genetics
  • Genetic Loci
  • Genome, Plant*
  • Genotype*
  • Genotyping Techniques / methods
  • High-Throughput Nucleotide Sequencing / methods*
  • Plant Breeding
  • Polymorphism, Single Nucleotide
  • Prunus persica / enzymology*
  • Prunus persica / genetics*
  • Sequence Analysis, DNA / methods*

Substances

  • DNA, Plant
  • CTGCAG-specific type II deoxyribonucleases
  • Deoxyribonucleases, Type II Site-Specific
  • GATC-specific type II deoxyribonucleases