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Dense blooms of toxin-producing Karenia brevis increase local surface ocean pH through CO2 uptake. To identify genes that may contribute to bloom-related environmental pH and pCO2 changes, transcriptomes with RNA from K. brevis Wilson cultures that had been acclimated to low CO2 (250 ppm) or recent CO2 (350 ppm) pCO2 levels were assembled. Among the annotated transcripts were PEPC, PPDK, and PEPCK enzymes found in the model C4 carbon fixation pathway. Previous studies have demonstrated that the enzymatic activity of PEPC, PPDK, and/or PEPCK in some algae species, including marine diatoms, is influenced by variations in dissolved inorganic carbon. We found significantly similar PEPC, PPDK, and PEPCK enzymes in the transcriptomes of K. brevis and two sister species Karenia papilionacea, and Karenia mikimotoi. One or more isoforms of PEPC were also identified in the transcriptomes of thirty additional photosynthetic phytoplankton species from nine phyla. Phylogenetic trees were constructed with neighbor joining and maximum likelihood techniques to characterize the evolutionary relationship among phytoplankton, terrestrial plant C4, and terrestrial plant C3 PEPC sequences. Based on the nucleotide trees constructed during this study, the Karenia PEPC transcripts were more closely related to the terrestrial C4 genes than the terrestrial C3 genes. Furthermore, PEPC phylogeny among phytoplankton closely resembles phylogenetic trees constructed with ribosomal RNA. This study confirmed that the toxin-producing dinoflagellates K. brevis, K. mikimotoi, and K. papilionacea express putative PEPC, PEPCK, and PPDK transcripts. [ABSTRACT FROM AUTHOR]

Imaging FlowCytobot (IFCB) combines video and flow cytometric technology to capture images of nano- and microplankton (∼10 to >100 μm) and to measure the chlorophyll fluorescence associated with each image. The images are of sufficient resolution to identify many organisms to genus or even species level. IFCB has provided >200 million images since its installation at the entrance to the Mission-Aransas estuary (Port Aransas, TX, USA) in September 2007. In early February 2008, Dinophysis cells (1–5 · mL−1) were detected by manual inspection of images; by late February, abundance estimates exceeded 200 cells · mL−1. Manual microscopy of water samples from the site confirmed that D. cf. ovum F. Schütt was the dominant species, with cell concentrations similar to those calculated from IFCB data, and toxin analyses showed that okadaic acid was present, which led to closing of shellfish harvesting. Analysis of the time series using automated image classification (extraction of image features and supervised machine learning algorithms) revealed a dynamic phytoplankton community composition. Before the Dinophysis bloom, Myrionecta rubra (a prey item of Dinophysis) was observed, and another potentially toxic dinoflagellate, Prorocentrum, was observed after the bloom. Dinophysis cell-division rates, as estimated from the frequency of dividing cells, were the highest at the beginning of the bloom. Considered on a daily basis, cell concentration increased roughly exponentially up to the bloom peak, but closer inspection revealed that the increases generally occurred when the direction of water flow was into the estuary, suggesting the source of the bloom was offshore. [ABSTRACT FROM AUTHOR]

In situanalysis of phytoplankton community structure was determined at five stations along the Texas Gulf coast using two instruments, the Fluoroprobe and FlowCAM. Results were compared with traditional methods to determine community structure (pigment analysis and microscopy). Diatoms and small nanoplankton (most likely haptophytes) dominated the phytoplankton community at all stations. Estimated chl concentrations for diatoms+dinoflagellates obtained via the Fluoroprobe were not significantly different for three of the five stations sampled when compared with HPLC-chemical taxonomy analysis, whereas the concentrations of green algal and cryptophyte chl were overestimated. The FlowCAM estimates of overall nanoplankton and microplankton cell abundance were not significantly different when compared with epifluorescence microscopy, and recorded images of phytoplankton cells provided a representative population of the phytoplankton community at each station. The Fluoroprobe and FlowCAM, when used in tandem, are potentially capable of determining the general characteristics of phytoplankton community structurein situand could be an important addition to biological observing systems in the coastal ocean. [ABSTRACT FROM AUTHOR]

Prochlorococcus is a ubiquitous marine oxyphotobacterium characterized by the presence of DV-chl a and b. In addition, the type strain Prochlorococcus marinus Chisholm et al. CCMP 1375 (or SS120), an isolate from the Sargasso Sea, contains low levels of an unusual phycoerythrin. Until now, it has been unclear if phycoerythrin occurs randomly within this systematic group and if the molecular characteristics of this phycoerythrin are restricted to this single strain. Here, we show that two additional Prochlorococcus strains from the Pacific Ocean also contain similar low levels of phycoerythrin. DNA sequence and phylogenetic analyses demonstrated that this phycoerythrin is very similar to the phycoerythrin of P. marinus SS120 and differs from the classic cyanobacterial phycoerythrins. In contrast, a third isolate from the Arabian Sea lacks phycoerythrin. Based on the DV-chl b:a ratio and 16S rRNA sequence data, we classify the two Pacific phycoerythrin-containing isolates as low-light-adapted strains and the Arabian Sea isolate as a high-light-adapted strain. Thus, we provide further evidence to link the physiology of an individual genotype and the presence or absence of functional phycoerythrin genes within the genus Prochlorococcus. [ABSTRACT FROM AUTHOR]

Differences in the halogenated natural products of Plocamium violaceum Farlow can be noted from different sites in the Monterey Bay region, California. Specimens from the northern region produce a set of alicyclic halogenated monoterpenes which are not synthesized by those from the southern region. Instead, the latter elaborates a set of acyclic halogenated monoterpenes not observed from the former. This situation is invariant regardless of life history froms or different seasons. Similarly, the composition of halogenated monoterpene structures from P. cartilagineum (L.) Dixon varies at different locations. From the data presented herein we conclude that at least two chemical types can be identified for P. violaceum based upon differences in their halogenated products. Thus we suggest that the north Monterey form can be dedignated as chemotype α and the southern one as chemotype β. [ABSTRACT FROM AUTHOR]

Multiple species of the genus Dinophysis produce diarrhetic shellfish toxins (okadaic acid and Dinophysis toxins, OA/DTXs analogs) and/or pectenotoxins (PTXs). Only since 2008 have DSP events (illnesses and/or shellfish harvesting closures) become recognized as a threat to human health in the United States. This study characterized 20 strains representing five species of Dinophysis spp. isolated from three US coastal regions that have experienced DSP events: the Northeast/Mid‐Atlantic, the Gulf of Mexico, and the Pacific Northwest. Using a combination of morphometric and DNA‐based evidence, seven Northeast/Mid‐Atlantic isolates and four Pacific Northwest isolates were classified as D. acuminata, a total of four isolates from two coasts were classified as D. norvegica, two isolates from the Pacific Northwest coast were identified as D. fortii, and three isolates from the Gulf of Mexico were identified as D. ovum and D. caudata. Toxin profiles of D. acuminata and D. norvegica varied by their geographical origin within the United States. Cross‐regional comparison of toxin profiles was not possible with the other three species; however, within each region, distinct species‐conserved profiles for isolates of D. fortii, D. ovum, and D. caudata were observed. Historical and recent data from various State and Tribal monitoring programs were compiled and compared, including maximum recorded cell abundances of Dinophysis spp., maximum concentrations of OA/DTXs recorded in commercial shellfish species, and durations of harvesting closures, to provide perspective regarding potential for DSP impacts to regional public health and shellfish industry. [ABSTRACT FROM AUTHOR]

Due to the increasing prevalence of Dinophysis spp. and their toxins on every US coast in recent years, the need to identify and monitor for problematic Dinophysis populations has become apparent. Here, we present morphological analyses, using light and scanning electron microscopy, and rDNA sequence analysis, using a ~2‐kb sequence of ribosomal ITS1, 5.8S, ITS2, and LSU DNA, of Dinophysis collected in mid‐Atlantic estuarine and coastal waters from Virginia to New Jersey to better characterize local populations. In addition, we analyzed for diarrhetic shellfish poisoning (DSP) toxins in water and shellfish samples collected during blooms using liquid‐chromatography tandem mass spectrometry and an in vitro protein phosphatase inhibition assay and compared this data to a toxin profile generated from a mid‐Atlantic Dinophysis culture. Three distinct morphospecies were documented in mid‐Atlantic surface waters: D. acuminata, D. norvegica, and a "small Dinophysis sp." that was morphologically distinct based on multivariate analysis of morphometric data but was genetically consistent with D. acuminata. While mid‐Atlantic D. acuminata could not be distinguished from the other species in the D. acuminata‐complex (D. ovum from the Gulf of Mexico and D. sacculus from the western Mediterranean Sea) using the molecular markers chosen, it could be distinguished based on morphometrics. Okadaic acid, dinophysistoxin 1, and pectenotoxin 2 were found in filtered water and shellfish samples during Dinophysis blooms in the mid‐Atlantic region, as well as in a locally isolated D. acuminata culture. However, DSP toxins exceeded regulatory guidance concentrations only a few times during the study period and only in noncommercial shellfish samples. [ABSTRACT FROM AUTHOR]

A new planktonic species of Prorocentrum is described from the Gulf of Mexico. First observed with the Imaging FlowCytobot, Prorocentrum texanum sp. nov. was characterized using LM, SEM, and TEM along with sequencing of the SSU, LSU, and ITS ribosomal regions and the mitochondrial cob and cox1 regions. P. texanum sp. nov. is a round to oval bivalvate dinoflagellate, with a prominent anterior, serrated solid flange on periflagellar a platelet and an opposing short, flat flange on the h platelet. The periflagellar area consists of 10 platelets. Both left and right valves have shallow round depressions and two-sized valve pores. The anterior ejectosome pore pattern differs between the left and right valve in relation to the periflagellar area and margins. Ten to eleven rows of tangential ejectosome pores are present on each valve. P. texanum sp. nov. has two varieties which exhibit distinct morphotypes, one round to oval (var. texanum) and the other pointed (var. cuspidatum). P. texanum var. cuspidatum is morphologically similar to P. micans in surface markings, but is smaller, and has a serrated periflagellar flange, and is genetically distinct from P. micans. Cytologically, P. texanum has two parietal chlo-roplasts, each with a compound, interlamellar pyrenoid, trichocysts, fibrous vesicles that resemble mucocysts, pusules, V- to U-shaped posterior nucleus, golgi, and tubular mitochondria. No genetic difference was found between the two varieties in the five genes examined. Phylogenetic analysis of the SSU, LSU, and ITS ribosomal regions place P. texanum sp. nov. as a sister group to P. micans. One isolate of P. texanum var . texanum produces okadaic acid. [ABSTRACT FROM AUTHOR]

Brachidinium capitatum F. J. R. Taylor, typically considered a rare oceanic dinoflagellate, and one which has not been cultured, was observed at elevated abundances (up to 65 cells · mL) at a coastal station in the western Gulf of Mexico in the fall of 2007. Continuous data from the Imaging FlowCytobot (IFCB) provided cell images that documented the bloom during 3 weeks in early November. Guided by IFCB observations, field collection permitted phylogenetic analysis and evaluation of the relationship between Brachidinium and Karenia. Sequences from SSU, LSU, internal transcribed spacer (ITS), and cox1 regions for B. capitatum were compared with five other species of Karenia; all B. capitatum sequences were unique but supported its placement within the Kareniaceae . From a total of 71,487 images, data on the timing and frequency of dividing cells was also obtained for B. capitatum, allowing the rate of division for B. capitatum to be estimated. The maximum daily growth rate estimate was 0.22 d. Images showed a range in morphological variability, with the position of the four major processes highly variable. The combination of morphological features similar to the genus Karenia and a phylogenetic analysis placing B. capitatum in the Karenia clade leads us to propose moving the genus Brachidinium into the Kareniaceae. However, the lack of agreement among individual gene phylogenies suggests that the inclusion of different genes and more members of the genus Karenia are necessary before a final determination regarding the validity of the genus Brachidinium can be made. [ABSTRACT FROM AUTHOR]

Immunological grouping, determined by affinities of polyclonal antibodies to surface antigens of intact cells, was used to characterize 19 clones of marine coccoid ultraplankton. The resulting cross-reactive antigen groups corresponded to pigment-groups as defined by HPLC analysis of chloroplast pigments (carotenoids and chlorophylls). Because immunological cross reactions are specific at the species levels in groups of algae having well defined morphological criteria, we suggest that immunilogical methods can be used to recognize algae presently indistinguishable by standard morphological criteria, and especially in oceanographic applications involving qualitative cell enumerations of the ultroplankton. [ABSTRACT FROM AUTHOR]