Introducing a novel 28S rRNA marker for improved assessment of coral
reef biodiversity
Abstract
Biodiversity monitoring based on DNA metabarcoding depends on primer
performance. Here, we develop a new metabarcoding primer pair that
targets a ~ 318 bp fragment of the 28S rRNA gene. We
validate the primer pair in assessing sponges, a notoriously challenging
group for coral reef metabarcoding studies, by using mock and natural
complex reef communities to examine its performance in species
detection, amplification efficiency, and quantitative potential. Mock
community experiments revealed a high number of sponge species (n = 94)
spanning a broad taxonomic scope (15 orders), limited taxon-specific
primer biases (only a single species exceeded a two-fold deviation from
the expected number of reads), and its suitability for quantitative
metabarcoding – there was a significant relationship between read
abundance and visual percent coverage of sponge taxa (R = 0.76). In the
natural complex coral reef community experiments, commonly used COI
metabarcoding primers detected only 30.9% of sponge species, while the
new 28S primer increased detection to 79.4%. These new 28S primers
detect a broader taxonomic array of species across phyla and classes
within the complex cryptobiome of coral reef communities than the
Leray-Geller COI primers. As biodiversity assessments using
metabarcoding tools are increasingly being leveraged for environmental
monitoring and guide policymaking, these new 28S rRNA primers can
improve biodiversity assessments for complex ecological coral reef
communities.