Viviana Peña

A Multilocus Species Delimitation Reveals a Striking Number of Species of Coralline Algae Forming Maerl in the OSPAR Maritime Area

Maerl beds are sensitive biogenic habitats built by an accumulation of loose-lying, non-geniculate coralline algae. While these habitats are considered hot-spots of marine biodiversity, the number and distribution of maerl-forming species is uncertain because homoplasy and plasticity of morphological characters are common. As a result, species discrimination based on morphological features is notoriously challenging, making these coralline algae the ideal candidates for a DNA barcoding study. Here, mitochondrial (COI-5P DNA barcode fragment) and plastidial (psbA gene) sequence data were used in a two-step approach to delimit species in 224 collections of maerl sampled from Svalbard (78°96’N) to the Canary Islands (28°64’N) that represented 10 morphospecies from four genera and two families. First, the COI-5P dataset was analyzed with two methods based on distinct criteria (ABGD and GMYC) to delineate 16 primary species hypotheses (PSHs) arranged into four major lineages. Second, chloroplast (psbA) sequence data served to consolidate these PSHs into 13 secondary species hypotheses (SSHs) that showed biologically plausible ranges. Using several lines of evidence (e.g. morphological characters, known species distributions, sequences from type and topotype material), six SSHs were assigned to available species names that included the geographically widespread Phymatolithon calcareum, Lithothamnion corallioides, and L. glaciale; possible identities of other SSHs are discussed. Concordance between SSHs and morphospecies was minimal, highlighting the convenience of DNA barcoding for an accurate identification of maerl specimens. Our survey indicated that a majority of maerl forming species have small distribution ranges and revealed a gradual replacement of species with latitude.

Biological importance of an Atlantic European maërl bed off Benencia Island (northwest Iberian Peninsula)

We studied the conservation status of a maerl bed off Benencia Island (NW Spain) over a 2-year period using SCUBA and dredging. The maerl bed, which includes a maerl beach, extended from the intertidal to subtidal zones (18 m), and occupied an area of 215 ha. It was composed of a pure unattached coralline algal deposit occasionally mixed with broken shells. The area estimated for the highest maerl cover (76–100%) was greater than reported in previous research. The living maerl layer reached 15 cm in depth and the living/dead maerl ratio was high (80–100% living maerl cover). The dominant maerl-forming species was Phymatolithon calcareum, although unattached plants of Mesophyllum sp. reaching 10 cm in diameter were found in two sites. Both maerl species are fruticose. The proportion of discoidal specimens of P. calcareum was related to depth. The associated flora comprised 137 species whose seasonal variations were very marked, with high floristic richness in summer (71 species) and low in winter (35 species). Crustose and turf-like species were the dominant floral components throughout the year, whereas the presence of other species was restricted to specific periods. We propose that Benencia Island be included in future Atlantic Iberian maerl bed conservation schemes.

MESOPHYLLUM SPHAERICUM SP. NOV. (CORALLINALES, RHODOPHYTA): A NEW MAËRL-FORMING SPECIES FROM THE NORTHEAST ATLANTIC1

Mesophyllum sphaericum sp. nov. is described based on spherical maërl individuals (up to 10 cm) collected in a shallow subtidal maërl bed in Galicia (NW Spain). The thalli of these specimens are radially organized, composed of arching tiers of compact medullary filaments. Epithallial cells have flattened to rounded outermost walls, and they occur in a single layer. Subepithallial initials are as long as, or longer than the daughter cells that subtend them. Cell fusions are abundant. Multiporate asexual conceptacles are protruding, mound‐like with a flattened pore plate, lacking a peripheral raised rim. Filaments lining the pore canal and the conceptacle roof are composed of five to six cells with straight elongate and narrow cells at their base. Carposporangial conceptacles are uniporate, protruding, and conical. Spermatangial conceptacles were not observed. Molecular results placed M. sphaericum near to M. erubescens, but M. sphaericum is anatomically close to M. canariense. The examination of the holotype and herbarium specimens of M. canariense indicated that both species have pore canal filaments with elongate basal cells, but they differ in number of cells (five to six in M. sphaericum vs. four in M. canariense). Based on the character of pore canal filaments, M. canariense shows similarities with M. erubescens (three to five celled). The outermost walls of epithallial cells of M. canariense are flared compared to the round to flattened ones of M. erubescens, the latter being widely accepted for the genus Mesophyllum. The addition of M. sphaericum as new maërl‐forming species suggests that European maërl beds are more biodiverse than previously understood.

Phylogenetic relationships of corallinaceae (Corallinales, Rhodophyta): taxonomic implications for reef-building corallines.

A new, more complete, five‐marker (SSU, LSU, psbA, COI, 23S) molecular phylogeny of the family Corallinaceae, order Corallinales, shows a paraphyletic grouping of seven well‐supported monophyletic clades. The taxonomic implications included the amendment of two subfamilies, Neogoniolithoideae and Metagoniolithoideae, and the rejection of Porolithoideae as an independent subfamily. Metagoniolithoideae contained Harveylithon gen. nov., with H. rupestre comb. nov. as the generitype, and H. canariense stat. nov., H. munitum comb. nov., and H. samoënse comb. nov. Spongites and Pneophyllum belonged to separate clades. The subfamily Neogoniolithoideae included the generitype of Spongites, S. fruticulosus, for which an epitype was designated. Pneophyllum requires reassesment. The generitype of Hydrolithon, H. reinboldii, was a younger heterotypic synonym of H. boergesenii. The evolutionary novelty of the subfamilies Hydrolithoideae, Metagoniolithoideae, and Lithophylloideae was the development of tetra/bisporangial conceptacle roofs by filaments surrounding and interspersed among the sporangial initials.

Seasonal patterns in the maërl community of shallow European Atlantic beds and their use as a baseline for monitoring studies

A study of three shallow Atlantic maërl beds (3–6 m depth) in NW Spain (Galicia) sampled beds every 1.5 months for a year. At each study site, temporal variation in the associated flora and two parameters, temperature and photosynthetic photon flux density (PPFD), were recorded in situ and means were calculated for each period. The range of mean periodic temperatures was 11.9–17.5°C, increasing in late spring–summer and decreasing to <12°C in winter. Mean PPFD was 24–160 µmol photons m−2 s−1; the maximum was recorded in spring (up to 571 µmol photons m−2 s−1). The variation in temperature and PPFD was correlated with seasonal changes in the flora. The composition of the associated flora at Benencia Island (Ría de Arousa) showed a strong correlation with day length while at Tambo Island (Ría de Pontevedra), it was correlated with temperature. A total of 127 species was recorded (in seven functional groups), annual algae being the most abundant group. The highest species richness and total percentage cover occurred in spring and summer, and the lowest in autumn and winter. Significant seasonal differences were detected at each site, particularly between summer and autumn–winter, due to variation in cover of the foliose species Ulva rigida and Dictyota dichotoma. We propose that future monitoring of European Atlantic maërl beds should include several study areas and two sampling seasons (preferably summer and late autumn–winter).

An integrative systematic approach to species diversity and distribution in the genus Mesophyllum (Corallinales, Rhodophyta) in Atlantic and Mediterranean Europe

For the first time, a comprehensive assessment of Mesophyllum species diversity and their distribution in Atlantic Europe and the Mediterranean Sea is presented based on molecular (COI-5P, psbA) and morphological data. The distribution ranges were redefined for the four species collected in this study: M. alternans, M. expansum, M. macroblastum and M. sphaericum. Mesophyllum sphaericum, which was previously known only from a single maerl bed in Galicia (NW Spain), is reported from the Mediterranean Sea. The known range of M. expansum (Mediterranean and Macaronesia) was extended to the Atlantic Iberian Peninsula. The occurrence of M. alternans was confirmed along the Atlantic French coast south to Algarve (southern Portugal). Mesophyllum lichenoides was only recorded from the Atlantic, whereas M. macroblastum appears to be restricted to the Mediterranean Sea. A positive correlation was observed between maximum Sea Surface Temperature (SSTmax) and the depth at which M. expansum was collected, suggesting that this species may compensate for higher SST by growing in deeper habitats where the temperature is lower. The latter indicates that geographic shifts in the distribution of coastal species as a result of global warming can possibly be mitigated by changes in the depth profile at which these species occur. Mesophyllum expansum, an important builder of Mediterranean coralligenous habitats, may be a good target species to assess its response to climate change.

Detection of gametophytes in the maerl-forming species Phymatolithon calcareum (Melobesioideae, Corallinales) assessed by DNA barcoding

Abstract Fertile gametangial plants of Phymatolithon calcareum, which are seldom reported in the Atlantic European coasts, were collected as encrusting, epilithic plants in a subtidal maerl bed in Brittany (France). Based on their morphological features, the plants were identified as P. calcareum. This identification was further confirmed by DNA barcodes using as a reference COI-5P sequences obtained from the neotype together with recent collections from the Atlantic European maerl beds. The reproductive structures were empty but they were regarded as mature female conceptacles. Compared to the two previous records of gametangial plants of P. calcareum for the Atlantic European waters, the uniporate conceptacles observed in this study are larger, and were collected at a different time of the year. To our knowledge, this is the first time that the occurrence of gametangial plants of P. calcareum is corroborated with molecular tools (DNA barcodes).

Phymatolithon lusitanicum sp. nov. (Hapalidiales, Rhodophyta): The Third Most Abundant Maerl-Forming Species in the Atlantic Iberian Peninsula

Abstract Phymatolithon lusitanicum is a new maerl species described based on an integrative systematic approach including molecular (COI-5P, psbA) and morphological data obtained from recent collections, as well as comparison of type material from the morphologically and ecologically alike NE Atlantic species P. lamii and P. laevigatum. Molecular analyses including type material of P. lamii and P. laevigatum were congruent in delimiting P. lusitanicum as an independent lineage from these crustose species. The three species shared a common external morphology of multiporate asexual conceptacles, but P. lusitanicum has been detected only unattached as maerl while P. lamii and P. laevigatum are crustose. Phymatolithon lusitanicum is particularly abundant in subtidal maerl beds of the Atlantic Iberian Peninsula (Galicia and the Algarve); however it has also been detected northwards in Ireland intertidally and in Western Mediterranean Sea (Alborán Sea, Balearic Islands) down to 64 m. Phymatolithon lusitanicum differs from other Phymatolithon species reported for the European coasts mainly by the external shape of the multiporate asexual conceptacles (pore plate flush with surface or slightly sunken without a conspicuous thick raised rim) and its unattached habit as maerl/rhodolith. In addition, the lectotype of Lithothamnion hamelii turned out to be conspecific to Phymatolithon calcareum, therefore this taxon is proposed as a heterotypic synonym of P. calcareum. Finally, our molecular analyses detected cryptic diversity within the European collections of Phymatolithon, while collections identified as P. lenormandii from Canada or P. repandum from New Zealand were resolved as unrelated to the remaining Phymatolithon. In the light of these results, it is clear that further work is necessary to resolve species diversity within the genus Phymatolithon and its relationship with related genera.

Phylogenetic analysis of rhodolith formation in the Corallinales (Rhodophyta)

Although the ecological importance of rhodolith (maerl, free-living coralline algae) beds is well-known, rhodolith-forming species have been neglected in molecular phylogenetic studies. This is the first molecular systematic study aimed at understanding whether the rhodolith habit is a fixed feature in lineages and determining the relationship (phylogenetic vs. environmental) between rhodolith and crustose habits. Phylogenetic relationships of rhodolith-forming species and encrusting coralline algae at generic and species levels were analysed using SSU rDNA and psbA sequences. Extensive sampling in the European North Atlantic, Pacific and Caribbean Mexico of Phymatolithon, Lithothamnion, Lithophyllum and Neogoniolithon taxa forming rhodoliths and crusts was accompanied by examination of type or topotype material. Phylogenetic reconstruction showed that Neogoniolithon contained a monophyletic group of rhodolith-forming species whereas other rhodolith-formers were closely related to encrusting forms in the genera Phymatolithon, Lithothamnion, Mesophyllum, Hydrolithon, Spongites and Sporolithon. DNA analysis showed that the crust-forming Lithophyllum cf. incrustans/dentatum also forms rhodoliths with a stone nucleus that occur on rocky shores. In contrast, species that form beds of non-nucleate rhodoliths (e.g. Neogoniolithon spectabile, N. strictum, Lithophyllum cf. incrustans/dentatum or sp. 1 and Phymatolithon calcareum) rarely form crusts. The rhodolith habit cannot be used to delimit species for taxonomic or identification purposes. Extensive taxonomic revision will be required to deal with problems such as the position of specimens identified as Lithophyllum margaritae in two unrelated lineages.

A Molecular and Morphological Study of Corallina sensu lato (Corallinales, Rhodophyta) in the Atlantic Iberian Peninsula

Abstract DNA barcodes and morphological observation were used to evaluate the number of species within the genus Corallina sensu lato in Atlantic Iberia and to identify a set of morphological characters that may discriminate them. Five species were detected: (i) C. officinalis, (ii) C. caespitosa, (iii) Ellisolandia elongata, (iv) Corallina sp. 1, and (v) Corallina sp. 2. The first three species were widespread whereas Corallina sp. 1 was only detected in southern Atlantic coasts of Iberia and Corallina sp. 2 occurred both in the Atlantic and the Mediterranean, nevertheless, with morphological and molecular differences. The widespread occurrence of the recently described C. caespitosa along Atlantic Iberia is substantiated with sequence data for the first time; it spans from the Bay of Biscay to Andalusia and the French Mediterranean. Molecular support is also provided for the presence of C. officinalis and E. elongata in Atlantic Iberia. Plants of C. officinalis were correctly identified attending to their external morphology, however two distinct species were detected under the two forms of C. elongata (“typical” and “elongated”) reported for Galicia in the literature (C. caespitosa and Ellisolandia elongata, respectively). Plant size, habit, branching, and shape of intergenicula seem the most informative morphological characters to discriminate among the various members of Corallina s.l. in Atlantic Iberia.