MOLECULAR PHYLOGENY OF ANTARCTIC PRASIOLA (PRASIOLALES,TREBOUXIOPHYCEAE) REVEALS EXTENSIVE CRYPTIC DIVERSITY1

Trebouxiophytes of the genus Prasiola are well known in Antarctica, where they are among the most important primary producers. Although many aspects of their biology have been thoroughly investigated, the scarcity of molecular data has so far prevented an accurate assessment of their taxonomy and phylogenetic position. Using sequences of the chloroplast genes rbcL and psaB, we demonstrate the existence of three cryptic species that were previously confused under Prasiola crispa (Lightfoot) Kützing. Genuine P. crispa occurs in Antarctica; its presence was confirmed by comparison with the rbcL sequence of the type specimen (from the Isle of Skye, Scotland). Prasiola antarctica Kützing is resurrected as an independent species to designate algae with gross morphology identical to P. crispa but robustly placed in a separate lineage. The third species is represented by specimens identified as P. calophylla (Carmichael ex Greville) Kützing in previous studies, but clearly separated from European P. calophylla (type locality: Argyll, Scotland); this alga is described as P. glacialis sp. nov. The molecular data demonstrated the presence of P. crispa in Maritime and Continental Antarctica. P. antarctica was recorded from the Antarctic Peninsula and Shetland Islands, and P. glacialis from the Southern Ocean islands and coast. Such unexpected cryptic diversity highlights the need for a taxonomic reassessment of many published Antarctic records of P. crispa. The results also indicate that marine species of Prasiola form a well‐supported monophyletic group, whereas the phylogenetic diversity of freshwater species is higher than previously suspected (at least three separate lineages within the genus include species living in this type of environments).     

Prasiola (Prasiolales,Trebouxiophyceae) in Japan: a survey of freshwater populations and new records of marine taxa

Recent collections from marine and freshwater locations have enabled the investigation of diversity of Prasiola in Japan. Sequence data from the rbc L and tuf A markers revealed the presence of three marine species and one freshwater species. Prasiola delicata was confirmed to occur on Daikokujima, Prasiola calophylla was found for the first time in Japan from Hokkaido, and a species within the P. meridionalis/linearis/stipitata complex was found on both Hokkaido and Daikokujima. Collections from a range of populations of freshwater Prasiola, identified here as P. japonica, were found to be conspecific and identical in rbc L and tuf A sequences to freshwater collections from Nepal, Korea, and China.     

MOLECULAR PHYLOGENY OF THE GREEN ALGAL ORDER PRASIOLALES (TREBOUXIOPHYCEAE,CHLOROPHYTA)1

The systematics of the Prasiolales was investigated by phylogenetic inference based on analyses of the rbcL and 18S rRNA genes for representatives of all four genera currently attributed to this order (Prasiococcus, Prasiola, Prasiolopsis, Rosenvingiella), including all type species. The rbcL gene had higher sequence divergence than the 18S rRNA gene and was more useful for phylogenetic inference at the ranks of genus and species. In the rbcL gene phylogeny, three main clades were observed, corresponding to Prasiola, Prasiolopsis, and Rosenvingiella. Prasiococcus was nested among species of Prasiola occurring in subaerial and supralittoral habitats. Trichophilus welckeri Weber Bosse, a subaerial alga occurring in the fur of sloths in Amazonia, was closely related to Prasiolopsis ramosa Vischer. The species of Prasiola were grouped into three well‐supported clades comprising (i) marine species, (ii) freshwater and terrestrial species with linear blades, and (iii) terrestrial species with rounded or fan‐shaped blades. Sequence divergence was unexpectedly low in the marine group, which included species with different morphologies. For the 18S rRNA gene, the phylogenetic analyses produced several clades observed for the rbcL gene sequence analysis, but, due to very little sequence variation, it showed considerably lower resolution for inference at the species and genus levels. Due to the low support of some internal branches, the results of the analyses did not allow an unambiguous clarification of the origin and the early evolution of the Prasiolales.     

THE PRASIOLALES (CHLOROPHYTA) OF ATLANTIC EUROPE: AN ASSESSMENT BASED ON MORPHOLOGICAL,MOLECULAR, AND ECOLOGICAL DATA,INCLUDING THE CHARACTERIZATION OF ROSENVINGIELLA RADICANS (KÜTZING) COMB. NOV.1

Despite a simple morphology and intensive studies carried out for more than two centuries, the systematics of the Prasiolales still presents several unsolved problems. The taxonomic relationships of several common species of Prasiolales, mostly from northern Europe, were investigated by a combination of morphological observations, culture experiments, and molecular analyses based on rbcL sequences. The results indicate that Rosenvingiella and Prasiola are separate genera. The capacity for production of tridimensional pluriseriate gametangia and the presence of unicellular rhizoids are the morphological features that discriminate Rosenvingiella from filamentous forms of Prasiola. The molecular data indicate that uniseriate filaments can be produced in at least three different species of Prasiola. The genetic diversity of uniseriate filamentous Prasiolales is higher than their simple morphology would indicate, and the provisional retention of Schizogonium Kützing 1843 as independent genus is recommended. The rbcL phylogeny confirms that Prasiola calophylla, P. crispa, and P. stipitata are distinct species, whereas P. stipitata and P. meridionalis are probably conspecific. Rosenvingiella polyrhiza is a strictly marine alga, and most records of Rosenvingiella in Europe are referable to Rosenvingiella radicans, proposed here as a new combination based on Ulothrix radicans Kützing 1849 . This is a primarily terrestrial alga that can occur from upper intertidal rock to locations situated hundreds of kilometers inland. The great confusion that has arisen in Europe between these two species in the last century is mostly due to misidentifications of marine populations of R. radicans.     

Prasiolales (Trebouxiophyceae,Chlorophyta) of the Svalbard Archipelago: diversity,biogeography and description of the new genera Prasionella and Prasionema

Species of Prasiolales (Trebouxiophyceae, Chlorophyta) are among the most common terrestrial and freshwater algae in polar regions. Comprehensive molecular studies of this group are available for Antarctica, but not yet for Arctic regions. We examined the diversity of the Prasiolales in the Svalbard Archipelago combining morphological observations of field-collected material, culture studies, molecular data (plastid rbcL and tufA sequences) and literature records. We confirmed the widespread occurrence of Prasiola crispa and P. fluviatilis, species recorded from Spitsbergen since the 19th century. Molecular phylogenetic analyses led to the discovery of two new genera of Prasiolales. Prasionema payeri is morphologically identical to filamentous stages of P. crispa, but represents an early-diverging lineage in the order. Prasionella wendyae is a colonial alga reproducing by aplanospores; its phylogenetic position is among the basal lineages of the order, but it could not be reliably reconstructed due to weak statistical support. The inclusion of P. wendyae in the prasiolalean phylogeny determined the paraphyly of Rosenvingiella, requiring the establishment of the new genus Rosenvingiellopsis for R. constricta. A poorly known species described from Spitsbergen, Ulothrix discifera, is transferred here to Rosenvingiella. Whereas some species of Prasiolales have bipolar distribution (P. crispa), others appear to be restricted to one or other of the poles. Our results suggest that polar regions are still a major repository of unknown algal diversity and highlight the importance of continued field surveys and the use of molecular data.     

Prasionema heeschiae sp. nov. (Prasiolales,Chlorophyta) from Campbell Island,New Zealand: first record of Prasionema in the southern hemisphere

A new species in the genus Prasionema (Prasiolales, Trebouxiophyceae) is described from Campbell Island, in the New Zealand subantarctic region, the first record of this genus in the southern hemisphere. Prasionema heeschiae sp. nov. is filamentous, uni- to predominantly biseriate, with disc-shaped cells, and an axial plastid with a central pyrenoid. It is anchored by an enlarged pigmented basal cell. There is evidence of reproduction by both spores and fragmentation. This species was found growing on a timber wharf above the high tide level. The only other species in the genus, P. payeri Heesch, M.Pazoutová & Rindi, was described from Spitzbergen growing on soil in a high nutrient environment. The bipolar distribution of Prasionema is discussed. Based on phylogenetic analyses that included sequence data from holotype material of southern filamentous Prasiolales, we reduce Rosenvingiella australis to synonymy with R. tasmanica.