Development of microsatellite markers for Lippia alba and related Lippia species

Mol Biol Rep. 2020 Jun;47(6):4911-4915. doi: 10.1007/s11033-020-05445-z. Epub 2020 Apr 23.

Abstract

Microsatellite primers were developed in Lippia alba complex to better understanding the origins and evolution of the species. We sought to increase the numbers of available simple sequence repeat (SSR) markers. We performed low-coverage (~ twofold) genomic DNA sequencing of a diploid accession and generated a de novo assembly comprising 175,572 contigs. Sixteen SSR loci were selected and of these 13 SSR loci were successfully amplified in 20 L. alba tetraploid accessions and in 12 other Lippia species. Only one SSR locus was monomorphic, whereas 12 loci were polymorphic, yielding one to nine alleles. The heterozygosity was similar among markers, with values of 0.274-0.485; the polymorphism information content values varied from 0.237 to 0.367. These markers were successfully amplified in related species with 85% of transferability on average. Thus, we demonstrate the utility of including a de novo assembly step to obtain SSR markers from low-coverage genomic datasets.

Keywords: Cross-amplification; NGS; Polyploid complex; Simple sequence repeat (SSR) markers; Verbenaceae.

MeSH terms

  • Alleles
  • Chromosome Mapping / methods
  • DNA Primers / genetics
  • DNA, Plant / genetics
  • Gene Frequency / genetics
  • Genotype
  • Heterozygote
  • High-Throughput Nucleotide Sequencing / methods
  • Lippia / genetics*
  • Microsatellite Repeats / genetics*
  • Polymorphism, Genetic / genetics
  • Sequence Analysis, DNA / methods

Substances

  • DNA Primers
  • DNA, Plant