Accessible Requires Authentication Published by De Gruyter September 4, 2014

Scaly green flagellates from Spanish Atlantic coastal waters: molecular, ultrastructural and pigment analyses

Aitor Alonso-González, Emma Orive, Helena David, Koldo García-Etxebarria, José Luis Garrido, Aitor Laza-Martínez and Sergio Seoane
From the journal Botanica Marina


Seven scale-bearing species of prasinophyceans (Chlorophyta) were studied in light and electron microscopy, pigment analysis, and molecular analysis of the small subunit (SSU) rDNA. Unialgal cultures were obtained from samples collected in the Nervión River estuary, Gulf of Biscay, Spanish Atlantic coast. Five of the species belong to the genus Pyramimonas (Prasinophyceae, Pyramimonaceae) and one each to the genera Mamiella (Mamiellophyceae, Mamiellaceae) and Nephroselmis (Nephroselmidophyceae, Nephroselmidaceae). The morphological features of all the strains analyzed agreed closely with the phylogenetic analysis, which in the case of the genus Pyramimonas presented several clusters corresponding to the subgenera Vestigifera (Pyramimonas orientalis, Pyramimonas moestrupii), Punctatae (Pyramimonas robusta), Pyramimonas (Pyramimonas propulsa), and Trichocystis (Pyramimonas grossii). Even though the phylogenetic relationship among these subgenera remains unclear, the results suggested that the Punctatae should remain an independent clade at least until more sequences or genes are analyzed. According to their accessory pigment composition, three groups were distinguished after their pigment ratios were calculated: prasinoxanthin-containing (Mamiella gilva), loroxanthin ester-containing (P. grossii and P. moestrupii), and siphonaxanthin-containing strains (Nephroselmis pyriformis and remaining Pyramimonas species). A high intraspecific variability was found, which highlights the need of an in-depth analysis and multiple technique approach to ensure accurate identification of nanoplanktonic microalgae.

Corresponding author: Emma Orive, Faculty of Science and Technology, Department of Plant Biology and Ecology, University of the Basque Country (UPV/EHU), 48940 Leioa, Spain, e-mail:


Alex Diez, Ricardo Andrade, and Jonh Arluzea helped in taking photographs with the electron microscope; Manuel Zapata, who, we are deeply sorry to say, passed away during this work, helped with the pigment analysis, a subject that he honored with his passionate research work throughout his career. Financial support for this research was provided by the Basque Government (project IT-481-10) and the Spanish Ministry of Economy and Competitiveness (projects CGL2010-19016 and GIC07/111-417-07) and the former Ministry of Education and Science (project CMT2006-04570/MAR). Grants from the University of the Basque Country (UPV/EHU) to H. David and from the Ministry of Education and Science to A. Alonso-González also supported this study. TEM work and genetic sequencing were performed in the unit of Analytical and High-Resolution Microscopy in Biomedicine, and the Sequencing and Genotyping Unit from SGIker services of the UPV/EHU, respectively.

Funding: Spanish Ministry of Economy and Competitiveness, (Grant/Award Number: ‘projects CGL2010-19016 and GIC07/111-417-07’), Spanish Ministry of Education and Science, (Grant/Award Number: ‘CMT2006-04570/MAR’), University of the Basque Country (UPV/EHU).


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Received: 2013-12-7
Accepted: 2014-8-5
Published Online: 2014-9-4
Published in Print: 2014-10-1

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