Identification of Earthworm Species in Uruguay Based on Morphological and Molecular Methods

Abstract

Molecular techniques could aid earthworm species identification, especially when morphological characters are not taxonomically informative, or difficult to discern. No previous study has investigated molecular-based methods for earthworm taxonomy in Uruguay. The present study aimed to make a first approach using DNA barcoding as a tool to smoothen the way towards determining the earthworm richness in Uruguay. This study was based on an earthworm collection, identified both by morphological characters and molecular techniques, from samplings from different agricultural soils in Montevideo and Paysandú, Uruguay. Adult individuals were identified by external morphology following available descriptions of regional earthworms. From each morphological group a representative sample was selected for genomic DNA extraction, mitochondrial COI region amplification and sequencing. Sequences obtained were subject to BLAST searches and compared to sequences available in GenBank. Eight out of 11 sequenced exotic species were fully identified and matched morphological characters and molecular information; two were less consistent, with lower sequence similarity percentage; and one could not be fully identified due to lack of close related sequences in GenBank. While most exotic species had representative sequences annotated in GenBank, this was only the case for one native species, highlighting the need to develop this important area at a regional level. This study could kick-start an innovative research program, since there are limited records of earthworm samplings in Uruguay and no identification of species by DNA sequences from national studies.