Field

For a full inventory of Moorea's marine invertebrate diversity, we propose a three year program that focuses on macro- (>1 cm) and meso- (>1mm) fauna of shallow reefs (<30m) and associated habitats. The objective is to document all animals greater than one millimeter. This represents a subjective but practical limit to individual-based surveys beyond which specialized methods or environmental sequencing (pooling individuals of different species) is typically used to assess microfaunal and microbial biodiversity. We will also conduct deep water surveys to the extent that budget and equipment permit, likely in partnership with IRD in New Caledonia, utilizing their RV Alis. We will adopt and further develop collecting methodologies employed and standardized by CReefs, a division of the Census of Marine Life, and partner with them as a regional node. We will use diverse collecting methods, ranging from hand sampling, to varied trap, suction, and extraction techniques. The MBP will be the primary test site for establishing protocols as proxies for estimating reef diversity across the tropics. Gustav Paulay will continue to lead this taxonomic component. From what we have learned in the pilot and because the marine invertebrate component is so large, we will have two full time persons to manage this aspect of the survey, one for macro- and one for mesobiota. Based on field experience at the Guam Marine lab and also in consultation with participants in other bioblitzes, we recognize the need to bring taxonomic experts to Moorea. Experts greatly enhance capacity to search for species in specialized microhabitats, sorting of collected material, and species identification. We expect to document a large portion of the macro- and meso-fauna for an estimated 5,000+ species.

Laboratory

Macro-fauna and much of the meso-fauna will be sorted live or fresh on site to morphospecies, photodocumented, subsampled for genetic analysis, and fixed for morphological study. Some bulk samples will be fixed first, but otherwise processed as above. Samples of morphospecies will be sent to appropriate expert for identification, or identified in house where appropriate expertise or literature is available. We will utilize the extensive, international taxonomic networks developed by FLMNH and NMNH, Paris. Morphological and tissue vouchers will be deposited at FLMNH, NMNH, Smithsonian, and/or other appropriate institutions. Morphological and sequence data will be used to reciprocally illuminate species limits and identification. Taxonomic, locality, genetic, and photographic information will be made available online through BioCode, BOLD, and FLMNH databases. These samples will also be linked with, identification work facilitated by, and genetic results compared with, other large-scale biodiversity surveys pursued at FLMNH, including CReefs (NW Hawaiian Islands), Scripps Line Islands Expedition, Marine Biodiversity Survey of Guam, and BIOTAS (SW Indian Ocean). Each of these surveys has resulted in 1,000+ species, and 100s to 1,000s of genetic samples. These collections will provide an Indo- Pacific-wide context and comparative data for assessing geographic spread and genetic connectivity of Moorea taxa. Sequencing efforts will be handled by the Smithsonian. Samples will be sequenced by major taxonomic group (mostly phyla), with primers and PCR parameters optimized for that group through past efforts (BioCode and other projects above) and further improved upon during the course of this project. COI will be used for most taxa, as this gene region has been consistently shown to be appropriate in most phyla of marine invertebrates, with other mitochondrial genes (16S, cytb, etc.) used as backup in the rare cases where COI is not successfully amplified. Rate of mitochondrial sequence evolution is too slow to allow separation of species among numerous sponge and cnidarian lineages. For these taxa we will work with the active Sponge Barcoding Project and Cnidarian Tree of Life (NSF) projects and use the most appropriate nuclear marker (ITS, other introns, etc).

Timeline:

For part of each year project staff will based on Moorea for sampling, than move to Florida for additional sample preparation and identification. Visiting experts will be hosted in Moorea. • Year 1: 6 month sampling effort, with invited experts focusing especially on mesofaunal sampling and identification. Morpho-taxonomic work started with processing and distribution of samples. Testing and optimization of fixation and PCR protocols.

2009: 3-6 month sampling effort, with invited experts especially specializing on selected sessile biota (sponges, cnidarians, bryozoans, ascidians, etc) and their commensal associates. Large scale sequencing of all acquired samples.
2010: 3 month sampling effort, with invited experts specializing on other relevant taxa. Completion of morphotaxonomic and genetic efforts.