Comparison of the biodiversity of epiphytic diatoms in the Euphrates-Tigris rivers using morphological and metabarcoding analyses

The Euphrates-Tigris catchment area is one of the major drainage basins in the Middle East but climate change, overuse of waters and subsequent salinization has brought this region to the brink of ecological devastation. The aim of this study was to investigate the biodiversity of epiphytic diatoms in Iraq, for the first time using a combination of morphological and metabarcoding methods. 47 samples were collected at six sampling sites along the Rivers Tigris and Euphrates in northern Basra, Southern Iraq, during the summer of 2019 (dry season) and winter of 2020 (rainy season). The composition of the epiphytic diatoms in each sample was compared with both methods. The microscopic-morphological analysis of the environmental samples resulted in the identification of 284 infrageneric taxa in 59 genera. For the metabarcoding analysis the V4 region of the 18S marker gene was used and resulted in 1454 ASVs (Amplicon sequence variants) comprising 54 genera and 108 species with several ASVs belonging to the same taxa. The non-metric multidimensional scaling (NMDS) plots for morphology showed a clear seasonal effect in the diatom community composition, while the NMDS plots for 18SV4 analyses did not show a seasonal effect in community composition for the same samples. The incomplete taxonomic reference database for the studied sites turned out to be the major limitation of the molecular approach. Nevertheless, the combination of morphological and molecular methods increased the detection and identification of the diatom assemblages and laid the foundation for understanding their biodiversity in this region.