Species delimitation methods reveal cryptic diversity in the Hypnea cornuta complex (Cystocloniaceae,Rhodophyta)

The simple and convergent morphologies of many red algae make these species difficult to identify using traditional morphological characters. Many cryptic species have been described in recent years based on molecular datasets, and this has led to the application of an integrative taxonomy approach in species delimitation. Here, we performed several species delimitation methods (mBGD, ABGD, SPN, PTP, GMYCs and GMYCm) based on two different loci (COI-5P and rbcL) in species of the Hypnea cornuta complex. These methods were combined with morphological and phylogenetic data, extensive sampling, analysis of topotype material, and historically relevant herbarium samples. Our findings demonstrate that the groups morphologically assigned to H. cornuta and H. stellulifera consist of five different cryptic species. H. cornuta is a polyphyletic taxon composed of three well-separated lineages, thus requiring sequencing of type or topotype specimens to determine which one is Hypnea cornuta sensu stricto. We have revealed that the distribution of H. stellulifera is limited to Asia, while the Brazilian specimens initially assigned to this species were clarified as a new endemic species: Hypnea cryptica sp. nov. Our results indicated that only an integrative approach combining several lines of evidence, including morphology, nomenclature history, molecular data, biogeography and ecology can correctly solve the taxonomic status of widely distributed cryptic species.     

A combined barcode and morphological approach to the systematics and biogeography of Laurencia pyramidalis and Laurenciella marilzae (Rhodophyta)

In the present study we undertook an integrative approach, using both morphological and molecular data (COI-5P + rbcL), to assess the presence of Laurencia pyramidalis in Lusitanian Macaronesia. We studied type material of L. pyramidalis from the herbarium of the Naturalis Biodiversity Center, the Netherlands, and designated a lectotype and syntypes. Vegetative and reproductive features of L. pyramidalis were observed and we included a specimen from the type locality in our molecular analyses. We also investigated the geographical distribution of Laurenciella marilzae, a species recently described from the Canary Islands. Barcode sequences (COI-5P and rbcL) were generated for L. pyramidalis from the type locality (Normandy, France), the Azores, Madeira and the Canary Islands, and for L. marilzae from its type locality (Tenerife, Canary Islands), the Azores and Brazil.     

A morphological and molecular assessment of the genus Sinotubimorpha (Halymeniaceae, Rhodophyta)

A critical reassessment of the morphological features of two closely related red algal genera,Grateloupia C.Agardh and Sinotubimorpha W.X.Li & Z.F.Ding (Halymeniaceae),pointed out that members of the t...     

A morphological and molecular assessment of the genus Prionitis J. Agardh (Halymeniaceae, Rhodophyta)

A critical reassessment of the morphological features of two closely related red algal genera, Grateloupia C. Agardh and Prionitis J. Agardh (Halymeniaceae), shows that members of the two genera share very similar reproductive (including the Grateloupia‐type auxiliary‐cell ampullae) and vegetative characters. Diagnostic features hitherto used for distinguishing these two genera, the texture of blades (lubricous to leathery in Grateloupia vs cartilaginous in Prionitis) and the position of reproductive structures (scattered over the entire blade in Grateloupia vs confined to particular portions of the blade in Prionitis), are continuous across some 75 species of both genera, thus making it difficult to draw a clear‐cut distinction between the two genera. In ribulose‐1,5‐bisphosphate carboxylase/oxygenase gene (rbcL) sequence analyses, the species of Grateloupia and Prionitis, including the two generitypes, constitute a large monophyletic clade in the Halymeniaceae. It is therefore proposed that Prionitis be included in the synonymy under Grateloupia and the appropriate combinations are proposed.     

Genetic diversity of Kappaphycus species (Gigartinales,Rhodophyta) in the Philippines

The mariculture of eucheumatoids (species of Kappaphycus and Eucheuma) in the Philippines has had a long history, dating back to the 1970s. Over this period, a number of varieties have been brought into domestication; some are now widely distributed and farmed in various regions of the country, but a significant number appear to have a more restricted distribution and are farmed only in certain areas. The taxonomy of many of these seaweed cultivars and their phylogenetic relationships still remain to be resolved at the specific and subspecific levels. In this study, two mitochondrial DNA markers, COI-5P region and cox2-3 intergenic spacer, were used to assess the genetic diversity of the farmed varieties and a few specimens collected from the wild. Analysis using haplotype networks revealed several new haplotypes for K. alvarezii, K. malesianus and K. striatus, mainly from specimens collected from eastern and southwestern regions of the Philippines. The inferred phylogenetic relationships based on both mtDNA markers resolved the identity of all the materials used in the study at the species level. We present molecular evidence that K. malesianus, in addition to K. alvarezii, K. cottonii, K. inermis, K. procrusteanus and K. striatus (and, hence, all currently recognized species of Kappaphycus) occurs in the Philippines. Collectively, these observations suggest that the Philippine archipelago has richer genetic diversity of farmed and wild Kappaphycus than do the other geographic regions, consistent with the hypothesis that the Philippines is, or is part of, the centre of Kappaphycus biodiversity in the world. These findings also reveal an untapped diversity that can potentially be exploited for improving the commercial production of these carrageenophytes.     

Pterosiphonia tanakae (Rhodomelaceae, Ceramiales), a new red algal species from Japan

The red alga Pterosiphonia tanakae Uwai et Masuda (Rhodomelaceae, Ceramiales) is established as a new species on the basis of material collected in Hiroshima Bay, the Inland Sea, Japan. It can be distinguished from the majority of previously described species of the genus by two key features, seven to 10 periaxial cells and the presence of weakly developed cortical cells. This species is further distinguished from P. pau-cicorticata Dawson by completely alternate-distichously branching and from Pjavanica (Martens) De Toni and P. spinifera (Kützing) Norris et Aken by the production of first-order laterals that bear branches of up to five orders, which results in the thallus having a widely spreading, fan-shaped appearance. The abundant production of vegetative trichoblasts is a further characteristic feature of P. tanakae.     

Delimitating cryptic species in the Gracilaria domingensis complex (Gracilariaceae,Rhodophyta) using molecular and morphological data

Species in the genus Gracilaria that display conspicuously flattened vegetative morphologies are a taxonomically challenging group of marine benthic red algae. This is a result of their species richness, morphological similarity, and broad phenotypic plasticity. Within this group, the Gracilaria domingensis complex is one of the most common, conspicuous, and morphologically variable species along the tropical western Atlantic Ocean. Previous research has identified that members of this complex belong to two distantly related clades. However, despite this increased phylogentic resolution, species delimitations within each of these clades remain unclear. Our study assessed the species diversity within this difficult complex using morphological and molecular data from three genetic markers (cox1, UPA, and rbcL). We additionally applied six single‐marker species delimitation methods (SDM: ABGD, GMYCs, GMYCm, SPN, bPTP, and PTP) to rbcL, which were largely in agreement regarding species delimitation. These results, combined with our analysis of morphology, indicate that the G. domingensis complex includes seven distinct species, each of which are not all most closely related: G. cervicornis; a ressurected G. ferox; G. apiculata subsp. apiculata; a new species, Gracilaria baiana sp. nov.; G. intermedia subsp. intermedia; G. venezuelensis; and G. domingensis sensu stricto, which includes the later heterotypic synonym, G. yoneshigueana. Our study demonstrates the value of multipronged strategies, including the use of both molecular and morphological approaches, to decipher cryptic species of red algae.     

Neosiphonia flavimarina gen. et sp. nov. with a taxonomic reassessment of the genus Polysiphonia (Rhodomelaceae, Rhodophyta)

Vegetative and reproductive development of Neosiphonia flavimarina gen. et sp. nov. (Rhodomelaceae, Ceramiales) from Bangpo on the western coast of Korea was investigated. This species is superficially similar to Polysiphonia, but differs distinctly from the latter in vegetative and reproductive structures. The plants attach by a solid disk composed of a dense cluster of rhizoids cut off from the pericentral cell wall, and bear erect indeterminate branches producing the lateral-branch initials from successive segments in a spiral arrangement. The procarps have a three-celled carpo-gonial branch. Spermatangial branches are formed on a primary branch of the trichoblasts, terminating in a single or occasionally two large, sterile cells. Tetra-sporangia are produced from the second pericentral cell adjacent to the trichoblast basal cell on indeterminate branches, and arranged spirally. Comparing several taxonomic characters among related genera, Neosiphonia occupies an independent phylogenetic position from Polysiphonia and leads to the conclusion that the genus may have a strong link with Fernandosiphonia which has a unilateral branching system. Relevant nomenclatural changes for several Polysiphonia species are also proposed.     

Australian Laurencia majuscula (Rhodophyta,Rhodomelaceae) and the Brazilian Laurencia dendroidea are conspecific

Morphological and molecular studies have been undertaken on two species of the red algal genus Laurencia J.V.Lamouroux: Laurencia majuscula (Harvey) A.H.S. Lucas and Laurencia dendroidea J.Agardh, both from their type localities. The phylogenetic position of these species was inferred by analysis of the chloroplast‐encoded rbcL gene sequences from 24 taxa. In all phylogenetic analyses, the Australian Laurencia majuscula and the Brazilian L. dendroidea formed a well‐supported monophyletic clade within the Laurencia sensu stricto. This clade was divided into two subclades corresponding to each geographical region; however, the genetic divergence between Australian L. majuscula and Brazilian L. dendroidea was only 0–1.35%. Examination of the type specimens and sequences of freshly collected samples of both Laurencia majuscula and L. dendroidea show the two to be conspecific despite their disjunct type localities.     

Taxonomic notes on Laurencia mariannensis (Rhodomelaceae, Rhodophyta)

The marine red alga Laurencia mariannensis Yamada (Rhodomelaceae, Ceramiales) is characterized by pale rose-red, softly fleshy, slender, terete axes (up to 600 μm in diameter), arising from a loosely entangled, stoloniferous basal system among species that have the following combination of features: the presence of longitudinally orientated secondary pit-connections between contiguous superficial cortical cells; the presence of projecting superficial cortical cells at the upper portions of branches; and the presence of lenticular thickenings in the walls of medullary cells, Furthermore, the presence of two or three corps en cerise per superficial cortical cell and one per trichoblast cell may characterize the species.     

Hypnea species (Gigartinales,Rhodophyta) from the southeastern coast of Brazil based on molecular studies complemented with morphological analyses,including descriptions of Hypnea edeniana sp. nov. and H. flava sp. nov.

The red algal genus Hypnea (Gigartinales) has a wide geographical distribution along tropical and subtropical coasts around the world. The relatively simple and plastic morphology, often influenced by the conditions of its habitat, complicates the identification of Hypnea species. Therefore, the number and status of some species remain in doubt. Molecular studies have been performed to supplement traditional studies based on morphology, mainly for Hypnea species occurring in Asia. In the present study, sequence data from the DNA barcode COI-5P for 114 samples from the southeastern coast of Brazil, indicated the occurrence of six taxa. Additionally, sequence data from the UPA and rbcL markers for representatives of each of those taxa confirmed the existence of six different species. After morphological analysis and comparison with sequences available in GenBank, these species were named as follows: H. aspera, H. cervicornis, H. cf. musciformis, H. spinella, and two new species, H. flava Nauer, Cassano & M.C. Oliveira and H. edeniana Nauer, Cassano & M.C. Oliveira. Hypnea cervicornis, often considered as a later synonym of H. spinella, should be considered as a distinct species based on morphology and divergence of the three molecular markers used. Hypnea aspera is a new record for the Atlantic Ocean.     

Taxonomic notes on Hawaiian Polysiphonia, with transfer to Neosiphonia (Rhodomelaceae, Rhodophyta)

After detailed observations of type material and other collections, five Hawaiian species of Polysiphonia Greville, nom. cons. are recognized to be species of Neosiphonia M. S. Kim et I. K. Lee; namely, Neosiphonia apiculata (Hollenberg) Masuda et Kogame, Neosiphonia beaudettei (Hollenberg) M. S. Kim et Abbott, comb. nov., Neosiphonia hawaiiensis (Hollenberg) M. S. Kim et Abbott, comb. nov., Neosiphonia profunda (Hollenberg) M. S. Kim et Abbott, comb. nov., and Neosiphonia rubrorhiza (Hollenberg) M. S. Kim et Abbott, comb. nov. These five species are ecorticate, having lateral branch initials and trichoblasts produced on successive segments, rhizoids separated from pericentral cells by a cross wall, three‐celled carpogonial branches (not seen in N. beaudettei and N. rubrorhiza), spermatangial branches arising on a primary branch of the trichoblasts, and tetrasporangia in a spiral series. Although certain characters were not available for some species, all other characters occur in a combination that is unique for members of Neosiphonia.     

Kallymenia crouaniorum (Kallymeniaceae,Rhodophyta), a new red algal species from the Laminaria hyperborea understorey community

In this paper we describe Kallymenia crouaniorum Vergés & Le Gall, sp. nov. (Kallymeniaceae), a new marine red alga from the north-eastern Atlantic Ocean. rbcL and LSU sequences of this species, previously misidentified in the field as Kallymenia reniformis, diverged from those of other Kallymenia species by at least 7.5% and 5.2%, respectively. Kallymenia crouaniorum also has a set of distinctive vegetative and reproductive characteristics, including a deeply lacerate frond, a short stipe, dentate margins, large cortical cells up to 110 μm in diameter, highly refractive stellate medullary cells with arms up to 1000 μm in length, and a monocarpogonial branch system. Molecular phylogenies inferred from rbcL and LSU data indicated, albeit with weak support, that this new species is a sister taxon of a lineage encompassing the generitype K. reniformis, as well as most species of Kallymenia included in the phylogenetic analysis. The main morphological characters that delineate monocarpogonial Kallymenia species are presented. Moreover, after reviewing the literature and several herbarium specimens, we found in the Weber-van Bosse Herbarium a specimen collected at Roscoff in August 1894 with the anatomical characters of the species described here, confirming that this newly described species has in fact been overlooked and is not a recent introduction.     

Diversity and distribution of Laurencia sensu stricto (Rhodomelaceae,Rhodophyta) from the Mexican Pacific,including L. mutueae sp. nov.

The alpha diversity of Laurencia sensu stricto, as well as the majority of the other algal groups along the Mexican Pacific, was underestimated because very few studies had incorporated the use of molecular markers in taxonomic studies, which would allow a more realistic estimation of diversity, incluing new records and new species. The records of Laurencia s.s. for the Eastern Pacific (California, USA to Chile) currently represent 23% of its richness worldwide. In this work, we proposed an updated census of the currently recognized species of Laurencia s.s. for the Mexican Pacific. This checklist reported the occurrence of 22 species that represent 16% of the 137 species worldly recorded. We further included taxonomic remarks, type localities, and a detailed distribution for each species. In addition, we proposed a new species for this region, Laurencia mutueae sp. nov., based on morphological observation as well as phylogenetic analyses. The distribution in the Mexican Pacific revealed that the highest records of species were for the sub‐temperate region, while the localities along the Mexican tropical Pacific were poorly recorded. The scarcity of floristic affinities between the eastern and the western and central parts of the North Pacific Ocean was hypothesized to be a consequence of a strong isolation and hard barriers, such as the water masses between these regions.     

Genetic differentiation and distribution of Pyropia acanthophora (Bangiales,Rhodophyta) in the Philippines

Pyropia acanthophora is a foliose Bangiales with widely known endemic populations in Indo-Pacific region. This alga has expanded its range recently as a consequence of introduction. In an attempt to explore the genetic diversity of Py. acanthophora within the Philippines and the impact of the introduction of the species from elsewhere, an examination of molecular differentiation and distribution was undertaken using the mitochondrial COI-5P and plastid rbcL gene sequences. The results revealed that the populations of Py. acanthophora in the Philippines exhibited high haplotypic and genetic diversities, and were found to be distinct from those previously reported as conspecific populations found in Taiwan, India, Japan, Hawaii, and from those introduced populations from Brazil. The network analyses as inferred from rbcL and from the combined COI-5P and rbcL genes showed evidence that the Philippine populations of Py. acanthophora exhibited a chaotic patchiness pattern characterized by a population with highly site-exclusive haplotypes, wide genetic variability and lack of local geographic patterns. The distribution of Py. acanthophora within the Philippines was also found to be greater than what was previously known, ranging from the extreme northwest to extreme northeast mainland Luzon coasts, including Camiguin Is., Cagayan. Understanding the genetic diversity and distribution of Py. acanthophora in the Philippines provides valuable information in relation to the conservation and effective resource management of native populations of Py. acanthophora in the tropical Asian region.     

Characterization of the edible red alga Meristotheca papulosa (Solieriaceae, Gigartinales) from Japan

The vegetative and reproductive morphology of the edible red alga Meristotheca papulosa (Montagne) J. Agardh (Solieriaceae) was reexamined based on material collected from various localities in Japan. Although the habit of the blades is variable according to the length and width of the axes, the frequency of branching and the abundance of proliferations, rbcL sequence analyses indicate their conspecificity. M. papulosa displays four distinctive reproductive features (presence of an auxiliary cell complex, occurrence of cystocarps on marginal proliferations and the blade surface (although very rare) in addition to the margins of axes, frequent production of spinose outgrowths on the pericarp and tetrasporangial initials typically basally attached to their parental cells) that have not been reported for M. papulosa from other areas. Although these features might warrant recognition of the Japanese entity as a separate species, a better understanding of their possible taxonomic value requires comparisons with M. papulosa from other geographic regions, including the type locality.     

Development of the red algal parasite Vertebrata aterrimophila sp. nov. (Rhodomelaceae,Ceramiales) from New Zealand

Parasitic red algae grow only on other red algae and have over 120 described species. Developmental studies in red algal parasites are few, although they have shown that secondary pit connections formed between parasite and host and proposed that this was an important process in successful parasitism. Furthermore, it was recorded that the transfer of parasite nuclei by these secondary pit connections led to different host cell effects. We used developmental studies to reconstruct early stages and any host cell effects of a parasite on Vertebrata aterrima. A mitochondrial marker (cox1) and morphological observations (light and fluorescence microscopy) were used to describe this new red algal parasite as Vertebrata aterrimophila sp. nov. Early developmental stages show that a parasite spore connects via secondary pit connections with a pericentral host cell after cuticle penetration. Developmental observations revealed a unique connection cell that grows into a ‘trunk-like’ structure. Host cell transformation after infection by the parasite included apparent increases in both carbohydrate concentrations and nuclear size, as well as structural changes. Analyses of molecular phylogenies and reproductive structures indicated that the closest relative of V. aterrimophila is its host, V. aterrima. Our study shows a novel developmental parasite stage (‘trunk-like’ cell) and highlights the need for further developmental studies to investigate the range of developmental patterns and host effects in parasitic red algae.