Identification of individual root-knot nematodes using low coverage long-read sequencing

PLoS One. 2021 Dec 1;16(12):e0253248. doi: 10.1371/journal.pone.0253248. eCollection 2021.

Abstract

Root-knot nematodes (RKN; genus Meloidogyne) are polyphagous plant pathogens of great economic importance to agriculturalists globally. These species are small, diverse, and can be challenging for accurate taxonomic identification. Many of the most important crop pests confound analysis with simple genetic marker loci as they are polyploids of likely hybrid origin. Here we take a low-coverage, long-read genome sequencing approach to characterisation of individual root-knot nematodes. We demonstrate library preparation for Oxford Nanopore Technologies Flongle sequencing of low input DNA from individual juveniles and immature females, multiplexing up to twelve samples per flow cell. Taxonomic identification with Kraken 2 (a k-mer-based taxonomic assignment tool) is shown to reliably identify individual nematodes to species level, even within the very closely related Meloidogyne incognita group. Our approach forms a robust, low-cost, and scalable method for accurate RKN species diagnostics.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • DNA, Helminth / genetics*
  • Female
  • High-Throughput Nucleotide Sequencing*
  • Plant Diseases / genetics
  • Plant Diseases / parasitology
  • Tylenchoidea* / classification
  • Tylenchoidea* / genetics

Substances

  • DNA, Helminth

Grants and funding

This project was internally funded by the University of Hull’s Energy and Environment Institute as part of a strategic THYME project.