The toxic spill of acid wastes rich in heavy metals/metalloids (AWHM) from the Aznalcóllar mine in April 1998, threatening the Doñana National Park, is considered to be the worst environmental disaster related to acute pollution in Spanish history. The aim of this work was to study the adaptation of microalgae (which play an important role as primary producers) from AWHM sensitivity to AWHM resistance by using the alga Scenedesmus intermedius as an experimental model. The Malthusian parameter (m) and the carrying capacity (K) were reduced by mud and soil samples collected from the affected area. A dose–effect analysis showed that fitness progressively diminished with increasing sample concentration. A fluctuation analysis demonstrated that AWHM-resistant cells arose by rare spontaneous mutations that occurred randomly prior to the incorporation of the AWHM. The rate of spontaneous mutation from AWHM sensitivity to AWHM resistance was 2.12×10−5 mutants per cell division. A competition experiment between wild-type AWHM-sensitive cells and AWHM-resistant mutants showed that in small populations the AWHM-resistant mutants are driven to extinction in the absence of selection for AWHM resistance. The resistant phenotypes are maintained in the absence of AWHM as the result of a balance between spontaneous mutation and natural selection, so that about 43 AWHM-resistant mutants per million cells are present in the absence of AWHM. Our experimental model suggests that mutation is essential for adaptation of microalgal populations to environmental changes. Rare spontaneous pre-adaptive mutation is enough to ensure the survival of microalgal populations in contaminated environments when the population size is large enough.

In order to elucidate the phylogeny and evolutionary history of the Bacillariaceae we conducted a phylogenetic analysis of 42 species (sequences were determined from more than two strains of many of the Pseudo-nitzschia species) based on the first 872 base pairs of nuclear-encoded large subunit (LSU) rDNA, which include some of the most variable domains. Four araphid genera were used as the outgroup in maximum likelihood, parsimony and distance analyses. The phylogenetic inferences revealed the Bacillariaceae as monophyletic (bootstrap support [ges ]90%). A clade comprising Pseudo-nitzschia, Fragilariopsis and Nitzschia americana (clade A) was supported by high bootstrap values ([ges ]94%) and agreed with the morphological features revealed by electron microscopy. Data for 29 taxa indicate a subdivision of clade A, one clade comprising Pseudo-nitzschia species, a second clade consisting of Pseudo-nitzschia species and Nitzschia americana, and a third clade comprising Fragilariopsis species. Pseudo-nitzschia as presently defined is paraphyletic and emendation of the genus is probably needed. The analyses suggested that Nitzschia is not monophyletic, as expected from the great morphological diversity within the genus. A cluster characterized by possession of detailed ornamentation on the frustule is indicated. Eighteen taxa (16 within the Bacillariaceae) were tested for production of domoic acid, a neurotoxic amino acid. Only P. australis, P. multiseries and P. seriata produced domoic acid, and these clustered together in all analyses. Since Nitzschia navis-varingica also produces domoic acid, but is distantly related to the cluster comprising the Pseudo-nitzschia domoic acid producers, it is most parsimonious to suggest that the ability of species in the Bacillariaceae to produce domoic acid has evolved at least twice.

The genus Pseudo-nitzschia includes several species capable of producing domoic acid, the causative agent of Amnesic Shellfish Poisoning. Some of these species have been recorded frequently in the Gulf of Naples. For one of the species, P. multistriata, which has been recurrently found in our sampling area since 1995, this is the first report for European waters. Here we provide further details on the fine structure of this species. Pseudo-nitzschia multistriata was the only one found to produce domoic acid among all the Pseudo-nitzschia species from the Gulf of Naples, and this finding raises the number of potentially toxic species in this genus to nine. Phylogenetic relationships among several Pseudo-nitzschia species were assessed using the hypervariable domains (D1–D3) of the large subunit (LSU) rDNA. The match between the phylogeny obtained and important taxonomic characters used in this genus are discussed. Results show that P. multistriata clusters with wider species lacking a central larger interspace in the raphe. Close genetic relationships were determined between P. fraudulenta and P. subfraudulenta, and between P. pungens and P. multiseries. Genetic differences among these pairs of species are comparable to those among isolates of P. pseudodelicatissima from the Gulf of Naples, indicating high intraspecific genetic diversity of Pseudo-nitzschia species in the relatively conserved LSU region. This could explain the problematic results obtained when testing a match between species-specific Pseudo-nitzschia LSU probes and our sequences.

Glutamate-l-semialdehyde (GSA) aminotransferase catalyses the final step in the C5 pathway converting glutamate to the tetrapyrrole precursor δ-aminolaevulinic acid. This enzyme is sensitive to gabaculine (2,3-dihydro-3-amino benzoic acid) and to 4-amino-5-fluoropentanoic acid (AFPA), which are irreversible, mechanism-based inhibitors of pyridoxal phosphate-dependent enzymes. Spontaneous mutants of Synechococcus PCC6301 resistant to these inhibitors contain altered enzyme that displays corresponding resistance to high concentrations of the inhibitor. The enzyme from strain GR6, resistant to both inhibitors, contains a three-amino-acid deletion at positions 5–7 and a Met248→Ile substitution. The enzyme from strain K40 resistant to AFPA but not to gabaculine, contains a Ser163→Thr substitution. GSA aminotransferases containing either the deletion or the substitution that are characteristic of the GR6 mutant were produced in Escherichia coli using the expression vector pMalc2. These engineered mutant enzymes were characterized in terms of their catalytic parameters and sensitivities to gabaculine and AFPA. Furthermore, maltose binding protein/aminotransferase fusion proteins were characterized spectrophotometrically to monitor the interaction of bound cofactor with diamino- and dioxo-compounds related to the substrate and both inhibitors. Results were compared with those for similarly produced recombinant wild-type, K40 and GR6 GSA aminotransferases. The engineered products with either the N-terminal deletion or the Met248→Ile substitution displayed catalytic efficiencies that were intermediate between the wild-type and GR6 or K40 enzymes. However, with respect to their absorption spectra, sensitivity to inhibitors and the reactivity of bound cofactor, they were essentially wild-type. These in vitro studies demonstrate that both changes in enzyme structure are necessary to obtain the distinctive properties of the GR6 aminotransferase, including resistance to high concentrations of gabaculine and AFPA.

A new diatom (Bacillariophyceae) genus and two new species are described from the arid region of the North American southwest. The new genus, Playaensis, and new species, P. circumfimbria and P. furtiva, are distinguished by frustule morphology and autecology. Playaensis possesses unique, narrowly lunate longitudinal ribs bordering the raphe and spathulate spines, but their evolutionary relationship to known genera is unclear. In Payton Lake, New Mexico, P. circumfimbria and P. furtiva are rare in the diatom community, and their geographic and geological extent are not known, but they may be restricted to alkaline, slightly saline lakes of the region.

Response of Aphanizomenon ovalisporum to certain environmental parameters was studied to gain a better understanding of the conditions which may have stimulated its autumnal bloom in Lake Kinneret. Optimal temperature for A. ovalisporum growth was 26–30 °C, resulting in growth rates of 0·2–0·3 day−1, similar to those observed in the lake. Maximal rate of CO2 fixation (assimilation numbers of 6–8 μg C μg−1 Chl h−1) was obtained at low irradiances (Ik of 40–100 μmol photons m−2 s−1), 200 μM Pi and low N[ratio ]Pi ratios. Growth was strongly affected by phosphorus availability, reaching a maximum at Pi concentrations above 40 μM. The high demand for phosphorus was indicated by an increase in alkaline phosphatase activity. The relative abundance of Pi in the cells increased by 4-fold in Pi-rich compared with Pi-limited cultures. Uptake of Pi was faster in Pi-depleted compared with Pi-sufficient cells. Maximal photosynthetic rates and K1/2(HCO-3) were 140–220 μmol O2 mg−1 Chl h−1 and 10–24 μM, respectively. At pH 7·0 the K1/2(CO2) was 2·2 and fell to 0·04 μM at pH 9·0. These data indicated that A. ovalisporum is a HCO-3 user, and can explain its high photosynthetic rates during the bloom, under high pH and low dissolved CO2 conditions. Na+ concentrations of about 5 mM were essential for A. ovalisporum growth at high pH approaching values in the lake.

Amphora angusta Gregory, Amphora ventricosa Gregory and Amphora macilenta Gregory have been misunderstood for the last 140 years. Gregory described these species from sublittoral sediments off the west coast of Scotland. Although the illustrations were excellent for their time (1857), it is impossible to be confident of identification using Gregory's paper, and subsequent authors have created such confusion that records of these taxa must be regarded as untrustworthy unless backed by photographic evidence. In this paper, the first to describe in detail the recently established genus Seminavis, we show that A. angusta is in fact a species of Amphora Ehrenberg, whereas A. ventricosa sensu Gregory represents two independent species of the genus Seminavis D. G. Mann, namely Seminavis ventricosa (Gregory) Garcia-Baptista (non S. ventricosa sensu Garcia-Baptista 1993) and Seminavis arranensis Danielidis & D. G. Mann, sp. nov. The form usually known as A. ventricosa is neither of these and requires a new name, Seminavis robusta Danielidis & D. G. Mann, sp. nov. The long forgotten and misclassified species A. macilenta Gregory is shown to be yet another Seminavis species, requiring a new combination as S. macilenta (Gregory) Danielidis & D. G. Mann, comb. nov.; contrary to previous claims, it is separate from A. ergadensis Gregory, which is a true Amphora. Valve and girdle ultrastructure, plastid arrangement and auxosporulation are described for Seminavis and resemble those in the Naviculaceae sensu stricto.

How are microphytobenthic biofilms adapted to the high incident irradiances and temperatures, low inorganic nutrient concentrations and high desiccation stresses on intertidal flats present in tropical environments? This study investigated biofilms subject to different environmental conditions in a range of tropical sites in Suva lagoon, Fiji. PAM fluorescence was used to measure photophysiological responses to the light climate. Biofilm colloidal carbohydrate, extracellular polymeric substances (EPS) and low molecular weight (MW) carbohydrate concentrations and diel carbohydrate production patterns were measured. Average biomass (Chl a) ranged from 15 to 36 mg m−2, and was highest in seagrass bed sediments, but biomass was not correlated with water column or sediment porewater nutrient concentrations. Biofilm photophysiology differed significantly along a combined gradient of light and nutrient availability, with Fv/Fm, relative ETRmax and Ek of biofilms highest in mangrove and intertidal main island sites and lowest in subtidal coral reef flats. Subtidal biofilms showed photoinhibition at irradiances >1000 μmol m−2 s−1. Significant correlations between Chl a and colloidal carbohydrate concentrations were present (except on intertidal sandflats), and tropical biofilms had higher ratios of colloidal carbohydrate and EPS to Chl a than temperate estuarine biofilms, probably due to a combination of high irradiance and low nutrient availability leading to the production of excess photoassimilates. The percentage of EPS present in the colloidal fraction was highest in coral sand biofilms (42%), which had the lowest nutrient concentrations, compared with other sites (25–32%). Intertidal biofilms predominantly consisted of large motile taxa and showed strong rhythms of vertical migration. During tidal emersion, high sediment temperatures (41 °C), irradiance (>2300 μmol m−2 s−1) and salinity (49‰) stimulated downward migration. In silty sediments, migration resulted in a reduction in photosynthetic activity during the midday period but, in sands with high light penetration (to a depth of >1700 μm), high production rates of EPS (18.2 μg carbo. μg Chl a−1 h−1) and low MW carbohydrate exudates (40.2 μg carbo. μg Chl a−1 h−1) occurred. Vertical migration, high Ek and high rates of photoassimilate dumping are all adaptations to living in the tropical intertidal zone. Seagrass and reef flat biofilms consisted of a diverse non-migratory flora of motile and non-motile taxa that were not subject to such extreme temperature and irradiance conditions. Low values of photosynthetic parameters and high colloidal and EPS content indicated that these biofilms were nutrient-limited.

A minute parasite of Neosiphonia poko (Hollenberg) Abbott from a shallow lagoon on the central-Pacific Johnston Atoll is described as Neotenophycus ichthyosteus Kraft et Abbott, gen. et sp. nov. The infective parasite cell first connects to a central-axial cell of the host, then emerges from between host pericentral cells at a node before dividing into a three- or four-celled primary axis. Epibasal cells of the parasite divide to form three pericentral cells whose derivatives produce a globular head on the basal cell and on which reproductive structures differentiate almost immediately. Trichoblasts on any life-history stage are completely lacking. Spermatangia are borne on mother cells across the whole thallus surface. Procarps consist of four pericentral cells that encircle a subapical fertile-axial cell in an ampullar configuration, one of the pericentral cells serving as the supporting cell and bearing a four-celled carpogonial branch and a single sterile cell. Diploidization results in a longitudinal/concave division of the auxiliary cell and formation of an arching linear series of inner gonimoblast cells, each dividing toward the thallus surface into gonimoblast filaments of very narrow, horizontally aligned cells terminated by initially monopodial, later by sympodial, carposporangia, the whole of the mature female gametophyte consisting of an amalgam of several cystocarps within a lax jacket of sterile gametophytic tissue. Tetrasporophytes are composed of lobes of pericentral-cell-derived filaments, each axial cell of which is ringed by three pericentral cells producing tetrahedral tetrasporangia enclosed by two pre-sporangial cover cells. Affinities of the new genus are discussed and comparison is made particularly to the enigmatic parasite Episporium centroceratis Möbius. It is concluded that relationships with any previously described tribe are so remote or obscure that the new tribe Neotenophyceae should be proposed for it.

This is the first description of a combination coccosphere of the heterococcolithophore species Coronosphaera mediterranea (Lohmann 1902) Gaarder in Gaarder & Heimdal (1977) and the holococcolithophore species Calyptrolithophora hasleana (Gaarder 1962) Heimdal in Heimdal & Gaarder (1980) (Figs 1, 2). A single specimen of this new combination coccosphere was found within 53000 analysed cells in one of 189 samples collected at the JGOFS time series station ALOHA, Hawaii, during 1993 to 1996. The specimen was in a sample collected during autumn 1996 at a water depth of 5 m at a temperature of 26·1 °C, salinity of 35·0 psu, phosphate of 0·002 μmol/kg and in nitrate-depleted water (for details see Cortés, 1998; Cortés et al., 2001).

Systematics of the red algal order Acrochaetiales and related taxa was investigated using combined small- and large-subunit nuclear ribosomal DNA (SSU and LSU rDNA, respectively) sequence data. These data were subjected to distance, parsimony and maximum likelihood analyses. The resulting phylogenies were congruent with previously published SSU results in that all included orders (Balbianiales, Batrachospermales, Nemaliales, Palmariales and Thoreales) were resolved as monophyletic except the Acrochaetiales, which consisted of two lineages (Acrochaetiales I and II). The Batrachospermales and Thoreales occupied equivocal positions as early diverging lineages, while the Balbianiales generally resolved as sister to an Acrochaetiales–Nemaliales–Palmariales (ANP) complex. Relationships among the four lineages of the ANP complex were not completely resolved, but detailed analyses weakly positioned Acrochaetiales II as sister to the Nemaliales, whereas Acrochaetiales I displayed a moderate to strong affiliation with the Palmariales. Acrochaetiales I included representatives of the genera Acrochaetium,Audouinella and Rhodochorton, whereas Acrochaetiales II had a number of acrochaetioid species including a representative of the genus Colaconema. Compared with published SSU phylogenies, bootstrap values within the two Acrochaetiales lineages increased substantially in combined SSU/LSU analyses. Based on these results, emended generic descriptions are provided for Acrochaetium, Audouinella, Colaconema and Rhodochorton, and a new family of acrochaetioid algae is described, the Colaconemataceae J. T. Harper et G. W. Saunders. The Acrochaetiaceae now includes the genera Acrochaetium, Audouinella and Rhodochorton, while the Colaconemataceae is considered monogeneric at this time. It is quite likely that additional genera will be recognized within the Colaconemataceae pending further investigation. Acrochaetiaceae is retained as the sole family of the Acrochaetiales, although the tenuous recognition of the Palmariales as distinct from this order is discussed, whereas Colaconemataceae is transferred to the new order Colaconematales J. T. Harper et G. W. Saunders.

Seventeen species of the mainly sand-dwelling genus Amphidinium are reported from south-eastern Australian waters. Three new species are described: A. boggayum, A. mootonorum and A. yuroogurrum. Additional features from previously described species are presented, including non-motile cell stages. Nine of these species are new records for Australia; however, these species are probably widespread and common. Two potentially ichthyotoxic species of Amphidinium were commonly found: A. carterae and A. cf. operculatum. A table of morphological characters is compiled as a step towards recognizing new characters with which to group species of this presently artificially defined unarmoured dinoflagellate genus.

Volcanic areas with highly acidic solfatara soils and temperatures of up to 56 °C are inhabited by the red algal genus Galdieria. We examined three highly acidic but non-volcanic habitats in the western part of the Czech Republic for the occurrence of this red alga. In soil samples from the National Nature Reserve of Soos we found, together with Euglena mutabilis, Pseudococcomyxa simplex and species of Chlorella, a new strain of Galdieria. In contrast to all other Galdieria strains described so far, the strain from Soos exhibited a low temperature optimum for growth of about 30 °C. Other properties, such as the substrate spectrum for heterotrophic growth, ultrastructure, fatty acid composition, thermostability of enzymes and the nitrogen source, showed no obvious differences from other strains of Galdieria. Within a phylogenetic tree based on 18S rRNA sequence data, the strain from Soos occupied a position at the base of the ‘Galdieria’-branch. Our findings indicate that the genus Galdieria is not restricted to volcanic and mining areas and that strains of Galdieria are able to compete successfully with green algae in habitats like Soos.

The Third European Phycological Congress (EPC3) will be held in Belfast from 21–26 July 2003. The following is an outline of the plenary lectures and symposia. Further details can be obtained from our website www.epc3.org (email: [email protected]). A full social programme will also be provided, and there will be mid-congress visits to attractions such as the Giant's Causeway and the Mountains of Mourne.