Chromium sequencing: the doors open for genomics of obligate plant pathogens

Biotechniques. 2018 Nov;65(5):253-257. doi: 10.2144/btn-2018-0019.

Abstract

It is challenging to sequence and assemble genomes of obligate plant pathogens and microorganisms because of limited amounts of DNA, comparatively large genomes and high numbers of repeat regions. We sequenced the 1.2 gigabase genome of an obligate rust fungus, Austropuccinia psidii, the cause of rust on Myrtaceae, with a Chromium 10X library. This technology has mostly been applied for single-cell sequencing in immunological studies of mammals. We compared scaffolds of a genome assembled from the Chromium library with one assembled from combined paired-end and mate-pair libraries, sequenced with Illumina HiSeq. Chromium 10X provided a superior assembly, in terms of number of scaffolds, N50 and number of genes recovered. It required less DNA than other methods and was sequenced and assembled at a lower cost. Chromium sequencing could provide a solution to sequence and assemble genomes of obligate plant pathogens where the amount of available DNA is a limiting factor.

Keywords: chromium 10X; fungal genomes; myrtle rust; rust fungi.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Basidiomycota / genetics*
  • Chromium / chemistry
  • DNA, Fungal / analysis
  • DNA, Fungal / genetics*
  • Gene Library
  • Genome, Fungal*
  • Genomics / methods*
  • Myrtaceae / microbiology*
  • Plant Diseases / microbiology*
  • Sequence Analysis, DNA / methods*

Substances

  • DNA, Fungal
  • Chromium