Fingerprint construction through genotyping by sequencing for applied breeding in Brassica rapa

Genome. 2022 Feb;65(2):105-113. doi: 10.1139/gen-2021-0021. Epub 2021 Oct 14.

Abstract

This study evaluated the genotyping by sequencing (GBS) protocol for fingerprinting Brassica rapa, and the data derived were more reliable than the re-sequencing data of B. rapa. Of the 10 enzyme solutions used to analyze the numbers of genotypes and single-nucleotide polymorphisms (SNPs) in B. rapa, five solutions showed better results, namely, A (HaeIII, 450-500 bp), E (RsaI+HaeIII, 500-550 bp), F (RsaI+HaeIII, 500-600 bp), G (RsaI+HaeIII, 'All' fragment), and J (RsaI+EcoRV-HF®, 'All' fragment). The five enzyme solutions showed less than 40% similarity in different individuals from various samples, and 90% similarity between two individuals from one sample. The E enzyme solution was the most suitable for fingerprinting B. rapa, revealing well-distributed SNPs in the whole genome. Of the 82 highly inbred lines and 18 F1 lines of B. rapa sequenced by GBS in the E enzyme solution, known parents of 10 F1 lines were verified, and male parents were discovered for 8 F1 lines that had only known female parents. This study provides a valuable method for screening parents for F1 lines in B. rapa for the efficient evaluation of GBS with varied library construction strategies.

Keywords: Brassica rapa; F1; F1 lines; applied breeding; fingerprinting; genotyping by sequencing; génotypage par séquençage; polymorphismes mono-nucléotidiques; single-nucleotide polymorphisms; sélection appliquée.

MeSH terms

  • Brassica rapa* / genetics
  • Chromosome Mapping
  • Genome, Plant
  • Genotype
  • Plant Breeding*
  • Polymorphism, Single Nucleotide