The AutoSpawner system - Automated ex situ spawning and fertilisation of corals for reef restoration.

The restoration of reefs damaged by global and local pressures remains constrained by the scale of intervention currently feasible. Traditional methods for ex situ sexual propagation of corals produce limited materials, typically of limited genetic diversity and only sufficient for small field trials. The development and validation of new technologies to upscale and automate coral propagation is required to achieve logistically and financially feasible reef restoration at ecologically relevant scales. To address the need for upscaled production of genetically diverse material for use in reef restoration we designed an automated system (the AutoSpawner) for harvesting, fertilising and washing gametes from tropical broadcast-spawning corals. The system includes a novel high density dynamic fertilisation process, which enables the production of large numbers of fertilised coral eggs (>7 million per night for highly fecund species) without any downstream negative effects on larval quality. The functionality of the system and the quality of the produced larvae was assessed using multiple species from two coral families (Acroporidae and Merulinidae) across a range of spawning and gamete characteristics. We present the schematics and protocols required for automated sexual propagation of high-quality coral larvae using this novel system; and demonstrate that the time demands, and labour costs, associated with traditional manual-based sexual propagation of corals can be reduced by up to 113-fold using the AutoSpawner.

A PCR-Based Assay Targeting the Major Capsid Protein Gene of a Dinorna-Like ssRNA Virus That Infects Coral Photosymbionts.

The coral-Symbiodinium association is a critical component of coral reefs as it is the main primary producer and builds the reef's 3-dimensional structure. A breakdown of this endosymbiosis causes a loss of the dinoflagellate photosymbiont, Symbiodinium, and/or its photosynthetic pigments from the coral tissues (i.e., coral bleaching), and can lead to coral mortality. Coral bleaching has mostly been attributed to environmental stressors, and in some cases to bacterial infection. Viral lysis of Symbiodinium has been proposed as another possible cause of some instances of coral bleaching, but this hypothesis has not yet been experimentally confirmed. In this study, we used coral virome data to develop a novel PCR-based assay for examining the presence and diversity of a single-stranded RNA (ssRNA) virus by targeting its major capsid protein (MCP) gene. Illumina sequence analysis of amplicons obtained with novel primers showed 99.8% of the reads had the closest taxonomic affinity with the MCP gene of the virus, Heterocapsa circularisquama RNA virus (HcRNAV) known to infect dinoflagellates, indicating that dinorna-like viruses are commonly associated with corals on the Great Barrier Reef. A phylogenetic analysis of MCP gene sequences revealed strong coral species specificity of viral operational taxon units (OTUs). This assay allows a relatively easy and rapid evaluation of the presence and diversity of this particular viral group and will assist in enhancing our understanding of the role of viral lysis in coral bleaching.