The effect of DNA degradation bias in passive sampling devices on metabarcoding studies of arthropod communities and their associated microbiota

PLoS One. 2018 Jan 5;13(1):e0189188. doi: 10.1371/journal.pone.0189188. eCollection 2018.

Abstract

PCR amplification bias is a well-known problem in metagenomic analysis of arthropod communities. In contrast, variation of DNA degradation rates is a largely neglected source of bias. Differential degradation of DNA molecules could cause underrepresentation of taxa in a community sequencing sample. Arthropods are often collected by passive sampling devices, like malaise traps. Specimens in such a trap are exposed to varying periods of suboptimal storage and possibly different rates of DNA degradation. Degradation bias could thus be a significant issue, skewing diversity estimates. Here, we estimate the effect of differential DNA degradation on the recovery of community diversity of Hawaiian arthropods and their associated microbiota. We use a simple DNA size selection protocol to test for degradation bias in mock communities, as well as passively collected samples from actual Malaise traps. We compare the effect of DNA degradation to that of varying PCR conditions, including primer choice, annealing temperature and cycle number. Our results show that DNA degradation does indeed bias community analyses. However, the effect of this bias is of minor importance compared to that induced by changes in PCR conditions. Analyses of the macro and microbiome from passively collected arthropod samples are thus well worth pursuing.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Arthropods / classification
  • Arthropods / genetics*
  • Arthropods / microbiology*
  • Biodiversity
  • DNA / analysis*
  • DNA / genetics*
  • DNA Barcoding, Taxonomic / methods*
  • DNA Barcoding, Taxonomic / statistics & numerical data
  • DNA Primers
  • Ecosystem
  • Hawaii
  • Metagenome
  • Metagenomics / methods
  • Metagenomics / statistics & numerical data
  • Microbiota / genetics*
  • Polymerase Chain Reaction / methods
  • Selection Bias

Substances

  • DNA Primers
  • DNA

Grants and funding

Rosemary Gillespie was funded by the National Science Foundation (NSF), DEB1241253.