Field
To complete the inventory of the island, samples will be taken from all elevation
zones. The following four sites, Mouaputa (830 m), Tohiea (1207 m), Mouapu (762m), and
Rotui (899 m) will be sampled from near sea level to the summit, where possible. Additional
low and mid elevation sites will be sampled to fill in any elevations, geographical areas, or
habitat types that were under sampled during the pilot study. In particular, we will target
sites around Fairurani (741m) and Tearai (770m) to the northeast and Mateotea (714m)
and Taufuapae (769m) to the northwest. Site selection will be done in consultation with the
plant and fungal teams. Collections will be carried out using both quantitative and
qualitative sampling methods. Insects, spiders, crustacea, mites, annelida, non-insect
hexapods, and myriapods will be processed. Quantitative methods will provide statistically
robust data (both numbers of species as well as abundance of individuals within species) for
comparisons between sites and to provide a baseline for future surveys. Rare species and
habitat types under-sampled quantitatively will be supplemented by standard qualitative
sampling methods. CDC backpack aspirator (vegetation surface, aerial), Berlese extraction
(forest litter/sedentary), and pitfall traps (forest litter/active) provide repeatable
quantitative samples. Malaise trap (flying/climbing), flight intercept (flying/falling), blacklight
(night flying), beating (vegetation), peeling barkexcavating wood (subcortical), sweep
net (vegetation surface), and hand examination (vegetation) provide diverse nonquantitative
samples. Teams of collectors will sample each site three times over the course
of a 12-month period. Rainy season collections will be weather dependent, with heavy
precipitation limiting high elevation sampling most. Each method will be evaluated to
determine the number of samples per site and parameters of each sample. Depending on
collections in year 1, selection of sites will be chosen for re-sampling in years 2 and 3. The
quantitative sampling methods employed will allow for comparisons between sites and
within sites over time. Additionally, environmental sequencing of mixed species samples
could reduce processing effort.
Laboratory
Specimens will be sorted, databased, identified (to lowest taxonomic group
possible), photographed, DNA-extracted, and mounted at the Gump Station immediately
following each collection. Fragile specimens from each sample will be prepared for
morphological identification on site (i.e., Lepidoptera and Miridae) with subsamples taken
for DNA extraction. Specimens selected for barcoding will be prepared, photographed, and
the whole abdomen will be removed and extracted nondestructively to allow for
amplification of associated Wolbachia (endosymbiotic bacteria) from the gut. The abdomens
will be removed from the lysis buffer, rinsed, and re-associated with the specimen. DNA
extractions will be PCR amplified at either the Gump Station or Berkeley and up to 5
specimens of each morphospecies will be sequenced. Taxonomic workers (invertebrate
coordinator, graduate students) working at Gump will identify specimens to species where
possible. Where species or generic identification is not possible, the workers will distribute
specimens to appropriate workers world-wide (see participants,
http://polynesia.berkeley.edu) for identification and/or formal species identification.
Voucher specimens and DNA extracts will be deposited in collections at the Essig Museum
(Berkeley), Bishop Museum (Honolulu), the NHM Paris, with a diagnostic collection housed
at Gump or elsewhere in French Polynesia. If and when long-term collection facilities are
available in French Polynesia, specimens will be repatriated. LUCID keys will be prepared for
several groups.
Timeline:
2008 – regular sampling of field sites, data processing, 1st cut
taxonomic identification, COI barcoding;
2009, 2010 – additional sampling, continued specieslevel identification and formal
description, COI barcoding.
