EAGER: Collaborative Research: Developing Transformation Technologies for Porphyra

The genome of the marine red alga Porphyra umbilicalis is being sequenced by the Joint Genome Institute. The sequence information will help scientists address many fundamental questions, because Porphyra spp. belong to an ancient eukaryotic lineage, are important human foods ( nori ), have complex life histories, and---even compared to other intertidal organisms--- possess an unusually stress-tolerant metabolism. Computer-based analyses of the new genomic data will be sufficient to address some research questions, but most studies (e.g., the basis of Porphyra\u27s tolerance to extreme drying or high light) will require experimental approaches based upon bioinformatics analyses. This project will develop the essential technology of stable genetic transformation in Porphyra to make such experimental work possible. The investigators will focus on transforming neutral spores of P. umbilicalis because these abundantly-produced spores lack a cell wall. This should make it possible to transform the cell by electroporation, among other approaches. Native Porphyra promoters of Porphyra genes will be used, based on information provided by the JGI sequencing project, and codon-optimized reporter genes will be synthesized. Spores will be treated with selective antibiotics to recover transformed sporelings, and the stability of the transformation will be assessed as sporelings mature to adults. The PIs and a postdoctoral associate will work across both participating laboratories to develop transformation technologies. The postdoctoral associate will investigate reproductive pathways or stress physiology during the development of transformation techniques. S/he will be well-prepared to make individual and collaborative advances with the Porphyra model system due to the comprehensive postdoctoral training and participation in the Porphyra NSF Research Coordination Network. This project will allow the scientific community to use the data from the whole genome sequencing project in experimental research on Porphyra, leading to fundamental advances in the areas of metabolism, evolution, and developmental biology

Hydrodynamic Regulation of Reproduction in Fucoid Algae: A Regional Model and Consequences for Population Structure

Fucoid algae dominate most rocky shores across the north Atlantic and contribute substantially to structuring of the coastal ecosystem. Reproduction in fucoid algae is sensitive to hydrodynamic conditions, resulting in high fertilization success because gamete release occurs only under calm conditions. These findings have important implications for asynchrony in gamete release between populations and the scale of population isolation. This study will 1) test a nascent model describing when successful fucoid reproduction can occur, 2) determine whether hybridization between Fucus vesiculosus and other fucoid algae occurs when gamete release is delayed by turbulent conditions, and 3) analyze whether genetic differentiation in F. vesiculosus is correlated with variable hydrologic conditions due to coastal topography. Quantitative data on gamete release at multiple sites on opposite sides of coastal points (and smaller features) will be collected. This will determine whether gamete release in fucoids occurs at different times over small spatial scales due to variable wave and wind exposures across opposite sides of the undulating coastline. Species-specific monoclonal antibodies to recognize fucoid sperm are available; they and the required pump/concentration apparatus have been tested successfully at Pemaquid Point. Local environmental data (e.g., wind speed, light levels, water motion) will be collected and compared with data from monitoring systems such as NOAA\u27s National Data Buoy Center stations. Maine shores have been stable for thousands of years; if the coastal F. vesiculosus is reproductively isolated in topographic fragments, population genetic structure may be affected. This will be tested with AFLP analysis of randomly collected adults. Storms delay gamete release from fucoid algae, and laboratory studies predict that natural hybridization will increase sharply if over-mature eggs participate in fertilizations. This will be tested in field studies of hybridization potential, achieved hybridization, and identification of hybrid position in the intertidal zone in F. spiralis versus F. vesiculosus (Maine) and F. serratus (an exotic) versus F. vesiculosus (Nova Scotia). The hypothesized environmental interaction with development as a regulator of hybridization and genetic structure in F. vesiculosus (and other fucoids) could have strong local and biogeographic effects on intertidal communities across the north Atlantic. Among the products of this research will be a robust regional model for fucoid reproduction as a function of relevant physical factors (i.e., wind speed and direction, wave height, light levels)

SGER: Investigation of Potential Co-Introduction of Fucus serratus and Littorina littorea to North America in 1800s

This research will apply new approaches and expertise to understanding the probable invasion of North American intertidal zones by the herbivorous snail Littorina littorea in the 1800s. The investigator developed the following hypothesis during her recent analyses of late 1700s to mid-1800s shipping records: Fucus serratus and Littorina littorea were co-introduced into North America from Britain via the dumping of intertidal rock ballast in ships arriving at Pictou Harbor during the massive emigration of nearly 40,000 Scots (and some Irish and English) in the late 1700s-mid-1800s. This hypothesis will be tested using innovative molecular techniques (i.e., assay of nuclear and mitochondrial loci with primers that have already been developed for population genetic and phylogeographic studies in both species). Snails and algae will be collected and screened from Pictou (Canada) and 3-4 of the best candidate sites from Britain, based on the frequency of arrivals of ships from different British ports near the time of the putative introductions. The investigator will attempt to determine whether genotypes have remained stable over time (mid-1800s compared to today) in Pictou with herbarium material of F. serratus from the 1800s. The broader impacts include: demonstrating that Pictou Harbor was an epicenter for marine invasions in the last century. This, in turn, should lead to discoveries of other co-introduced species and confirm and extend our theoretical understanding of the trajectory of marine invasions based on different life histories and dispersal strategies of the species (e.g., L. littorea produces larvae; F. serratus produces rapidly attaching zygotes). The proposed SGER should lead to a full, collaborative proposal to pursue the co-introduction and broader multi-species invasion questions within a year

Mechanisms by Which Marine Algae Respond to Environmental Variables Affecting Reproductive Success

Many marine species reproduce by external fertilization. This mode of reproduction might be expected to result in low levels of fertilization and recruitment because of the dilution of eggs and sperm by the turbulent sea. Recent work, however, demonstrates that fucoid algae (i.e., brown seaweed\u27s that are important components of rocky, intertidal zones and estuaries) sense water motion and release eggs and sperm only when the sea is calm. Nearly 100% of their eggs are fertilized. The proposed experiments will determine how these algae sense water motion and whether similar responses are found in several other species that are harvested commercially or are important nuisance species. Studies of the reproductive tissues with electron microscopic and physiological techniques will establish the basis of the mechanical forces that expel eggs and sperm from adults and how these are blocked by high water motion. Field studies of natural communities in New England estuaries will complement the laboratory work. These studies will advance understanding of the mechanisms by which marine algae respond to environmental variables that affect reproductive success. This information should be useful to basic scientists, marine engineers, coastal zone managers, and mariculturists

Acquisition of Laser Scanning Confocal Microscope for Biological and Materials Research

Biological and Materials research at the University of Maine will be strongly impacted by the acquisition of a Laser Scanning Electron Microscope as a result of this NSF-MRI award. The Leica confocal unit, along with an upright and inverted microscope and digital camera will form a multi-user facility for campus researchers working with a range of biological and materials problems. Initially, 13 faculty members from 8 academic departments have projects planned for the instrument. The microscope will be the first of its kind on the University of Maine campus. A wide range of research problems will be attacked through use of this instrument in conjunction with existing instrumentation. Examples include: Biological research projects involve understanding the maintenance of bone structure through mapping of the distribution of proteins, the reproduction of algae in troubled marine ecosystems, and bacterial or viral diseases of fish. Additionally, improved understanding of fundamental microbe-plant symbiosis and wood decay processes will allow for future applied research to attack economically and socially important problems. The development of biofilms for sensors of biological and chemical warfare agents will be aided through film characterization using this instrument. This $10 million effort is a University/private industry/Department of Defense partnership. Materials-related research includes determination of morphology and fracture of wood-based composite material, microfracture characterization of cement-based materials, and characterization of paper roughness. This work, along with environmental scanning electron microscopy and X-ray microtomography, is focused on the measurement of microstructural mechanisms of material behavior and its improvement through subsequent processing changes. The ultimate benefit will include more efficient use of natural resources, better performance and lowered product costs

A Revolutionary Model to Improve Science Education, Teachers, and Scientists

To meet many modern global challenges, we need to promote scientific and technical literacy. The U.S. National Science Foundation (NSF) supports a “revolutionary” program to connect science education at all levels, from elementary through graduate school. The authors demonstrate how Maine has benefited from this program. They describe the University of Maine’s NSF-funded “GK-12 STEM” program, which placed graduate and advanced undergraduate science and technology students in elementary, middle, and high school classrooms; provided equipment for the schools; and offered training and professional development for the partner teachers. The authors urge the state, universities, and school districts to continue to use this model to increase science literacy and research capacity

GK-12: NSF Graduate Teaching Fellows in K-12 Education at the University of Maine

Eight districts in central Maine that comprise the Penobscot River Educational Partnership (PREP); four of them, including Maine Indian Education, partners in a current GK-12 project, have joined with the University of Maine to form Fellow-teacher teams to introduce K-12 students to experiments, field trips, and discussions in areas such as chemistry, climate change, marine sciences, molecular biology, geology, food sciences, and ecology. The program is: a) helping teachers and students reach the State of Maine\u27s legislatively-mandated standards for Science & Technology (the Maine Learning Results), b) strengthening Fellows\u27 communication and teaching skills, c) providing professional development for Teachers, d) enriching science for K-12 students, e) providing young male and female role models of SMET professionals to children in grades 3-11, and f) strengthening contacts between GK-12 science faculty and K-12 districts. The K-12 students are monitoring water chemistry and species diversity and abundance in cooperating federal wildlife refuges in areas near them. These shared monitoring activities link classes throughout the entire scope of the project. The spatially and temporally distributed data enables the teams to introduce interesting analyses and discussions across partner classes interacting through videoconferences. Each Fellow works intensively with two teachers in PREP and with a teacher from eastern Maine (Washington & Hancock Counties), western Maine (Madison), or southern Maine (Damariscotta, site of the University of Maine\u27s marine sciences laboratory). The power of Maine\u27s network of ATM classrooms, is being used to expand the Fellows\u27 role modeling and introduce Fellows to a variety of teaching styles. The broader impacts of the project include strengthened backgrounds in science and attendance at the Maine summer Science Camp for the cooperating teachers. The K-12 districts\u27 benefits include the enriched learning of their students and access to the equipment from microscopes to thermal cyclers that is necessary to meet the goals of the Learning Results, but which many districts lack. The University of Maine is benefiting from K-12 students who come to the University better prepared in science and is fulfilling its mission as a Land Grant/Sea Grant institution to serve both the state of Maine and the nation as a whole

Algae as nutritional and functional food sources: revisiting our understanding.

Global demand for macroalgal and microalgal foods is growing, and algae are increasingly being consumed for functional benefits beyond the traditional considerations of nutrition and health. There is substantial evidence for the health benefits of algal-derived food products, but there remain considerable challenges in quantifying these benefits, as well as possible adverse effects. First, there is a limited understanding of nutritional composition across algal species, geographical regions, and seasons, all of which can substantially affect their dietary value. The second issue is quantifying which fractions of algal foods are bioavailable to humans, and which factors influence how food constituents are released, ranging from food preparation through genetic differentiation in the gut microbiome. Third is understanding how algal nutritional and functional constituents interact in human metabolism. Superimposed considerations are the effects of harvesting, storage, and food processing techniques that can dramatically influence the potential nutritive value of algal-derived foods. We highlight this rapidly advancing area of algal science with a particular focus on the key research required to assess better the health benefits of an alga or algal product. There are rich opportunities for phycologists in this emerging field, requiring exciting new experimental and collaborative approaches.AGS & KEH thank the Biotechnology and Biological Sciences Research Council (BBSRC BB/1013164/1) of the UK for funding. The University of Dundee is a registered Scottish charity, No. SC015096. PP is supported by IDEALG in the frame of the stimuli program entitled “Investissements d’avenir, Biotechnologies-Bioressources” (ANR-10-BTBR-04-02). The open access fee was supported by NSF-OCE-1435021 (MLW), DIC project 1823-06 (MEC), Maine Sea Grant (NOAA) 5405971 (SHB), NSF #11A-1355457 to Maine EPSCoR at the University of Maine (SHB), and the listed funding to AGS and PP

Algae as nutritional and functional food sources: revisiting our understanding.

Global demand for macroalgal and microalgal foods is growing, and algae are increasingly being consumed for functional benefits beyond the traditional considerations of nutrition and health. There is substantial evidence for the health benefits of algal-derived food products, but there remain considerable challenges in quantifying these benefits, as well as possible adverse effects. First, there is a limited understanding of nutritional composition across algal species, geographical regions, and seasons, all of which can substantially affect their dietary value. The second issue is quantifying which fractions of algal foods are bioavailable to humans, and which factors influence how food constituents are released, ranging from food preparation through genetic differentiation in the gut microbiome. Third is understanding how algal nutritional and functional constituents interact in human metabolism. Superimposed considerations are the effects of harvesting, storage, and food processing techniques that can dramatically influence the potential nutritive value of algal-derived foods. We highlight this rapidly advancing area of algal science with a particular focus on the key research required to assess better the health benefits of an alga or algal product. There are rich opportunities for phycologists in this emerging field, requiring exciting new experimental and collaborative approaches.AGS & KEH thank the Biotechnology and Biological Sciences Research Council (BBSRC BB/1013164/1) of the UK for funding. The University of Dundee is a registered Scottish charity, No. SC015096. PP is supported by IDEALG in the frame of the stimuli program entitled “Investissements d’avenir, Biotechnologies-Bioressources” (ANR-10-BTBR-04-02). The open access fee was supported by NSF-OCE-1435021 (MLW), DIC project 1823-06 (MEC), Maine Sea Grant (NOAA) 5405971 (SHB), NSF #11A-1355457 to Maine EPSCoR at the University of Maine (SHB), and the listed funding to AGS and PP