Four microalgae species were evaluated for their bioremediation capacity of anaerobic digestion effluent (ADE) rich in ammonium nitrogen, derived from a biogas plant. Chlorella vulgaris, Chlorella sorokiniana, Desmodesmus communis and Stichococcus sp. were examined for their nutrient assimilation efficiency, biomass production and composition through their cultivation in 3.7% v/v ADE; their performance was compared with standard cultivation media which consisted in different nitrogen sources, i.e., BG-11NO3 and BG-11ΝΗ4 where N-NO3 was replaced by N-NH4. The results justified ammonium as the most preferable source of nitrogen for microalgae growth. Although Stichococcus sp. outperformed the other 3 species in N-NH4 removal efficiency both in BG-11NH4 and in 3.7% ADE (reaching up to 90.79% and 69.69% respectively), it exhibited a moderate biomass production when it was cultivated in diluted ADE corresponding to 0.59 g/L, compared to 0.89 g/L recorded by C. vulgaris and 0.7 g/L by C. sorokiniana and D. communis. Phosphorus contained in the effluent and in the control media was successfully consumed by all of the species, although its removal rate was found to be affected by the type of nitrogen source used and the particular microalgae species. The use of ADE as cultivation medium resulted in a significant increase in carbohydrates content in all investigated species.

BackgroundTerrestrial, freshwater and marine green algae constitute the large and morphologically diverse phylum of Chlorophyta, which gave rise to the core chlorophytes. Chlorophyta are abundant and diverse in freshwater environments where sometimes they form nuisance blooms under eutrophication conditions. The phylogenetic relationships among core chlorophyte clades (Chlorodendrophyceae, Ulvophyceae, Trebouxiophyceae and Chlorophyceae), are of particular interest as it is a species-rich phylum with ecological importance worldwide, but are still poorly understood. In the Mediterranean ecoregion, data on molecular characterization of eukaryotic microalgae strains are limited and current knowledge is based on ecological studies of natural populations. In the present study we report the isolation and characterization of 11 green microalgae strains from Greece contributing more information for the taxonomy of Chlorophyta. The study combined morphological and molecular data.ResultsPhylogenetic analysis based on 18S rRNA, internal transcribed spacer (ITS) region and the large subunit of the ribulose-bisphosphate carboxylase (rbcL) gene revealed eight taxa. Eleven green algae strains were classified in four orders (Sphaeropleales, Chlorellales, Chlamydomonadales and Chaetophorales) and were represented by four genera; one strain was not assigned to any genus. Most strains (six) were classified to the genus Desmodesmus, two strains to genus Chlorella, one to genus Spongiosarcinopsis and one filamentous strain to genus Uronema. One strain is placed in a separate independent branch within the Chlamydomonadales and deserves further research.ConclusionsOur study reports, for the first time, the presence of Uronema in an aquatic environment up to 40 °C and reveals new diversity within the Chlamydomonadales. The results from the ITS region and the rbcL gene corroborated those obtained from 18S rRNA without providing further information or resolving the phylogenetic relationships within certain genera, due to the limited number of ITS and rbcL sequences available. The comparison of molecular and morphological data showed that they were congruent. Cosmopolitan genera with high worldwide distribution inhabit Greek freshwaters.

The genus Dunaliella belongs to the division Chlorophyta and is known for its ability to survive in highly saline environments. Dunaliella is an important source of carotenoids, especially beta-carotene and has a wide range of applications. In this study, we aimed to isolate and identify Dunaliella strains from active and abandoned saltworks in Greece. Four seasonal samplings were carried out in seven active saltworks and two samplings were performed in an abandoned saltwork throughout the year 2020. Strains were characterized based on their morphological and phylogenetic traits, whilst their beta-carotene potential were evaluated. Fifteen (15) Dunaliella strains were isolated and classified into nine species based on morphological and morphometrical features. The isolated strains were assigned to different species such as D. parva, D. granulata, D. minuta, D. terricola, D. asymmetrica, D. bioculata, D. viridis, D. minutissima, and D. polymorpha. The results of the phylogenetic analysis indicate the formation of distinct clades among different Dunaliella species and suggest that morphological and morphometrical features may not always align with the phylogenetic position of species in the Dunaliella clade. Strains were found to produce a low amount of beta-carotene under default laboratory conditions. This study comprises the first phylogenetic inference of several Dunaliella species and highlights a gap on molecular data for Dunaliella spp. We provide valuable information on the diversity of Dunaliella strains in the saltworks of Greece, which can be used for further research and biotechnological applications.

In the large and morphologically diverse phylum of Chlorophyta, new taxa are discovered every year and their phylogenetic relationships are reconstructed by the incorporation of molecular phylogenetic methods into traditional taxonomy. Herein, we aim to contribute to the photosynthetic microorganisms’ diversity knowledge in the Mediterranean area, a relatively unexplored ecoregion with high diversity. Based on a polyphasic approach, 18 Chlorophyta isolates were investigated and characterized. Morphological characteristics and ultrastructure, the phylogeny based on 18S rRNA gene (small subunit ribosomal RNA), 18S–28S internal transcribed spacer (ITS region), and the ribulose-1,5-bisphosphate carboxylase/oxygenase large subunit region (rbcL gene), support establishing four new genera (Nomia, Ava, Akraea, Lilaea) and five new species (Spongiosarcinopsis limneus, N. picochloropsia, Av. limnothalassea, Ak. chliaropsychia, and L. pamvotia) belonging to orders Sphaeropleales, Chlorellales, and Chlamydomonadales. For some of them, this is the first report of their occurrence in specific aquatic environments.

No abstract