ENDER /ECOGEM/CYBER collaborative project: PROJECT TITLE: Asparagopsis taxiformis (a marine red alga) as an indicator of climate change.

Location: Kiholo Bay, Hawaii (initial site) & Kaneohe Bay (Oahu); Archipelago later in project. Introduction: Asparagopsis taxiformis or limu kohu is a highly valued edible marine red alga in Hawaii. It was so prized by the Hawaiians for its peppery flavor that only ali`i (Hawaiian royalty) were allowed to eat it (Abbott, 1984). The alga is found throughout the world in tropical marine environments and more recently, its appearance in the Mediterranean Sea has earned it the unwelcomed label of invasive species (Andreakis et al., 2007). The species has also been examined for its potential as a source of pharmaceutical agents (G. Genovese et al, 2009; del Val et al, 2001; McConnell and Fenical, 1977; Woolard, Moore, and Roller, 1979). Early studies indicate that extracts of this alga contain anti-Leishmania compounds and antibacterial halogenated compounds. The early Hawaiians recognized the disease fighting components of the red alga for there are reports of the use of a seaweed poultice against coral cuts. Found on edges of reef in areas of constant water motion, the plant has largely disappeared from regions where it was once very abundant. The reason for the decrease in abundance is not fully understood. There appears to have been at least three separate introductions of Asparagopsis taxiformis in the Hawaiian Islands (Sherwood, 2008). Assessing 56 samples from the gametophyte stage of the alga, Dr. Sherwood was able to successfully sequence the cox2-3 spacer region for 36 samples and the COI gene for 42 of the samples from the mitochondrial DNA. Her analyses indicated that there are 3 lineages of the red alga in Hawaii. Based on archival samples and distribution analysis, lineage 2, which is found in the Northwestern Hawaiian Islands as well as the Main Hawaiian Islands, is most likely the original native lineage with close ties to the species clade reported for the Indo-Pacific, central Mediterranean, and Southern Portugal. Lineage 1 predominates in the Main Hawaiian Islands and is also known on the Pacific coast of Central America. Lineage 1 appears to be supplanting Lineage 2 in the Main Hawaiian Islands. Lineage 4 is confined to Waikiki and Alan Moana areas of Honolulu on the southern shores of Oahu. It has only been known since 1991. Lineage 3 in Dr. Sherwoods study contained A taxiformis not found in Hawaii but found in Caribbean waters, Brazil, Lebanon, and the Canary Islands. We have no current information on whether the genetically different A. taxiformis clades in Hawaii have different growth characteristics, differences in taste and defense mechanisms to herbivorous fish and invertebrates. We have no information on how these difference clades adapt to environment. Hawaiian oral history indicates that limu kohu was harvested at a particular time of year depending on the region and the color indicated whether the limu was high or low in iodine (red indicating low iodine). The harvest was best a few days after a heavy rain which made the limu grow really long (Kamaui Aiona, M.S. thesis, Botany at UH Manoa, 2003.) J Padilla-Gamino and R.C. Carpenter examined two populations of A. taxiformis, one from Santa Catalina Island, California, and one from Kaneohe Bay, Oahu, Hawaii. They found that both populations had the same high temperature tolerances for maximum net photosynthesis, but different cold temperature tolerances. The California strain was more tolerant of temperature fluxes than the Hawaiian strain (Padilla-Garmino and Carpenter, 2007). Planned Pilot Study: 1. Genetically type the limu kohu in Kiholo Bay and Kaneohe Bay 2. Create benthic habitat maps of the limu kohu and other benthic algae at these sites. 3. Assess growth rates of the algae under different growth conditions (increased or decreased nutrients, changes in salinity, changes in temperature and light)

4. Assessment of halogenated compounds, antimicrobial compounds in algae. Determine whether different growth conditions affect production of natural products. 5. Determine effect of natural products on seaweed-herbivore interactions. 6. Develop transcriptomic studies. Identify indicator genes. 7. Field studies to correlate with field conditions. 8. Enlarge studies to include the entire Hawaiian Archipelago. Members of Team: Jo-Ann Leong (ECOGEM), Jim Beets (ENDER), Maria Haws (ECOGEM), Bob Borris (ECOGEM), Alison Sherwood (ECOGEM), Donna Delparte (CYBER) Students/Research Assistants: Mindy Mizobe, Caitlin Kryss, Rebecca Most

Benchmarks: 1. Assemble team: August 2011 2. Collect limu kohu samples, photographs, maps of benthos (Kiholo Bay, Kaneohe Bay) August-Nov, 2011. 3. Genotyping of samples: preliminary analysis of morphology and genotype, sites. Sept.-Dec. 2011. 4. Set up and conduct in vitro limu growth experiments Sept 2011 to Sept. 2012 5. Preliminary results ready for publication, grant proposal with objectives for no. 5,6,7, and 8. June 2012 Potential Funding Sources: 1. NSF:
a. b. Plant, Fungal and Microbial Developmental Systems-The Plant, Fungal and Microbial Development programmatic area supports research that addresses developmental processes in plants from algae to angiosperms, microbes and fungi. (Div. of Integrated Organismal Systems) The goal of this joint NSF-JST program is to advance novel biological knowledge in metabolomics in the areas of energy and the environment, and to foster greater collaborative interactions between Japanese and U.S. scientists in these priority areas. The focus of METABOLOMICS will be on plants, microbes, and algae and eligible research areas will include but will not be limited to:

Capture of all major metabolites Development of standards and annotation of unknown metabolites Identification of specialized metabolites of potential value

In recent years, metabolomics has matured to the point where it is now possible to consider cataloging the complete profiles of small molecules in cells. Such profiling is critically important because these small molecule metabolites are the end products of gene expression and represent the highresolution biochemical phenotype of the cell, tissue, and organism. Key goals of metabolomics include 1) chemical annotation, i.e. determining the chemical structure of each molecule, 2) biological annotation, i.e. connecting each metabolite to a specific enzyme, biochemical pathway, or biological process, and 3) metabolomic annotation, i.e. the distribution of each metabolite in different cells of an organism which includes spatial and temporal information as well as concentration.

2.

USDA: National Institute of Food and Agriculture

a. Alaska Native-Serving and Native Hawaiian-Serving Institutions Education Grants Program The purpose of this program is to promote and strengthen the ability of Alaska Native-Serving Institutions and Native Hawaiian-Serving Institutions to carry out education, applied research, and related community development programs. NIFA intends this program to address educational needs, as determined by each institution, within a broadly defined arena of food and agricultural sciencesrelated disciplines. Priority will be given to those projects that enhance educational equity for underrepresented students; strengthen institutional educational capacities; prepare students for careers related to the food, agricultural, and natural resource systems of the United States; and maximize the development and use of resources to improve food and agricultural sciences teaching programs.

3.

State Legislature
a. Native Hawaiian limu cultivation project b. Invasive species task force