The appearance of photosynthetic eukaryotes (algae and plants) dramatically altered the Earth's ecosystem, making possible all vertebrate life on land, including humans. Dating algal origin is, however, frustrated by a meager fossil record. We generated a plastid multi-gene phylogeny with Bayesian inference and then used maximum likelihood molecular clock methods to estimate algal divergence times. The plastid tree was used as a surrogate for algal host evolution because of recent phylogenetic evidence supporting the vertical ancestry of the plastid in the red, green, and glaucophyte algae. Nodes in the plastid tree were constrained with six reliable fossil dates and a maximum age of 3,500 MYA based on the earliest known eubacterial fossil. Our analyses support an ancient (late Paleoproterozoic) origin of photosynthetic eukaryotes with the primary endosymbiosis that gave rise to the first alga having occurred after the split of the Plantae (i.e., red, green, and glaucophyte algae plus land plants) from the opisthokonts sometime before 1,558 MYA. The split of the red and green algae is calculated to have occurred about 1,500 MYA, and the putative single red algal secondary endosymbiosis that gave rise to the plastid in the cryptophyte, haptophyte, and stramenopile algae (chromists) occurred about 1,300 MYA. These dates, which are consistent with fossil evidence for putative marine algae (i.e., acritarchs) from the early Mesoproterozoic (1,500 MYA) and with a major eukaryotic diversification in the very late Mesoproterozoic and Neoproterozoic, provide a molecular timeline for understanding algal evolution.

The Cyanidiales is a group of asexual, unicellular red algae, which thrive in acidic and high temperature conditions around hot springs. These unicellular taxa have a relatively simple morphology and are currently classified into three genera, Cyanidium, Cyanidioschyzon and Galdieria. Little is known, however, about the biodiversity of Cyanidiales, their population structure and their phylogenetic relationships. Here we used a taxonomically broadly sampled three-gene data set of plastid sequences to infer a robust phylogenetic framework for the Cyanidiales. The phylogenetic analyses support the existence of at least four distinct Cyanidiales lineages: the Galdieria spp. lineage (excluding Galdieria maxima), the Cyanidium caldarium lineage, a novel monophyletic lineage of mesophilic Cyanidium spp. and the Cyanidioschyzon merolae plus Galdieria maxima lineage. Our analyses do not support the notion of a mesophilic ancestry of the Cyanidiales and suggest that these algae were ancestrally thermo-acidotolerant. We also used environmental polymerase chain reaction (PCR) for the rbcL gene to sample Cyanidiales biodiversity at five ecologically distinct sites at Pisciarelli in the Phlegrean Fields in Italy. This analysis showed a high level of sequence divergence among Cyanidiales species and the partitioning of taxa based on environmental conditions. Our research revealed an unexpected level of genetic diversity among Cyanidiales that revises current thinking about the phylogeny and biodiversity of this group. We predict that future environmental PCR studies will significantly augment known biodiversity that we have discovered and demonstrate the Cyanidiales to be a species-rich branch of red algal evolution.

Amoxicillin is a widely used penicillin-like antibiotic, and due to its presence in several effluents of Italian STPs, its environmental fate along with its toxicity toward simple organisms have been investigated in model conditions. The present study shows that under abiotic conditions both hydrolysis and direct photolysis could be responsible for the transformation and removal of amoxicillin in aquatic environment, especially in slightly basic media. Quantum yields for the solar direct photolysis have been calculated along with kinetic constants and half-life times. Indirect photolysis experiments in the presence of natural photosensitizers such as nitrate ions and humic acids indicate that nitrate ions have no influence on the photodegradation rate of amoxicillin, while humic acids are able to enhance it. Standard batch experiments have been also performed under biotic conditions. The results indicated that also biodegradation on activated sludge is an effective pathway through which amoxicillin can be removed from the aquatic environment. Rate constants for biodegradation and adsorption have been calculated by applying simple pseudo-first-order kinetic models. Algal bioassays indicate that, in the range of concentrations from 50 ng/L to 50 mg/L, amoxicillin is not toxic toward eucariotic organisms such as the Chlorophyceae Pseudokirkneriella subcapitata and Closterium ehrenbergii and the Bacillariophyceae Cyclotella meneghiniana, but it shows a marked toxicity toward the Cyanophyta Synechococcus leopolensis.

A bstract: The Cyanidiophyceae are a group of unicellular organisms that diverged from ancestral red algae around 1.3 billion years ago. Present-day species are restricted to hot springs and geothermal habitats from around the world. Because of discontinuous geothermal environments, the distribution patterns and dispersal modes of the cyanidiophycean species are poorly understood. Iceland is the third largest island in the Atlantic Ocean and has intense underground volcanic activity that generates broad hydrothermal areas with different ecological conditions that are excellent for thermoacidophilic microfloral development. We analyzed populations to address the Icelandic cyanidiophycean biodiversity and dispersal. A global rbch phytogeny showed two main populations inhabiting Iceland, Galdieria sulphuraria and G. maxima. Their areas of distribution are not completely superimposed because they coexisted only in New Zealand, Kamchatka (Russia), Japan, and Iceland. Because of the strong monophyly of Icelandic species with Japanese and Russian species, we hypothesized an origin and dispersion of Icelandic G. suphuraria and G. maxima from northeastern Asia. On the basis of network analysis of rbch haplotypes, it is likely that the southwestern region of Iceland is the diversity center of both G. sulphuraria and G. maxima.

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Campi Flegrei is a large volcanic area situated northwest of Naples, Italy. Two archeological sites, the Sybil's Cave and the Piscina Mirabilis, are artificial caves dug in the yellow tuff and used during antiquity for various purposes. This paper describes for the first time the algal biodiversity of these caves and determines whether environmental factors such as light intensity and humidity are influential in species distribution. A total of twenty-two algal species were identified by molecular methods (18S rDNA); the largest group was Cyanobacteria (eleven species), followed by algae Chlorophyta (six), Rhodophyta (two) and Bacillariophyta (two). Cluster analysis of algal distribution in the caves in relation to light and humidity showed no relevant differences in algal distribution between the two caves. Three different algal groups were identified. The first one includes strains strictly dependent on low humidity, a second cluster was mainly associated with sites where humidity is not a severe constraint, and a third group, mainly represented by filamentous cyanobacteria, is probably dependent on high humidity, since it was detected only at Piscina Mirabilis.