A critical perspective on interpreting amplicon sequencing data in soil ecological research

Lauren V.AlteioaJoanaSénecaaAlbertoCanarinibRoeyAngelcJanJansadKseniaGusevaaChristinaKaiseraAndreasRichteraHannesSchmidta

Received 19 April 2021, Revised 2 July 2021, Accepted 8 July 2021, Available online 10 July 2021.


Microbial community analysis via marker gene amplicon sequencing has become a routine method in the field of soil research. In this perspective, we discuss technical challenges and limitations of amplicon sequencing and present statistical and experimental approaches that can help addressing the spatio-temporal complexity of soil and the high diversity of organisms therein. We illustrate the impact of compositionality on the interpretation of relative abundance data and discuss effects of sample replication on the statistical power in soil community analysis. Additionally, we argue for the need of increased study reproducibility and data availability, as well as complementary techniques for generating deeper ecological insights into microbial roles and our understanding thereof in soil ecosystems. At this stage, we call upon researchers and specialized soil journals to consider the current state of data analysis, interpretation, and availability to improve the rigor of future studies.