Enhancing amphibian biomonitoring through eDNA metabarcoding.

Area of Science:

• Ecology
• Genetics
• Conservation Biology

Background:

• Biodiversity surveying is enhanced by environmental DNA (eDNA) metabarcoding, valued for efficiency and sensitivity.
• This technique is crucial for monitoring elusive species, including rare and endangered amphibians.

Purpose of the Study:

• To establish a robust metabarcoding pipeline for amphibian monitoring.
• To evaluate primer performance for eDNA-based amphibian detection.

Main Methods:

• Primer design and in silico PCR simulation.
• Laboratory validation using target DNA amplification.
• Field validation through mesocosm experiments.

Main Results:

• The Am250 primer, targeting the mitochondrial 16S gene, showed superior performance in taxonomic resolution and detection ability.
• Am250 achieved an 83.8% species amplification rate and 75.4% accurate identification for Chinese amphibians in silico.
• eDNA detection persisted for weeks post-organism removal in mesocosms, with primer read abundance influencing biomass estimation.

Conclusions:

• A comprehensive framework involving in silico, in vitro, and mesocosm validation is essential for selecting optimal eDNA metabarcoding primers.
• Primer selection strategy should be tailored to monitoring groups to maximize species detection and result credibility.