Efficient coralline algal psbA mini barcoding and High Resolution Melt (HRM) analysis using a simple custom DNA...

Area of Science:

• Marine Biology
• Molecular Systematics
• Phycology

Background:

• Coralline algae form vital maerl and rhodolith habitats supporting rich biodiversity.
• These ecosystems face threats from calcium carbonate harvesting and trawling.
• Morphological identification of maerl-forming species is unreliable, necessitating molecular methods.

Purpose of the Study:

• To develop a simplified DNA extraction, amplification, and sequencing method for coralline algae.
• To enable robust and efficient identification of maerl species and other coralline algae.
• To reduce the time and resources required for molecular systematics of these organisms.

Main Methods:

• A novel, easy DNA preparation method for coralline algae.
• Quantitative Polymerase Chain Reaction (qPCR) using a new psbA assay.
• High-Resolution Melt (HRM) analysis coupled with qPCR.

Main Results:

• The DNA preparation method yielded sufficient quality for qPCR and sequencing.
• The psbA qPCR assay identified six species and revealed two new Operational Taxonomic Units (OTUs).
• qPCR-HRM successfully differentiated closely related species, Lithothamnion erinaceum and L. cf. glaciale.

Conclusions:

• The developed DNA preparation and qPCR technique offers a cost-effective and time-efficient approach for coralline algae identification.
• This method is expected to invigorate research in coralline algae molecular systematics.
• Accurate identification is crucial for the conservation of threatened maerl and rhodolith habitats.