USAP-DC

Traditional models of oceanic food chains have consisted of photosynthetic algae (phytoplankton) being ingested by small animals (zooplankton), which were ingested by larger animals (fish). These traditional models changed as new methods allowed recognition of the importance of bacteria and other non-photosynthetic protozoa in more complex food webs. More recently, the wide-spread existence of mixotrophs (organisms that can both photosynthesize and ingest food particles) and their importance as microbial predators has been recognized in many oceanographic areas. In the Southern Ocean, the only two surveys of mixotrophs have suggested that there may be seasonal differences in their importance as predators. During the long polar night (winter), the ability of mixotrophs to ingest particulate food may aid in their survival thus ensuring a sufficient population in spring to support a phytoplankton bloom once photosynthesis rates can increase. Thus mixotrophs may provide a critical early food source upon which zooplankton and larger animals depend on for growth and reproduction. This project will advance understanding of mixotroph diversity and their ecological impact within the Southern Ocean microbial food web. Specifically, efforts will be focused on mixotrophy in the western Antarctica peninsula region during the austral spring and autumn when there are likely to be changes in the relative importance of photosynthesis and ingestion to mixotrophs. The project will provide research opportunities for undergraduate and graduate students and a post-doctoral researcher. There will be real-time outreach from the Southern Ocean to the public via blogs and interviews, and to high school art students through an established program that blends science and art education. Despite traditional views of protists as either "phototrophic" or "heterotrophic," there are many photosynthetic protists that consume prey (mixotrophy). Mixotrophy is a widespread phenomenon in aquatic systems and phytoplankton groups with known mixotrophic species, notably chrysophytes, cryptophytes, prymnesiophytes, prasinophytes and dinoflagellates, are present and often abundant in Antarctic waters. However, in the Southern Ocean, the presence of mixotrophic phytoflagellates has been surveyed only twice prior to this project: in the Ross Sea during Austral spring 2008 and summer 2011. The primary goals of the project are to gain better understanding of mixotroph diversity and their ecological impact with respect to the Southern Ocean microbial food web. The contribution of mixotrophs to primary production and bacterial consumption is likely linked to the taxonomic composition of the community and the abundance of particular species. Abundances of novel mixotrophic species will be evaluated via qPCR, which will be coupled with assessments of rates of feeding and photosynthesis with the goal of describing how active mixotrophs direct the movement of carbon through food webs. These experiments will help the determination of how viable and widespread mixotrophy is as a nutritional strategy in polar waters and give direct information on the currently unknown diversity of mixotrophic taxa under different environmental conditions occurring in austral spring and autumn. Furthermore, the methods will simultaneously yield information on the whole communities of protists - mixotrophic, phototrophic and heterotrophic. In addition, a method to examine aspects of the taxonomic and functional diversities of the bacterivorous/mixotrophic community will be employed. A thymidine analog (BrdU) will be used to label DNA of eukaryotes feeding on bacteria. The BrdU-labeled eukaryotic DNA will be isolated using immunoprecipitation. High-throughput sequencing of the labeled DNA (bacterivores) versus unlabeled community DNA will determine the diversity of bacterivorous mixotrophs relative to other microeukaryotes. Flow cytometric sorting based on chlorophyll to focus on mixotrophic species. These approaches will elucidate a gap in current knowledge of the influence of microbial interactions in the Southern Ocean under different conditions. This award reflects NSF's statutory mission and has been deemed worthy of support through evaluation using the Foundation's intellectual merit and broader impacts review criteria.

Person	Role
Sanders, Robert	Investigator and contact
Gast, Rebecca	Co-Investigator
Jeffrey, Wade H.	Co-Investigator

Antarctic Organisms and Ecosystems

Award # 1744767

USAP-1744767_1

Deployment	Type
LMG 19-04	ship expedition
NBP 22-05	ship expedition
NBP19-10	ship expedition

None in the Database

Repository	Title (link)	Format(s)	Status
R2R	LMG1904 expedition data	Not Provided	exists
Figshare	Companion datasets to Diversity of microbial eukaryotes along the West Antarctic peninsula in austral spring.	Excel; ASCII	exists
R2R	Expedition Data of NBP1910		exists
NCBI	NBP1910_protist_community_RNA Raw sequence reads; NBP2205_protist_community_RNA Raw sequence reads will be made available here after processing is completed	None	exists
R2R	Expedition Data of NBP 2205	None	exists

1. Grattepanche, J.-D., W.J. Jeffrey, R.J. Gast, R.W. Sanders. 2022. Diversity of microbial eukaryotes along the West Antarctic peninsula in austral spring. Frontiers in Microbiology 13 (doi:10.3389/fmicb.2022.844856)