Amplicon sequence variant-based meiofaunal community composition revealed by DADA2 tool is compatible with species...

Area of Science:

• Marine Biology
• Molecular Ecology
• Bioinformatics

Background:

• Morphological identification of meiofauna is challenging and time-consuming.
• Molecular methods like DNA barcoding offer an alternative for species identification.

Purpose of the Study:

• To implement the amplicon sequence variant (ASV) concept for meiofaunal species composition analysis.
• To develop a reliable 18S rRNA gene metabarcoding reference library for meiofauna.

Main Methods:

• PCR amplification of the 18S rRNA gene from meiofaunal samples.
• High-throughput sequencing on the Illumina platform.
• Bioinformatic processing using DADA2 software to generate ASV libraries.
• Clustering of ASVs at a 3% dissimilarity threshold.
• BLASTN annotation of the reference library.

Main Results:

• A 470-490 bp 18S barcode was successfully amplified and sequenced.
• The quality-filtered (q≤25) ASV library contained 461 ASVs, demonstrating high uniqueness (average 1.34 ASVs/cluster).
• The reference library accurately represented expected meiofaunal taxa.

Conclusions:

• ASV-based metabarcoding is a viable method for meiofaunal species identification.
• The developed 18S rRNA gene reference library provides a robust tool for meiofaunal community analysis.
• This approach enhances the accuracy and efficiency of meiofaunal biodiversity assessments.