Afterward, the 1:1 propylenoxide-resin mixture was left overnight to evaporate at room temperature, then fresh pure resin was added, and the samples were put for 4 h on a rotator. Dehydrated cells were embedded in 3:1, 2:1 and 1:1 propylenoxide-Spurr’s resin (PolySciences, Niles, IL, USA) for 24 h at 4 ☌ each step.
#SCAPPLE ON THE WEST COAST SERIES#
After washing as previously, the samples were dehydrated at room temperature in a graded acetone series followed by 2 × 30 min of propylenoxide treatment at 4 ☌. The algal cells, encapsulated in solidified agarose blocks, were fixed in 3% ( v/ v) glutaraldehyde (PolySciences, Niles, IL, USA) in 50 mM sodium cacodylate buffer (pH 7) for 4 h at room temperature, then washed 3 × 15 min with the same buffer and post-fixed in 1% ( w/ v) osmium tetroxide overnight at 4 ☌. Cells were collected by gentle centrifugation and embedded in 2% w/ v low melting point agarose (ABT, Madrid, Spain), by mixing 10 μL of algal pellet with 10 μL molten agarose. Sample Collection, Strains Isolation, and Growthįor Transmission Electron Microscopy (TEM), a mixed sample of each strain, from cultures of 10 days, 1 month, and 4 months after inoculation, was prepared. This study aims to characterize in detail the diversity of green algae, previously isolated from different habitats in Greece, by combining molecular phylogeny, morphology, and ultrastructural data. Greece is a country in the Mediterranean with an immoderate level of diversity and endemism of species, a fact which has led to the frequent recording of new taxa of photosynthetic microorganisms. Some studies have as central content the biotechnological potential of green algae. Furthermore, in several studies, molecular cloning techniques are used to identify microalgae in freshwater ecosystems.
The plethora of studies is limited to morphological descriptions of microalgae and/or surveys on the ecological status of ecosystems. Culture-dependent approaches to studying the molecular diversity and phylogeny of microalgae derived from Mediterranean aquatic environments are limited. Nevertheless, the Mediterranean region is considered as a relatively unexplored area regarding the diversity of microalgae. For some of them, this is the first report of their occurrence in specific aquatic environments.Īccording to Singh and Saxena, territories around the Mediterranean Sea are more suitable for algae cultivation than those used in industry owing to the favorable climate.
pamvotia) belonging to orders Sphaeropleales, Chlorellales, and Chlamydomonadales. 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. Based on a polyphasic approach, 18 Chlorophyta isolates were investigated and characterized. Herein, we aim to contribute to the photosynthetic microorganisms’ diversity knowledge in the Mediterranean area, a relatively unexplored ecoregion with high diversity. 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.
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