Protocol for the Molecular Identification of Autotrophic Euglenids in Environmental Samples.

Area of Science:

• Microbial Ecology
• Molecular Biology

Background:

• Eukaryotic microbial biodiversity is largely underexplored, with universal methods failing to capture full diversity.
• Autotrophic euglenids are often overlooked due to habitat and genetic complexities, rendering universal primers inadequate.

Purpose of the Study:

• To develop and validate a customized workflow for enhanced assessment of autotrophic euglenid biodiversity.
• To improve the accuracy and resolution of euglenid diversity and distribution studies.

Main Methods:

• Utilized group-specific primers targeting the V2 region of the 18S rDNA.
• Optimized biomass harvesting and DNA extraction protocols.
• Applied high-throughput sequencing and bioinformatics analysis.

Main Results:

• Successfully validated the workflow on 190 samples from 16 Polish water bodies.
• Achieved improved detection and a clearer understanding of euglenid diversity and distribution.
• Provided a comprehensive step-by-step protocol for euglenid biodiversity assessment.

Conclusions:

• The customized workflow significantly enhances the accuracy of euglenid biodiversity assessments.
• This approach offers a more effective solution for studying previously underexplored microbial groups.
• The protocol facilitates detailed insights into euglenid distribution and ecological roles.