A novel genetic strategy to enable rapid detection of rare non-native alleles.

Established invasive species represent one of the most harmful and challenging threats to native biodiversity, necessitating methods for Early Detection and Rapid Response. Cryptic invasions are particularly challenging and often require expensive and time-consuming molecular surveys which limits their usefulness for management. We present a novel application of the Fluidigm SNP-Type Assay to identify rare non-native alleles that significantly reduces the cost and time to generate diagnostic results. We demonstrate the efficacy of this method using experimental Fluidigm pools (99% accuracy) and sequence data (96% accuracy). We apply our novel methodology to an endangered population of California tiger salamanders in Sonoma County where two individual non-native tiger salamander hybrids have previously been detected since 2008. We screened 5805 larvae in 387 sample-pools containing 15 larvae each. We did not detect any non-native hybrids in the population, a result that was verified with sequence data, though we strongly recommend additional years of sampling to confirm hybrid absence. Our success with a challenging, large-genome amphibian suggests this method may be applied to any system, and would be particularly useful when it is necessary for conservation practitioners to rapidly identify rare taxa or genes of interest.