Genotyping of single spore isolates of a Pasteuria penetrans population occurring in Florida using SNP-based markers

J Appl Microbiol. 2017 Feb;122(2):389-401. doi: 10.1111/jam.13345. Epub 2016 Dec 14.

Abstract

Aims: To generate single spore lines of a population of bacterial parasite of root-knot nematode (RKN), Pasteuria penetrans, isolated from Florida and examine genotypic variation and virulence characteristics exist within the population.

Methods and results: Six single spore lines (SSP), 16SSP, 17SSP, 18SSP, 25SSP, 26SSP and 30SSP were generated. Genetic variability was evaluated by comparing single-nucleotide polymorphisms (SNPs) in six protein-coding genes and the 16S rRNA gene. An average of one SNP was observed for every 69 bp in the 16S rRNA, whereas no SNPs were observed in the protein-coding sequences. Hierarchical cluster analysis of 16S rRNA sequences placed the clones into three distinct clades. Bio-efficacy analysis revealed significant heterogeneity in the level virulence and host specificity between the individual clones.

Conclusions: The SNP markers developed to the 5' hypervariable region of the 16S rRNA gene may be useful in biotype differentiation within a population of P. penetrans.

Significance and impact of the study: This study demonstrates an efficient method for generating single spore lines of P. penetrans and gives a deep insight into genetic heterogeneity and varying level of virulence exists within a population parasitizing a specific Meloidogyne sp. host. The results also suggest that the application of generalist spore lines in nematode management may achieve broad RKN control.

Keywords: Pasteuria penetrans; SNP; biotype; heterogeneity; host; root-knot nematode; specificity; spore.

MeSH terms

  • Animals
  • DNA, Bacterial / genetics
  • Florida
  • Genotype
  • Pasteuria / genetics*
  • Pasteuria / isolation & purification*
  • Phylogeny
  • Plant Roots / parasitology
  • Polymorphism, Single Nucleotide
  • RNA, Ribosomal, 16S
  • Solanum lycopersicum
  • Spores, Bacterial
  • Tylenchoidea / genetics
  • Tylenchoidea / microbiology*
  • Virulence

Substances

  • DNA, Bacterial
  • RNA, Ribosomal, 16S

Associated data

  • GENBANK/KT923061
  • GENBANK/KT923062
  • GENBANK/KT923063
  • GENBANK/KT923064
  • GENBANK/KT923065
  • GENBANK/KT923066