bioRxiv preprint doi: https://doi.org/10.1101/452110; this version posted October 26, 2018. The copyright holder for this preprint (which was not certified by peer review) is the author/funder. All rights reserved. No reuse allowed without permission. PINNULARIA BAETICA SP. NOV. Pinnularia baetica sp. nov. (Bacillarophyceae): a new diatom species found in an alkaline mountain lagoon in the south of Europe (Granada, Spain) DAVID FERNÁDEZ-MORENO1,2, ANA T. LUÍS3,4, PEDRO M. SÁNCHEZ- CASTILLO2 1Dnota Medio Ambiente, Granada, C/Baza, 6, Poligono Juncaril, 18220 Albolote. Granada, Spain. 2Department of Botany, Campus Fuentenueva, University of Granada, 18071 Granada, Spain. 3Universidade de Aveiro, Department of Geosciences, Geobiotec – Geobiosciences, Technologies and Engineering, Campus de Santiago, P–3810–193 Aveiro, Portugal 4CESAM Associated Lab – Department of Biology, Campus de Santiago, 3810-193 Aveiro, Portugal corresponding author's e-mail: [email protected] Phylum Ochrophyta Caval.-Sm. (Cavalier-Smith 1995) Class Bacillariophyceae Haeckel emend. Medlin & Kaczmarska (Medlin & Kaczmarska 2004) Subclass Bacillariophycidae Round (Round et al. 1990) Order Naviculales (Bessey 1907 sensu emend) Family Pinnulariaceae D.G. Mann, 1990 Genus Pinnularia C.G. Ehrenberg, 1843 Pinnularia baetica Fernández Moreno & Sánchez Castillo sp. nov 1 bioRxiv preprint doi: https://doi.org/10.1101/452110; this version posted October 26, 2018. The copyright holder for this preprint (which was not certified by peer review) is the author/funder. All rights reserved. No reuse allowed without permission. Abstract A new benthic freshwater diatom species belonging to the genus Pinnularia was found in Laguna Seca of Sierra Seca in the north of the province of Granada, Spain. Pinnularia baetica sp.nov. is proposed as a new species based on observations under light (LM) and scanning electron microscopy (SEM) and its special ecology typical of a calcareous lagoon. The most similar taxa to P. baetica is P. atlasii and with more differences P. infirma and the last two were studied through material obtained in lagoons of northern Morocco. Although there are similarities in the morphological characters of the frustule, it was possible to verify through LM and SEM micrographs, evident differences between P. baetica and the other two taxa; on the one end, P. baetica has a panduriform shape more pronounced than P. infirma and bigger size. On the other hand, the absence of spines in P. baetica and the more convergent striation at the poles are the main differences with P. atlasi. Keywords: alkaline mountain lagoon, diatoms, Morocco, Pinnularia atlasi, Spain, spines, Sardinia Introduction The south of Spain has the richest, most varied and best preserved natural heritage of wetlands in Spain and the European Union. The Andalusian wetlands present a great 2 bioRxiv preprint doi: https://doi.org/10.1101/452110; this version posted October 26, 2018. The copyright holder for this preprint (which was not certified by peer review) is the author/funder. All rights reserved. No reuse allowed without permission. diversity of ecological types and constitute fundamental areas for the conservation of Biodiversity The most important studies of benthic lacustrine communities from southern Spain are those of REED (1995), SÁNCHEZ-CASTILLO (1988), SÁNCHEZ CASTILLO (1993), LINARES-CUESTA & SÁNCHEZ-CASTILLO (2007) and SÁNCHEZ- CASTILLO et al. (2008). In addition to LINARES-CUESTA (2003) who made an inventory of the epiphytic and epilithic diatom communities of the lagoons from Sierra Nevada, as well as some works done in Doñana lakes, where some new species were described (BLANCO et al. 2013). Many of the reamaining diatom records were assigned to lothic systems (MARTÍN FARFÁN, 2016). This older literature does still not reflect all the diatom diversity in the south of Spain, as long as there is not any record in many of the more than 200 wetlands found it in Andalucia. The species from Pinnularia EHRENBERG genus are part of a complex taxonomical group of great diversity, with 708 taxa accepted according to the last revision made in AlgaeBase (GUIRY & GUIRY 2018). Despite of the high diversity of this genus, in the south of Spain is very scarce and the most important populations are in Sierra Nevada Mountain and in acidic rivers as Rio Tinto. Traditionally the species from Pinnularia genus have been reported as living in extreme environmental conditions, like the freezing temperatures of the Antartic region where Pinnularia catenaborealis (PINSEEL et al. 2016), Pinnularia sofia (VAN DE VIJVER et al. 2004), Pinnularia obaesa and Pinnularia australorabenhorstii (VAN DE VIJVER 2008), and several other species of Pinnularia (VAN DE VIJVER & ZIDAROVA 2011; ZIDAROVA et 3 bioRxiv preprint doi: https://doi.org/10.1101/452110; this version posted October 26, 2018. The copyright holder for this preprint (which was not certified by peer review) is the author/funder. All rights reserved. No reuse allowed without permission. al. 2012) are living well adapted. They can also be present in the surrounding streams of mining areas, characterized by high metal concentrations and very low pH, as it is the case of Pinnularia aljustrelica (LUÍS et al. 2012) and Pinnularia paralange-bertalotii (FUKUSHIMA et al. 2001). What all of them have in common is the oligotrophic characteristics of their waters (VAN DE VIJVER et al. 2012). Other relevant aspect, is that some of the described species of the Antarctic region can form colonies through the presence of marginal spines (VAN DE VIJVER et al. 2004; PINSEEL et al. 2016), although most of the Pinnularia species have isolated life forms (ROUND et al. 1990). Although chain formation by spines affects many biological processes, such as sexual interactions, predation and nutrient uptake (e.g. FRYXELL & MILLER 1978; PAHLOW et al. 1997), it is not clear the main function in other taxa of the Pinnularia species (PINSEEL et al. 2016). Among those that have spines it is important to highlight Pinnularia atlasi DARLEY (DARLEY 1990). In that study, DARLEY did not show the presence of spines, however in a posterior study carried out in Sardinia by LANGE-BERTALOT et al. (2003), spines were detected in the valvar margin from other population of P. atlasi, mostly concentrated in the central area and disappearing towards the ends of the valve. Pinnularia infirma KRAMMER & LANGE-BERTALOT (1985), a second taxon used for comparison was firstly described in central Europe, in a locality situated at 250 m of altitude in Franken, Germany. It was also found in Ouiouane, Morocco, together with P. atlasi. No ultrastructural data was provided and consequently, no reference to the presence of spines was given. This study is part of a larger one that intends to know the diatomological diversity of the coastal zone from the lagoons of Andalusia (southern Spain) (unpublished results). 4 bioRxiv preprint doi: https://doi.org/10.1101/452110; this version posted October 26, 2018. The copyright holder for this preprint (which was not certified by peer review) is the author/funder. All rights reserved. No reuse allowed without permission. According to European algal framework directive (POKAINE et al. 2016), studies using diatoms are normally focused in riverine communities and not so much in lacustrine ones. Thus, this is why this study is crucial to improve the knowledge on diatom communities from lagoons, specially alkaline ones. The main objective of this work is the description of a new species from the genus Pinnularia, Pinnularia baetica sp. nov. A comparison will be made regarding the frustule morphology of the population found in the province of Granada (South of Spain) both in Light Microscopy (LM) and in Scanning Electron Microscopy (SEM), with P. atlasi described by DARLEY, obtained directly from (locotype)the 2 sites of northern Morocco (Ouiouane and Guedrouz) where DARLEY originally obtained the type population. In spite of type material was asked, it was not possible to received. It will also be compared with the description done by LANGE-BERTALOT et al. (2003), in the Island of Sardinia because DARLEY’S description did not include ultrastructural analysis. Methods Sampling The epipelic diatom communities of sediments samples came from the littoral zone of three lagoons, one from South Europe (Spain) and two from North Africa (Morocco) (Table 1, Fig.1): 1) The Laguna Seca (Granada, Spain), that is located at the north of the Natural Park of Sierra de Castril (Complex of Baetic Mountains). It is dominated by outcrops of limestone and dolomites originated in the Upper Jurassic. The Laguna Seca is at 2000 m of altitude, and is small pond located in a shallow 5 bioRxiv preprint doi: https://doi.org/10.1101/452110; this version posted October 26, 2018. The copyright holder for this preprint (which was not certified by peer review) is the author/funder. All rights reserved. No reuse allowed without permission. basin, filled with rainwater and snowmelt; it is usually dry at the beginning of the dry season. This material was collected in 2007. The Ouiouane (33°07´52.58´´ N, 5°20´36.72´´ W) and Guedrouz (31°26´12.45´´ N, 7°28´06.82´´ W) were problematic to sampling, because the coordinates were not shown in the original P. atlasi described by DARLEY. We did a search on Google earth to find Guedrouz lagoon from the Darley´s description “Timenkar plates, at 2200 m of altitude in the central High Atlas in the region of Marrakech”, finally with the help of people from the high Atlas we found the lagoon. The Ouiouane lagoon was chosen on google earth with a selection of several lagoons close to the city of Azrou at the mountains, as long as Darley described the location close to this city. With the help of field microscope. we distinguished the typical panduriform outline and shape of Pinnularia atlasi at the littoral zone of Ouiouane lake, The other lagoons and lakes did not have the presenceof this diatom species.