Improving quantitative laboratory analysis of phycobiliproteins to provide high quality validation data for ocean color remote sensing algorithms

Identification and characterization of phytoplankton communities and their physiology is a primary aim of NASA’s PACE satellite mission. The concentration and composition of phytoplankton pigments modulate the spectral distribution of light emanating from the ocean, which is measured by ocean color satellites, and thus provide critical information on phytoplankton community composition and physiological parameters. One diagnostic class of pigments not routinely well-characterized is the phycobiliproteins (PBPs), and NASA has a requirement to collect and distribute high quality in situ data in support of data product validation activities for ocean color missions. Phycobiliproteins are light-harvesting proteins that are the predominant photosynthetic pigments in some classes of phytoplankton including cyanobacteria, such as Synechococcus, Trichodesmium, and Microcystis. With the advance of hyperspectral ocean color sensors such as on PACE (expected to launch in late 2022), it is essential that we implement routine analysis of PBPs that satisfies several considerations: reproducible, high extraction efficiency for a variety of environments, and Suitable for large scale analysis. Published techniques for PBP analysis vary in recommendations for: collection, extraction, disruption mode, and analysis; evidence suggests the variation in results may depend at least in part on the species and even strain(s) of interest. Experiments that tested variations in these parameters have drawn very different conclusions regarding extraction efficiency and reproducibility. Cyanobacteria are more difficult to extract than other PBP-containing algae such as cryptophytes, but can be important primary producers. We used a cryptophyte (Rhodomonas salina) and cyanobacterium (Synechococcus sp.) to compare extraction efficiencies of water samples concentrated via centrifugation to filtered samples using two different extraction buffers (phosphate and asolectin-CHAPS). Samples were analyzed on a fluorometer configured for PC and PE detection. The results have important implications for collection and storage of samples for routine analysis; some previous studies (although not all) have suggested that filtered samples have a much lower extraction efficiency than whole water samples.