The effects of a harmful alga on bivalve larval lipid stores

Marine invertebrates often have complex life histories that include a swimming planktivorous larval stage, at which time they are vulnerable to a variety of stressors, including those associated with nutritional stress and harmful algal blooms. Lipid stores have been shown to be especially important for post-metamorphic survivorship and growth in a variety of marine invertebrates. We investigated the effects of the harmful brown tide alga Aureococcus anophagefferens on the lipid stores and growth of larvae of the hard clam (northern quahog, Mercenaria mercenaria), a dominant bivalve in many western Atlantic bays and estuaries. M. mercenaria was the dominant bivalve in Great South Bay, Long Island, until the mid-1970s, but very few larvae are presently found in these waters. Recent brown tide blooms have been hypothesized to pose a barrier to recovery of M. mercenaria populations and hinder recent restoration efforts by negatively affecting clam larvae. To test whether a diet of the brown tide alga affects the accumulation of beneficial lipid stores, we fed larvae one of three diets representing equal biovolumes of Isochrysis galbana, a nutritious control alga; A. anophagefferens, the brown tide alga for which nutritional quality is not presently known; or a mixture of the two. Larvae fed only brown tide had significantly less lipid stores than those in the other dietary treatments. In addition, brown tide negatively affected larval size. We also tested for evidence of tradeoffs between larval growth and lipid stores, predicting that when the diet was less nutritious as in the brown tide treatments, larval size and lipids would be negatively correlated. In contrast, we found that larvae fed a mixed algal diet or only A. anophagefferens showed a significant positive correlation between lipid stores and size, suggesting that some larvae were simply better at obtaining food and associated nutrients. Larval success likely depends on a complex interplay between genetic and environmental factors. Our study suggests that poor nutrition associated with a harmful alga can have negative effects on larval size and lipids stores, which in turn are mediated by the inter-individual variability in the ability to grow and accumulate necessary lipid stores. Phytoplankton quality is likely to be important for the sustainability of bivalve populations even when it primarily impacts the larval phase; and a diet of brown tide algae may have lasting legacies for juveniles and adults.     

DESCRIPTION AND CHARACTERIZATION OF THE ALGAL SPECIES AUREOUMBRA LAGUNENSIS GEN. ET SP. NOV. AND REFERRAL OF AUREOUMBRA AND AUREOCOCCUS TO THE PELAGOPHYCEAE1

The Texas brown tide alga (strain TBA-2) is described as Aureoumbra lagunensis Stockwell, DeYoe, Hargraves, et Johnson, gen. et sp. nov. Pigment composition, chloroplast structure, and 18s ribosomal RNA gene sequence data indicate that A. lagunensis and the east coast brown tide alga Aureococcus anophagefferens (originally placed in the Chrysophyceae) belong in the class Pelagophyceae. The new genus Aureoumbra with A. lagunensis as the type species differs from Aureococcus in 18s ribosomal RNA gene sequence, pyrenoid form, nitrogen physiology, and possession of basal bodies. The genus Aureococcus is placed in the order Pelagomonadates and family Pelagomonadaceae while ordinal placement of Aureoumbra is deferred.     

The small subunit of ribulose-1,5-bisphosphate carboxylase is plastid-encoded in the chlorophyll c-containing alga Cryptomonas Φ

The gene for the small subunit of ribulose-1,5-bisphosphate carboxylase (Rubisco) is located in the large single-copy region of the plastid genome of the chlorophyll c-containing alga Cryptomonas . The coding sequence is 417 base pairs long, encoding a protein of 139 amino acids, considerably longer than most other small subunit proteins. It is found 83 base pairs downstream from the gene for the large subunit and is cotranscribed with it. An 18 base pair perfect inverted repeat is located 8 base pairs beyond the termination codon. Sequence analysis shows the gene to be more closely related to cyanobacterial and cyanelle small-subunit genes than to those of green algae or land plants. This is the first reported sequence of a Rubisco small-subunit gene which is plastid-encoded and it exhibits a number of unique features. The derived amino acid sequence shows extensive similarity to a partial amino acid sequence from a brown alga, indicating that this gene will be of major interest as a probe for the small subunit genes in other algae and for determining possible evolutionary ancestors of algal plastids.     

Utility of rbcS gene as a novel target DNA region for brown algal molecular systematics

The usefulness of molecular phylogenetic studies has increased remarkably as the quantity and quality of available DNA sequences has increased. When compared with the progress that has occurred in angiosperms and animals, there have been relatively few target DNA regions identified for use in taxonomic studies of brown algae. Therefore, in this study, we developed a new set of primers to amplify Rubisco small subunit (rbcS) gene sequences and determined the rbcS gene sequences of various species of brown algae including those belonging to Dictyotales, Ectocarpales, Fucales and Sphacelariales. The level of sequence variations in the rbcS gene varied according to the brown algal lineages. When focusing on the relationship of species within the genus Sargassum, the rbcS gene sequences provided useful information regarding the phylogenetic relationship among sections of the subgenus Bactrophycus. Based on the broad applicability and phylogenetic utility of the rbcS gene, we suggest that the sequence be used as a new target region for the molecular systematics of brown algae.     

CELL CYCLE REGULATORS IN THE FLORIDA RED TIDE DINOFLAGELLATE, GYMNODINIUM BREVE

The diel cycle is a key regulator of the cell cycle in many dinoflagellates, and may play a rate limiting role in bloom formation. Diel phasing of the cell cycle in the Florida red tide dinoflagellate, Gymnodinium breve Davis was previously described in our laboratory. In cultures grown on a 16:8 light:dark cycle, S-phase began 6–8 h into the light phase, and mitosis followed 12–14 h later. The dark/light "dawn" transition was found to provide the diel cue that serves to entrain the G. breve cell cycle. However the cell cycle mechanisms and regulators acted upon by this cue are poorly understood in dinoflagellates. The cell cycle regulatory complex, CDK1-cyclinB, is therefore currently being investigated. Cyclin dependent kinase (CDK) was first identified in G. breve using two approaches: (1) identification of a 34 kDa protein immunoreactive to an antibody raised against a conserved amino acid sequence unique to the CDK protein family (PSTAIR) and (2) inhibition of the cell cycle by olomoucine, a selective CDK inhibitor. Several approaches are currently being employed in order to describe its partner, cyclin B: (1) PCR on genomic DNA with primers deduced from known cyclin box sequences, (2) G. breve expression library screening with an antibody raised against the fission yeast cyclin B (3) western blot analysis on whole protein extracts and cyclin B immunoprecipitated proteins. Current work focuses on the differential expression of the cyclin B homologue in G. breve during its cell cycle and its relation to diel cycle control.     

Enhanced Tolerance Against Salt-Stress and Freezing-Stress of Escherichia coli Cells Expressing Algal bbc1 Gene

The expression of the eukaryotic bbc1 (breast basic conserved) gene (the bbc1 gene of the marine green alga Chlamydomonas sp. W-80 strain) enhanced the tolerance against salt-stress and freezing-stress in E. coli cells. The expression of the BBC1 protein in the E. coli cells carrying the algal bbc1 gene and that in the Chlamydomonas W-80 cells were examined by Western blotting analysis. The result suggests that the eukaryotic BBC1 protein expressed in the E. coli cells has a protective function against the cellular dehydration. Received: 24 April 2000 / Accepted: 21 August 2000     

Variety of cell cycle patterns in the alga Scenedesmus quadricauda (Chlorophyta) as revealed by application of illumination regimes and inhibitors

In the course of the cell cycles of synchronous cultures of the chlorococcal alga Scenedesmus quadricauda, the following were monitored: total protein and RNA accumulation as a measure of growth processes, the timing of the commitment points at which the cells trigger the sequence of reproductive processes (DNA replication, nuclear and cellular division) and the course of the reproductive processes. The synchronous cultures were grown either under various lighting regimes, or in the temporary presence of specific inhibitors of either proteosynthesis (cycloheximide) or DNA replication (5-fluorodeoxyuridine). By adjusting the length of the light period, the cell cycle could be manipulated. Cell cycle patterns could be altered to give different numbers of sequences of reproductive processes. The extent of their mutual overlap could be influenced and the number of daughter cells produced could be altered. Schematic illustrations of various cell cycle patterns and comparisons with those of higher plants and other algal species are presented.     

The role of hypersalinity in the persistence of the Texas 'brown tide' in the Laguna Madre

A brown tide bloom of the alga Aureoumbra lagunensis was presentwithout interruption in the Laguna Madre of Texas from January1990 through October 1997. This is the longest continual phytoplanktonbloom of which we are aware. Although the factors leading tothe initiation of this bloom have been well documented, thefactors contributing to its persistence are still being investigated.Two physical characteristics of the Laguna Madre may play animportant role: the long turnover time for waters in this coastallagoon (–1 year) and the hypersaline conditions that usuallyexist (40–60 PSU) due to evaporation exceeding precipitation.In this study, we examined the effects of salinity on the growthrates of the brown tide alga and on the growth of one of itsprotozoan grazers. Historical data from before the onset ofthe brown tide provide evidence for the suppression of microzooplanktonpopulations and mesozooplankton growth caused by hypersalinity.The brown tide alga will grow in a remarkably wide range ofsalinities ranging from 10 to 90 PSU. Maximum growth rates areachieved at salinities ranging from 20 to 60 PSU. One commongrazer on the brown tide alga, the heterotrophic dinoftagellateOxyrrhis marina, was found to grow more slowly under hypersalineconditions. The normally hypersaline conditions of the LagunaMadre may, therefore, favor the brown tide alga over other phytoplanktonspecies that do not grow well under hypersaline conditions,and also suppress the growth and feeding rates of potentialgrazers.     

Microbial herbivory on the brown tide alga, Aureococcus anophagefferens: results from natural ecosystems, mesocosms and laboratory experiments

Experiments were conducted with natural plankton assemblages from two areas in Great South Bay (GSB) and the Peconic Bays Estuary System, NY, to compare the rates of growth and pelagic grazing mortality of Aureococcus anophagefferens with co-occurring phytoplankton. We hypothesized that A. anophagefferens would experience low mortality rates by microbial herbivores (relative to feeding pressure on other algae) thus providing it with a competitive advantage within the phytoplankton community. In fact, substantial rates of mortality were observed in nearly every experiment in our study. However, mortality rates of A. anophagefferens were less than intrinsic growth rates of the alga during late spring and early summer in Great South Bay, resulting in positive net growth rates for the alga during that period. This timing coincided with the development of a brown tide in this estuary. Similarly, growth rates of the alga also exceeded mortality rates during bloom development in natural plankton assemblages from the Peconic Bays Estuary System held in mesocosms. In contrast to the situation for A. anophagefferens, growth rates of the total phytoplankton assemblage, and another common picoplanktonic phytoplankter (Synechococcus spp.), were frequently less than their respective mortality rates. Mortality rates of A. anophagefferens in both systems were similar to growth rates of the alga during later stages of the bloom. Laboratory studies confirmed that species of phagotrophic protists that consume A. anophagefferens (at least in culture) are present during brown tides but preference for or against the alga appears to be species-specific among phagotrophic protists. We conclude that two scenarios may explain our results: (1) protistan species capable of consuming the brown tide alga were present at low abundances during bloom initiation and thus not able to respond rapidly to increases in the intrinsic growth rate of the alga, or (2) the brown tide alga produced substance(s) that inhibited or retarded protistan grazing activities during the period of bloom initiation. The latter scenario seems less likely given that significant mortality of A. anophagefferens was measured during our field study and mesocosm experiment. However, even a minor reduction in mortality rate due to feeding selectivity among herbivores might result in a mismatch between growth and grazing of A. anophagefferens that could give rise to significant net population growth of this HAB species. Either scenario infers an important role for trophic interactions within the plankton as a factor explaining the development of brown tides in natural ecosystems.     

Isolation of a full-length mitotic cyclin cDNA clone CycIIIMs from Medicago sativa: Chromosomal mapping and expression

Cyclins in association with the protein kinase p34^cdc2and related cyclin-dependent protein kinases (cdks) are key regulatory elements in controlling the cell division cycle. Here, we describe the identification and characterization of a full-length cDNA clone of alfalfa mitotic cyclin, termed CycIIIMs. Computer analysis of known plant cyclin gene sequences revealed that this cyclin belongs to the same structural group as the other known partial alfalfa cyclin sequences. Genetic segregation analysis based on DNA-DNA hybridization data showed that the CycIIIMs gene(s) locates in a single chromosomal region on linkage group 5 of the alfalfa genetic map between RFLP markers UO89A and CG13. The assignment of this cyclin to the mitotic cyclin class was based on its cDNA-derived sequence and its differential expression during G2/M cell cycle phase transition of a partially synchronized alfalfa cell culture. Sequence analysis indicated common motifs with both the A- and B-types of mitotic cyclins similarly to the newly described B3-type of animal cyclins.     

THE RED ALGAL EPIPHYTES MICROCLADIA COULTERI AND M. CALIFORNICA (RHODOPHYCEAE,CERAMIACEAE). I. TAXONOMY,LIFE HISTORY AND PHENOLOGY1

In the northeast Pa@, there are two epiphytic species of the genus Microcladia Grev. Microcladia coulteri Ham. grows on a wide range of red algal and occasionally brown algal basiphytes whereas M. californica Farl. grows only on the brown alga Egregia menziesii (Turn.) Aresch. Although both are obligate epiphytes in the field, they complete their life history in culture in the absence of their respective basiphytes. Along the coast of central California, M. californica exhibits distinct seasonal patterns in reproduction whereas M. coulteri does not. In addition to previously documented daferences in cystocarp development and branchlet patterns, the two taxa can be delineated by differences in spore sizes, timing of spore attachment and morphology of the primary rhizoid. The haploid chromosome number for both species ranges from 14-16. Attempts to hybridize the two species have been unsuccessful. In addition, restriction fragment length polymorphism analyses of the plastid DNA of the two species provides further evidence that Microcladia coulteri and Microcladia californica represent distinctive species.     

Genomics of brown algae: current advances and future prospects

The analysis of the complete genome sequence of the filamentous brown alga Ectocarpus provided many new insights into brown algal biology and has improved our understanding of how this organism has adapted to its seashore environment. Since the publication of the genome sequence in 2010, numerous studies have continued the analysis of the constituent genes or have combined genome data with experimental work, allowing progress in several key areas, including metabolism and reproductive biology. Ectocarpus will continue to be exploited as a model organism in future years, but genomic approaches should also be extended to additional brown algal species in order to fully exploit the diversity of this phylogenetic group and to facilitate the application of new knowledge to economically important seaweeds such as kelps.     

SEQUENCE AND PHYLOGENY OF THE PSAB GENE OF PYLAIELLA LITTORALIS(PHAEOPHYTA)1

The psaB gene codes for one of two highly conserved P700 chlorophyll a apoproteins of photosystem I. This gene was cloned from the brown alga, Pylaiella littoralis (L.) Kjellm., and its primary sequence was determined. The inferred amino acid sequence of the P. littoralis protein was compared to homologous sequences from land plants, green algae, and a cyanobacterium. The psaB protein sequence is very conserved in all the examined taxa, and an unrooted phylogenetic tree, generated from a distance matrix, shows that the P. littoralis gene is closer to that of the cyanobacterium Synechocossus sp. PCC 7002 than are those of green algae, land plants, and Euglena gracilis.     

Microbial load in mass cultures of green algaeScenedesmus acutus and its processed powder

Microbial contamination in cultures of the alga,Scenedesmus acutus raised in outdoor open tanks and also in the processed powder of the alga was monitored; The total bacterial population increased with time during the growth period of six days. When a combination of molasses and carbondioxide was employed as carbon source for this alga, the bacterial load increased to 10 colony forming units/ml. Yeast, molds and also coliforms were quantitated. Drum-drying the algae drastically reduced the bacterial load and storing the algal powder for a period of over 3 months did not increase the bacterial load. Pathogens likeSalmonella andStaphylococcus were not detectable either in the open cultures or in the drumdried algal powder. Although there are not set standards available in literature on the permissible level of the microbial contamination in algal biomass for use in foods, the microbial load appears to be within the limits of permissible levels stipulated by Indian Standard Institution standards for baby foods.     

Brandtia ciliaticola gen. et sp. nov. (Chlorellaceae,Trebouxiophyceae) a common symbiotic green coccoid of various ciliate species

Many freshwater protists harbor unicellular green algae within their cells and these host‐symbiont relationships slowly are becoming better understood. Recently, we reported that several ciliate species shared a single species of symbiotic algae. Nonetheless, the algae from different host ciliates were each distinguishable by their different genotypes, and these host‐algal genotype combinations remained unchanged throughout a 15‐month period of sampling from natural populations. The same algal species had been reported as the shared symbiont of several ciliates from a remote lake. Consequently, this alga appears to play a key role in ciliate‐algae symbioses. In the present study, we successfully isolated the algae from ciliate cells and established unialgal cultures. This species is herein named Brandtia ciliaticola gen. et sp. nov. and has typical ‘Chlorella‐like’ morphology, being a spherical autosporic coccoid with a single chloroplast containing a pyrenoid. The alga belongs to the Chlorella‐clade in Chlorellaceae (Trebouxiophyceae), but it is not strongly connected to any of the other genera in this group. In addition to this phylogenetic distinctiveness, a unique compensatory base change in the SSU rRNA gene is decisive in distinguishing this genus. Sequences of SSU‐ITS (internal transcribed spacer) rDNA for each isolate were compared to those obtained previously from the same host ciliate. Consistent algal genotypes were recovered from each host, which strongly suggests that B. ciliaticola has established a persistent symbiosis in each ciliate species.