ABSTRACTS 1

1 2 PALATABILITY AND CHEMICAL DEFENSES DINOFLAGELLATE GENOMICS: RESULTS OF MACROALGAE IN THE ANTARCTIC FROM AN EST APPROACH PENINSULA Bachvaroff, T. R.1,∗, Herman, E. M.2 & Amsler, C. D.1,∗, Amsler, M. O.1, McClintock, J. B.1, Delwiche, C. F.1 Iken, K. B.1, Hubbard, J. M.1 & Baker, W. J.2 1Cell Biology and Molecular Genetics, University of 1Department of Biology, University of Alabama at Maryland, College Park, MD 20742; 2Soybean Genomic Birmingham, Birmingham, AL 35294-1170; 2Department Improvement Laboratory, USDA/ARS, Beltsville, MD of Chemistry, University of South Florida, Tampa, FL 20705, USA 33620, USA Dinoflagellates are enigmatic protists with odd nu- We examined palatability of 37 species of nonen- clear features, interesting plastid gene arrangements crusting macroalgae from the Antarctic Peninsula. and a proclivity for endosymbiotic relationships. Rel- This represents approximately 30% of the entire atively little molecular work has been done on di- antarctic macroalgal flora and 75% of the 49 noflagellates, and only a handful of genes have been nonencrusting species we collected. Organic extracts characterized in these organisms. We have begun an from most species were also prepared and mixed Expressed Sequenced Tag (EST) project with the aim into artificial foods. We examined palatability using of collecting plastid targeted but nuclear encoded feeding bioassays with three common, macroalga- genes from peridinin-containing dinoflagellates. This consuming animals (an omnivorous antarctic rock- provides an opportunity to understand the integra- fish, Notothenia coriiceps; an omnivorous sea star, tion of endosymbiont genes into the host cell. Our se- Odontaster validus; and a herbivorous amphipod, Gon- quencing effort has produced about 1000 unique ESTs dogenia antarctica). Thallus pieces from 23 of 34 from a Gonyaulax polyedra cDNA library obtained from macroalgal species tested with the fish (68%) were David Morse. Of the 1535 total ESTs (72%, >400 bp), rejected. Of the 23 species rejected as thallus, or- 466 sequences were identifiable by BLAST with a bit ganic extracts of 16 were bioassayed using the fish score above 50. The most abundant transcript is the with 9 (56%) unpalatable. Thallus pieces from 21 of peridinin–chlorophyll-binding protein, which consti- 36 macroalgal species tested with the sea star (58%) tuted 3% of the clones. More recently, we began se- were rejected. Of the 21 species rejected as thal- quencing from an Amphidinium carterae cDNA library lus, organic extracts of 20 were bioassayed using the made in our laboratory. This provides insight into ex- sea stars and 14 (70%) were unpalatable. Overall, pression patterns and is a resource for other workers 28 of the 37 species assayed as thallus (76%) were in the field. Several clearly identifiable plastid genes rejected by either or both the fish and sea stars. have been found and preliminary analysis confirms The amphipod assay was not suitable for use with the chromophyte, and therefore, tertiary origin of the thallus but was utilized with organic extracts of 23 peridinin plastid. macroalgal species that were rejected as thallus by either or both the fish and sea stars. Of these, 14 (61%) of the species’ extracts were rejected by 3 the amphipods. Unpalatability was highest among ALGAL STIMULATION OF THE BERKELEY PIT the brown algae examined with only an ephemeral, LAKE SYSTEM ectocarpoid species not rejected as thallus out of Bartkowiak, B. & Mitman, G. G.∗ 10 species tested. Of the remaining nine brown Montana Tech University of Montana, 1300 West Park algal species, six of seven tested were also un- Street, Butte, MT 59701, USA palatable as extracts, including all the ecologi- cally dominant, perennial species in the area. We This study was conducted to evaluate four species conclude that unpalatability to herbivores is com- of algae (Chromulina freiburgensis Dofl., Chlorella ellip- mon in antarctic macroalgae and that chemical soidea Gerneck, C. vulgaris Beijerinck, and Chlamy- defenses may play an important role in the un- domonas acidophila Negoro) isolated from an acid pit palatability of many algal species (NSF OPP9814538, lake for their bioremediative potential. The Berkeley OPP9901076). Pit Lake system, located in Butte, MT, USA is an aban- 2 ABSTRACTS doned open-pit mine. As a result of acid rock drainage tinued agricultural and residential inputs of fertilizers the water in this pit is very acidic and contains high and pesticides will likely exacerbate the cyanobacterial concentrations of dissolved metals. First, optimal nu- dominance leading to further reductions in aquatic trient levels for each species of algae were determined. health and aesthetic values. Previous chemical treat- Levels of nutrients tested include: 5, 10, 15 mg N/L, ment and macrophyte removal have achieved limited and 0, 3, 6 mg P/L. The experiment was conducted success, and might have altered algal community dy- with sterile filtered Berkeley Pit water. Next, each namics. Remediation approaches that might improve species of algae were grown under optimal nutrient water quality include: reducing upstream inputs via levels and dissolved metal removal potentials were sediment traps or lagoons; reducing in-lake nutri- examined, using Inductively-Coupled Plasma–Atomic ents via sediment removal; reducing residential inputs Emission Spectrometry (ICP–AES). In addition, field via improved septic/sanitation systems; and shoreline sampling was preformed to gain a better understand- vegetation filter strips. ing of the microbial community within the pit. Finally, field parameters including, light, temperature, dis- solved oxygen, specific conductivity, redox potential, 5 and turbidity were examined to determine the actual conditions for microbial growth within the Berkeley DESMIDS AND DINOFLAGELLATES OF pit. ECUADOR Carty, S.∗ & Hall II J. D. Department of Biology, Heidelberg College, Tiffin, OH 4 44883, USA PHYTOPLANKTON COMMUNITY The freshwater flora of Ecuador is poorly known. COMPOSITION IN THE TRI-LAKES AREA OF This report provides preliminary information about CENTRAL WISCONSIN, USA the freshwater Pyrrhophyta and two families in the Bell, R. A. Chlorophyta. There has been some work on diatoms Department of Biology, University of Wisconsin – Stevens but other groups remain unstudied. In June 2001, Point, Stevens Point, WI 54481, USA samples were collected as whole water or by using a 10-mm plankton net from 52 locations in the inte- Tri-Lakes (Upper and Lower Camelot, Sherwood, Ar- rior (Amazonian) and Andean regions and examined rowhead) in Adams County, WI, USA are man-made within hours using a Swift field microscope. Nine ad- impoundments draining substantial agricultural lands ditional samples from Ecuador have been provided and surrounded by considerable shoreline residen- by Dr Miriam Kannan. Amazonian sites consisted of tial development. The planktonic algal community, as oxbow lakes along the Rio Shiripuno, and small ponds sampled from June to November 2000, was sparse-to- and lakes along the road from the river to Coca. An- moderately dense, fairly diverse (69 genera from six dean sites were principally in the Cajas National Park divisions basin-wide), and unremarkable in taxonomic near Cuenca. Genera in the Closteriaceae include composition. All sites sampled displayed the general Closterium, Gonatozygon, Spinoclosterium, and Penium. algal successional trends expected from northern- Desmidiaceae include Arthrodesmus, Bambusina, Cos- temperate, mildly eutrophic waters. These included marium, Desmidium, Euastrum, Hyalotheca, Micrasterias, sparse but taxonomically diverse communities in the Pleurotaenium, Spondylosum, Staurastrum, Staurodesmus, spring; a late spring pulse of diatoms; a late summer Triploceras, and Xanthidium. Pyrrhophyta include Cer- pulse of green algae; and a steadily increasing com- atium, Gymnodinium, Peridinium, Sphaerodinium, and ponent of Cyanobacteria leading to their community Woloszynskia. dominance by the end of the growing season. Upper Lake Camelot (55 genera) best represented this pat- tern. Lower Lake Camelot (53 genera) had a large green algal pulse but only a small diatom pulse. Lake 6 Sherwood was the most taxonomically diverse body PHORMIDIUM RETZII (OSCILLATORIALES): (63 genera) and had the most extreme pulses of di- GENOTYPIC VARIATION AND PHENOTYPIC atoms and greens. Lake Arrowhead had the lowest PLASTICITY taxonomic diversity (39 genera), was the most domi- Casamatta, D. A.1,∗, Vis, M. L.1 & Sheath, R. G.2 nated by Cyanobacteria, and had only minor pulses of 1Department of Environmental and Plant Biology, Ohio diatoms and greens. The algal communities indicate University, Athens, OH 45701; 2Provost Office, California a mesotrophic to slightly eutrophic lake status. Con- State University, San Marcos, CA 92096, USA ABSTRACTS 3

Phormidium retzii is one of the most commonly en- Life on earth originated in a wet world and remained countered lotic taxa in North America and has been there for millions of years until the first terrestrial in- recorded from a wide range of habitats worldwide. vaders made their initial forays into a new and un- The genetic variability of this cosmopolitan, fresh- exploited terrestrial habitat. Although cyanobacteria, water cyanobacterium was assessed using gene se- and perhaps fungi as well, were important participants quences and random amplified polymorphic DNA in the conquest of the land, it was the evolution green (RAPD) markers. Strains consistent with the mor- algae into the initial land plants that started amaz- phological species circumscription were utilized from ing evolution and diversification the terrestrial flora British Columbia, Canada to Rio Claro, Costa Rica, and fauna with which we are now familiar. Given the and from Rhode Island to Washington State, USA. paradigm that life arose in the sea, it is perhaps surpris- In addition, some strains were from geographically ing to find that freshwater green algae – not marine close (<25 miles apart) sites. The 16S rRNA gene was forms – were the successful conquerors, i.e. the algae sequenced for eight of the 12 strains. Sequence ho- that gave rise to the land plants. The first land colo- mology ranged from 90.95 to 98.71. The nine RAPD nizers were, of necessity, primary producers (i.e. food primers utilized yielded a total of 133 distinct bands for organisms that arrived or evolved later) and were from the 12 strains. The strains were variously related likely to have been cyanobacteria (blue-green algae), and showed great variation in pairwise genetic dis- followed by green algae (at least four separate inva- tances (0.23–0.83). Given the lack of sequence similar- sions) and fungi, which also made a successful assault ity, P.retzii as presently circumscribed most likely rep- on land together with algae in the form of lichens. The resents several cryptic species not clearly distinguish- symposium will survey recent studies on these groups able with light microscopy of morphological charac- and their habitats, and describe how these colonizers ters. This conclusion may explain the lack of correla- changed and were changed by adaptations to the new, tion between geographic proximity and genetic simi- dry world. It will explore the question: ‘Why did only larity. To assess the level of environmentally induced one group of terrestrial green algae give rise to the phenotypic plasticity of this taxon, one strain was land plants?’ The success of these colonizers is rele- subjected to three different current velocity and nu- vant to all subsequent terrestrial life (including our- trient regimes (low, medium, and high) employing selves), as well as the search for extraterrestrial life, six stream mesocosms. ANOVAs and Bonferroni mul- which, if it exists, is presumed to have begun in an tiple comparison tests were performed with cell vol- aquatic environment. umes, ratios of cell length : width and percent cover data. Low nutrient treatments elicited significantly (P < 0.05) greater cell volumes and percent cover 8 than did high treatments. Furthermore, current ve- locity did not show any significant effect on cell PHYLOGEOGRAPHY OF morphology. As such, it appears that a single strain BATRACHOSPERMUM HELMINTHOSUM (RHODOPHYTA) IN NORTH AMERICA is capable of wide morphological variability in re- ∗ sponse to some environmental conditions. The re- Chiasson, W. B., Machesky, N. J. & Vis, M. L. sults of these two studies have great implications Department of Environmental and Plant Biology, Ohio in the utilization of cyanobacterial morphospecies University, Athens, OH 45701, USA concepts. The freshwater red alga, Batrachospermum helmi- nthosum, is distributed primarily in streams of east- ern North America. Although it is often an abun- dant and conspicuous component of the algal flora 7 for many streams, little is known about the genetic re- GREEN ALGAL CONQUEST OF THE LAND: lationship among populations throughout its range. MANY CONQUESTS, ONE VICTORY? The purpose of our study was to elucidate geographic Chapman R. L.1,∗, Delwiche, C. F.2 & patterns among populations to gain insight into the McCourt, R. M.3 biogeographic distribution of this species and possi- 1Department of Biological Sciences, Louisiana State ble modes of dispersal. Individuals were sampled in University, Baton Rouge, LA 70803-4110; 2Department 13 stream segments as follows: four locations in Ohio, of Cell Biology and Molecular Genetics, University two locations in Michigan and one location each in of Maryland, College Park, MD 20742-5815; Indiana, North Carolina, Tennessee, Louisiana, 3Academy of Natural Sciences, Philadelphia, Rhode Island, Massachusetts and Connecticut. The PA 19103, USA mitochondrial spacer region (371 bp) between the 4 ABSTRACTS

COX2 and COX3 genes was chosen because it is vari- 10 able among individuals. This region was sequenced NEW SYSTEMATIC INSIGHTS IN THE for 15 individuals from each location. Thirteen hap- CERAMIUM SINICOLACOMPLEX : RESURRECTION lotypes were identified among the 13 locations with OF C. INTERRUPTUM S. & G. (CERAMIACEAE, most locations having one or two haplotypes. Two of RHODOPHYTA) the haplotypes were dominant being recorded from Cho, T. O.1,∗, Fredericq, S.2, Murray, S. N. & six of the streams. The Indiana, Louisiana and first Boo, S. M.3 Michigan population had a unique haplotype. The 1Department of Biology, University of Louisiana, Lafayette, second Michigan and fourth Ohio populations shared LA 70504, USA; 2Department of Biology, California State a unique haplotype. The Tennessee population had University, Fullerton, CA 92834, USA; 3Department of four haplotypes, three of which were closely related Biology, Chungnam National University, Daejon 305-764, but the fourth haplotype was not and linked to the Korea haplotypes from Michigan and Ohio. The relation- ships among these populations are very complex but Setchell & Gardner (1924) provided a taxonomic there may have been a recent dispersal event via birds treatment for the genus Ceramium from lower Califor- among various streams. nia and the Gulf of California, Mexico, in which they described several new species, including C. sinicola S. & G. described from Ensenada, BCN and C. interruptum S. & G. described from the vicinity of La Paz, BCS. The 9 latter was later reduced to variety rank in C. sinicolaby CHARACTERIZATION OF MACROALGAL Dawson (1950), a taxonomic decision that has subse- EPIPHYTES ON THALASSIA TESTUDINUM IN quently been widely adopted. Phylogenetic analyses TAMPA BAY, FLORIDA inferred from three molecular markers (chloroplast- Cho, T. O.1,∗, Fredericq, S.1 & Yates, K. K.2 encoded rbcL, the RUBISCO spacer, and nuclear 1Department of Biology, University of Louisiana at encoded SSU rDNA) from recent collections from Lafayette, Lafayette, LA 70504-2451; 2US Geological the Pacific North-west (California, Oregon) and the Survey 600, 4th Street South, St. Petersburg, FL 33701, Gulf of California reveal a well supported assem- USA blage of three corticated taxa: C. codicolaJ. Ag. 1894, C. sinicola and C. sinicola var. interruptum. Sequence di- Blooms of seagrass epiphytes have potentially impor- vergence values among the three taxa are sufficient tant economic and ecological consequences in Tampa to warrant separate species ranking; hence, we rein- Bay, one of the Gulf of Mexico’s largest estuaries. state C. interruptum as a widely distributed epiphyte As part of a Tampa Bay pilot study to monitor the for the region. C. sinicolais more closely related to impact of environmental stresses, precise character- C. codicolathan to C. interruptum, with the former two ization of epiphyte diversity is required for efficient restricted to the host Codium. The molecule-based re- management of affected resources, and thus may be lationships are congruent with evolutionary trends in used as a rational basis for assessment of ecosystem cortication pattern and attachment mode. health. Fixed monitoring sites were selected north of Port Manatee, composed of dense and sparse Tha- 11 lassia testudinum seagrass sites. A total of 13 epiphytic PHYTOPLANKTON OF A MISSISSIPPI RIVER species encompassing green, brown and red macroal- WATERSHED IN COASTAL LOUISIANA gae were manually collected in May 2001 from dense Ciugulea, I. I.1, Waters, D. A.2,∗, Day J. R. J. W.3 & seagrass beds versus nine species from sparse beds. Epi- Chapman, R. L.2 phytes only collected in the dense beds were Enteromor- 1Department of Botany and Microbiology, University of pha flexuosa, Sphacelaria rigidula, Ceramium byssoideum, Bucharest, Romania; 2Department of Biological Sciences, and Herposiphonia tenella; epiphytes only occurring 3Department of Oceanography and Coastal Sciences, in the sparse beds were Griffithsia and Stylonema al- Louisiana State University, Baton Rouge, LA, USA sidii. A correlation seems to emerge among attach- ment mode of epiphyte to host, presence of corti- In July–August 2001, phytoplankton samples were col- cation and epiphyte length. A main goal of this on- lected from a coastal marsh south of New Orleans, going study is the determination of indicator species LA. This wetland area is usually cut off from the Mis- for both healthy and stressed seagrass bed environ- sissippi River by levees and other flood control devices, ments using both taxonomic and gene-sequencing but periodically the Caernarvon Diversion is opened techniques. to allow fresh river water to flow into the marsh. As ABSTRACTS 5 part of a larger project, the present study seeks to 13 determine the affect the periodic input of freshwa- EFFECTS OF SAMPLING SCALE AND ANALYSIS ter has on phytoplankton dynamics in the Caernarvon METHOD ON PERCEPTIONS OF Diversion watershed. The first phase of the project - PHYTOPLANKTON SPECIES ASSOCIATIONS collecting and identifying algae during the summer Davis, B. E.1,∗, Interlandi, S. J.2, Kilham, S. S.2 & when the Caernarvon Diversion is kept closed - has Theriot, E. C.1 been completed. A total of 168 species from 76 genera 1Integrative Biology, University of Texas at Austin, Austin, representing 8 phyla were identified and digital im- TX 78712; 2School of Environmental Science, Engineering, ages recorded. Little or no previously published data and Policy, Drexel University, Philadelphia, PA 19104, on the taxonomy and ecology of phytoplankton from USA this area are available. Sampling in spring 2002, when the diversion is open, will allow comparative analy- Phytoplankton associations generally are identified by sis of phytoplankton composition and productivity. nonrandom co-occurrence of phytoplankton species, In a collaborative study, the phytoplankton data will and are considered to be a result of similar species be combined with information on nutrients, chloro- response to properties of the environment. Because phyll a, turbidity, salinity, etc. and used in a model to an association is characterized by co-occurrence of demonstrate how the ecosystem is affected by the di- species across samples, or synchrony of the abundance version. Also, a database of digital images is available at of species over time, we proposed that the percep- http://www.chapman.lsu.edu/digitalalgae. In addi- tion of phytoplankton association is subject to spa- tion to the taxonomic identification data and images, tial and temporal sampling scale and analysis method. the site will eventually include physico-chemical and We investigated this using an intensive phytoplankton GIS data on the collection sites. Supported in part by dataset from three lakes of the Greater Yellowstone a grant from the Office of Sea Grant Development at Ecosystem, with particular concentration on Asteri- LSU. onella formosa (AF) and Aulacoseira subarctica (AS). We compared numerical analyses, including Fager anal- ysis of recurrent groups, rank correlation, and other 12 measures of synchrony, for differences in assessment ORIGIN AND EARLY EVOLUTION OF PLANT of co-occurrence. The analyses were conducted on the BODY SYMMETRY AND GRAVITY RESPONSES entire dataset, and then the dataset was reduced to Cook, M. E.1,∗ & Graham, L. E.2 assess the effect of different spatial or temporal sam- 1Department of Biological Science, Illinois State University, pling regimes on apparent association. While the two Campus Box 4120, Normal, IL 61790-4120; 2Department diatom species significantly co-occurred across sam- of Botany, University of Wisconsin, 430 Lincoln Drive, ples, assessment of synchrony was affected by analysis Madison, WI 53706-1381, USA method and sampling regime. AF and AS abundance were positively correlated across lakes and within each Land plant bodies exhibit both apical–basal and radial lake, but peaks in abundance of the two species did not symmetry, and they are able to detect and respond significantly coincide over time. In addition, species to gravitational forces. These attributes were, likely abundance were negatively correlated across depths, important factors in the success of earliest plants on and we argue that this is significant to the autecology land. This study focuses on features of charophycean of each species. These results indicate that the percep- green algae likely to have been pre-adaptive to early tion of phytoplankton association may simply be an establishment of plant symmetry and gravitational re- artifact of the limits of our understanding of the dis- sponses, though most modern charophyceans occupy tribution and physiology of the organisms. aquatic habitats where the buoyancy of water counter- acts the effects of gravity. Trait mapping suggests that even the earliest-divergent modern members of the 14 streptophyte clade have bodies whose symmetry de- A GUIDE TO THE SEAWEEDS OF FLORIDA parts significantly from the spherical condition, and Dawes, C. J.1,∗ & Mathieson, A. C.2 that cellular mechanisms defining aspects of radial 1Department of Biology, University of South Florida, symmetry and polarized tip growth originated early. Tampa, FL 33620; 2Department of Plant Biology, Genes, cell biological approaches, and taxa are iden- University of New Hampshire, Durham, NH 03824, USA tified for which further exploration is likely to illu- minate early evolution of plant body symmetry and A review of Florida seaweeds indicates a highly di- gravity responses. verse flora within a large coastal zone (over 700 km) 6 ABSTRACTS spanning more than 7◦ of latitude, containing tropi- use of resources, anchor themselves effectively and in cal, subtropical, and temperate communities. Surpris- so doing stabilize the substrate and permit the for- ingly, there have been relatively few detailed floristic mation of modern soils, and maintain metabolic ac- studies of Florida seaweeds, except for the Florida Keys tivity for long periods of time even in the absence and on the east coast. The patterns are a reflection of of rainfall and surface moisture. Molecular phyloge- the early predominance of winter visitors and lack of netic analyses show the order Charales is the sister resident phycologists. The seaweeds of the west coast group to all land plants, with the Coleochaetales sis- of Florida were virtually ignored until the 1950s. We ter to the land plant/Charales lineage. Thus, in a estimate that the state’s seaweed flora consists of at very real sense, the embryophytes are ‘drier algae.’ least 670 taxa including 219 green, 97 brown, 349 red Many groups of green algae live in the terrestrial en- algae, and at least 3 species of Vaucheria. Such a compi- vironment, but only one of these – the land plants – lation is significantly higher than the numbers of taxa has radiated into a wide range of habitats. It is not recorded for Florida by Taylor (1960), as well as those known why the land plant lineage has been so suc- for North Carolina (i.e. Schneider & Searles, 1991), cessful. Traits that are likely to have played a role in and Caribbean reefs (Littler & Littler, 2000). The ma- their success include cell wall biochemistry, desicca- jority of Florida seaweeds have tropical/Caribbean tion resistance and tolerance, structural complexity, affinities, although a significant temperate compo- and various reproductive strategies. In all probability, nent also occurs, presumably reflecting the influence the success of the land plant lineage was not the result of the northern floras of the Gulf of Mexico and south- of a single ‘key innovation’, but an emergent prop- eastern coast of the US. The tropical nature of the erty resulting from complex interactions among these state’s flora is supported by the R/P and R + C/P in- and other features of the lineage. Comparative study dices of Feldman (1937) and Cheney (1997) with these of the properties of diverse aquatic and terrestrial al- ratios being 3.6 and 5.8, respectively. Florida’s sea- gae can be used to identify properties that were im- weed diversity is enhanced by the extensive salt marsh portant in the colonization of the land and how they and seagrass communities of the west coast, plus the interact. coral reef habitats of the Florida Keys. The present re- view summarizes several important historical studies, conspicuous geographic patterns, the type of commu- nities, and the structure of a guide to the seaweeds 16 of Florida. The guide will also serve as a historical DIVERSITY OF FUSIFORM reference for possible anthropogenic impacts in the MICROCHLOROPHYTE ALGAE FROM ITASCA future. STATE PARK, MINNESOTA Dimmer, S. K.∗, Dean, M., Fawley, M. W. & Fawley, K. P. Department of Biological Sciences, North Dakota State 15 University, Fargo, ND 58105, USA ONE SMALL STEP: WHY DID THE CHAROPHYTES HAVE THE RIGHT STUFF? Through the use of molecular techniques, several Delwiche, C. F.1,∗, Karol, K. G.1 & McCourt, R. M.2 different types of fusiform and twisted-fusiform mi- 1Cell Biology and Molecular Genetics, University of crochlorophyte algae (Monoraphidium, Ankistrodesmus, Maryland, College Park, MD 20742; 2Department of etc.) have been characterized from samples taken Botany, Academy of Natural Sciences, Philadelphia, PA from lakes and ponds of Itasca State Park, MN. PCR– 19103, USA RFLP analysis of 18S ribosomal RNA genes was used to categorize 29 different isolates from the Itasca State The origin and evolution of land plants (i.e. em- Park lakes into nine different types. The 18S riboso- bryophytes) represents one of the seminal events in mal DNA sequences were determined for one isolate the history of life on earth. When the land plant lin- from each type for phylogenetic analysis. Light mi- eage took hold upon the continents, there were pro- croscopy was used so that morphological character- found changes in the global environment including istics as well as molecular characteristics can be com- dramatic modification of the erosion regime, corre- pared and contrasted. The morphological characteris- lated changes in marine nutrient inputs, and striking tics were consistent for each PCR–RFLP type. Further fluctuations in atmospheric chemistry. These changes discussion will include comparisons of the distribution took place not only because of the ability of land and diversity of these organisms from Itasca State Park plants to survive on land, but also because their struc- to those from Arrowwood National Wildlife Refuge, ture and physiology permitted them to make efficient ND. ABSTRACTS 7

17 land use, trophic change, and sedimentation using a whole-basin approach to reconstruct loading history A TOTAL EVIDENCE APPROACH TO INFERRING of nutrients, sediments, heavy metals, and organics. THE PHYLOGENETIC RELATIONSHIPS WITHIN Dating chronologies based on 210Pb inventories indi- AULACOSEIRA (BACILLARIOPHYTA) ∗ cated both cores recovered a sediment sequence dat- Edgar, S. & Theriot, E. ing from pre- and post-European settlement (c. 1850) Section of Integrative Biology, University of Texas at in the St. Croix River basin. Select cores were sub- Austin, Austin, TX 78713, USA jected to magnetic susceptibility, loss-on-ignition, and Phylogenies provide the context in which hypothe- diatom microfossil analysis. Sedimentary increases in ses about character evolution can be tested. But the magnetic susceptibility were indicative of increased phylogenetic understanding of many diatom taxa, in- erosion and transport of ferromagnetic mineral grains cluding some of the more spatially and temporally due to initiation of settlement, logging, and agricul- ubiquitous genera, is insufficient to fruitfully pursue tural activities in the basin. A three-fold increase in questions about character evolution within them. Con- sediment accumulation began in the mid-1800s in the sequently, we have estimated the phylogeny of Aula- northern basin and by 1900 in the southern basin. coseira using a combination of nucleotide sequence Diatom accumulation increased 20–50-fold since set- data and data derived from morphological charac- tlement with a shift from benthic- to planktonic- teristics of the silica cell wall. In addition to the use dominated assemblages. Simultaneous with the assem- of traditional ‘discrete’ or ‘qualitative’ morphologi- blage shift were the introduction and establishment of cal characters, we have assessed the use in our cladis- many planktonic diatoms considered ubiquitous indi- tic analyses of ‘continuous’ or ‘quantitative’ charac- cators of eutrophy. The fossil diatom assemblages were ters, including descriptors of the ontogenetic trajec- further analyzed using weighted-averaging calibration tories of these characters over a species’ life cycle (size and reconstruction of historical water column total range). Also, molecular characters gathered from a phosphorus (TP). Reconstructed TP values showed chloroplast genome marker (rbcL) as well as a nuclear that water column nutrient values have increased 2.5– genome marker (18S rRNA) were used simultaneously 3-fold since presettlement times. Presettlement values in the cladistic estimation of the phylogeny of approxi- of about 0.02 mg/L TP were found in both cores with mately 12 species of Aulacoseira. In order to have more TP increases beginning c. 1910 and especially dra- thorough taxon sampling for a better estimate of phy- matic increases after World War II. Modern recon- logeny, we included several fossil species, as well as structed TP values (about 0.055 mg/L) were similar taxa where cultures were unavailable, by utilizing the to TP concentrations reported from monitoring dur- morphologically based characters despite the absence ing the last few decades; however, the river was clearly of molecular characters for these taxa. impacted well before monitoring efforts were begun.

18 19 POST-EUROPEAN SEDIMENTATION AND PHOSPHATASE EXPRESSION BY CHLORELLA NUTRIENT LOADING IN LAKE ST. CROIX: A VULGARIS (CHLOROPHYCEAE) IS MEDIATED BY NATURAL IMPOUNDMENT ON THE ST. CROIX INTERNAL PHOSPHORUS LEVELS AND RIVER, USA EXTERNAL pH Edlund, M. B.1,∗, Triplett, L.1,2 & Engstrom, D. R.1 Evens, T. J.∗ & Koenig, R. 1St. Croix Watershed Research Station, Science Museum of US Department of Agriculture, Agricultural Research Minnesota, 16910, 152nd St. N., Marine on St. Croix, Service, Southern Regional Research Center, New Orleans, MT 55047; 2Department of Geology, Limnological LA 70124, USA Research Center, University of Minnesota, Minneapolis, MT 55455, USA Phosphorus has been implicated as one of the causative agents of the eutrophication of lacustrine, The final 37 km of the St. Croix River were natu- estuarine and coastal waters. One of the major inputs rally dammed by the Mississippi River to form Lake of phosphorus into these waters comes from agricul- St. Croix approximately 9500 years ago. The St. Croix tural nonpoint sources in the form of dissolved or- River is currently regarded as having ‘exceptional ganic phosphorus (DOP). In order to better under- resource value’, as one of the least impacted large stand how DOP may directly impact the microalgal Midwest river systems. Twenty-four 2-m piston cores community structure, it is necessary to understand the were recovered in 1999–2001 from Lake St. Croix sub- physiology of DOP uptake and utilization. To this end, basins to identify post-European settlement signals of cultures of Chlorella vulgaris were grown in custom pho- 8 ABSTRACTS tobioreactors in acid (pH 5.5) and alkaline (pH 7.5) Very little is known about the diversity and distribu- media under phosphate replete and starved condi- tion of freshwater unicellular coccoid green algae. tions in order to determine the types and triggers of The simple morphology and small size of these or- phosphatase expression by this organism. Analysis of ganisms make reliable identification extremely diffi- differential phosphatase expression during phosphate cult. Current hypotheses suggest that species of coc- stress indicates that cultures of C. vulgaris grown under coid algae are widely distributed, opportunistic organ- alkaline conditions derepressibly express two alkaline isms that are always present in aquatic systems. These phosphatases, while cultures grown under acid con- hypotheses are based on the assumption that phy- ditions weakly express only one alkaline phosphatase. toplankton communities include only one or a few Analysis of internal and external phosphorus levels, species of coccoid algae. We examined the diversity whole-cell phosphatase activity and in-gel activity of and distribution of green-colored autosporic coccoid protein extracts indicate that alkaline phosphatase ex- algae in Itasca State Park in Minnesota and Arrow- pression is further mediated by internal, as opposed wood National Wildlife Refuge in North Dakota us- to external, phosphorus levels. ing molecular techniques and light microscopy. Our preliminary results indicate a higher level of diversity of coccoid green algae than has been reported in re- 20 gional flora. Contrary to current hypotheses, commu- BIODIVERSITY AND ASSOCIATION OF nities of coccoid green algae vary among the different DINOFLAGELLATES IN CORAL REEF RUBBLE, lakes. CARRIE BOW CAY, BELIZE Faust, M. A. Smithsonian Institution, 1210 Silver Hill Road, Suitland, MD 20746, USA 22 The species diversity and distribution of benthic di- DIVERSITY OF COCCOID ALGAE IN noflagellates are illustrated from the back reef sand MINNESOTA AND NORTH DAKOTA LAKES ∗ habitats at Carrie Bow Cay. Sand appears an ideal en- Fawley, M. W. , Fawley, K. P. & Hoffman, M. J. vironment for nurturing a variety of species. Sand sup- Department of Biological Sciences, North Dakota State ported blooms of toxic, nontoxic, and parasitic assem- University, Fargo, ND 58105, USA blages of dinoflagellates. Morphology of dinoflagel- lates is illustrated in SEM and LM pictures; including The coccoid algae (a.k.a ‘little green balls’) are im- the life cycle of a parasitic dinoflagellate’s vegetative portant primary producers in freshwater communi- stage, sporogenetic stages, and the morphology of the ties, but very little is known about their diversity. feeding organelle. The feeding organelle of the pri- This lack of knowledge results from the extremely mary dinoflagellate cysts is a hold fast and a peduncle- small size and simple morphology of these organisms, like infestation tube, a ‘sucker organelle’, that inserts which makes identification by light microscopy es- itself into copepod eggs. The parasitic dinoflagellate sentially impossible. However, molecular techniques continues feeding on crustacean eggs since it is the allow differentiation of taxa at various levels and only food source during vegetative cell division within can ultimately lead to species identification. We have the cyst. This is the first observation of the presence isolated several hundred cultures of coccoid algae of parasitic dinoflagellates in shallow coral reef wa- from lakes and ponds in Itasca State Park in north- ter. The study attempts to provide new knowledge on western Minnesota and Arrowwood National Wildlife dinoflagellate associations and morphology of sand- Refuge in central North Dakota. Many of these isolates welling species in the microscopic food web of shallow have been examined by light microscopy, 18S rDNA warm tropical waters. Even, sand-inhabiting inverte- PCR–RFLP analysis, and 18S rDNA sequence analy- brates are not exempt from parasitic dinoflagellates. sis. In addition, some taxa with multiple isolates have been examined by rbcL and ribosomal RNA ITS se- quence analysis. Our results indicate a high level of 21 diversity, including possible new species of Chlorella, DIFFERENCES AMONG COCCOID GREEN Muriella, Pseudomuriella and Mychonastes, among oth- ALGAL COMMUNITIES IN MINNESOTA AND ers. We will discuss the results of our analyses, includ- NORTH DAKOTA LAKES ing implications for identification of taxa and higher Fawley, P. K.∗ & Fawley, M. W. level systematics. Supported by National Science Department of Biological Sciences, North Dakota State Foundation Grants DBI-00703867, MCB-0084188 and University, Fargo, ND 58105, USA DEB-0128952. ABSTRACTS 9

23 We investigated whether sexual and apomictic fronds ASSESSING THE BIODIVERSITY OF differed in their spatial distribution by sampling for MONORAPHIDIUM USING 18S rDNA vertical differences in intertidal height and horizontal SEQUENCES patchiness. Fronds were sampled when reproductive Fawley, M. W.1, Fawley, K. P.1 & Dean, M. L.2,∗ and cultured for life history. We found sexual vari- 1Department of Biological Sciences, North Dakota State ants were significantly aggregated at lower intertidal University, Fargo, ND 58105; 28715 W. Mallard Ct., heights while apomicts were found throughout the in- Franklin, WI 53132, USA tertidal range. Sexual fronds were strongly seasonal and reproduced October–December whereas apomic- The taxonomy of the genus Monoraphidium is unclear tic fronds reproduced throughout the year. These re- due in part to the absence of morphological features sults parallel the large-scale geographic distribution to clearly distinguish one species from another. Phy- with apomicts inhabiting marginal areas, and sug- toplankton samples collected from lakes in the Ar- gest dispersal capability may also facilitate geographic rowwood National Refuge in eastern North Dakota parthenogenesis. were found to contain several morphological species of Monoraphidium. Eighteen Monoraphidium isolates 25 were examined with light microscopy and six morpho- MACROALGAE OF THE HAWAIIAN ISLANDS: 23 logical species were identified. PCR–RFLP of the 18S ADDITIONAL STREAM SEGMENTS rDNA was used to type the isolates. Following digestion Filkin, N. R.1,∗, Sherwood, A. R.2 & Vis, M. L.1 by Hae III and Taq I, the 18S rDNA PCR–RFLP patterns 1Department of Environmental and Plant Biology, Ohio indicated 10 different types. Presently, the 18S rDNA University, Athens, OH 45701; 2Department of Botany, product is being sequenced for each of the 10 types. By University of Hawaii, Manoa, HI, USA examining morphological characters and 18S rDNA sequences, congruence between morphology and se- The Hawaiian Islands are located 3700 km from the quence data may be compared. Also, because there nearest continental land mass and as such provide is a lack of morphological characters defining Mono- an interesting research opportunity to study fresh- raphidium species, diversity within the 18S rDNA se- water organisms intolerant of seawater. The freshwa- quences may aid in the taxonomy of the genus and ter biota have been only partially studied with little its place within the Chlorococcales. Supported by Na- research on the stream macroalgae. The only previ- tional Science Foundation Grants MCB-0084188 and ous study to focus solely on stream macroalgae, re- DBI-0070387. ported 25 new species to the Hawaiian Islands from 34 stream segments sampled. The purpose of our study was to expand the sampling of streams to better 24 determine macroalgal distributions and species rich- SPATIAL AND TEMPORAL DISTRIBUTION OF ness. Twenty-three additional stream segments (seven LIFE HISTORY VARIANTS IN THE RED ALGA, on Oahu, eight on Kauai and eight on Hawaii) were MASTOCARPUS PAPILLATUS sampled for macroalgae. The physical and chemical Fierst, J. L.∗, Kubler, J. & Dudgeon, S. R. parameters of each stream were measured. Stream California State University Northridge, 18111 Nordhoff segments ranged greatly in size from 1.2 to 40 m St., Northridge, CA 91330, USA in width. Water temperature was relatively uniform (about 21 ◦C) but other chemical parameters differed We investigated small-scale spatial structure in Masto- from site to site (pH 5.5–8.9, specific conductance 20– carpus papillatus, a red alga with sexual and apomictic 200 mS/cm). Mean species richness per stream seg- life-history variants. Spatial structure and habitat par- ment was 3.9 with one to eight species collected per titioning can facilitate the coexistence of ecologically segment. Ninety populations of 44 infrageneric taxa similar organisms, and in M. papillatus a large scale were identified from the Cyanobacteria (20), Chloro- geographic spatial division is well documented. Vari- phyta (18), Rhodophyta (3) and Chrysophyta (3). The ants of M. papillatus display geographic parthenogen- most abundant taxa were Spirogyra sp. 2, Audouinella esis where sexual fronds are common south of San pygmaea and Phormidium retzii. All three of these species Francisco Bay and apomicts dominate north of San are cosmopolitan. Twenty-two of the taxa are new Francisco Bay. Although geographic parthenogenesis records for streams in the Hawaiian Islands. The large in M. papillatus is well-documented small-scale spatial percentage (50%) of new taxa suggests that more re- structure was not previously known due to the similar search is needed to fully catalog the Hawaiian stream appearance of sexual and apomictic upright fronds. macroalgal diversity. 10 ABSTRACTS

26 compare the effects of different radiation sources or THE ECOLOGICAL SIGNIFICANCE OF various ozone depletion scenarios. However, no data MYCOSPORINE-LIKE AMINO ACIDS IN ALGAE are available from which BWFs for partitioning can be Franklin, L. A. determined, as experimental techniques have varied Smithsonian Environmental Research Center, P.O. Box 28, widely. These BWFs are essential for predicting the ef- Edgewater, MD 21037, USA fects of increased UV-B on other trophic levels, and for analyzing the cost and benefit of synthesizing pro- The links among mycosporine-like amino acids tective compounds. Using a polychromatic approach, 14 (MAAs), algal physiological ecology, and the envi- we are developing BWFs for the partitioning of C 3 ronment may be viewed from two sides. The UV- in Thalassiosira pseudonana ( H) at photosynthetically absorption spectra of MAAs make them prime candi- saturating irradiance. We are comparing these func- dates for UV-screening compounds, and indeed, nu- tions to BWFs calculated from simultaneous measure- merous studies have shown that a high concentration ments of primary productivity. We have found consis- of MAAs is correlated with various degrees of protec- tent differences in sensitivity to UV exposure among tion of cellular processes from UV damage. Thus, it the pools, and are testing these differences for signif- might be said that the structure of terrestrial, eulit- icance using a variety of model and calculation ap- toral and sublittoral algal communities depends in proaches. part on their ability to synthesize effective quantities of MAAs. Corollaries to this hypothesis might include the proposition that those species incapable of syn- thesizing MAAs will be excluded from UV-containing 28 environments, and that UV is necessary to trigger MAA A SURVEY OF THE OFFSHORE MARINE synthesis. However, a number of studies have demon- MACROALGAE FROM THE NORTH-WESTERN strated that triggers of MAA synthesis need not in- GULF OF MEXICO HARD BANK COMMUNITIES clude exposure to UV, but may include the relative Fredericq, S.1,∗, Cho, T. O.1, Gavio, B.1, Gurgel, C. availability of nitrogen, osmotic stress, or high levels F.1, Hickerson, E.2, Lin, S. M.1, Lopez-Bautista, J.1, of photosynthetically active radiation. Furthermore, Phillips, N.1, Viguerie, M. & Wysor, B.1 the response to these factors may vary as a function of 1Department of Biology, University of Louisiana at other environmental conditions. These issues will be Lafayette, Lafayette, P.O. Box 42451, LA 70504-2451; discussed with respect to elucidating the master switch 2Flower Garden Banks National Marine Sanctuary, Bryan, for MAA synthesis. TX 77803, USA

The upper north-western Gulf basin is characterized 27 by a relatively shallow but well-developed continen- BIOLOGICAL WEIGHTING FUNCTIONS FOR tal shelf with an extensive system of mid- and outer THE EFFECT OF UV RADIATION ON CARBON shelf hard banks formed on unique tectonically cre- PARTITIONING IN MICROALGAE ated salt diapirs of varying origin and composition. Franklin, L. A.∗ & Neale, P. J. Observations done by our laboratory while partici- Smithsonian Environmental Research Center, P.O. Box 28, pating in monitoring cruises to the National Marine Edgewater, MD 21037, USA Sanctuaries of the E and W Banks of the Flower Gar- dens (TX), Stetson Banks (TX), Sonnier Banks (LA) UV radiation clearly inhibits phytoplankton photosyn- and dredging cruises off Louisiana reveal a surpris- thesis, with direct effects on both photosystem II and ingly different floristic composition among the sites. the enzyme, Rubisco, that catalyses the first reaction of Several range extensions, new macroalgal records for the photosynthetic carbon reduction cycle. The subse- the Gulf of Mexico, and new species imply a much quent partitioning of fixed carbon into macromolec- more complex algal community than previously doc- ular pools (carbohydrates, lipids, and proteins) is also umented. Our ongoing project includes the develop- affected by UV exposure, although relative changes in ment of a modern taxonomic, phylogenetic and mor- allocation are not well-documented and vary consider- phological database essential to informed manage- ably. The wavelength specificity of inhibition of pho- ment for preservation of biological diversity. It also tosynthesis has been described by spectral weighting provides a framework of seasonal algal composition functions, or biological weighting functions (BWFs) against which long-term trends and anomalies in algal that combine the effect of UV radiation with interac- distribution and health of the Gulf of Mexico can be tive effects of longer wavelengths. BWFs permit one to assessed at future dates. ABSTRACTS 11

29 the structure of Mediterranean marine ecosystems fol- NEW INSIGHTS IN THE SYSTEMATICS OF THE lowing its 1984 invasion. The southern California inoc- PHYLLOPHORACEAE (GIGARTINALES, ulation of C. taxifolia is believed to have resulted from RHODOPHYTA) the release of aquarium specimens. Besides C. taxifolia, Fredericq, S.1,∗, Lopez-Bautista, J. M.1 & Moe, R. L.2 other species of Caulerpa being sold for aquarium 1Department of Biology, University of Louisiana at use also may have the potential to invade temper- Lafayette, Lafayette, LA 70504-2451; 2University ate waters. As a first step towards making this deter- Herbarium, VLSB 2465, University of California, Berkeley, mination, the availability (% frequency) of Caulerpa CA 94720-2465, USA species being sold in southern California for aquar- ium use was ascertained. Fifty retail saltwater aquar- The Phyllophoraceae, consisting of about 100 species ium stores were visited in three southern California worldwide, stands out in exhibiting a wide spectrum of counties between November 2000 and August 2001. unique life history types that makes it unusually inter- Sixteen Caulerpa taxa were identified from the retail esting for assessing the phylogenetic importance of re- aquarium outlets. Caulerpa species were sold in 52% productive traits relative to classification criteria. Type of these stores. C. taxifolia, ‘Mediterranean form’, was of life history and position of the reproductive struc- offered for sale in 10% of the visited stores; C. serrulata tures on the thallus have traditionally formed the basis var. hummii (18%), C. racemosa (14%), and C. racemosa for separating 11 genera in the Phyllophoraceae; how- var. lamourouxii (14%) were the most commonly sold ever, phylogenetic analyses inferred from three sets species. These data indicate that the aquarium indus- of DNA sequences [chloroplast-encoded rbcL, nuclear try is bringing into the region other species of Caulerpa large-subunit ribosomal RNA gene (LSU rDNA), and besides C. taxifolia. Some of these species may also have internal transcribed spacer regions (ITS) of nuclear the potential to invade temperate waters. ribosomal DNA], instead indicate a lack of correla- tion between type of life history and phylogenetic rela- tionships among the established taxa. This lack of cor- 31 relation dramatically challenges all of the traditional PHYLOGENETIC ANALYSES OF NUCLEAR 18S taxonomy, and we will present a revised classification rRNA GENE SEQUENCES INDICATE THAT THE for the family that downplays life-history features. The MASTOPHOROIDEAE (CORALLINALES, study will answer the question which morphological RHODOPHYTA) IS A POLYPHYLETIC TAXON features can be used as meaningful indicators of phy- Gabel, J.∗, Bailey, J. C. & Freshwater, D. W. logenetic relationships in the Phyllophoraceae. The Department of Biological Sciences and Center for Marine results will also be addressed in light of global biogeo- Science, UNC-Wilmington, 601 S. College Road, graphic hypotheses for the family. Wilmington, NC 28403, USA

Nuclear small subunit ribosomal RNA (18S rRNA) 30 gene sequences were determined for six species THE DIVERSITY AND AVAILABILITY OF representing three genera classified within the CAULERPA SPECIES FOUND IN RETAIL Mastophoroideae and also for two species of Amphi- AQUARIUM OUTLETS IN SOUTHERN roa (Lithophylloideae). These data were combined CALIFORNIA, USA with previously published 18S rRNA sequences for 37 Frisch, S. M.∗ & Murray, S. N. other coralline species and analyzed to: (1) determine Department of Biological Science, California State the phylogenetic position of the Mastophoroideae University, Fullerton, CA 92834, USA within the Corallinales; and (2) examine relationships among Hydrolithon, Metamastophora, Neogoniolithon, Biological invasions are recognized as a serious threat and Spongites. Trees derived from parsimony and maxi- to marine biodiversity. Within the last 2 years, inva- mum likelihood analyses of these data indicate that the sive populations of Caulerpa taxifolia were found in Mastophoroideae is not monophyletic. Instead, our southern California’s coastal waters as well as in New data suggest that group is polyphyletic and includes South Wales, Australia. In addition, two other sea- species belonging to three (or more) distinct evo- weeds (Undaria pinnatifida and Caulacanthus ustulatus) lutionary lineages. The nongeniculate genus Neogo- also appear to have recently invaded southern Cali- niolithon is resolved as sister to the geniculate sub- fornia’s warm temperate waters. The introduction of family Corallinoideae. Hydrolithon is resolved as sis- C. taxifolia has attracted much attention because this ter to the geniculate genus Metagoniolithon, an Aus- exotic seaweed is thought to have significantly altered tralian endemic. In turn, Spongites is positioned at the 12 ABSTRACTS base of a clade including Hydrolithon/Metagoniolithon ales, Rhodymeniales and Nemastomatales will be il- as well as species classified in the Lithophylloideae. lustrated. Finally, Metamastophora is placed in a well supported, but isolated, position as sister to all other taxa bear- ing uniporate tetrasporangial conceptacles. Our re- 33 sults imply that some reproductive and vegetative char- GRATELOUPIA TURUTURU (HALYMENIACEAE, acters among the suite of features used to delimit RHODOPHYTA): THE CORRECT IDENTITY OF the Mastophoroideae were acquired independently THE INVASIVE SPECIES IN THE ATLANTIC and are not homologous. Significantly, our results KNOWN AS GRATELOUPIA DORYPHORA AS suggest that genicula are nonhomologous structures INFERRED FROM MOLECULAR AND that evolved independently in four separate coralline MORPHOLOGICAL EVIDENCE ∗ taxa and from four nongeniculate ancestors belong- Gavio, B. & Fredericq, S. ing to distinct evolutionary lineages. On the basis of Department of Biology, University of Louisiana at Lafayette, our findings Porolithon pachydermum is transferred to P.O. Box 42451, Lafayette, LA 70504-2451, USA Hydrolithon as H. pachydermum (Foslie). Grateloupia doryphora (Montagne) Howe, originally de- scribed from Peru, has repeatedly been reported as an 32 invasive species in Atlantic and Mediterranean waters. NEW INSIGHTS IN THE Various attempts to explain this species’ route of in- CRYPTONEMIALES–RHODYMENIALES vasion have, so far, been unsatisfying. In the present COMPLEX AND RESURRECTION OF THE paper, we provide new evidence on the basis of com- ALLIED RED ALGAL ORDER NEMASTOMATALES parative rbcL sequence analysis and morphology that KYLIN 1925 AS INFERRED FROM rbcL this invasive species in the NE and NW Atlantic cor- SEQUENCE ANALYSIS AND COMPARATIVE responds to G. turuturu Yamada, originally described REPRODUCTIVE MORPHOLOGY from Japan. The true origin of this alga follows a well- Gavio, B. recognized trend of invasive marine organisms that Department of Biology, University of Louisiana at Lafayette, have on various occasions colonized the Atlantic and P.O. Box 42451, Lafayette, LA 70504-2451, USA Mediterranean Sea from Pacific NE Asia.

In a global molecular phylogeny of florideophycean red algae inferred from chloroplast-encoded rbcL se- 34 quence analysis, a major monophyletic assemblage EFFECTS OF OUTGROUP SELECTION ON comprises the Cryptonemiales (=Halymeniales), the PHYLOGENETIC HYPOTHESES OF THE Rhodymeniales, the Schizymeniaceae (Schizymenia, Ti- HETEROKONT ALGAE tanophora, Platoma) and the Nemastomataceae (Nemas- Goertzen, L. R.1,∗ & Theriot, E. C.2 toma, Predaea). The phylogenetic significance of the 1Department of Biology, Indiana University, Bloomington, auxiliary cell and its interaction with the fertilized egg IN 47405; 2Texas Memorial Museum, 2400 Trinity St., cell in this assemblage is discussed in relation to estab- University of Texas, Austin, TX 78705, USA lished and newly proposed classification schemes. The order Nemastomatales Kylin 1925 is reinstated and Nuclear ribosomal SSU and chloroplast rbcL data for emended to contain the nonprocarpic Schizymeni- heterokont algae and potential outgroups were ana- aceae and Nemastomataceae. Unifying characters of lyzed separately and together under parsimony. A se- the Nemastomatales include fertilized carpogonia that ries of taxon sampling experiments were performed. may establish fusions with carpogonial nutritive cells Traditional heterokont classes (e.g. diatoms, phaeo- prior to the formation of septate connecting filaments, phytes, etc.) were monophyletic in most analyses of and simple gonimoblasts developing outwardly from either dataset and in both together. Relationships auxiliary cells or from connecting filaments in their among classes and of heterokonts to outgroup taxa vicinity. The auxiliary cell is a transformed vegeta- are sensitive to taxonomic sampling. Stability of het- tive intercalary cell (Sebdeniaceae), that becomes sur- erokont algae relationships was analyzed by deletion of rounded by either clusters of nutritive cells (Nemas- outgroup classes, starting with those presumably most tomataceae), involucral filaments (Schizymeniaceae) distant from the heterokonts to those closest. In only or by three-dimensional ampullary filaments (Haly- three analyses using SSU data were the heterokonts meniaceae including the Corynomorphaceae), or is monophyletic: with all available outgroup taxa, with part of a procarp (Rhodymeniales). The homology of the outgroup being hyphochytrids alone or with the outward gonimoblast initiation and maturation into outgroup being hyphochytrids plus Developayella. For a simple ball of carposporangia in the Cryptonemi- rbcL data, heterokonts were monophyletic when all ABSTRACTS 13 available rhodophytes were used, when any combi- was found between bacterial and phytoplankton diver- nation of rhodophytes tested were used, and when sity (Shannon–Weaver Index) in this bog, suggesting rhodophytes plus bacteria were used. Any addition of the two microbial communities are linked. The proto- cryptophyte and/or haptophyte sequence resulted in plankton, however, were relatively uniform through- a nonmonophyletic heterokont algae. In both cases, out the ice-free period. The ice-covered period may distribution of heterotrophic stramenopiles among be treated as a fifth phase. When snow accumulates the heterokont algae changed as different outgroups over the ice-covered bog, light penetration is blocked were used. Combined data was similarly equivocal and photosynthesis shut off. Heterotrophic protozoa about monophyly of the heterokont algae. The litera- assume dominance in the plankton. ture suggests that this result is obtained regardless of whether parsimony or likelihood methods are used, 36 and that is because taxon sampling affects all inference procedures. We suggest not only that more molecu- PHYLOGENY OF THE GRACILARIACEAE lar data be obtained, but that morphological evidence (GRACILARIALES, RHODOPHYTA) INFERRED for monophyly of the heterokonts be re-examined. Fi- FROM rbcL SEQUENCE ANALYSIS: TAXONOMIC IMPLICATIONS nally, a community effort needs to be made to syn- ∗ chronize character sampling across different datasets Gurgel, C. F. D. to increase the number of taxa sampled and to create Department Botanica,ˆ CCS, Institute de Biologia, parallel datasets for all data types. Universidade Federal do Rio de Janeiro, Ilha do Fundao,ˆ Rio de Janeiro 21941-900, RJ, Brazil; Department of Biology, University of Louisiana at Lafayette, P.O. Box 35 42451, Lafayette, LA 70504-2451, USA SPECIES DIVERSITY OF MICROBIAL COMMUNITIES IN A NORTHERN TEMPERATE This research promotes the correlation of molecu- HUMIC LAKE lar phylogenies and morphological data with bio- Graham, J. M.∗, Kent, A. D., Lauster, G. H., geographic hypotheses for the economically impor- Yannarell, A. C., Graham, L. E., Kratz, T. K. & tant agarophyte family Gracilariaceae. The generic Triplett, E. W. concepts are evaluated on the basis of chloroplast- Limnology Laboratory, Center for Limnology, University of encoded rbcL sequences from 150+ specimens world- Wisconsin, 680 N. Park St., Madison, WI 53706-1492, wide. The results confirm the monophyly of the fam- USA ily and identifies three large clades, one of which corresponds to the ancestral, antiboreal genera Cur- Species richness and species diversity of bacterioplank- diea and Melanthalia, one to Gracilariopsis, and one ton, phytoplankton and protoplankton were exam- to Gracilaria sensu lato which identifies at least nine ined in Crystal Bog, a humic lake in northern Wiscon- distinct independent evolutionary lineages, includ- sin, as part of an NSF sponsored Microbial Observa- ing Hydropuntia. The species currently attributed to tory Program. Crystal Bog is part of the North Temper- Hydropuntia comprise two well-supported clades with ate Lakes Long-Term Ecological Research site. Auto- different biogeographic patterns, one composed of mated ribosomal intergenic spacer analysis (ARISA) Indo-Pacific species and the other of atlantic species. was used to assess bacterial diversity. Phytoplankton At least the three most basal clades within Gracilaria and protoplankton were enumerated in a settling sensu lato deserve to be considered distinct genera: chamber with an inverted microscope. Four distinct (1) a new genus centered around G. chilensis and phases in the ice-free season of the bog can be rec- G. tenuistipitata; (2) Hydropuntia sensu stricto encom- ognized based on microbial populations. The first passing pacific species (G. urvilleii, G. eucheumatoides, phase extends from ice-out until about the first week G. edulis), and (3) a new genus composed of atlantic of April. During this phase three species of dinoflag- species currently placed in Hydropuntia (G. cornea, ellates come to dominate the phytoplankton, with G. crassissima, G. usneoides). Cystocarp features within Glenodinium quadridens representing 60% of total phy- the Gracilaria senso lato clades are more phyloge- toplankton biovolume. By mid-April the spring di- netically informative than do male characters. The noflagellate blooms collapse. The second phase is a henriquesiana-type of spermatangial concepacle ap- period of instability as a series of species dominates peared independently at least four times. The textorii- the phytoplankton. In mid-July dinoflagellates again type of male apparatus is represented in two distinct dominate the third phase, and Peridinium limbatum clusters of Gracilaria. Several unknown and poorly de- alone represents 90% of the bog phytoplankton. The scribed species are being described, resurrected from fourth phase is a stable period as P.limbatum slowly synonymy or re-circumscribed. G. textorii seems to be declines in late summer and fall. A strong correlation part of a major complex of flat species in the Pacific 14 ABSTRACTS

Ocean. The current number of Gracilariaceae is un- In the photosynthetic dinoflagellate Dinophysis, pig- derestimated in the Western Atlantic due to conver- ment composition and ultrastructural characters gence in habit type and the apparent homoplasy in place the origin of the chloroplast with the crypto- vegetative and reproductive anatomy. phytes. Presented here is the first molecular data from the plastid of Dinophysis. We cloned and sequenced the coding regions of psbA and small subunit (SSU) 37 rDNA from four populations of D. acuminata and from THREE NEW SPECIES OF GRACILARIOPSIS D. acuta. These data were used to test two hypotheses (GRACILARIALES, RHODOPHYTA) FROM THE about Dinophysis plastid evolution: (1) The plastid is NORTH-WESTERN ATLANTIC of cryptophyte origin; and (2) The plastid is a perma- Gurgel, C. F. D.1,2,∗ & Fredericq, S.2 nent replacement. The psbA-coding region was iden- 1Department Botanica,ˆ CCS, Institute de Biologia, tical among the populations of D. acuminata and be- Universidade Federal do Rio de Janeiro, Ilha do Fundao,ˆ tween this species and D. acuta. On the other hand, Rio de Janeiro 21941-900, RJ, Brazil; 2Department of the SSU rDNA showed polymorphisms, both between Biology, University of Louisiana at Lafayette, P.O. Box species and among populations. Phylogenetic analysis 42451, Lafayette, LA 50704-2451, USA shows that both psbA and SSU rDNA sequences firmly place the Dinophysis spp. plastid within the crypto- A recent molecular phylogenetic study inferred from phytes, confirming its origin through plastid replace- chloroplast-encoded rbcL sequence analyses based on ment. The polymorphic SSU rDNA sequences group 20 specimens of Gracilariopsis. Dawson 1949 worldwide together and are sister to a cryptophyte ultraplankter confirmed the monophyly of the genus and revealed (OCS20). The SSU rDNA sequence heterogeneity be- the existence of three new Caribbean species (Gurgel tween species (about 1%) is comparable to the hetero- etal. J. Phycol., submitted). Gracilariopsis sp. 1 collected geneity among different isolates of D. acuminata, sug- in the Mexican Gulf of Mexico is sister to G. tenuifrons. gesting these are population-level differences and not Gracilariopsis sp. 2 collected in Caribbean Panama indicative of different plastid captures by Dinophysis. (Colon´ City) and Venezuela (Paraguana Pen´ınsula) Interestingly, a second class of cloned coding regions is morphologically closely related to G. panamensis. was also isolated from each population. These psbA Gracilariopsis sp. 3 collected in Venezuela (La Vela and SSU rDNA sequences were evolutionarily more de Coro) is distinct from all other terete and stringy divergent and specifically related to florideophyte red species that characterize the genus by being a flat- algae. Given the mixotrophic habit of Dinophysis, the foliose taxon; it also shows a basal topology with un- possibility arises that this second class may have come certain phylogenetic relationship to an undetermined from red algal preys in the food vacuoles of our single- species from Japan and China and to G. heteroclada cell isolates. These results highlight the importance of from the Philippines. The lack of clear-cut vegetative sampling multiple genes and populations in determin- and reproductive features among the species coupled ing the complex evolutionary history of the Dinophysis with a high degree of thallus plasticity may account plastid. for the fact that genetically distinct species were not previously recognized as taxonomically distinct. Our molecular and morphological studies reveal that the 39 diversity of Gracilariopsis species in the western Atlantic BIOASSESSMENT OF STREAM-WATER QUALITY and probably in the rest of the world is currently un- USING BENTHIC AND PLANKTONIC ALGAE derestimated. Morphologically detailed descriptions COLLECTED ALONG AN URBAN INTENSITY for all three new species are provided. GRADIENT IN THE EASTERN CORNBELT PLAINS ECOREGION, OHIO, USA Hambrook, J. A. 38 US Geological Society, 6480 Doubletree Ave., Columbus, THE PLASTID OF DINOPHYSIS OH 43229-1111, USA (DINOPHYCEAE): PHYLOGENETIC EVIDENCE FOR A PERMANENT REPLACEMENT Algae possess many attributes that make them ideal Hackett, J. D.1,∗, Maranda, L.2 & Bhattacharya, D.1 organisms to employ in water-quality investigations. 1Department of Biological Sciences, Center for Comparative They are ubiquitous, ecologically important, and sen- Genomics, University of Iowa, Iowa City, IA 52242; sitive to a broad range of stressors; moreover, they re- 2Graduate School of Oceanography, University of Rhode spond rapidly to changes in water quality. As part of the Island, Narragansett, RI 02882-1197, USA USGS National Water Quality Assessment (NAWQA) ABSTRACTS 15

Program, two methods of analyzing stream-dwelling ent diffusing substrates is needed. To test a new ap- algae were used to assess relations among nutrient proach for nutrient diffusion, Petri dishes were filled concentrations, algae, and overall biological integrity with agar and the nutrient of choice. A Whatman of streams along a gradient from agricultural to ur- glass microfiber filter (GF/F) was attached to the top ban land cover. One method is based on algal pro- of the dish, which served as the medium through duction (chlorophyll a, biomass, and 48-h measures which the nutrients diffused and as a uniform surface of dissolved oxygen), whereas the other is based on area for periphyton colonization. Before testing in a algal community assemblages (species abundance, stream impacted by acid mine drainage, laboratory biovolume, and autecology). Quantitative periphyton studies were conducted to simulate field conditions. and phytoplankton samples were collected from 30 The Petri dishes readily diffused the nutrients and stream reaches in small watersheds (drainage area 7.7– were durable to the acid conditions. Six replicates with 188.7 km2) in south-west Ohio, during July 2000. Phy- a control, nitrogen (0.5 M), phosphorus (0.5 M), and toplankton Chl a ranged from 0.5 mg/L in an urban nitrogen + phosphorus (0.5 M of each) Petri dishes area to 43 mg/L in an agricultural basin, whereas the were deployed in four AMD impacted streams for the epilithic periphyton Chl a ranged from 2 mg/m2 in month of February. Samples were analyzed for chloro- an urban area to 142 mg/m2 in an agricultural basin. phyll a, ash-free dry mass (AFDM), and taxa present. Spearman correlations were used to indicate relations The chlorophyll a analyses showed the phosphorus between the physical, chemical, and biological factors treatment to be significantly (P < 0.05) greater than and measures of algal production. Multivariate analy- all other treatments and the nitrogen treatment was sis of algal species abundance and metrics will be used significantly (P < 0.05) lower. However, these trends to characterize gradients of algal community response were not seen in the AFDM results with no difference relative to chemical and landscape factors. Results will among treatments detected. Seasonal assessment of be discussed in relation to the influence of urban land- this approach is currently under way. use intensity and the US Environmental Protection Agency nutrient criteria proposed for this region. The 41 extent to which the algal production and community structure results are useful for bioassessment of stream IMPACTS OF REDUCED SALINITY ON health will be evaluated with regard to understand- SEAGRASSES IN INDIAN RIVER LAGOON ing community responses to organic enrichment, dis- Hanisak, M. D. solved oxygen, specific conductance, light availability, Harbor Branch Oceanographic Institution, Fort Pierce, FL pesticide concentrations, and the monitoring of wa- 34946, USA tershed management practices. From November 1993 to November 1995, a study was conducted in Indian River Lagoon (IRL) to deter- 40 mine relationships of light attenuation, water qual- A NEW METHOD FOR STUDYING NUTRIENT ity, and seagrass at sites representing a range of water LIMITATION OF PERIPHYTON: A CASE STUDY quality conditions. There were considerable interan- FROM ACID MINE DRAINAGE STREAMS nual differences at the sites. The most striking event Hamsher, S. E.1,∗, Casamatta, D. A.1, Filkin, N. R.1, occurred at a station in northern Banana River, an McClintic, A. S.1, Chiasson, W. B.1, Verb, G. R.2 & area of IRL considered the least anthropogenically Vis, M. L.1 impacted. This station had a well-developed, multi- 1Department of Environmental and Plant Biology, Ohio species seagrass bed (Halodule wrightii and Syringodium University, Athens; 2Department of Biology, Ohio Northern filiforme, with small quantities of Halophila engleman- University, OH, USA nii). Throughout the first year, salinity was stable (90% of weekly values ranged from 26 to 32 ppt). Begin- Acid mine drainage (AMD) affects about 16 900 km of ning in November 1994, the cover, shoot density, and streams in the Appalachian region, but little is known biomass of all three species declined precipitously due about the biology of these habitats. Typically, these to sharply reduced salinities associated with high rain- habitats have low primary productivity most likely due fall and run-off. Salinities during the following year to metal stress. However, these streams are also very low varied little and remained low (90% of weekly values in nutrients. Traditionally, nutrient limitation studies ranged from 13.5 to 19.8 ppt). The lack of recovery of periphyton have been conducted employing ter- to earlier salinity levels was due to the long residence racotta pots. Recently, research has concluded that time (about 1 year) in that portion of IRL. Seagrass this methodology may have inherent flaws due to vari- cover, shoot density, and biomass reached minima in able porosity. Therefore, development of new nutri- May 1995. By August 1995, Halodule began to recover, 16 ABSTRACTS but nearly 8 years later, Syringodium has not recovered those found for the ITS and other gene regions previ- to 1994 levels. ously used by researchers investigating species bound- aries at the intraspecific level in symbiotic, terrestrial fungi. In addition to ITS 1, 2 and the 5.8 s gene re- 42 gions, I am currently using the 5’ end of the EF1a 18S-BASED TAXONOMY AND INTRASPECIFIC coding region to construct intraspecific genealogies ITS SEQUENCE VARIATION IN THE MARINE for H. irritans. By comparing these genealogies to each FUNGAL ENDOSYMBIONT HALOGUIGNARDIA other and to the geographic distribution of samples, I IRRITANS INFECTING CYSTOSEIRA OSMUNDACEA aim to determine if more than one genetic species is ALONG THE CALIFORNIAN COAST present within the morphological species H. irritans. Harvey, J. B. J. Department of Ecology and Evolutionary Biology, A316 Earth and Marine Sciences, University of California, Santa 44 Cruz, CA 95064, USA SYNCHROTRON-BASED INFRARED IMAGING OF EUGLENA GRACILIS SINGLE CELLS Abstract withdrawn Hirschmugl, C.1,∗, Bunta, M.1, Holt, J.1, Giordano, M.2, Skilirov, A.3 & Strickler, J. R.4 1 43 Department of Physics, University of Wisconsin-Milwaukee, Milwaukee, WI 53211, USA; INTRASPECIFIC GENETIC VARIATION IN 2Faculty of Sciences, Institute of Marine Sciences, University H. IRRITANS, A FUNGAL ENDOSYMBIONT OF of Ancona, Via Brecce Bianche, 60131 Ancona, Italy; MARINE BROWN ALGAE ON THE NORTH 3Advanced Analysis Facility; 4WATER Institute, University AMERICAN PACIFIC COAST of Wisconsin-Milwaukee, Milwaukee, WI 53211, USA Harvey, J. B. J. Department of Ecology and Evolutionary Biology, A316 Infrared (IR) microspectroscopy is used to examine Earth and Marine Sciences, University of California, Santa and identify the chemistry in individual living cells of Cruz, CA 95064, USA Euglena gracilis. The spectra provide relative concen- trations of lipids, proteins and carbohydrates in the The marine ascomycete genus Haloguignardia occurs specimen. Diffraction-limited, spatially resolved IR im- endophytically in members of the marine brown al- ages of single cells of the microalga were measured us- gal family Sargassaceae globally. This example of en- ing synchrotron radiation as a bright IR source. These dosymbiosis has been morphologically described: the contour maps reveal the distributions of the proteins, fungal component internally infects the algal host re- lipids and carbohydrates including the phospholipids, sulting in prolific cell growth, forming galls composed sugars and paramylon. We compare the images with chiefly of host algal cells but containing fungal repro- the biochemistry for E. gracilis. We also present the ductive structures and vegetative hyphae. H. irritans IR images for two different cells, which were exposed induces the formation of galls in the brown algae to different culture conditions prior to the measure- Cystoseira osmundacea and Halidrys dioica along the Pa- ments. The overall trends observed for the two speci- cific coast from Oregon to Baja California, Mexico. mens are in qualitative agreement with the hypothesis Using culture-independent molecular techniques, I that there was a significantly reduced concentration sequenced the 18S rDNA gene region for H. irritans of nitrogen in the culture as a function of time. Im- and generated a 18S-based taxonomy consistent with portantly, this study demonstrates the potential to ex- the current taxonomy for this morphological species. amine changes in the chemistry of living cells while In order to study intraspecific genetic variation in modifying environmental stimuli. H. irritans, I have sequenced the ITS rDNA (ITS 1, 2 and the 5.8 s) regions for five separate gall-tissue sam- ples from Santa Rosa Island in southern California and for five samples from Monterey and Carmel in 45 central California. Intraspecific DNA sequence varia- CELL SIGNALING IN THE tion in the ITS regions of H. irritans reveals consistent CHEMOACCUMULATION RESPONSE OF THE sequence divergence between sites sampled. The fun- COLONIAL GREEN ALGA gal ITS regions for H. irritans total 613 bp in length ASTREPHOMENE GUBERNACULIFERA and contain 40 synapomorphic characters for a total Hoops, H. J.∗ & Gifford, S. M. of 6.5% variation in informative loci between southern Biology Department, SUNY-Geneseo, 1 College Circle, and central Californian sites. This value is similar to Geneseo, NY 14454, USA ABSTRACTS 17

A. gubernaculifera can grow on acetate even in the dark. ing on their MAA content. The first group (I) includes Interestingly, it displays a well-developed chemoaccu- mainly deep-water species, typically lacking MAAs. The mulation response to added acetate. We suggest that second group (II), algal species found in a broad range this chemoaccumulation results from a receptor-based of water depths (eu- and sublittoral), which are able signaling pathway because the alga responds to the to flexibly synthesize and accumulate MAAs. The third structurally similar molecule propionate even though group (III) includes supra- and eulittoral taxa, which the latter compound cannot support growth. Based always contain high MAA concentrations. In labora- on suggestions about chemotactic signaling pathways tory studies, we showed that taxa of group II and III in other eukaryotes and postulated phototactic re- responded in three different ways based on MAA accu- sponse of some algae, we hypothesized that the signal- mulation when exposed to different radiation condi- ing pathway involves heterotrimeric G-proteins. The tions (PAR, PAR + UVA, PAR + UVA + UVB). Either effectiveness of the chemoresponse was determined they: (a) exhibit highest total MAA concentration un- by measuring the diameter of the chemoaccumula- der the full artificial spectrum; (b) increase their MAA tion cloud after addition of 10 mL aliquots of me- concentration after exposure to PAR and PAR + UVA dia containing acetate to colonies swimming in me- or (c) MAA concentration declines after exposure to dia without acetate. Neomycin, which inhibits the G- the full spectrum. Our studies have indicated that protein dependent activity of phospholipase C, com- when coupled with UVR, exposure to temperature pletely inhibited chemoaccumulation at 100 mM, and fluctuations ranging from 0 to 10 ◦C also affect MAA markedly reduced the density of the accumulation biosynthesis. response at 10 mM with no obvious changes in gen- eral colonial motility. EGTA, a calcium chelator, abol- ished the chemoresponse. Mastoporan, a G-protein activator, caused accumulation even in the absence of acetate. These results support the hypothesis that chemoaccumulation in A. gubernaculifera uses a G- protein mediated, calcium-dependent, cell-signaling 47 pathway. AN EXAMINATION OF THE EFFECTS OF A RESERVOIR, LAKE ASHTABULA, ON THE PERIPHYTIC ALGAE IN THE SHEYENNE RIVER, 46 NORTH DAKOTA MAA SYNTHESIS AND ACCUMULATION IN Jaskowiak, M. A.∗, Fawley, K. P. & Fawley, M. W. POLAR MACROALGAE ARE CONTROLLED BY Department of Biological Sciences, North Dakota State ABIOTIC FACTORS University, Fargo, ND 58105, USA Hoyer, K.1,∗, Karsten, U.2 & Wiencke, C.1 1Alfred Wegener Institute for Polar and Marine Research, The Sheyenne River is a third order stream that drains Am Handelshafen 12, D-27570 Bremerhaven; 2Institute of a significant portion of central and eastern North Aquatic Ecology, University of Rostock, Dakota. In 1950, the Baldhill Dam was built and as a Albert-Einstein-Strasse 3, D-18075 Rostock, Germany result, the reservoir, Lake Ashtabula, was formed. The periphytic algae in this river were extensively studied Mycosporine-like amino acids (MAAs) are regarded from 1997 to 1999. During this research, the effects as powerful sunscreens protecting the algae against of differences in environmental variables on the algal harmful UV radiation. The MAA protection efficiency communities were assessed with Canonical Correspon- was tested in algal samples by measuring the opti- dence Analysis (CCA). In this analysis, the periphytic mum quantum yield of photosynthesis using pho- algal communities in the downstream sites appeared tosystem II fluorescence. It could be demonstrated to be quite different from those communities found that the recovery of photosynthesis after exposure to upriver of Lake Ashtabula. Other research on differ- enhanced UV radiation is faster in individuals with ent rivers has shown profound changes in the algal high MAA content. MAAs can be synthesized in sev- community after a dam was built along the river. Re- eral polar macroalgae in response to different radia- dundancy Analysis (RDA) was used to determine if tion conditions. Although MAA induction patterns are the sites downstream are more statistically similar with very species-specific, some similarities can be found. Lake Ashtabula than with the upstream sites. Finally, Field studies indicate that plants from different growth CCA was used to examine which environmental vari- habitats providing distinct radiation climate can be ables explain the most variance in Lake Ashtabula and grouped into three physiological categories depend- the downstream sites. 18 ABSTRACTS

48 49 RELATIONSHIP BETWEEN PERIODIC ESTIMATING DIVERGENCE TIMES WITHIN THE RESUSPENSION EVENTS AND PLANKTONIC CHAROPHYTA: A BAYESIAN APPROACH USING DIATOM COMMUNITY STRUCTURE IN LAKE FOSSIL AND DNA SEQUENCE DATA MICHIGAN: A FIELD AND LABORATORY Karol, K. G.1,∗, McCourt, R. M.2 & Delwiche, C. F.1 INVESTIGATION 1Department of Cell Biology and Molecular Genetics, Julius, M. L. University of Maryland – College Park, College Park, MD Department of Biological Sciences, St. Cloud State 20742; 2Department of Botany, Academy of Natural University, St. Cloud, MN, USA Sciences, 1900 Benjamin Franklin Parkway, Philadelphia, PA 19103, USA Lake Michigan provides an ideal location for com- paring episodic physical forcing events (storms) on Morphological and molecular studies have advanced phytoplankton processes and the more persistent sea- our understanding of charophycean green algal re- sonal variability of phytoplankton communities. This lationships. A recent multigene phylogenetic analysis is due to the duration and extent of the highly tur- has identified the Charales as the closest living rela- bid, recurrent coastal plume (RCP) in the lake dur- tives of land plants with the Coleochaetales sister to ing the winter/early spring. Although the RCP can the Charales/land plant lineage. With a robust phy- coincide with the initiation of the basin wide spring logeny now in hand, it is possible to explore phylogeny- diatom bloom, linkages between the duration and in- dependent questions that were previously difficult to tensity of the plume and the prominent role of light assess. Estimating the divergence time of the land availability in regulating Lake Michigan phytoplank- plant lineage is one such question. The most recent ton growth during the spring isothermal period have time estimate for the colonization of land by plants been postulated, but not verified. The concurrent is 1061 ± 109 million years ago and 703 ± 45 million physical and biological events provide a novel oppor- years ago for the divergence of vascular plants and tunity to examine phenomena associated with the RCP bryophytes, a result much older than that found in affecting the distribution and abundance of species in the fossil record (roughly 470 million years ago). Sev- Lake Michigan’s planktonic diatom flora. In this study, eral problems are inherent in estimating divergence planktonic diatom assemblages from pre, post and ac- times using DNA sequence data; these include the as- tive spring plume events were examined from stations sumption of a molecular clock and choosing from a along Lake Michigan’s southern shoreline. Species sparse fossil record for an external calibration date. abundance changed rapidly during storm events. Sed- Unlike most algae, a rich fossil record exists for the iment resuspension via storm activity created a sub- Charales in the form of calcified oospores termed gy- optimal growth environment. Post-storm event di- rogonites. Representative gyrogonites that can be at- atom phytoplankton communities were floristically tributed to all six of the extant genera in the Characeae distinct from prestorm event communities, with rest- have been identified with reasonable accuracy. These ing cell-forming taxa playing a significant role in these six calibration points, identified from a closely related community-restructuring periods. Laboratory simu- lineage of land plants, can be used in conjunction with lations of resuspension events using Lake Michigan a multigene DNA dataset to estimate the time that the sediments were conducted under a variety of en- land plant and Charales lineages diverged from their vironmental conditions. Parameters varied included common ancestor. day length, temperature, and silica. The resulting assemblages were quantitatively counted. A statisti- 50 cally significant relationship was identified between PUBLIC OUTREACH MATERIALS REGARDING day length and the vegetative growth of many rest- HARMFUL ALGAL BLOOMS AND THEIR ing cell-forming diatom species. When day length POSSIBLE EFFECTS ON HUMAN HEALTH was calculated for poststorm event field data, it re- Kirkpatrick, B.1,∗, Fleming, L. E.2, Stephan, W. B.2, vealed a high correlation between poststorm event Backer, L.3, Clark, R.4, Squicciarini, D.2, Weisman, communities and those predicted by the labora- R.2 & Van De Bogart, G.2 tory simulations. Timing of storm events and lat- 1Mote Marine Laboratory, 1600 Ken Thompson Parkway, itudinal position of the aquatic system are then Sarasota, FL 34236; 2NIEHS Center, University of Miami, important elements for consideration when pre- 4600 Rickenbacker Causeway, Miami, FL 33149; 3Center dicting diatom phytoplankton community structure for Disease Control, 1600 Clifton Road NE, Atlanta, GA due to the relationship these factors have with day 30333; 4Florida Department of Health, 4052 Bald Cypress length. Way, Tallahassee, FL 32399, USA ABSTRACTS 19

The potential interactions between marine and fresh- are Cyclotella bodanica var. glabriscula and C. bodanica water harmful algal blooms (HABs) and humans are var. oligactis. Great Lakes populations of these taxa are increasing. Humans are exposed through food, drink- significantly different from the original populations ing water, and recreational and occupational water use described from alpine lakes in Austria and Switzer- to an increasing number of organisms and their tox- land and merit taxonomic separation. Many of the ins. Nevertheless, the amount of clinical and epidemi- endemic taxa are no longer present in the modern ologic research concerning acute and chronic human Great Lakes assemblages. The precise time of their health effects from the HAB organisms and their tox- disappearance from the system is not know, but modi- ins is relatively sparse. At the same time, the public fications in Great Lakes diatom populations clearly co- is increasingly aware of and interested in the poten- incided with the development of substantial European tial dangers associated with exposure to HABs. Public settlement of the Great Lakes region. C. americana, for health authorities and researchers must respond to example, was always limited to Lakes Erie and On- these public health concerns. The development of ap- tario but disappeared from both of these lakes prior propriate educational and outreach materials based to 1900. With the disappearance of these taxa, came a on limited scientific databases is the challenge of in- number of introductions to the lakes. This work rep- forming the public concerning the possible human resents the first attempt to document these floristic health effects of HAB organisms and their toxins, and changes. their prevention. Educational materials developed by a group of researchers and public health personnel for general HABs, Florida Red Tide Toxins (brevetox- 52 ins), Cyanobacteria, and Ciguatera are discussed, as well as methods for their dissemination. SYSTEMATICS OF COCCOID GREEN ALGAE: MORPHOLOGY VS. 18S rRNA GENE PHYLOGENY Krienitz, L.1,∗, Wolf, M.1, Hegewald, E.2 & 51 Hepperle, D.1,3 ENDEMISM AND INVASION IN THE GREAT 1Leibniz-Institute of Freshwater Ecology and Inland LAKES DIATOM COMMUNITY Fisheries, D-16775 Stechlin-Neuglobsow; 2Institute of Kreuger, N.∗ & Julius, M. L. Chemistry and Dynamics of the Geosphere, Research Centre Department of Biological Sciences, St. Cloud State Juelich, D-52425 Juelich; 3Albrecht-von-Haller Institute of University, St. Cloud, MN, USA Plant Sciences, University of Goettingen, D-37037 Goettingen, Germany Population dynamics and health for individual Great Lakes diatom species is explored. Information dealing Classical approaches using morphological characters with how these taxa compete with introduced species for the circumscription of taxa in coccoid green algae and rapid environmental changes in modern times do not adequately reflect the phylogenetic relation- is examined. Diatomists have identified a number of ships. Phylogenetic trees based on 18S rRNA gene se- these taxa, but discussion is limited with little or no quence analysis provide new insights into the system- information covering the ecological range of the taxa atics of these algae. We give a review and discuss the over time. Some of these include Cyclotella americana content of orders of coccoid green algae and evalu- Fricke; C. bodanica var. stellata Skvortzow; and Stephan- ate the usefulness of some traditionally used diacritic odiscus superiorensis Theriot. A number of taxa endemic features. Whereas the content of the orders Chloro- to the Great Lakes undescribed in the literature also coccales and Chlorellales needs to be reduced, the or- exist. These are species wedged into taxonomic cat- der Sphaeropleales comprises more families than in egories from the European taxonomic system. Re- the traditional circumscription. According to molecu- cent advances in the understanding of diatom species lar biological investigations the Neochloridaceae, Se- boundaries, suggest these names are inappropriately lenastraceae and Scenedesmaceae formerly classified used for a number of taxa identified in paleolimno- in the Chlorococcales and later into the Chlorellales logical investigations of the Great Lakes. This prob- belong to the Sphaeropleales. The Hydrodictyaceae, lem developed because taxonomic information out- formerly a member of the Protosiphonales, are also side of the European flora is not readily available to to be grouped in the Sphaeropleales. Some members researchers, and because the focus of paleolimnolog- of the Ulotrichales, which propagate by binary fission ical studies is primarily environmental reconstruction (e.g. Nannochloris and Catena) cluster together with au- minimizing the time that can be spent unraveling tax- tosporine coccoid taxa within the Trebouxiophyceae. onomically troublesome species. Examples of these With the example of members of the Selenastraceae, 20 ABSTRACTS

Oocystaceae and Chlorellacea it is shown that the mor- terial in water from in vivo spectroscopy (filter pad phology of cells as well as the formation of coenobia, technique) will also be discussed. spines, mucilage and incrustations cannot be used to separate genera.

54 PRESERVATION OF NOSTOCACEAN 53 HORMOGONE MOTILITY IN DESICCATED UV-ACCLIMATION OF CULTURED CALCIUM CARBONATE AGAR PATCHES PHYTOPLANKTON EXPOSED TO NATURAL Lazaroff, N. AND ENHANCED UVB RADIATION AND Foundation for Microbiological Analysis, Vestal, NY METHODOLOGICAL ASPECTS OF MAA 13850, USA QUANTIFICATION Laurion, I.∗ & Roy, S. Filamentous nitrogen fixing cyanobacteria are resis- Institut des Sciences de la Mer, Universite du Quebec a tant to desiccation, nutrient depletion, adverse tem- Rimouski, 310 Allee des Ursulines, Rimouski, Que., perature and challenging photic environments. This Canada is generally thought to account for their role as pio- neering forms of life on denuded strata. It has also lent Bloom-forming dinoflagellates often synthesize high credence to the possibility that cyanobacteria with pro- concentrations of mycosporine-like amino acids longed viability may have been able to survive condi- (MAAs) as a protection from high PAR and UV radia- tions of interplanetary space to implement a pansper- tion, which may afford them a competitive advantage mian hypothesis for the origin of life on earth. Pro- over other species. Photoprotection and long-term longed retention of viability had been noted in our acclimation to natural and enhanced UVB radiation collection of myxotrophically grown strains of nosto- were examined during 4 weeks in three dinoflagellates caceans cultured in liquid or solid media containing and one diatom. Semi-continuous, nutrient-replete calcium carbonate. Such media permitted satisfactory cultures were maintained outdoors under natural recovery and regrowth after storage at low light in- light, with or without additional UVB (from lamps). tensities for 2 years or more. Moreover, even the old- The photosynthetic performance (XE-PAM fluorome- est calcium carbonate cultures contained motile hor- try) and the concentration and composition of MAAs mogonia when observed microscopically. This phe- and photoprotective carotenoids (PPCs; comprising nomenon can be used to advantage in sending cul- xanthophylls) were followed weekly. A great variety tures to other laboratories. Not only are organisms of MAAs was found in the dinoflagellates (up to 10, maintained during passage through the postal system identified by cochromatography and LC–MS). Large but since the cultures formed hormogonia readily, differences between sp. (and between two strains of it was possible to produce preservable homogeneous the same sp.) were observed. In most cases, the com- agar lawns that generated hormogonia as needed for position and concentration of MAAs and PPCs in- demonstration or experiment. A practical technique creased under enhanced UVB. Growth rate and pho- consists of seeding axenic strains of Nostoc species tosynthetic performance were sometime lower under grown in liquid shake culture under cool-white fluo- enhanced UVB, while in other cases photoacclimation rescent illumination into solid media containing 0.7% seemed to occur. The least affected sp. was Alexan- sucrose, 0.05% finely divided calcium carbonate and drium tamarense who responded most rapidly to en- 1.4% purified agar. The preparations are incubated hanced UVB. The algae also responded to changes under red fluorescent illumination to produce several in natural incident radiation. Some methodological crops of hormogonia in successive cycles of develop- aspects of MAA quantification were also investigated ment, within the agar. The heavily grown lawns are cut (with four species of dinoflagellates). Our results in- into disks or squares, then transferred to dishes and dicate how freezing (liquid-N) likely breaks the cells, dried in desiccators with anhydrous calcium sulfate. mobilizing the water-soluble MAAs in the filter moist The dry patches produce swarms of hormogonia in 2 (otherwise highly packaged in fresh cells), while the or 3 h, depending upon the rate of rehydration. The organelles containing the pigments remain relatively patches are facile tools for purifying cultures through intact. The implications of standard sample handling the isolation of hormogonia and studying cyanobac- on the evaluation of MAA concentration in algae and terial physiology by the rapid response of motile hor- on the estimation of the absorption of suspended ma- mogonia. ABSTRACTS 21

55 56 FATTY ACID AND STEROL COMPOSITION OF A LIPID BIOMARKER ANALYSIS OF THE TOXIC KARENIA BREVIS BLOOM IN THE GULF OF DINOFLAGELLATE PFIESTERIA PISCICIDA: MEXICO DISTRIBUTION OF STEROLS AND FATTY ACIDS Leblond, J. D.1,∗ & Chapman, P. J.2 WITHIN THE CLASS DINOPHYCEAE 1Department of Biology, P.O. Box 60, Middle Tennessee Leblond, J. D.1,∗ & Chapman, P. J.2 State University, Murfreesboro, TN 37132; 2US 1Department of Biology, P.O. Box 60, Middle Tennessee Environmental Protection Agency, Gulf Ecology Division, State University, Murfreesboro, TN 37132; 2US National Health Effects and Environment Research Environmental Protection Agency, Gulf Ecology Division, Laboratory, 1 Sabine Island Dr, Gulf Breeze, FL 32561, National Health Effects and Environment Research USA Laboratory, 1 Sabine Island Dr, Gulf Breeze, FL 32561, USA In the Gulf of Mexico, recurring algal blooms caused by Karenia brevis (formerly known as Gymno- Within US waters, regular blooms of harmful dinoflag- dinium breve) have significant adverse health and eco- ellates occur in the Gulf of Mexico and Chesapeake nomic impacts. K. brevis is one member of a small Bay regions. Although the causes of blooms are not group of dinoflagellates, related morphologically and fully understood, events in Gulf of Mexico waters have by DNA-based phylogenetic analysis, that synthesize been recorded for over 30 years, and are almost ex- the carotenoid, gyroxanthin diester, in place of the clusively caused by the photosynthetic dinoflagellate, more widely distributed peridinin. While this novel Karenia brevis. Conversely, blooms in the Chesapeake photopigment has been proposed as a biomarker, es- Bay region, caused by the heterotrophic dinoflagellate pecially for remote-sensing imaging technologies, to Pfiesteria piscicida, appear to be a relatively new phe- detect the emergence of K. brevis blooms, other chem- nomenon, and are hypothesized to be linked to an icals such as sterols and triglycerides, respectively, with increase in nutrient-rich runoff from chemical plants potential to report the distribution and physiological and livestock farms along tributaries leading into the condition of K. brevis are required. Recent work from Chesapeake. Despite the environmental, economic, our laboratories characterizing the lipids of dinoflag- and human health relevance of P.piscicida, there ap- ellates has confirmed that K. brevis, together with those pears to be no published work on the characterization few close relatives lacking peridinin, possesses a rela- of its lipid composition. The objective of this study tively simple sterol profile comprised of two unusual was, therefore to examine the sterol and fatty acid primary 4-methyl sterols, designated ED and NED, composition of cultured P.piscicida isolates as a pre- each with an ergosterol-type side chain. A recent di- cursor to eventual field studies of blooms of this or- noflagellate bloom in the waters of the north-west Gulf ganism. GC–MS analysis of trimethylsilyl ether deriva- of Mexico near the Gulf Breeze EPA laboratory pro- tives of sterols in free sterol and sterol ester fractions vided an opportunity to examine the usefulness of revealed that the overwhelming majority of sterols these sterols and other lipids as indicators of K. brevis was found as free sterols. Furthermore, free sterols in phytoplankton communities. Lipid extracts of fil- were found to resemble those of other dinoflagel- tered bloom samples, fractionated to separate free lates, with the dominant compound being the com- and esterified sterols, were examined by GC–MS of mon dinoflagellate sterol, dinosterol; a number of trimethylsilyl ether derivatives. ED and NED were the other 4-methyl sterols common to other dinoflagel- major sterols found in all bloom samples. Fatty acids lates were also identified. The fatty acids of P.piscicida found in lipid fractions containing membrane phos- were found to be primarily associated with a fraction pholipids, chloroplast-associated glycolipids, and stor- containing cellular membrane phospholipids; small age triglycerides, respectively, differed significantly. amounts of the recently described highly unsaturated The glycolipid fraction was found to contain octade- fatty acids, octacosaoctaenoic acid [28 : 8(n-3)] and oc- capentaenoic acid [18 : 5(n-3)], a fatty acid commonly tacosaheptaenoic acid [28 : 7(n-6)], were observed in associated with dinoflagellates. The phospholipid frac- this fraction. The dominant fatty acid (approximately tion was found to contain small amounts of the re- 40–60% of the phospholipid fatty acids) was docosa- cently described highly unsaturated fatty acids, oc- hexaenoic acid [22 : 6(n-3)]. Unlike K. brevis (see ac- tacosaoctaenoic acid [28 : 8(n-3)] and octacosahep- companying poster), P.piscicida does not appear to taenoic acid [28 : 7(n-6)]. Fatty acids from the triglyc- possess significant amounts of unusual sterols or fatty eride fraction were more abundant than those associ- acids which may serve as potential biomarkers for this ated with glycolipids or phospholipids. organism. 22 ABSTRACTS

57 bryophyceae, characterizing the early evolution and diversification of land plants depends on understand- THE TRANSITION FROM ALGAE TO ing phylogenetic relationships among the less exten- EMBRYOPHYTES: CHLOROPLAST sively studied green algal members of the Charophyta. PHYLOGENOMIC EVIDENCE (II) ∗ Relationships within the Charophyta have previously Lee, J. & Manhart, J. R. been based on morphology and inferred through phy- Department of Biology, Texas A&M University, College logenetic analyses of molecular data that include ri- Station, TX 77843-3258, USA bosomal DNA and several genes from the chloroplast The transition of plant life from aquatic algae to land and mitochondrial genomes. To this point, however, plants was one of the major events in the history of few phylogenetic analyses of nuclear, protein-coding life. However, in hypothesizing the evolutionary path genes have been conducted. We are attempting to iso- of the transition, limited shared phenotypic charac- late and sequence actin-coding regions from represen- ters in aquatic algae and land plants (embryophytes) tative members of the Charophyta. Actin is a highly have been a major hinderance. Chloroplast genomes conserved protein that is ubiquitous among eukary- contain characters useful in tracing evolutionary his- otes; these features make actin an ideal subject for tories. Embryophyte chloroplast genomes are distin- comparative sequence analysis. A major component guished from algal cpDNAs by the presence of over of the cytoskeleton, actin plays a number of impor- 20 group II introns and three ribosomal protein oper- tant roles in plant cell processes such as cell division ons (rpl23, clpP and 3Œrps12 operons). These phy- and cytoplasmic streaming. Hence, in addition to be- logenomic features indicate a phylogenetic relation- ing potentially useful for resolving organismal rela- ship of charophytes and embryophytes. In addition tionships, the evolution of the actin protein itself is to these operons and introns, the evolution of rRNA intrinsic to the study of morphological evolution and and psbB operon evolution of streptophyte lineages development within green plants. will be incorporated with major biological phenotypic features to produce a phylogenetic tree. Basal em- bryophytes, the antithetic hypothesis, monophyly of embryophytes, and paraphyly of charophytes will be 59 discussed. Strepotophytes are classified into three ma- NUMEROUS TRANSITIONS TO LAND IN GREEN jor groups (basal streptophytes, mid-divergent strepto- PLANTS: THE ‘OTHER’ LAND PLANTS phytes and late divergent charophytes-embryophytes). Lewis, L. A. Department of Ecology and Evolutionary Biology, University of Connecticut, 75 North Eaglevile Road, Storrs, CT 06269, USA 58 ACTIN PHYLOGENY IN THE CHAROPHYTA In addition to the more conspicuous embryophyte Lewandowski, J. D.∗ & Delwiche, C. F. group, numerous other green plant lineages have Department of Cell Biology and Molecular Genetics, made successful transitions to land. Phylogenetic anal- University of Maryland – College Park, College Park, MD yses using molecular data indicate that these lineages 20742, USA are primarily derived from freshwater ancestors, and include free-living and lichen photobiont algae in the Photosynthetic green plants, the Viridiplantae, are class Trebouxiophyceae, and several distinct lineages composed of two distinct lineages, the Chloro- of free-living algae in the class Chlorophyceae. In all phyta and Charophyta. The Chlorophyta include cases, these independently derived land plants must the Chlorophyceae, Trebouxiophyceae, Ulvophyceae possess adaptations for environments with reduced and most prasinophytes. The Charophyta include water availability, oxidative stress, and different light the Mesostigmatales, the traditional charophycean regimes than experienced by their aquatic ancestors. green algal orders Chlorokybales, Klebsormidiales, In the most extreme cases, these algae are living in very Zygnematales, Coleochaetales and Charales, and land arid habitats. Given the monophyly of green plants plants, referred to here as Embryophyceae. The Em- and the evolution of independent terrestrial lineages bryophyceae are embedded within the paraphyletic from aquatic ancestors, these lineages represent nat- charophycean green algae, so land plants are, there- ural experiments in the transition of green photosyn- fore best considered a specialized group of green al- thetic organisms to terrestrial living, and offer a rich gae adapted to terrestrial life. Due to the close evo- source of taxa that can be compared experimentally lutionary relationship between green algae and Em- with the embryophytes. ABSTRACTS 23

60 and tetrasporophytes. Branching pattern was char- CHLORORESPIRATION IN GREEN ALGAE acterized by the Strahler method, in which termi- ISOLATED FROM DESERT CRUSTS nal branches are counted as primary branches, and Lewis, L. A.∗, Cardon, Z. G. & Tyser, D. higher order branches are counted where lower order Department of Ecology and Evolutionary Biology, branches meet. Since Ceramium shows dichotomous University of Connecticut, EEB, 75 North Eagleville Road, branching, this method can be used to characterize Storrs, CT 06269, USA the degree in which branching is dichotomous. The angle of branching was also measured. Female game- Photosynthetic organisms enduring extreme temper- tophytes had a higher ratio of primary to secondary atures, low water availability, or high light require pho- branches (3.6–5.1) than male gametophytes (2.5) and toprotective mechanisms to prevent sustained damage tetrasporophytes (2.5–2.6), which have a similar ratio to photosynthetic machinery. Green microalgae liv- to each other. This indicates that males and tetrasporo- ing in desert crust communities of the south-western phytes have a more dichotomous pattern of branching US experience all these environmental stresses, yet while female gametophytes have more proliferative photophysiological studies of green algae in the lit- primary branches. The differences in branching pat- erature have focused on only a handful of common tern appear to be related to the location of reproduc- aquatic and marine species. We are examining the vari- tive cells, which occur along the surface of branches ation in green algal photoprotective mechanisms that in male gametophytes and tetrasporophytes, but occur is the result of natural selection acting independently near the apices in female gametophytes. The branch- in multiple lineages of highly diverse desert green al- ing angle of female and male gametophytes is simi- gae (Chlorophyta) within the classes Chlorophyceae lar to each other, and greater than that of tetrasporo- and Trebouxiophyceae. We have found that unusu- phytes. Therefore, gametophytes and sporophytes dif- ally extensive dark reduction of the plastoquinone fer in branching angle. pool is a prominent photophysiological feature among these desert algae; this reduction may be linked with 62 enhanced chlororespiration. Recently, chlororespira- APOGLOSSEAE TRIB. NOV. (DELESSERIACEAE, tion in higher plants has been linked through mu- RHODOPHYTA) BASED ON TWO MOLECULAR tant analysis to control of the carotenoid synthetic DATASETS AND CYSTOCARP DEVELOPMENT pathway, heat stress, and starch metabolism among Lin, S.-M.1,∗, Fredericq, S.2 & Hommersand, M. H.3 other pathways, though the function of chlororespi- 1Research Department, National Museum of Marine ration remains controversial. Given that green algae Biology & Aquarium, 2 Houwan Road., Checheng, and higher plants are monophyletic, analysis of poten- Pingtung 944, Taiwan; 2Department of Biology, University tial chlororespiration in desert green algae may help of Louisiana at Lafayette, Lafayette, LA 70504-2451; decipher the evolution of the chlororespiratory pro- 3Department of Biology, University of North Carolina at cess as well as its potential role in photoprotection in Chapel Hill, Chapel Hill, NC 27599-3280, USA desert habitats. The genus Apoglossum J. Agardh has traditionally been 61 placed in the tribe Delesserieae. Our morphologi- BRANCHING MORPHOLOGY OF cal observations on Apoglossum and the ‘Delesseria’ GAMETOPHYTES AND TETRASPOROPHYTES species belonging to the resurrected genus Paraglos- OF CERAMIUM CODICOLA IS RELATED TO sum from the Southern Hemisphere show that they PHASE AND REPRODUCTIVE STRUCTURES possess a distinct procarp. Apoglossum has two one- Lewis, R. J.∗ & Lanker, M. D. celled sterile groups that divide after fertilization and Department of Biology, Wheaton College, Wheaton, IL Paraglossum consists of two one-celled sterile groups 60187, USA that do not divide after fertilization, but in which the nuclei divide to produce four to eight nuclei in the Most red algae with a Polysiphonia-type life history undivided sterile groups. In contrast, the procarp in are regarded as having isomorphic phases. However, D. sanguinea, the type species of the tribe Delesserieae, some show minor deviations in morphology, includ- consists of two 2–6-celled sterile groups before fertil- ing differences in branching reported in female ga- ization and the number of sterile cells double after fer- metophytes and tetrasporophytes of Ceramium.By tilization. The development of the carposporophyte in sampling populations of Ceramium codicola, an epi- both Apoglossum and Paraglossum is shown to be unique phyte of Codium fragile, it was possible to character- in the subfamily Delesserioideae. The pit connections ize the morphology of female and male gametophytes between the segments of the gonimoblast filaments 24 ABSTRACTS broaden without fusion and secondary carposporan- tory of the development of the phragmoplast may re- gia are produced. This pattern is absent in other mem- main in the analysis of the gene sequences for this bers of the Delesserioideae, but has been reported for group of proteins. the tribes Myriogrammeae and Schizoserideae in the new subfamily Phycodryoideae Lin, Fredericq et Hom- 64 mersand. Based on both LSU rDNA and rbcL sequence analyses, the Delesserieae is restricted to species of MOLECULAR SYSTEMATICS OF THE Delesseria and Membranoptera from the North Atlantic SUBAERIAL GREEN ALGAL FAMILY and North Pacific Oceans. In contrast, the species of TRENTEPOHLIACEAE (CHLOROPHYTA; Apoglossum and Paraglossum form a distinct cluster hav- TRENTEPOHLIALES) INFERRED FROM 18 SSU ing a center of distribution in the Southern Hemi- rDNA SEQUENCES 1,∗ 3 2 sphere with only a few species, such as A. ruscifolium, Lopez-Bautista, J. M. , Chapman, R. L. , Rindi, F. 3 reaching the Northern Hemisphere. In this study, de- & Guiry, M. 1 tails of postfertilization development leading to the Department of Biology, University of Louisiana at 2 cystocarp are illustrated for the type species of Apoglos- Lafayette, Lafayette, LA 70504-2451, USA; Department sum and Paraglossum and a new tribe, the Apoglosseae, of Biological Sciences, Louisiana State University, Baton 3 is recognized in the Delesseriaceae. Rouge, LA 70803-1705, USA; Department of Botany, Martin Ryan Institute, National University of Ireland, 63 Galway, Ireland PHRAGMOPLASTINS, THE TRENTEPOHLIALES, Representatives of the monophyletic Trentepohliales AND THE EVOLUTION OF THE CYTOKINESIS are widely distributed in the tropics, subtropics, and IN GREEN PLANTS temperate regions worldwide. They grow in soil, or are ∗ Lopez-Bautista, J. M.1, & Chapman, R. L.2 epilithic, epiphytic or endophytic. The family com- 1Department of Biology, University of Louisiana at prises approximately 70 species placed in at least Lafayette, Lafayette, LA 70504-2451; 2Department of four genera (Trentepohlia, Cephaleuros, Phycopeltis and Biological Sciences, Louisiana State University, Baton Stomatochroon), with Trentepohlia sensu lato (includ- Rouge, LA 70803-1705, USA ing Printzina and Physolinum) accounting for half of the species in this family. PCR amplification and se- Green plants have been shown to have a common an- quencing of the 18 SSU rDNA of 18 isolates of the cestor and to contain two lineages: the Chlorophytes Trentepohliaceae were used to assess the monophyly and the Streptophytes, both encompassing represen- of the genus Cephaleuros and to determine the phylo- tatives of the original ‘green algae’. The phragmoplast- genetic relationships among species of Trentepohlia mediated cell division characterizes the land plants in sensu lato. Distance, Parsimony, and Maximum Likeli- the Streptophyte lineage and some basal green algal hood analyses indicate that Trentepohlia sensu lato is orders (Coleochaetales, Charales and Zygnematales). basal and includes two species recently transferred to However, a well-developed phragmoplast-type cell di- Phycopeltis (P.umbrina) and Printzina (P.lagenifera). In vision has been documented in two subaerial green contrast, Cephaleuros is a derived monophyletic clade. algae (Cephaleuros parasiticus and Trentepohlia odorata) Analysis of isolates of C. virescens, from the USA, Tai- belonging to the order Trentepohliales, an order wan, and South Africa indicate that this taxon may that molecular sequence data place unequivocally consist of different species sharing a convergent mor- within the Chlorophytes (rather than Streptophytes). phology. The results of this study have implications for Is the phragmoplast-mediated cell division of the the taxonomy of the genera. Trentepohliales a case for homology or nonhomol- ogy? In order to reveal the answer to this question, we are exploring the potential phylogenetic informa- 65 tion inferred from gene sequences of phragmoplas- BIODIVERSITY AND POTENTIAL USE OF tin, a dynamin-like protein which has been demon- MARINE MACROALGAE FROM THE OFFSHORE strated to be associated with cell plate formation dur- OIL PLATFORMS IN THE GULF OF MEXICO ing the phragmoplast-mediated cytokinesis in land Lopez-Bautista, J. M.1,∗, Fredericq, S.1, plants. Primers were designed based on an available Chapman, R. L.2 & Waters, D. A.2 phragmoplastin sequence from soybean, and yielded 1Department of Biology, University of Louisiana at PCR amplifications from the green algae (Trentepohlia Lafayette, Lafayette, LA 70504-2451; 2Department of and Cephaleuros), a bryophyte (Bazzania) and land Biological Sciences, Louisiana State University, Baton plants (Glycine and Arabidopsis). The evolutionary his- Rouge, LA 70803-1705, USA ABSTRACTS 25

In the north-western Gulf of Mexico, more than 3000 California and Oregon. S. rhodymenioides Joly et Alveal oil and gas platforms are supporting a significant com- 1969 is reinstated for the entity from Chile. The range munity of marine organisms in an area wherein nat- of S. phyllophoroides (J. Agardh) Millar 1990 described ural hard substrata are scarce. This preliminary study from northern NSW, Australia, is expanded to accom- is designed to provide information on the algal com- modate the taxon from South Australia referred to as ponent in terms of species diversity of marine red, S. interrupta. S. leptophylla J. Agardh 1885 is a distinct brown and green macroalgae and cyanobacteria col- South Australian species. A new name needs to be onizing these artificial reefs. The vertical, horizon- provided for the New Zealand taxon. The genus also tal and seasonal distribution and relative abundance comprises a new entity (sterile) from Alaska, and a new are being documented. Taxonomic results are be- (sterile) species from offshore Louisiana. Stenogramme ing evaluated for potential use of some of the algae is a well-defined monophyletic genus, but its posi- present as sources of natural products for the biotech- tion within the Phylophoraceae remains equivocal. nological industry. The island character of this steel archipelago in the north-western Gulf of Mexico pro- vides an ideal environment to test biogeographical re- lationships with adjacent floras. This research is sup- 67 ported in part by the Minerals Management Service, US Department of the Interior. PHYSIOLOGICAL AND BIOCHEMICAL RESPONSES OF TWO SEAWEEDS TO NUTRIENT OR LIGHT DEPRIVATION Lourenco,¸ S. O.∗ & Berges, J. A. 66 School of Biology and Biochemistry, Queen’s University of CHARACTERIZATION AND POSITION OF THE Belfast, Belfast, Northern Ireland, UK GENUS STENOGRAMME IN THE PHYLLOPHORACEAE (GIGARTINALES, The algae Fucus vesiculosus (brown) and Enteromor- RHODOPHYTA) pha compressa (green) were cultured in long-term ex- Lopez-Bautista, J. M.1,∗, Fredericq, S.1, Lindstrom, periments in which natural seawater without nutri- S. C.2, Nelson, W. A.3 & Ramirez, M. E.4 ent enrichment was used, or in which the plants 1Department of Biology, University of Louisiana at were kept in complete darkness. Measurements of Lafayette, Lafayette, LA 70504-2451, USA; 2Department variable fluorescence (F v/F m), an indicator of pho- of Botany, #3529-6270, University Blvd., University of tosynthetic competence, showed a gradual decrease British Columbia, Vancouver, BC, Canada V6T 1Z; of F v/F m in nutrient-deprivation experiments, vary- 3Museum of New Zealand Te Papa Tongarewa, P.O. Box ing from 0.64 to 0.25 (E. compressa) and from 0.65 467, Wellington, New Zealand; 4Seccion Botanica, Museo to 0.40 (F. vesiculosus) after 6 months of cultivation. Nacional de Historia Natural, Santiago, Chile F. vesiculosus was virtually unaffected after more than 2 months in darkness, showing values of F v/F m around The red algal genus Stenogramme was described by 0.65 throughout the experiments, but cultures of Harvey (1840) for entities having flattened, dichoto- E. compressa collapsed after 45 days in darkness. In both mously branched thalli with cystocarps borne in nar- species lower concentrations of chlorophyll a resulted row, linear sori resembling an interrupted midrib on of nutrient-deprivation experiments, decreasing three blades, and with isomorphic tetrasporophytes with times in F. vesiculosus and 25 times in E. compressa by scattered circular to irregular tetrasporangial ne- the end of the experiments. F. vesiculosus cultured in mathecia. In the current literature, the most com- darkness showed 50% more chlorophyll a than the mon species listed is Stenogramme interrupta (C. agardh) control, while plants of E. compressa kept in dark and Montagne ex Harvey 1848, said to have a worldwide in light showed similar values of chlorophyll a during distribution in temperate seas. DNA sequences from the first 30 days of cultivation. The tissue-N content three molecular datasets (chloroplast-encoded rbcL, of both species decreased dramatically in nutrient- and nuclear-encoded ITS and LSU rDNA) instead re- deprivation experiments, establishing variations in veal that the genus is more species-rich than is cur- C : N ratio from 20 : 1 to 79 : 1 (F. vesiculosus) and from rently appreciated, and that several species go under 8.6 : 1 to 75 : 1 (E. compressa). The algae showed small the name S. interrupta. The latter species, described variations in C : N ratio throughout the experiments from Cadiz, Spain, is found to be restricted to the East- in darkness. Such as previously identified for phyto- ern Atlantic. The name Stenogramme californica Harvey planktonic species, the variations in the physiologi- 1840, the type species described from the Pacific coast cal responses exhibited by the seaweeds tested suggest of North America, is reinstated for members from that these conventional diagnostics of limitations may 26 ABSTRACTS be misleading in natural algal assemblages, which are very widespread in distribution (including common composed by mixed species. Gel electrophoretic is cur- species such as Spirogyra, Zygnema, Cosmarium, and rently being performed to identify possible proteins Staurastrum). Thallus shape ranges from simple, ob- that may respond specifically to these experimental long unicells (saccoderm desmids, or Mesotaeni- conditions. aceae), to unbranched filaments of cells with smooth walls (Zygnemataceae), to elaborately ornate and deeply incised unicells or filaments made of such cells 68 (placoderm desmids, in four families). Prior analy- DISTRIBUTION AND MORPHOLOGY OF ses of sequences of the chloroplast-encoded gene rbcL DIATOMS IN HIGH ALTITUDE STREAMS ON (1354 bp, or 95% of the gene) sampled from all six KAUAI (HAWAII) families showed that two of the traditional families Main, S. P. (saccoderms and filamentous forms) are paraphyletic, Biology Department, Wartburg College, Waverly, IA 50677, i.e. they do not form two independent clades, al- USA though the component genera may together consti- tute a monophyletic group. In contrast, three fami- Diatom samples collected from Kawaikoi Stream differ lies of placoderm desmids formed three monophyletic in species composition and relative abundance from groups within a larger clade. New analyses using par- those in Kokee Stream. Kuwaikoi collects drainage tial sequences from the mitochondrial gene coxIII from the Alakai Swamp, which developed on a 1200– (608 bp, or 76% of the gene) for 21 species of con- 1500 m elevation ridge of Mt. Waialeale extending at a jugating green algae from all six families supported right angle to the NE winds. Kokee lies about 2000 m the conclusions of the rbcL analysis, although boot- further west at about 1100 m elevation. Frustulia and strap support for each gene along was weak for many Eunotia species characteristic of low conductivity wa- branches. The coxIII sequences are more variable ters predominate. Greater diversity of Achnanthes, Nav- than rbcL and have proportionately more parsimony- icula, Pinnularia and Surirella taxa occurs in the some- informative sites (43% vs. 34%, respectively). Com- what more conductive waters of Kokee. The morpho- bining the two genes improved the resolution of the logical species observed bear a close, but not exact, resulting tree: (1) bootstrap support increased for resemblance to mainland taxa described in the litera- all branches, particularly in regards relationships of ture. Morphological variations are documented using placoderm desmids; and (2) monophyly of a clade light and scanning electron microscopy. containing filamentous and saccoderm desmids was strongly supported. We conclude that phylogeny of conjugating green algae will be much improved by 69 sampling from additional genes. PHYLOGENY OF THE CONJUGATING GREEN ALGAE (ZYGNEMATALES AND DESMIDIALES) BASED ON PLASTID AND MITOCHONDRIAL 70 GENE SEQUENCES THE ROOTS OF LAND PLANTS: RECENT McCourt, R. M.1,∗, Park, J. 2, Hewitt, D. A.3 & RESEARCH ON EARLY DIVERGING LINEAGES Karol, K. G.4 IN THE EVOLUTION OF HIGHER DRIER ALGAE 1Department of Botany, Academy of Natural Sciences, McCourt, R. M.1, Karol, K. G.2 & Delwiche, C. F.2,∗ Philadelphia, PA 19103; 2Department of Biology, 1Department of Botany, Academy of Natural Sciences, University of Pennsylvania, Philadelphia, PA 19104; Philadelphia, PA 19103; 2Cell Biology & Molecular 3Farlow Herbarium, Harvard University, Cambridge, MA Genetics, University of Maryland, College Park, MD 02138; 4Cell Biology and Molecular Genetics, University of 20742, USA Maryland, College Park, MD 20742, USA Recent multigene phylogenetic analyses of charo- The conjugating green algae (Order Zygnematales phycean algae sensu Mattox and Stewart have ad- and Desmidiales) are charophyceans, that is, they are vanced our understanding of the relationships of the among the green algae most closely related to land green algae that are most closely related to land plants. plants. Compared to the five other groups of charo- Several clades of green algae are monophyletic with phyceans (Mesostigma, Klebsormidiales, Chloroky- land plants (i.e. liverworts, mosses, nonvascular and bales, Coleochaetales, Charales), the conjugating vascular plants that arose from an aquatic ancestor) greens are by far the most diverse taxonomically to the exclusion of other green algae: Mesostigma (6 families, about 55 genera, 3–4000 species) and (a flagellate unicell); the Chlorokybales (small pack- ABSTRACTS 27 ets of cells); Klebsormidiales (unbranched filaments); coid), Tribonematales (filamentous), and Vaucheri- Zygnematales (conjugating unicells, filaments, and ales (siphonous) are not monophyletic. The major- colonies); Coleochaetales (filamentous and parenchy- ity of species are divided between two clades defined matous thalli); and Charales (branched filamentous by the presence or absence of a bipartite cell wall. thalli with complex nodal structure). The morpholog- Also, several families, particularly those placed in the ical and molecular diversity of the extant members of Mischococcales, are resolved as polyphyletic. Results this nearly exclusively freshwater clade is remarkable. indicate that coccoid and filamentous life forms may Moreover, the taxon diversity within clades is highly have arisen independently at least three times, and asymmetric, having yielded several distinctive albeit some coccoid and filamentous life forms are resolved small groups of green algae, as well as one of the as closely related. For example, Heterococcus (fil- major evolutionary success stories on earth, viz. land amentous) is more closely related to Mischococcus plants. The most recent analysis (Karol etal. (2001) and Pseudopleurochloris (coccoid) than it is to other Science 294 : 2351–2353) of four genes from plastid, mi- members of the Tribonematales. Our data strongly tochondrial, and nuclear compartments strongly sup- support the contention that Bumilleriopsis and Pseu- port the sister relationships of Charales (stoneworts) dobumilleriopsis are congeneric with Bumilleria. The to the most primitive land plants. The analysis also siphonous xanthophytes including Asterosiphon, Botry- shows that within this clade there has been a trend to- dium, and Vaucheria probably do not form a mono- ward increasing complexity of thallus form (from uni- phyletic group although robust bootstrap support for cells, to filaments, to branching and parenchymatous this conclusion is low. Our results indicate that the thalli) and reproduction (absence of sexual reproduc- generic, familial, and ordinal classification of the Xan- tion to iso- or anisogamy, to oogamy). Although the thophyceae requires major revision. major clades mentioned above are well supported by molecular analyses, relationships among these groups 72 are less well-resolved, especially for the Klebsormidi- ales, Chlorokybales, and Mesostigma. This paper re- PHYLOGENETIC RELATIONSHIPS AMONG PEDIASTRUM sPP. AND HYDRODICTYON sPP. views the morphological and molecular evidence for ∗ the relationships among and within these groups. McManus, H. A. & Lewis, L. A. More sequence data from a broader sampling of taxa Department of Ecology and Evolutionary Biology, at the base of the charophyceans (including perhaps University of Connecticut, 75 North Eagleville Road, U-43, some algae not now known to be charophytes) should Storrs, CT 06269, USA provide a clearer picture of the evolution of this group Within the class Chlorophyceae, the family Hydrod- and the reasons for its success. ictyaceae (order Sphaeropleales) consists of taxa that form flat or net-like coenobia and reproduce asexu- 71 ally by way of biflagellated zoospores. Two taxa within PHYLOGENETIC ANALYSIS OF THE this family, Pediastrum and Hydrodictyon, share many MISCHOCOCCALES, TRIBONEMATALES, AND features during development, especially the manner VAUCHERIALES (XANTHOPHYCEAE) INFERRED of daughter colony formation. The colonies of Pedi- FROM 18S rRNA GENE SEQUENCES astrum differ from Hydrodictyon in that growth is pla- McElhinney, A.1,∗, Bailey, J. C.1 & Andersen, R. A.2 nar resulting in two-dimensional colonies, while three- 1Department of Biological Sciences and Center for Marine dimensional nets are formed in Hydrodictyon. Stud- Science, UNC-Wilmington, 601 S. College Road, ies of the Chlorophyceae using morphological and Wilmington, NC 28403; 2Bigelow Laboratory for Ocean ultrastructural data, as well as molecular sequence Sciences, W. Boothbay Harbor, ME 04575, USA data, have supported the close relationship of Pedi- astrum and Hydrodictyon and they have remained in The Xanthophyceae have traditionally been classified the family Hydrodictyaceae throughout major clas- using a phenetic classification system based upon veg- sification revisions. However, in these studies only etative cell structure. Species belonging to the class single species of Pediastrum (P.duplex) and Hydrodic- are placed in one of six orders corresponding to tyon (H. reticulatum) were included, therefore the ex- amoeboid, coccoid, filamentous, flagellate, palmel- act relationship of these two taxa could not be ex- loid, or siphonous organization. Nuclear 18S rRNA plored. Preliminary molecular data from a second gene sequences were determined for 29 species and species of Pediastrum, P.boryanum, indicate that Hydro- aligned with 10 previously determined sequences. Par- dictyon may be derived from Pediastrum. In this anal- simony and maximum likelihood trees inferred from ysis, H. reticulatum resolves as sister taxon to P.duplex these data imply that the orders Mischococcales (coc- with P.boryanum the ancestral taxon. Further molec- 28 ABSTRACTS ular studies of the family Hydrodictyaceae allow ex- dissimilarity) was dramatic and associated with differ- ploration of the relationships between Pediastrum ences in abiotic variables along the estuarine gradient. spp. and Hydrodictyon spp., and assist in determining whether they are monophyletic. Preliminary analyses 74 include H. reticulatum and H. africanum and several species of Pediastrum, P.duplex, P.boryanum, P.tetras, CHARACTERIZATION OF sufB SEQUENCES FROM NONGREEN PHOTOAUTOTROPHS P.biradiatum and P.simplex. An understanding of the ∗ phylogenetic relationships among the species of Pe- Misner, I. J., Bailey, J. C. & Freshwater, D. W. diastrum and Hydrodictyon will allow a more complete Department of Biological Sciences and Center for Marine comparison of colony formation. Science, UNC-Wilmington, 601 S. College Road, Wilmington, NC 28403, USA

Partial sequences for the hypothetical chloroplast 73 frame 24 (ycf24) were determined for 13 species of al- RELATING PHYTOPLANKTON DYNAMICS TO gae representing the classes Bacillariophyceae, Chrys- ENVIRONMENTAL FORCING IN THE LOWER ophyceae, Pelagophyceae, Pinguiophyceae, Prymne- ST. JOHNS RIVER ESTUARY: A MULTIVARIATE siophyceae, Raphidophyceae, and Xanthophyceae. In ASSESSMENT many databases ycf24 is considered an ATP-binding ∗ Millie, D. F. , Pigg, R. J., Bendis, B. J. & cassette (ABC) transporter. Our analyses of trans- Steidinger, K. A. lated amino acid sequences demonstrate that these 2Florida Marine Research Institute, Fish and Wildlife sequences lack Walker A and B boxes, a conserved Conservation Commission, 100 8th Avenue, St. Petersburg, ABC signature sequence motif, and transmembrane FL 33701, USA domains that characterize ABC transporters. Our data, as well as those of others, convincingly indicate that The St. Johns River, a 300-mile riverine and estu- ycf24 is not an ABC transporter and is instead homolo- arine system located along the north-eastern coast- gous to the sufB gene first described from prokaryotes. line of Florida, has undergone extensive eutroph- The function of the sufB gene product in eukaryotic ication through point and nonpoint source nutri- plastids is not known with certainty but it likely plays ent inputs. Moreover, recent reports of sudden fish a key role in iron–sulfur (2Fe−2S) cluster formation. kills and a high incidence of fish with lesions sug- Our data, unpublished results and database searches gest the potential for harmful algal blooms. As part imply that sufB is plastid encoded in the ‘red’ plas- of a NOAA-funded project involving the characteri- tid lineage, but has been transferred to the nucleus zation of water quality parameters in relation to wa- or lost within the streptophyte lineage. Phylogenetic ter inflows and nutrient inputs and the development analyses of all available eucaryotic sufB sequences in- of species-specific markers/probes for instrumental- dicate that this gene is a useful marker for tracing the based monitoring efforts, we used primer-based mul- ancestry of plastids and their hosts. For example, our tivariate analyses to examine the relationship between sufB analyses indicate that the 35-Kb circular plastid abiotic variables and both total and phylogenetic- genome of Plasmodium falciparum is a member of the group chlorophyll a concentrations (derived from ‘red’, not ‘green’, plastid lineage and was acquired via photopigments and ChemTax matrix factorization of secondary endosymbiosis. Our results also support the diagnostic carotenoids) during 2001. Seven sampling hypothesis that, despite their similarities, the plastids sites (identified through principle components ordi- of prymnesiophytes and heterokont algae were inde- nation of physical/chemical parameters as ranging pendently obtained by separate secondary endosym- from oligo- to mesohaline) were sampled intensively biotic events. over 2-week periods on a seasonal basis. Chlorophyll a concentrations typically ranged from 5 to 35 µg/L with the greatest concentrations occurring at the oligo- 75 haline sites. Phytoplankton assemblages were domi- ARE ‘GREEN TIDES’ HARMFUL ALGAL nated by diatoms, cryptophytes, and cyanobacteria, BLOOMS? ALLELOPATHIC PROPERTIES OF and together typically comprised up to 90% of the to- EXTRACTS FROM ULVA FENESTRATA AND tal chlorophyll a. Temporal variability in phytoplank- ULVARIA OBSCURA ton assemblages followed seasonal trends impacted by Nelson, T. A.1,∗, Lee, D.1, Smith, B. C.2 & Prins, R.1 meteorological and hydrological forcing. Spatial vari- 1Department of Biology, Seattle Pacific University, Seattle, ability in phylogenetic-group abundance (illustrated WA 98119-1997; 2School of Aquatic and Fishery Sciences, through multidimensional scalar ordination of sample University of Washington, Seattle, WA 98195, USA ABSTRACTS 29

Toxic properties are rarely associated with macroalgal experiments. Incubator experiments alone were used blooms. A few studies have demonstrated inhibition to measure nutrient uptake rate, chlorophyll a and b of invertebrate larval development by extracts from content, and protein content (as determined by far- Ulva spp. and we have observed substantial discol- UV and bicinchoninic acid (BCA) techniques). In- oration of the seawater in small bays following mass creasing nutrient concentration caused significantly desiccation-induced mortality of Ulvaria obscura.We more growth in the seawater table experiment but examined the effects of extracts from Ulva fenestrata only a trend toward greater growth in the incuba- and U. obscura on enzyme systems, larval oyster devel- tor. There was no significant interspecific difference opment, Fucus gardneri zygote development, epiphytic in growth rate in the seawater table experiment, but diatom growth, and growth of Ulva and Ulvaria. Ex- Ulva grew significantly faster than Ulvaria in the in- tracts from Ulvaria inhibited trypsin activity, but not cubator experiment. Tissue nitrogen increased signif- pepsin or amyloglucosidase activity when tested at pH icantly with increasing nutrient content in the growth optimized for each enzyme system (pH 8, 2, and 5.5, medium in the incubator experiment and showed a respectively). When tested at neutral to slightly alka- similar (albeit not significant) trend in the seawater line pH (7 or 8), Ulvaria extracts also inhibited amy- table experiment. Ulvaria had higher tissue nitrogen, loglucosidase activity. ‘Aged’ extract caused greater protein, and chlorophyll a concentrations than Ulva enzyme inhibition than fresh extract. Maximum in- in all experiments. The ratio of chlorophyll b : a was hibition occurred in 6-h-old-extract. Oyster larval de- also higher in Ulvaria than in Ulva. Far-UV and BCA velopment was inhibited by extracts from both algae, techniques yielded equivalent protein concentration although the visible effects were much greater when in Ulva but protein concentration determined by BCA the larvae were exposed to extracts of Ulvaria. Fucus was 2.5 times higher in Ulvaria than when determined zygote development was inhibited by extracts of both by far-UV. Nutrient uptake rates were similar between species, although the effect was measurably greater in both species. DMSP concentration was not affected sig- extracts from Ulvaria. Epiphytic diatom growth and nificantly by nutrient regime, but [DMSP] was higher growth of Ulva and Ulvaria were inhibited by extracts in Ulvaria than in Ulva. These results help explain from both species. Thus, extracts from both Ulva fen- previous observations of these species. Ulvaria gener- estrata and U. obscura have allelopathic properties, al- ally grows more slowly than Ulva because it requires though the impacts are more widespread for Ulvaria more nitrogen to make chlorophyll and protein but extracts. These properties could alter competitive in- has similar nitrogen uptake rates. Ulvaria is also shade teractions by inhibiting germination or development adapted, while Ulva can acclimate to various condi- of algal and animal competitors. Multiple toxins are tions. probably involved, since the two species tested cause visibly different responses. 77 THE EFFECTS OF THALLUS TOUGHNESS AND 76 NUTRITIVE QUALITY ON PALATABILITY OF THE EFFECTS OF NITROGEN REGIME ON ANTARCTIC MACROALGAE GROWTH, [CHN], [DMSP], [CHLOROPHYLL], Peters, K. J.1,∗, Amsler, C. D.1, Amsler, M. O.1, [PROTEIN] AND NITROGEN UPTAKE RATE IN McClintock, J. B.1, Iken, K. B.1 & Baker, W. J.2 ULVARIA OBSCURA AND ULVA FENESTRATA 1Department of Biology, University of Alabama at Nelson, T. A.1,∗, Van Alstyne, K. L.2 & Ribarich, H.1 Birmingham, Birmingham, AL 35294-1170; 2Department 1Department of Biology, Seattle Pacific University, Seattle, of Chemistry, University of South Florida, Tampa, FL WA 98119; 2Shannon Point Marine Center, 1900 33620, USA Shannon Point Road, Anacortes, WA 98221, USA Palatability of macroalgae can be affected by many dif- Ulva fenestrata and Ulvaria obscura are major con- ferent factors including thallus toughness, nutritional stituents of ulvoid macroalgal blooms in the north- quality (carbohydrates, lipids, proteins, ash), and the eastern Pacific. These blooms have been associated presence of deterrent secondary metabolites. Palata- with a variety of environmental problems and are bility bioassays were performed using two different thought to be caused by increased nitrogen concentra- antarctic algal grazers (the antarctic rockfish Notothe- tion. We grew these algae at several nutrient concen- nia coriiceps and the sea star Odontaster validus). These trations in incubator and seawater table experiments. data were then compared to toughness and chemical Growth rate and the concentrations of DMSP, C, H, composition data. Toughness was determined using a and N in the tissues were determined for both kinds of penetrometer and a wide range of thallus toughness 30 ABSTRACTS was observed in individuals of differing species as well formation may explain why most multicellular algae as of the same species. Some of the tougher individu- remain recalcitrant to cryopreservation, and lead to als required more than 30 N of force to cut through protocols that allow the cryopreservation of a broader the thallus while others required less than 1 N to break range of organisms. through. Each algal species is also being tested for per- cent composition of carbohydrates, lipids, proteins, as 79 well as ash. It is hypothesized that the algal species with greater nutritional quality and/or less thallus tough- PROTIST ORIGIN OF THE ULVOPHYCEAE ness are more likely to be acceptable foods to con- (CHLOROPHYTA) REVEALED BY SSU RDNA sumers (NSF OPP9814538, OPP9901076). ANALYSES OF MARINE COCCOID GREEN ALGAE Proschold,¨ T.∗†, Surek, B., Marin, B. & 78 Melkonian, M. SURVIVAL OF CHLAMYDOMONAS SUBSEQUENT Botanical Institute I, University of Cologne, Gyrhofstr. 15, TO CRYOPRESERVATION IS PREVENTED BY A D-50931 Koln,¨ Germany SUBSTANCE RELEASED FROM DAMAGED CELLS Piasecki, B.1,∗, Fan, Yang1, Diller, K. R.2 & Traditionally, brackish and marine coccoid green al- Brand, J. J.1 gae are placed as isolated species of common treboux- 1Molecular Cell and Developmental Biology, 2Department iophycean or chlorophycean genera (e.g. Chlorella, of Biomedical Engineering, University of Texas at Austin, Chlorococcum); exclusively the marine genera Chlorocys- Austin, TX 78712, USA tis and Halochlorococcum are characterized by quadri- flagellated zoospores and a ‘Codiolum-stage’ (char- Storage of algae in liquid nitrogen offers a convenient acteristic for the Ulvophyceae) in their life histo- way to permanently preserve many cultures with mini- ries. In Chlorocystis cohnii, sexual reproduction could mum genetic drift. We have successfully cryopreserved only be induced by presence of the tube-dwelling di- over 1400 strains in the Culture Collection of Algae at atom Berkeleya rutilans. Phylogenetic analyses of the the University of Texas (UTEX), but over one-fourth SSU rDNA sequences of 32, mostly marine, coccoid of UTEX cultures remain recalcitrant. Our previous or sarcinoid strains confirmed the monophyly of the studies demonstrate that unicellular algae frozen at Ulvophyceae. The class contain 11 independent well- a high cell density (>2000000 cells/mL) do not sur- supported lineages; of which eight contain coccoid vive cryopreservation well. We now show that viability species. (1) The basal branch of the ulvophytes is during cryopreservation of Chlamydomonas reinhardtii the Oltmannsiellopsis clade (in agreement with the or- UTEX #89 at high cell densities is not affected by der Oltmannsiellopsidales emend.) which contains the physical proximity of cells. Instead, when cells the quadriflagellated monads (O. viridis and O. uni- are damaged, a small organic molecule(s) is pro- cellularis) and the type species of the coccoid genus duced and released into the medium preventing cry- Halochlorococcum (H. marinum). (2) The other species opreservation of other cells in the culture. Healthy of Halochlorococcum (except H. saccatum) formed to- C. reinhardtii cultures were treated with extracts pre- gether with Chlorocystis and Chlorella salina a signifi- pared from damaged cells or culture medium sepa- cant clade corresponding to the order Chlorocystidales rated from damaged cells, then cryopreserved by a in emended form. (3) Gomontia polyrhiza is closely re- standard method (Crutchfield etal. (1999) Eur J Phy- lated to Urospora penicelliformis within the Acrosipho- col 34: 43–52). Viability was measured after thawing by nia group (Acrosiphoniales). (4) The sarcinoid Tri- the ability of a cell wall to preclude Evan’s blue dye, chosarcina polymorpha and T. mucosa, and the freshwa- and was confirmed by quantitative agar plating. The ter planctonic Helicodictyon planctonicum are member inhibitory substance from these fractions appears to af- of the Ulothrix clade together with filamenteous taxa. fect living cells only during cryopreservation, but can (5) The type species of Pseudoneochloris (P.marina) be produced from cells by a variety of treatments. The form together with three unidentified marine coc- inhibitor is heat-stable, but heating cells to 80 ◦C pre- coid strains an independent lineage within the Ul- cludes its production. The substance is stable when vophyceae (called Pseudoneochloris clade). (6–8) Ig- separated from cellular particulate matter, but grad- natius tetrasporus (isolated from soil), the strain CCMP ually becomes inactive in the presence of cell mem- 250 originating from desert soil and the marine strain brane fractions. The inhibitor is a water-soluble and CCMP 1293 isolated from Palau Islands represent heat-stable organic compound(s), with a molecular three single lineages within the ulvophytes. From mass less than or equal to 3500. Characterizing the our results, the class Ulvophyceae can be subdivided inhibitory substance and the pathway that leads to its taxonomically into at least 11 orders (including the ABSTRACTS 31

Ulvales, Cladophorales and Dasycladales) with the Algal and seagrass community composition were mon- largest biodiversity at the coccoid level. itored biennially along three transect lines in Belize †Present address: Department of Molecular and Cell from 1997 through 2001. Two sites were on South Wa- Biology, Brown University, Providence, RI 02912, USA. ter Caye. The first was on the seaward side, between the reef crest and the caye. The second was on the landward side of the caye. The third site was on Long 80 Caye, Glover’s Reef Atoll. This transect line ran from a coral rubble filled embayment, through a small chan- CALCIFICATION AND DECALCIFICATION nel (formed by hurricane Hattie in 1960, bisecting DURING EPITHALLIAL CELL TURNOVER IN Long Caye), and into a seagrass meadow in the in- CORALLINE RED ALGAE terior of the atoll. After a 38-year hiatus, hurricanes 1,∗ 2 Pueschel, C. & Wegeberg, S. struck Belize in 1998, 2000 and 2001, giving us the 1 Department of Biological Sciences, State University of New opportunity to examine changes in plant community York at Binghamton, Binghamton, NY 13902, USA; structure before and after the storm sequence. Min- 2 Marine Biological Laboratory, Strandpromenaden 5, imal interannual changes were seen at South Water 3000 Helsingor, Denmark Caye. The seagrasses at Long Caye were nearly elim- inated along the transect line due to burial by sand. Epithallial cells of the coralline red algae are charac- Peak density of Thalassia testudinum at Long Caye was terized by unusual structural specialization, which in- over 60 shoots/m2 in 1997 and less than 2 shoots/m2 clude deep invaginations of the distal cell surface, and in 1999 and 2001. Halodule wrightii and Syringodium by unique development, which culminates in senes- filiforme were both completely eliminated between cence, shedding, and replacement of the cells. Elec- 1997 and 1999. Padina and Dictyota spp. flourished in tron microscopic study of epithelial cell differentia- the embayment following the hurricane, but had been tion in morphologically and taxonomically disparate virtually absent prior to the sequence of storms. Fu- species suggests that the unusual features of epithelial ture monitoring will be conducted to follow recovery cell structure and development stem from the fact that of the meadow and look for other long-term changes. these dynamics occur within a calcified matrix. Distal wall ingrowths begin to form on the initial cells, cells whose cleavage eventually gives rise distally to new ep- ithelial cells. After the distal wall ingrowths form, the 82 overlying crosswall becomes rich in organic material. SEQUENCE DIVERSITY OF THE GENE tufA IN For this organic wall material to be deposited into the COLEOCHAETE: AN ALGAL MODEL FOR existing crosswall, the wall must first be decalcified; HORIZONTAL GENE TRANSFER ∗ therefore, the presence of abundant organic material Ricker, J. M. , Lewandowski, J. D. & Delwiche, C. F. in the crosswall provides a marker of localized decal- Cell Biology and Molecular Genetics, University of cification. We propose that the location and time of Maryland – College Park, College Park, MD 20742-5815, origin of distal wall ingrowths indicate a connection USA between the ingrowths and two coordinated processes: localized secretion of wall material, and decalcifica- Studying the organellar gene transfer of tufA in tion of the overlying cell wall in preparation for the Coleochaete serves as a model system for understand- movement of the young epithelial cell into a new loca- ing the effect of functional horizontal gene transfer on tion relative to the surrounding calcified matrix. The sequence conservation and divergence. Previous work large plasmalemmal surface area associated with the indicates that tufA, which codes for protein synthesis distal wall ingrowths allows for a greater abundance elongation factor TU, is encoded only in the chloro- of membrane-associated components, such as proton plast genome in most green algae, red algae, and glau- pumps, that could drive localized cell wall decalcifica- cocystophytes, but only in the nuclear genome of em- tion. bryophytes. In the charophytes, the lineage of green al- gae that gave rise to embryophytes, tufA is present var- iously within the chloroplast and/or nuclear genome. 81 This implies that tufA was probably transferred to the INTERANNUAL VARIATION IN CORAL REEF nucleus from the chloroplast after the charophytes di- PLANT COMMUNITIES OF CENTRAL BELIZE verged from other green algae but before the diver- Rhodes, M. J. & Nelson, T. A.∗ sification of the group. The chloroplast copy has un- Department of Biology, Seattle Pacific University, Seattle, dergone three separate fates within the charophytes: WA 98119-1997, USA maintained in Charales, divergent in Coleochaetales, 32 ABSTRACTS and lost in Zygnematales. Previous research indicates greater at 29 ◦C than 24 ◦C, but maximal growth rates the presence of multiple nuclear copies and a highly were reduced approximately 50–90% at temperatures divergent chloroplast encoded tufA in Coleochaete or- of 32 and 34◦C, respectively. The most sensitive clone, bicularis. This research aims to isolate the chloroplast- RD04, stopped dividing after two divisions at 29 ◦C. encoded tufA from a representative panel of Coleochaete The growth rate of JR02F2 and RD01 reached a max- in order to characterize patterns of conservation in the imum at PAR levels greater than 100 mE/m2/s and divergent chloroplast tufA. Putative chloroplast copies remained within a range of 0.75–0.85 divisions/day of tufA have been isolated through PCR and subse- up to 390 mE/m2/s. quent molecular cloning. By comparing the DNA se- quences from copies of the chloroplast-encoded tufA from a representative panel of Coleochaete, we hope 84 to gain insight into patterns of sequence conservation and from this infer the current function of the diver- PHOTOPROTECTIVE COMPOUNDS IN gent chloroplast tufA. RICE-FIELD CYANOBACTERIA Sinha, R. P.∗ & Haeder, D.-P. Institute for Botany 1, Friedrich-Alexander University, Staudtstr. 5, D-91058 Erlangen, Germany 83 EFFECTS OF ENVIRONMENTAL PARAMETERS Cyanobacteria are cosmopolitan in distribution, form- ON THE GROWTH CHARACTERISTICS OF ing a prominent component of microbial populations SYMBIODINIUM SPP. ISOLATED FROM CORALS in both aquatic as well as terrestrial ecosystems. They Rogers, J. E.1,∗, Davis, R. H.2 & Oliver, L. M.1 are the dominant microflora in rice-fields, where they 1US Environmental Protection Agency, Office of Research significantly contribute to fertility as a natural biofer- and Development, National Health and Environmental tilizer. The fluence rates of UV-B radiation impinging Effects Research Laboratory, Gulf Ecology Division, 1 on the natural habitats is of major concern since being Sabine Island Dr, Gulf Breeze, FL 32561; 2UWF/EPA photoautotrophic organisms, cyanobacteria depend Science Training in Ecology Program, Gulf Breeze, FL, USA on solar radiation as the primary source of energy. UV-B radiation causes reduction in growth, survival, Symbiodinium spp. were isolated from Porites porites pigment and protein content, heterocyst frequency (JR02F2 and RD03), Montipora capitata (JR12A7), and fixation of carbon and nitrogen in cyanobacte- Madracis mirabolis (RD02), Montastrea faveolata ria. However, a number of cyanobacteria synthesize (RD04), Pocillopora damicornis (JR13E1), and an water soluble colorless mycosporine-like amino acids unknown coral (RD01). Growth rates and the distri- (MAAs) and the lipid soluble yellow-brown colored bution between motile gymnodinoid cells and sessile sheath pigment, scytonemin to counteract the dam- coccoid cells were determined in 200 µL cultures in aging effects of UV-B. Anabaena sp., Nostoc commune, 96-well dilution plates. Clones of each isolate were Scytonema sp. and Lyngbya sp. were isolated from rice- maintained in L1 medium (prepared with 34 ppt ster- fields in India and screened for the presence of pho- ile Gulf of Mexico water), at 26 ◦C with illumination toprotective compounds. Spectroscopic and biochem- provided at 100–150 mE/m2/s PAR (400–700 nm) ical analyses revealed the presence of shinorine, a during a 10 : 14 (light : dark) photoperiod. Under bisubstituted MAA containing both glycine and ser- these conditions, generation times of nearly once ine group with an absorption maximum at 334 nm in per 24 h were observed for clones, JR02F2, RD01 and all cyanobacteria except Lyngbya sp. There was a circa- RD02, in log growth. Clone RD04 had a generation dian induction in the synthesis of this compound by time of approximately 5 days. The fastest growing UV-B. Polychromatic action spectra for the induction clones produced the highest percentage of motile of MAAs in Anabaena sp. and N. commune show the in- cells (>90) during the daylight portion of the diurnal duction to be UV-B dependent peaking at 290 nm. An- cycle. Growth characteristics of selected clones were other photoprotective compound, scytonemin, with examined over a range of salinities, temperatures and an absorption maximum at 386 nm (also absorbs sig- PAR intensities. Salinities ranging from 25 to 45 ppt nificantly at 300, 278, 252 and 212 nm) was detected in had little if any affect on growth rate. At salinities all cyanobacteria except Anabaena sp. In addition, two less than 25 ppt significant decreases in growth were unidentified, water-soluble, yellowish and brownish generally observed; no growth was common at 5 and compounds with an absorption maximum at 315 nm 10 ppt. In most cases inhibitory effects of temperature were recorded only in Scytonema sp. The spectral char- were observed at temperatures greater than 29 ◦C. For acteristics of these compounds neither matches with example, generation times for JR02F2 were slightly MAAs nor with scytonemin and seems to be a novel ABSTRACTS 33 compound acting as an effective UV screens. In con- MAAs shinorine, palythine, MAA 331, asterina-330, clusion, a particular cyanobacterium having photo- and porphyra-334 present in the organisms examined protective compounds may be a potent candidate as were generally best extracted at 45 ◦C in 25% aque- biofertilizers for crop plants. ous MeOH. In Porphyra sp. and natural freshwater phytoplankton, the mean total MAA concentrations obtained with this protocol were, respectively, about 85 13 and 3 times higher than when extracted in 100% ◦ PHYLOGENY OF KLEBSORMIDIUM MeOH at 4 C. Depending on the organism exam- (KELBSORMIDIALES; CHAROPHYTA) BASED ON ined, both MeOH concentration and temperature af- rbcL , atpB , coxIII AND nad5 SEQUENCE DATA fected extraction efficiency and final MAA concentra- Snyder, S. W.∗, Karol, K. G. & Delwiche, C. F. tion. Our results stress the need for a priori testing the 2106 H. J. Patterson Hall, Building 073, University of influence of these variables to assure that the highest Maryland, College Park, MD 20742-5815, USA concentration is obtained without altering the qual- itative MAA composition. Based on these results, di- The charophycean order Klebsormidiales consists of rect comparison of MAA concentrations reported in two unbranched filamentous lineages, the fresh wa- the scientific literature should be done with caution. ter Entransia and the semiterrestrial Klebsormidium. Molecular data have supported the close relationship Klebsormidiales have with land plants. A monograph exists for the European species of Klebsormidium; this 87 monograph provides a cladistic analysis of morpho- TRACING INVASIONS OF CAULERPA TAXIFOLIA logical characteristics for eight Klebsormidium species, WITH MOLECULAR MARKERS but a species level molecular phylogenetic analysis Stam, W. T.1,∗, Meusnier, I.2, Destombe, C.2, has not yet been performed for the group. We have Valero, M.2 & Olsen, J. L.1 obtained 50 strains of Klebsormidium, representing 11 1Departement of Marine Biology, Centre for Ecological and morphological species, both from nature and from Evolutionary Studies, University of Groningen, the culture collections. Phylogenetic analyses using rbcL, Netherlands; 2Labaratoire de Gen´ etique´ et Evolution des atpB, coxIII and nad5 gene sequence data from these Populations Veg´ etales,´ UPRESA CNRS 8016, Universitede´ strains and 40 outgroup sequences suggest that several Lille-1, France traditional Klebsormidium species may not be mono- phyletic. Comparative sequence data from the rDNA ITS have proven very useful in species identification and identi- fying major biogeographic groups of Caulerpa taxifolia. 86 However, the low variability of the sequence, and the A NOTE OF CAUTION ON USING ABSOLUTE need to screen hundreds of samples has limited its METHANOL CONCENTRATIONS AND LOW utility for population level studies. Here we report TEMPERATURES TO QUANTIFY on the performance of two new markers suitable for MYCOSPORINE-LIKE AMINO ACIDS IN ALGAE population level screening: the chloroplast rDNA 16S Sommaruga, R.∗ & Tartarotti, B. intron-2 and intersimple-sequence-repeat fingerprints Institute of Zoology and Limnology, University of (ISSRs). A new analysis of ITS1 insertion–deletion pat- Innsbruck, Technikerstr. 25, 6020 Innsbruck, Austria terns was also conducted using all available sequences. A number of new insights have emerged. First, the in- Mycosporine-like amino acids (MAAs), a family of UV- vasive aquarium strain (Mediterranean, California) is absorbing compounds, are widespread among many clearly from Brisbane, but the Brisbane populations marine and freshwater organisms, however, no stan- themselves may be the result of an introduction from dard method to analyze these compounds is available. Northern Australia. Second, an analysis of the new Consequently, protocols to extract MAAs largely dif- data in conjunction with an ITS1-insertion–deletion fer, for example, in the type of solvent, concentra- analysis further suggests that the Mediterranean pop- tions, temperatures, and times used. In this study, ulations may be the result of not one, but two sepa- we tested whether the concentrations of MAAs are rate introductions. Third, intrapopulation genetic di- affected by extraction at different temperatures and versity between invasive Mediterranean and ‘native’ methanol (MeOH) concentrations. Natural assem- Australian populations revealed the occurrence of blages of freshwater phytoplankton and the marine two divergent and widespread clades. The first clade alga Porphyra sp. were used as test organisms. The grouped nontropical invasive populations with coastal 34 ABSTRACTS inshore populations of Australia while the second clus- that have been raised in support of a single plastid tered all offshore/oceanic populations so far stud- origin. ied. C. taxifolia, therefore exists as a complex of in- dependent ecotypes that probably represent nascent 89 species. Fourth, despite our finding of nine distinct ULTRASTRUCTURE AND PHYLOGENY OF A nuclear and five distinct chloroplast profiles, strong NEW SPECIES BELONGING TO THE linkage disequilibrium was found in most specimens, PRYMNESIOPHYCEAE which indicates a predominance of asexual reproduc- ∗ Tanczos, M. A.1, , Bailey, J. C.1 & Andersen, R. A.2 tion. However, nucleo-cytoplasmic recombination was 1Department of Biological Sciences and Center for Marine detected in one case, supporting hybridization both Science, UNC-Wilmington, 601 S. College Road, within and between populations. Wilmington, NC 28403; 2Bigelow Laboratory for Ocean Sciences, W. Boothbay Harbor, ME 04575, USA

88 Ultrastrucural and molecular data were used to ex- THE CASE FOR A SINGLE-PLASTID ORIGIN amine a new alga (CCMP2000) collected from the REVISITED: CONVERGENT EVOLUTION IN Arabian Sea. Nonmotile vegetative cells are typically ORGANELLAR GENE CONTENT oblong to ovate, solitary and lack a gelatinous enve- Stiller, W. J.1,∗, Reel, D. C.1 & Johnson, J. C.2 lope. Ultrastructural analyses of these cells revealed 1Department of Biology, Howell Science Complex N108, the presence of a nucleus located in the posterior por- 2Institute for Coastal and Marine Resources, East Carolina tion of the cell. A single large Golgi body with numer- University, Greenville, NC 27858, USA ous cisternae is located anterior to the nucleus as are numerous profiles of mitochondria possessing tubu- Three eukaryotic lineages generally are believed to lar cristae. Two peripheral plastids are present hav- have plastids that are primary in origin; that is, de- ing immersed pyrenoids that are traversed by one or scended directly from a cyanobacterial endosymbiont. more thylakoids. In some cells, the plastids are con- The recovery of these plastids as a monophyletic group spicuously lobed forming finger-like extensions. Non- in most molecular phylogenetic analyses, along with motile vegetative cells possess reduced mature and similarities in genome content and protein targeting immature flagella as well as a haptonema that are mechanisms, have been cited as strong evidence in 1 micron long or less. A distinct internal covering support of the hypothesis of a single endosymbiotic and external wall surround the cell protoplasm. The origin of all plastids. Although these data indeed are outer wall is apparently composed of simple scales, but consistent with a single plastid origin, they also are the composition (organic or mineralized?) of these consistent with the proposition of multiple endosym- scales is unknown. The outer wall may form a pro- biotic origins. Each hypothesis requires certain evolu- tuberance, or ‘foot’, at the posterior end of non- tionary assumptions in order to be reconciled with all motile cells. Interestingly, many vegetative cells are in- existing data; at present, it is unclear which of these fected by a labyrinthulid. Flagellate stages have been assumptions most likely reflect the historical process observed and possess roughly equal length flagella that gave rise to plastid diversity. Here we examine sim- and a haptonema. Phylogenetic analyses of nuclear ilarities in gene content among representatives of the 18S rRNA and plastid-encoded rbcL genes indicate three primary plastid lineages, using as a control the CCMP2000 is most closely related to Phaeocystis. How- genome of a mitochondrion that almost certainly orig- ever, CCMP2000 does not fit the description of Phaeo- inated as an independent endosymbiotic association. cystis, and therefore, probably warrants recognition as To minimize metabolic constraints on gene retention a new taxon. we focus on two datasets, ribosomal protein and trans- fer RNA genes, neither of which is tied directly to 90 specific organellar functions. Analyses of all possible THE EFFECTS OF BIOGENICALLY ACTIVE pair-wise comparisons among the three plastids and COMPOUNDS ON CYCLOTELLA MENEGHINIANA mitochondrion indicate that genomic similarities are (BACILLARIOPHYTA) most consistent with convergent evolution due to con- Tedrow, O.∗, Julius, M. L. & Schoenfuss, H. L. straints on gene loss, rather than with hypothesized Department of Biological Sciences, St. Cloud State shared evolutionary histories. We find no evidence University, St. Cloud, MN, USA of phylogenetic signal in the pattern of gene loss overlying this convergence. In light of these results, Biogenically active compounds have been discovered we address other lines of evidence and arguments in many surface water samples in Europe and North ABSTRACTS 35

America. Wastewater effluent is identified as a major of the earliest valve formation, appears to be about source of contamination and found to revert much the size of a single areola. Early rimoportulae (labiate of the metabolic products of these biologically active processes) are interspersed between central areolae compounds back to their original form. This presen- and are simple oval openings. Internally these form tation explains methodology for determining the ef- their labiate lip-like structures just before cribrum for- fects of these contaminants through a newly developed mation. Ridges form on the external side of the basal bioassay, examining the physiological response of a di- layer in a honeycomb pattern and produce the walls atom to these compounds. Diatoms represent an im- of the loculate structures. The ridges expand outward portant aspect of the primary production community. until cribrum formation is initiated. Small protuber- They are a desirable food source over other mem- ances grow out from the edge of the ridge-like struc- bers of the primary production community through tures and then become anvil-shaped as they grow to- storage of photosynthetically produced sugars in the ward the center of the pores and form the ‘roofs’ of form of lipids rather than starch. Therefore, members the loculi. In external view of the mature valve, the of higher trophic levels selectively feed on diatoms pores are smoothly covered over except for a ring of when present with other members of the primary pro- C-shaped slits. These cribra are flush with the surface duction community. This study examines the effects of the valve. At about the time the cribra are com- of select biogenically active compounds on the gross pleting, rotae are formed within the porelli of the morphology and physiological development of the di- pseudocelli by extensions from two or three sides. atom species Cyclotella meneghiniana. Clearly, unicellu- The quadrangular T. formosum develops along much lar protists such as diatoms are not susceptible to these the same pattern. Differences are the less numerous, contaminants in a manner directly analogous to that more centrally located rimoportulae, which possess expressed in vertebrates. However, many of these com- external tubes and the formation of highly domed pounds are lipophilic making them particularly effec- cribra. tive in entering the diatom cell membrane. Compar- ing cell density, lipid composition, fatty acid concen- trations, and the electron microscope ultra-structure 92 in control and exposed cultures achieve this study’s experimental objectives. This diatom was selected be- THE CHLOROPLAST GENOME SEQUENCE OF cause it commonly occurs in most freshwater environ- CHLOROKYBUS ATMOPHYTICUS: EVIDENCE ments and has been the subject of other toxicological THAT CHAROPHYCEAN GREEN ALGAE FROM studies. An adequate literature base also exists for eval- AN EARLY DIVERGING LINEAGE ADAPTED TO TERRESTRIAL LIFE uating results of this experiment. The species grows ∗ rapidly and is easy to maintain in culture. The use of Turmel, M. , Otis, C., De Cambiaire, J.-C., Pombert, a phytoplankton species as a test organism comple- J.-F. & Lemieux, C. ments other investigations involving vertebrate mod- Departement´ de biochimie et de microbiologie, Universite´ els at the apex of the trophic hierarchy by considering Laval, Quebec,´ Canada G1K 7P4 effects at the base of the food web. Land plants and green algae belonging to the Charo- phyceae sensu Mattox and Stewart form the Strep- tophyta, a lineage sister to the Chlorophyta which 91 comprises most or all of the other green algae. A DEVELOPMENT OF VALVES IN THE MARINE third green plant lineage, at the base of the split of DIATOM GENUS TRIGONIUM the Chlorophyta and Streptophyta, is possibly rep- Tiffany, M. A. resented by the green alga Mesostigma viride; how- Department of Biology, San Diego State University, San ever, this remains controversial because some phylo- Diego, CA 92182, USA genetic analyses place Mesostigma within the Strepto- phyta. Five orders have been recognized in the Charo- Trigonium arcticum and T. formosum, marine diatoms phyceae: the Chlorokybales, Klebsormidiales, Zygne- epiphytic on Ectocarpus sp., were collected from the matales, Coleochaetales, and Charales. Based on its sublittoral off Point Loma, California. Forming valves sarcinoid thallus, the only species in the Chlorokybales, of these two sympatric species were studied using Chlorokybus atmophyticus, has long been regarded as scanning electron microscopy. The earliest develop- the earliest-diverging member of the Charophyceae. ing valve of T. arcticum encountered is a porous basal This rare species has been collected only in terres- layer of silica, triangular, with pseudocelli just starting trial habitats. In the present study, we have deter- to form at each corner. The central annulus, the site mined the complete chloroplast DNA (cpDNA) se- 36 ABSTRACTS quence (149 681 bp) of Chlorokybus to gain insight into 94 its phylogenetic position and also to better understand PHYLOGENETIC ANALYSES OF THE SPECIES how the chloroplast genome evolved in the Strepto- OF CHLOROSARCINOPSIS AND phyta. We have found that Chlorokybus cpDNA closely NEOCHLOROSARCINA (CHLOROPHYCEAE) resembles Mesostigma cpDNA in showing a high degree Watanabe, S.1,∗, Mitui, K.2, Nakayama, T.2 & Inouye, of ancestral features. It displays a quadripartite struc- I.2 ture, encodes 138 genes (only two extra genes com- 1Department of Education, Toyama University, Toyama pared to Mesostigma cpDNA), and contains a single 930-8555; 2Institute of Biological Sciences, University of intron (in trnL(UAA)). The gene order in Chloroky- Tsukuba, Ibaraki 305, Japan bus cpDNA is similar to that of Mesostigma cpDNA; we estimate that 14 inversions would be required The genus Chlorosarcinopsis forms sarcinoid packages to convert the gene order of one alga into that of of ?@ cells and reproduces by forming naked biflag- the other. Phylogenetic analyses of combined chloro- ellate zoospores. The genus Neochlorosarcina is distin- plast protein sequences from Mesostigma, Chlorokybus, guished from Chlorosarcinopsis in having a thin cell other streptophytes, and a number of chlorophytes wall around zoospores. We analyzed phylogenetic rela- will be presented. Overall, our results support the tionships of seven species of Chlorosarcinopsis and four idea that the Chlorokybales represent an early diverging species of Neochlorosarcina using 18S rDNA sequence lineage. data. The species of the Chlorosarcinopsis were sepa- rated into two clades, and those of Neochlorosarcina formed a monophyletic branch, which was far from the 93 Chlorosarcinopsis clades. Groover and Bold (1969) in- RECIRCULATING CULTURE FOR vestigated physiological abilities of species of Chlorosar- CHONDRACANTHUS EXASPERATUS cinopsis (which included species of Neochlorosarcina), Waaland, J. R. and showed that the species were roughly divided Department of Botany, University of Washington, Seattle, into three groups by usage of nitrogen and carbon WA 98195, USA components. These groups correspond two clades of Chlorosarcinopsis and a clade of Neochlorosarcina. This The red alga Chondracanthus exasperatus is a source of result gave a phylogenetic meaning to the differences the phycocolloid carrageenan as well as an ingredi- in the physiological abilities, and supported the sepa- ent referred to as ‘intralamellar gel’ in a recently de- ration of Neochlorosarcina from Chlorosarcinopsis based veloped cosmetic formula (US Patent 6136 329). The on a morphological attribute. high value of the cosmetic product has sparked re- newed interest in cultivation of this species. Previous cultivation methods for this species include open wa- 95 ter culture on nets and immersed cultivation in tanks DIGITAL ALGAE: A BOON TO RESEARCH AND supplied with flow-through pumped seawater. The in- TEACHING stallation of a high capacity seawater supply, pump- Waters, D. A.1,∗, Ciugulea, I. I.2 & Chapman, R. L.1 ing and drain system is a major cost for flow through 1Department of Biological Sciences, Louisiana State systems. Recirculating or re-use seawater systems that University, Baton Rouge, LA, USA; 2Department of Botany minimize seawater turnover may offer significant cost and Microbiology, University of Bucharest, Romania savings over single-pass, flow through seawater sys- tems. In this research several options for minimizing As part of a larger coastal ecology project, we have ini- seawater use have been tested: recirculating batch cul- tiated a web-accessible, digitized phytoplankton image ture in which nutrient replenished (carbon dioxide and taxonomy database of algae collected in a coastal and mineral nutrients) natural or artificial seawater is wetland south of New Orleans, Louisiana. Examin- used with minimal turnover and spray culture in which ing phytoplankton dynamics is a large component of plants are suspended in air saturated with nutrient re- the project, titled ‘Pulses—The Importance of Pulsed plenished natural or artificial seawater medium. Small Physical Events for Watershed Sustainability in Coastal volume (<2 L), single-plant bioreactors and larger Louisiana’ (a joint NSF/EPA/USDA Water and Water- multiplant, 20, 80 and 320 L (sea water volume) im- shed program). A total of 168 species of algae from 76 mersion and spray systems have been developed and genera in 8 phyla have been identified from the phyto- tested. Results from these systems will be presented. plankton samples, and 130 high-resolution (300 dpi) Research supported by Washington Sea Grant, Wash- digital images taken. These taxonomic identification ington Biotechnology Center and Soliv International data are included in a database with collection infor- Corporation. mation (i.e. location, date). Ecological data (i.e. tur- ABSTRACTS 37 bidity, salinity, and nutrients such as nitrate and nitrite, Mycosporine-like amino acids (MAAs) are a class of ammonium, phosphorous, and silica) will be added UV-absorbing compounds synthesized by a variety of to the database. Having this information available on marine and freshwater algae and cyanobacteria with the web is an important resource for continued re- absorption maxima between 310 and 360 nm. To date, search in coastal Louisiana and perhaps other regions ∼25 different MAAs have been characterized with sev- with similar ecosystems and hydrology. The database eral new compounds identified each year. A major can also be a useful tool for teaching high school and difficulty in the analysis of MAAs is the lack of com- college students. Microscopic phytoplankton are no- mercial standards. Thus far, MAAs have been identi- toriously difficult to identify, and the images can be fied by their retention time and UV absorption after used in conjunction with keys to help students con- separation via high performance liquid chromatog- firm their algal determinations. Students would also raphy (HPLC). Complementing the HPLC method be able to see the main features of different groups of with the use of mass spectrometry (LC–MS) offers algae and learn some ecological aspects of the wetland greater sensitivity and reduces the hindrance standard habitat where they were collected. Please visit the web unavailability presents by providing detailed molec- site at chapmanlab.lsu.edu/digitalalgae. This project ular information (molecular weight and fragmenta- was supported in part by a grant from the Office of tion patterns). The molecular weight information Sea Grant Development at LSU. adds one more level to our ability to characterize MAAs and any new UV-absorbing compounds discov- ered. The fragmentation patterns obtained via LC–MS 96 also contributes diagnostic information for structure ALGAL POLYUNSATURATED FATTY ACIDS: elucidation and identification. LC–MS analysis has FOOD, PHEROMONES AND FOUL ODOUR been used to examine UV-absorbing compounds in Watson, S.1,2,∗ & Janzen, L.2 methanolic extracts from Phaeocystis antarctica. When 1Environment Canada, NWRI, 867 Lakeshore Road, samples were analyzed using isocratic HPLC elu- Burlington, Ontario L7R 4A6; 2Department Biological tion, several UV-absorbing peaks were eluted. How- Sciences, University of Calgary, 2500 University Drive ever, molecular weight and fragmentation patterns of N.W. Calgary, Alberta T2N 1 N4, Canada some UV-absorbing compounds were not commiser- ate with those expected for MAAs. Additionally, pa- Algal polyunsaturated fatty acids (PUFAs) are known lythenic acid was detected rather than mycosporine- to be high-energy, often-essential resources to freshwa- glycine : valine as had been previously reported for ter aquatic food webs. On the other hand, high PUFA this organism. Gradient elution provided further sep- cell content in some algal taxa has been linked with the aration into MAA and ‘unknown’ groupings. The un- production of derivatives that may act as pheromones, known UV-absorbing component has longer retention allelogens or toxins. It has been known for some time times and different mass spectral characteristics with that these compounds function in marine chemical wavelength maxima between 315 and 325 nm. This ecology, but recent evidence indicates that they may study benefited immensely by the addition of MS tech- play similar roles in freshwater ecosystems. This paper niques, as many of the compounds would have re- presents field and laboratory studies of planktonic and mained either misidentified or entirely unidentified. periphytic communities from freshwater systems, and The increased analytical capability afforded by LC–MS shows that these communities are rich sources of cer- allows us to further investigate the structural diversity tain PUFAs and derivatives which may function in both and biochemistry of MAAs aiding our study of the role positive and negative foodweb interactions. This also MAAs play as sunscreen agents. has important implications for surface water quality, as these compounds are potent sources of rancid fishy odours. 98 MYCOSPORINE-LIKE AMINO ACIDS IN 97 HARMFUL MARINE MICROALGAE THE USE OF LIQUID Wulff, A.∗ & Appelgren, M. CHROMATOGRAPHY–MASS SPECTROMETRY Department of Marine Ecology, Marine Botany, Goteborg (LC–MS) IN THE IDENTIFICATION AND University, Sweden CHARACTERIZATION OF MYCOSPORINE-LIKE AMINO ACIDS (MAAS) Two current environmental issues are the increas- Whitehead, K.∗ & Hedges, J. I. ing occurrence of harmful algal blooms and the School of Oceanography, Box 355351, University of increasing level of ultraviolet radiation. The objec- Washington, Seattle, WA 98105, USA tive of this ongoing project is to investigate whether 38 ABSTRACTS there is a coupling between these two phenomena, southern Oman is rich with representatives of Codium, i.e. to test the hypothesis that UV-B radiation func- Scinaia, Galaxaura, Melanothamnus, and the families tions as a selective pressure favoring surface-blooming Gelidiaceae and Dictyotaceae. Several representative toxic/harmful marine microalgal species capable of marine algae from southern Oman will be depicted producing UV-absorbing compounds (mycosporine- and discussed. like amino acids, MAAs). A series of experiments were designed to test if the light environment in- duces the production of MAAs. Treatments were PAR 100 with and without the addition of UV-B radiation. Pre- THE CENTRAL AMERICAN ISTHMUS: liminary results show a different response for dif- IMPLICATIONS FOR INTRASPECIFIC ferent species. Most species increased their produc- PHYLOGENY AND BIOGEOGRAPHY OF A tion of MAAs when exposed to an increased PAR in- PANTROPICAL GREEN ALGA tensity (300 µmol photons/m2/s) and in, e.g. Alexan- Wysor, B. drium ostenfeldii and Gyrodinium aureolum a further in- Department of Biology, University of Louisiana at Lafayette, crease was observed upon exposure to UV-B radiation. P.O. Box 42451, Lafayette, LA 70504-2451, USA The content and composition of MAAs for a num- ber of potentially harmful marine microalgae will be An intraspecific phylogenetic study was undertaken shown. In addition, the outcome of small changes in to resolve the evolutionary relationship of isolates of the HPLC method used will be discussed. the green alga Phyllodictyon anastomosans (Harv.) Kraft et Wynne that occur on Atlantic and Pacific coasts of the Central American Isthmus. Patterns of vicari- ance related to the emergence of the Central Ameri- 99 can Isthmus were evident, but numerous examples of HIGHLIGHTS OF RECENT COLLECTIONS OF recent trans-oceanic and trans-isthmian dispersal ob- MARINE ALGAE FROM THE SULTANATE OF scured the underlying pattern. This study, one of the OMAN first studies to assess the impact of the emergence of Wynne, M. J. the Central American Isthmus in seaweed phylogeog- Department of Ecology and Evolutionary Biology, raphy, provided an ideal opportunity to estimate the University of Michigan, Ann Arbor, MI 48109, USA rate of sequence evolution using a single time point. Using this newly calibrated molecular clock the tim- Collections of benthic marine algae made over the ing of an historical introduction across the isthmus is past 3 years on the Dhofar coast of the Sultanate shown to be concordant with a shallow water breach of Oman have continued to provide a more com- of the isthmus dated at approximately 2.3–2.0 million plete picture of the floristic affinities of this up- years ago. This work contributes to a growing body of to-now poorly explored region. This Algal Biodi- literature that suggests marine algae are fairly success- versity Project of Oman was funded by the British ful at dispersing over long distances in recent times. Government’s Darwin Initiative grant for the ‘Sur- vival of Species’ and managed by HTS Develop- ment Ltd., of the UK. A rich and floristically com- 101 plicated flora is emerging, with connections to sub- COMPARATIVE PHYLOGEOGRAPHY OF tropical and warm temperate regions. Many new al- RETICULATE CLADOPHORALEAN ALGAE gal taxa have been discovered and described, and Wysor, B.1,∗, Kooistra, Wiebe H. C. F.2 & Fredericq, numerous new records for Oman or for the Indian Suzanne1 Ocean have been reported. Some species newly re- 1Department of Biology, University of Louisiana at ported for Oman (and the Indian Ocean) had previ- Lafayette, P.O. Box 42451, Lafayette, LA 70504-2451, ously been known from Japan (Distromium decumbens, USA; 2Stazione Zoologica ‘Anton Dohrn’, Villa Kallymenia crassiuscula, Dudresnaya japonica, and Chon- Communale I, I-80121 Naples, Italy dria crassicaulis). Sporochnus pedunculatus and Pedobe- sia simplex had been known from the North Atlantic The study of species distributions requires well- and the Mediterranean and are new records for the In- defined species concepts and well-resolved species dian Ocean. Some new species of red algae have been relationships. Ideally, the biogeography of taxa will described, such as a Dipterocladia and a Plocamium. be concordant with their genetic signatures identi- Some brown algal novelties have also been discovered, fied by molecular markers. For groups with poorly including new species of Turbinaria and Jolyna, and a defined species (or generic) concepts, taxon sam- new genus of Chordariaceae. The macroalgal flora of pling can be highly problematic because morpholog- ABSTRACTS 39 ical plasticity may deter sample collection of a species mentally benign form of commerce. However, finfish when it expresses alternative morphologies. This may and shellfish aquaculture operations are a source of give the impression of a disjunct species distribution an effluent with high concentrations of dissolved inor- when it is actually continuous across a particular geo- ganic nutrients (N, P). To prevent eutrophication, the graphic range. Likewise, morphological convergence EPA is developing stringent guidelines for the release can obscure biogeographic patterns, but with the op- of N and P into coastal waters. An integrated recir- posite effect. To avoid taxon sampling problems in culating aquaculture system, coupling the growth of a phylogeographic study of the green alga Phyllodic- seaweed and fish, can solve these problems for urban tyon anastomosans (Harv.) Kraft et Wynne, a species aquaculture facilities; not only is the effluent remedi- known for its morphological variability, we included ated but an additional multiproduct, high-value crop taxa across four genera (Boodlea G. Murray & De can be generated. One tank-based (on land) marine Toni, Cladophoropsis Børgesen, Phyllodictyon J. E. aquaculture operation is GreatBay Aquaculture, LLC Gray, Struveopsis Rhyne et H. Robinson) often char- (Portsmouth, NH), a land-based hatchery and grow- acterized by overlapping character states. The inclu- out facility for high value summer flounder and cod. sion of isolates outside the taxonomic boundaries of Our work is to develop an integrated finfish/seaweed P.anastomosans allowed us to extend earlier phylogeo- recirculating aquaculture system (RAS) suitable for graphic studies of the closely related C. membranacea urban aquaculture. Our RAS system will integrate (C. agardh)Børgesen and compare patterns between the culture of summer flounder and native species two closely related taxa. Results suggest that the Cen- of seaweed (i.e. Porphyra). BRVAS students are work- tral American Isthmus represents a formidable but ing along side undergraduate and graduate students permeable barrier to dispersal. in the construction and operation of these systems. There are at least seven recognized species of Porphyra in the north-east. We have begun mesoscale evaluation 102 of P.leucosticta, since it may be a good candidate for the food (sushi) and for r-phycoerythrin industries. THE AQUACULTURE OF The mass culture techniques (in both free culture PORPHYRA LEUCOSTICTA (RHODOPHYTA) FOR and attached to nets) for this Porphyra species are de- AN INTEGRATED FINFISH/SEAWEED veloping. We will report on the mass seeding technolo- RECIRCULATING AQUACULTURE SYSTEM IN gies that we have developed and the specific growth AN URBAN APPLICATION rates of P.leucosticta at the BRVAS culture facilities. Yarish, C.∗, He, P., Carmona, R., Liu, S., Kraemer, G., Neefus, C., Chopin, T., Nardi, G., Curtis, J., Lonergan, S. & Trupp, P. Department of Ecology and Evolutionary Biology, 103 University of Connecticut, One University Place, Stamford, MOLECULAR CHARACTERISTICS OF CAULERPA CT 06901-2315, USA; Fisheries College, Shanghai (CHLOROPHYTA) POPULATIONS IN TAIWAN Fisheries University, Shanghai 200090, China; Yeh, W.-J.∗, Lin, S.-C. & Chen, G.-Y. Environmental Research Institute, University of Department of Aquaculture, National Pingtung University Connecticut, The Longley Building, 270 Middle Turnpike, of Science and Technology, Pingtung 912, Taiwan Route 44, U-210, Storrs, CT 06269-3210, USA; Biology Department, State University of New York, Purchase, NY, Tropical and subtropical Caulerpa species are epi- USA; Department of Plant Biology, Office of Biometrics, demic to northern and southern Taiwan seashore. University of New Hampshire, Durham, NH 03824, USA; We here describe the molecular characteristics of Department of Biology, Center for Coastal Studies and rRNA gene, including the internal transcribed spacers Aquaculture, The University of New Brunswick, Saint (ITS) in this genus and a comparison was made with John, New Brunswick E2L 4L5, Canada; GreatBay associated species sequences registered in the Gen- Aquaculture, LLC, 153 Gosling Road, Portsmouth, NH Bank. A total of nine specimens (C. racemosa var. mi- 03801, USA; Bridgeport Regional Vocational Aquaculture crophysa, macrophysa, peltata, laetevirens, C. serrulata and School, 60 St. Stephen Road, Bridgeport, CT 06605, USA C. elongata) were investigated. Samples were collected and distinguished first from morphological character- Aquaculture represents an excellent opportunity to istics. The genomic DNA was individually extracted help rejuvenate blighted coastal urban areas on the using CTAB protocol. To amplify the 18S rRNA, ITS1, north-east coast. Aquaculture requires relatively little 5.8S rRNA, ITS2 and 28S rRNA sequences, a primer space, often acquired at reduced cost in ungentrified pair NS7 and ITS4, sequences located in nuclear rRNA city areas, and can represent an attractive, environ- region, was used. The range of 980–1100 bp DNA frag- 40 ABSTRACTS ment from genomic DNA of individual plant was gen- plastid from a haptophyte though a tertiary plastid erated from polymerase chain reaction (PCR). Ampli- replacement; (3) ‘Form II’ rubisco replaced the an- fied fragments were sequenced automatically with ABI cestral rbcL after the divergence of the peridinin-, sequencer. The sequences were aligned with those of and fucoxanthin-containing dinoflagellates; and (4) other Caulerpa species retrieved from the GenBank. we confirm that the plastid of P.foliaceum originated The phylogenetic tree was constructed using Clustal from a Stramenopiles endosymbiont. method. The comparison of whole sequence and 18S rRNA and ITS1 region among these specimens shows that the interspecies divergence within C. racemosa var. 105 microphysa, macrophysa, peltata and laetevirens is larger THE SINGLE, ANCIENT ORIGIN OF CHROMIST than those of intraspecies of C. taxifolia, C. serrulata PLASTIDS and C. elongata. It is supposed that they (C. racemosa var. Yoon, H. S.1, Hackett, J. D.1, Pinto, G.2 & microphysa, macrophysa, peltata and laetevirens) should Bhattacharya, D.1,∗ be in the position of different species in Caulerpa genus 1Department of Biological Sciences and Center for from the present rRNA sequences analysis. The se- Comparative Genomics, University of Iowa, 239 Biology quences of 5.8S region are shown more conserve than Building, Iowa City, IA 52242, USA; 2Dipartimento di that of 18S region. The identity of ITS1 and ITS2 Biologia vegetale, Universita Federico II, Via Foria 223, regions in present species are low, comparing with 80139 Napoli, Italy C. taxifolia, C. maxicana. Phylogenetic comparisons suggest that plastid pri- mary endosymbiosis, in which a single-celled protist 104 engulfs and ‘enslaves’ a cyanobacterium, likely oc- curred once and resulted in the primordial alga. This THE MONOPHYLETIC ORIGIN OF THE photosynthetic cell diversified, through vertical evo- PERIDININ-, AND lution, into the ubiquitous green (Chlorophyta) and FUCOXANTHIN-CONTAINING red (Rhodophyta) algae, and the more scarce Glau- DINOFLAGELLATE PLASTID THROUGH cophyta. However, some modern algal lineages have TERTIARY REPLACEMENT ∗ a more complicated evolutionary history involving a Yoon, H. S. , Hackett, J. D. & Bhattacharya, D. secondary endosymbiotic event, in which a protist Department of Biological Sciences, University of Iowa, Iowa engulfed an existing eukaryotic alga (rather than a City, IA 52242, USA cyanobacterium), which was then reduced to a sec- The dinoflagellates contain diverse plastids of uncer- ondary plastid. Secondary endosymbiosis explains the tain origin. To determine the origin of the peridinin- majority of algal biodiversity, yet the number and and fucoxanthin-containing dinoflagellate plastid, we timing of these events is unresolved. Here we ana- sequenced the plastid-encoded psaA, psbA, and rbcL lyzed a five-gene plastid dataset to show that a diverse genes from various red and dinoflagellate algae. The group of chlorophyll c-containing protists compris- psbA gene phylogeny, which was made from a dataset ing cryptophyte, haptophyte, and stramenopiles algae of 15 dinoflagellates, 22 rhodophytes, five crypto- (Chromista) share a common plastid that most likely phytes, seven haptophytes, seven stramenopiles, two arose from a single, ancient (about 1260 million years chlorophytes, and a glaucophyte as the outgroup, sup- ago) secondary endosymbiosis involving a red alga. ports monophyly of the peridinin-, and fucoxanthin- This finding is consistent with C. monophyly and impli- containing dinoflagellates, as a sister group to the cates secondary endosymbiosis as a driving force in haptophytes. The monophyletic relationship with the early eukaryotic evolution. haptophytes is recovered in the psbA + psaA phy- logeny, with stronger support. The rubisco tree uti- lized the ‘Form I’ red algal type of rbcL and included 106 fucoxanthin-containing dinoflagellates. The dinoflag- ACCUMULATION OF TRIACYLGLYCEROLS IN ellate + haptophyte sister relationship is also recov- HAEMATOCOCCUS PLUVIALIS IS CORRELATED ered in this analysis. Peridinium foliaceum is shown to WITH THAT OF ASTAXANTHIN ESTERS group with the diatoms in all the phylogenies. Based Zhekisheva, M.∗, Boussiba, S., Khozin-Goldberg, I., on our analyses of plastid sequences, we postulate Zarka, A. & Cohen, Z. that: (1) the plastid of peridinin-, and fucoxanthin- Microalgae Biotechnology Laboratory, The Jacob Blaustein containing dinoflagellates originated from a common Institute for Desert Studies, Ben-Gurion University of the ancestor; (2) the ancestral dinoflagellate acquired its Negev, Sede-Boker Campus 84990, Israel ABSTRACTS 41

The chlorophyte Haematococcus pluvialis accumulates during HPLC preparation as a result of pH shifts, as large quantities of astaxanthin under stress conditions. well as the ecological implications of the presence, Under either nitrogen starvation or high light, the concentration, and forms of these compounds (see proportion of each picogram of astaxanthin was ac- J Phycology 1999; 35, for relevant papers). This sym- companied by that of 5 or 3–4 pg of fatty acids, re- posium will provide opportunities for intercalibration spectively. In both cases, the newly formed fatty acids, of laboratories involved in MAA analyses, suggestions consisted mostly of oleic (up to 34% of fatty acids regarding standardization of extraction protocols, as in comparison to 13% in the control), palmitic and well as results from field- and laboratory-based studies. linoleic acids, were deposited mostly in triacylglyc- erols. Furthermore, the enhanced production of oleic acid was linearly correlated with that of astaxanthin. 108 Astaxanthin which is mostly monoesterified, is de- MYCOSPORINE-LIKE AMINO ACID posited in globules made of triacylglycerols. We sug- INTERCALIBRATION EFFORT USING gest that the production of tailor-made oleic acid-rich REPLICATE SAMPLES ∗ triacylglycerols on the one hand and the esterifica- Zimba, P. V.1, & Boue, S.2 tion of astaxanthin on the other hand, enable the oil 1USDA/ARS, P.O. Box 38, Stoneville, MS 38776; globules to maintain the high content of astaxanthin 2USDA/ARS, 1100 Robert E. Lee Blvd., New Orleans, LA esters. 70124, USA

The measurement of mycosporine-like amino acids 107 (MAAs) has several inherent problems, including the MYCOSPORINES: DETECTION lack of commercial standards for identification and METHODOLOGIES AND ASSESSMENT quantification. This symposium provided the oppor- Zimba, P. V.1,∗ & Wulff, A.2 tunity to coordinate analyses with six laboratories ac- 1USDA/ARS, P.O. Box 38, Stoneville, MS 38776, USA; tively involved in MAA research. Two samples were 2Department of Marine Ecology, Goteborg University, P.O. provided to each laboratory and included freeze-dried Box 461, Goteborg, Sweden nori (Porphyra sp.) and a freeze-dried (filtered) sam- ple of the cyanobacteria Microcystis aeruginosa Kutzing.¨ Mycosporine-like amino acids (MAAs) are found in a Each laboratory provided extraction methodologies, variety of prokaryotic and eucaryotic algae, as well as chromatograms of the identified peaks, as well as es- higher plants, fungi, and animals. These compounds timates of the concentration of each analyte. All labo- function as a photoprotective sunscreen to prevent ratories were able to identify major chromatographic ultra-violet light damage. MAAs may thus be one of the components of the samples (Porphyra: shinorine, por- competitive advantages that facilitated development phyra, mycosporine 2-glycine, asterina, palythine; Mi- ◦ of ozone (by oxytrophs), and thereby may be a com- crocystis: shinorine, porphyra). Sequential cold (4 C ◦ petitive advantage for the proliferation of cyanobacte- for 18 h) and hot (45 C for 2 h) extractions of the ria and other harmful algal species. Numerous diffi- same sample resulted in differing analyte recovery. culties exist with assessment of MAAs, including iden- Several currently unidentified compounds were ob- tification of the compounds, conversion of isomers served in freshwater samples.