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.     

Reinstatement of Grateloupia subpectinata (Rhodophyta, Halymeniaceae) based on morphology and rbcL sequences

Morphological observations and molecular analyses of the north‐western Pacific species of the red algal genus Grateloupia (Halymeniaceae) indicate the presence of an entity, which is somewhat similar in gross morphology to G. asiatica Kawaguchi et Wang but is distinguished from the latter species by some morphological features. These include: (i) a somewhat fleshy texture; (ii) wider and much thicker (4.5–10 mm wide and up to 1300 μm thick) axes, of which an inner cortex consists of more (6–9) cells; (iii) generally longer (up to 17 cm), marginal and surface proliferations that are clearly constricted (terete) at bases; and (iv) much elongated, oblong auxiliary cells. Phylogenetic analysis using the ribulose‐l,5‐bisphosphate carboxylase/ oxygenase (rbcL) gene of G. asiatica and the alga in question shows them to be distantly related and strongly supports the differentiation of these two entities at the species level. Judging from the literature, this entity is actually Grateloupia subpectinata Holmes, which has been placed into synonymy under G. asiatica [as G. filicina (Lamouroux) C. Agardh] or G. prolongata J. Agardh in previous reports, and therefore the Holmes name is reinstated.     

Grateloupia turuturu (Halymeniaceae,Rhodophyta) is the correct name of the non-native species in the Atlantic known as Grateloupia doryphora

Grateloupia doryphora (Montagne) Howe, originally described from Peru, has repeatedly been reported as an invasive species in Atlantic and Mediterranean waters. Various attempts to explain this species' route of introduction have been unsatisfactory. New evidence from comparative rbcL sequence analysis and morphology suggests that this adventive species in the NE and NW Atlantic corresponds with G. turuturu Yamada, originally described from Japan. This provenance follows a well-recognized trend of invasive marine organisms that have colonized the Atlantic Ocean and Mediterranean Sea from Pacific NE Asia.     

A COMPARATIVE STUDY OF THE RED ALGA GRATELOUPIA FILICINA (HALYMENIACEAE) FROM THE NORTHWESTERN PACIFIC AND MEDITERRANEAN WITH THE DESCRIPTION OF GRATELOUPIA ASIATICA, SP. NOV.

Morphological observations and molecular analyses of the red alga Grateloupia filicina (Halymeniaceae) from two geographically distant regions, eastern Asia (Japan and northern China) in the northwestern Pacific and Italy in the Mediterranean, reveal the presence of two distinct entities. Morphologically, the eastern Asian entity differs substantially from the Italian entity in the following ways: 1) thin and soft thalli with wider axes, 2) denser medullary filaments, 3) scattered reproductive structures over the entire thallus, and 4) a mature auxiliary cell that is oval and slightly larger than other ampullary cells. Phylogenetic analysis based on the ribulose-1,5-bisphosphate carboxylase/oxygenase gene ( rbc L) sequences revealed that the eastern Asian and Italian entities are phylogenetically far apart, strongly supporting the differentiation of these two entities at the species level. The eastern Asian entity is therefore described as a new species, Grateloupia asiatica. This species can be distinguished from most known species of Grateloupia that have widely flattened thalli by its compressed to narrowly flattened axes with numerous pinnate proliferations and from a few species with similar thalli by a particular combination of features, including a gelatinous texture, mostly simple and narrower axes, a thinner cortex, and the absence of catenate proliferations.     

Research note: First report of the Japanese species Grateloupia lanceolata (Halymeniaceae, Rhodophyta) from California, USA

The Japanese red alga Grateloupia lanceolata (Okamura) Kawaguchi was discovered in southern California at Santa Catalina Island in spring 2003 and April 2008 and in central California at the mouth of the Elkhorn Slough in Moss Landing in May, June and July of 2008. The morphology of thalli from both localities agrees with published figures. Sequences from the rbc L gene and the nuclear marker, internal transcribed spacer-1 from Californian G. lanceolata were identical to those from two specimens of G. lanceolata introduced to the Thau Lagoon, Mediterranean France and a specimen from Japan. It is likely that the import of oysters for mariculture played a role in its introduction into California.     

Taxonomic notes on the Halymeniaceae (Gigartinales,Rhodophyta) from Japan. III. Synonymization of Pachymeniopsis Yamada in Kawabata with Grateloupia C. Agardh*

The taxonomic status of the red algal genus Pachymeniopsis (Halymeniaceae) was evaluated based on a morphological study of the type species Pachymeniopsis lanceolata (Okamura) Yamada in Kawabata (=Aeodes lanceolata Okamura). Features used to separate Pachymeniopsis from allied genera such as Aeodes or Grateloupia were found to be inconsistently expressed within even single populations, and P. lanceolata is considered to be best placed in Grateloupia. The two remaining species, P. yendoi and P. elliptica, are judged to be conspecific, with P. elliptica also best placed in Grateloupia. The genus Pachymeniopsis is, therefore, considered synonymous with Grateloupia. The transfer of P. lanceolata to Grateloupia is proposed as Grateloupia lanceolata (Okamura) Kawaguchi, comb. nov. The reinstatement of Grateloupia elliptica Holmes, which includes Pachymeniopsis yendoi as a synonym, is also proposed.     

Grateloupia Serra sp. nov. H. W. Wang & Y. Lou (Halymeniaceae,Rhodophyta): a new species previously confused with Grateloupia asiatica in China

A new Grateloupia species from Luxun park, Qingdao Province, North China, was discovered during recent investigations and named Grateloupia serra H. W. Wang &; Y. Lou sp. nov. Morphological observations showed that: (1) the thalli were purple to dark red, cartilaginous and mucilaginous in texture, 15–45?cm in height; (2) the surface of thalli was covered with numerous proliferous branchlets and proliferous branchlets that were dentate when on the main axes; (3) the thalli were 450–550?µm thick, a cortex consisted of 6–8 layers of oblong or rounded cells and a medulla covered by compact medullary filaments; (4) the carpogonial branch was 6-celled and the auxiliary-cell branch was 5-celled, they were typical Grateloupia-type; (5) the internal structure of mature tetrasporangia were cruciately divided, oblong or square in shape. The morphological differences were supported by molecular analyses based on ribulose-1, 5-bisphosphate carboxylase/oxygenase gene (rbcL) sequences. Sequences of four G. serra sp. nov. samples were embedded into the Grateloupia clade and showed no pairwise divergence.     

Cultivation of Grateloupia acuminata (Halymeniaceae,Rhodophyta) by regeneration from cut fragments of basal crusts and upright thalli

Regeneration of Grateloupia acuminata Okamura from fragments of basal crusts and young upright thalli was studied in culture. The carpospores and tetraspores develop into basal crusts and these in turn produce upright thalli. When the crusts and the young upright thalli were cut into fragments, the fragments of basal crusts regenerated into new crusts, whereas those of upright thalli formed adventitious filaments. For cultivation in the sea, the cut fragments of basal crusts were inoculated onto oyster shells and synthetic twines of Nori-net and cultured for about one month in the laboratory. The oyster shells and the twines with regenerated crusts were then transferred into the sea from September to December. Many upright thalli developed and grew well during the cold season of the year (December to March).Author for correspondence     

Observations on the fine structure of spermatozoids and vegetative cells of the green alga Golenkinia

Spermatozoids and vegetative cells of the green alga Golenkinia minutissima Iyengar et Balakrishnan have been examined by light and electron microscopy. The biflagellate spermatozoids are of a somewhat specialised type, elongated with the nucleus attached to the flagellar bases, and containing a reduced chloroplast without pyrenoid or eyespot. The flagellar apparatus and root system has been examined in detail and is compared with that found in other green algae. The flagella are of a previously unknown type; they contain only one central microtubule—possibly non-functional—but they move in an apparently normal way. Present knowledge about flagellar roots in green algae has been assembled in a table, showing that the cruciate root has now been found in 10 genera, belonging to almost as many families. Exceptions are Oedogonium, which contains a modification of this type, and the Charales, which are very different. During spermatogenesis in Golenkinia each spermatozoid is surrounded by a wall which disappears at maturity. This fact may prove to be of taxonomic value.

The spines on the vegetative cells are composed of regularly arranged longitudinal fibrils, possibly cellulose, attached to the inner part of the two-layered cell wall. The content of the vegetative cell is typically chlorococcalean.     

THE NEW GENUS YONAGUNIA KAWAGUCHI ET MASUDA (HALYMENIACEAE,RHODOPHYTA), BASED ON Y. TENUIFOLIA KAWAGUCHI ET MASUDA SP. NOV. FROM SOUTHERN JAPAN AND INCLUDING Y. FORMOSANA (OKAMURA) KAWAGUCHI ET MASUDA COMB. NOV. FROM SOUTHEAST ASIA1

Yonagunia Kawaguchi et Masuda, gen. nov. (Halymeniaceae, Rhodophyta) is proposed to accommodate a new species, Yonagunia tenuifolia Kawaguchi et Masuda and the species currently known as Prionitis formosana (Okamura) Kawaguchi et Nguyen. Based on auxiliary cell ampullar features, Yonagunia is included in the group of genera with the simplest type of ampulla (the Grateloupia type) that comprises Dermocorynus, Grateloupia, Kintokiocolax, Phyllymenia, and Zymurgia. However, Yonagunia differs from these genera in the behavior of cells in the ampullar filaments immediately after diploidization, most cells of the primary and secondary filaments simultaneously dividing to form grape‐like clusters of small globular cells that subsequently elongate and produce involucral filaments to laxly surround the maturing carposporophyte. Yonagunia is resolved by our rbcL gene sequence analyses as one of five monophyletic clades within the Halymeniaceae (an Aeodes/Pachymenia, a Polyopes, a Carpopertis/Cryptonemia/Halymenia, a Yonagunia, and a Grateloupia clade) that is positioned as sister to the Grateloupia clade. Carpogonial branch apparatuses are identified as a potential taxonomic significance on the same level as auxiliary cell ampullae.     

帚状蜈蚣藻的修订研究——基于形态观察和rbcL序列分析

通过形态观察和分子分析相结合的方法,对青岛（模式标本产地）产的帚状蜈蚣藻（Grateloupia fastigiata Li et Ding）进行了重新鉴定,结果表明:①藻体为深紫红色,高9-26 cm,基部呈圆柱状,1-2回不规则羽状分枝,皮层厚度为85-160 μm,由8-11层细胞构成;助细胞生殖枝丛为典型的Grateloupia型,果胞枝生殖枝丛主枝由6个细胞组成,辅助细胞生殖枝丛主枝由5个细胞组成（6cpb-5auxb型）,与亚洲蜈蚣藻（G.asiatica Kawaguchi et Wang）一致。②基于rbcL基因序列构建的系统树显示:本研究的10个帚状蜈蚣藻样本与青岛产的亚洲蜈蚣藻之间无碱基差异,形成独立的进化支,与大连石槽、石庙子产的亚洲蜈蚣藻的碱基差异均为1 bp（0.08%）,与日本、韩国产的亚洲蜈蚣藻的碱基差异分别为2 bp（0.16%）、3 bp（0.24%）,均属于种内差异,与Grateloupia clade I中蜈蚣藻属内其他种的碱基差异为18（1.47%）-76 bp（6.19%）,均属于种间差异。通过形态观察和rbcL基因序列分析,认为帚状蜈蚣藻与亚洲蜈蚣藻为同一种,将帚状蜈蚣藻作为亚洲蜈蚣藻的异名。     

SYSTEMATICS OF GRATELOUPIA FILICINA (HALYMENIACEAE,RHODOPHYTA), BASED ON rbcL SEQUENCE ANALYSES AND MORPHOLOGICAL EVIDENCE,INCLUDING THE REINSTATEMENT OF G. MINIMA AND THE DESCRIPTION OF G. CAPENSIS SP. NOV.1

Grateloupia filicina (C. Agardh) Lamouroux, originally described from the Mediterranean Sea, has long been considered a textbook example of a marine red alga with a cosmopolitan distribution. An rbcL‐based molecular phylogeny, encompassing samples covering the entire geographic distribution of the species, revealed a plethora of “cryptic” species, whereby the presence of genuine G. filicina is limited to the Mediterranean basin. The phylogeny revealed a strong biogeographic imprint, with specimens from temperate regions resolved in clades composed of species inhabiting the same geographic region. Presence of widely divergent morphologies in the temperate clades indicated that several lineages have converged independently to a G. filicina‐type morphology. Tropical representatives are resolved in a single clade with very uniform G. filicina‐type morphology and pairwise sequence divergences that are lower than the average divergence observed in temperate lineages. This, combined with a lack of clear geographic structure among the tropical lineages, may indicate a more recent divergence with long‐range dispersal capacities. Violations to the biogeographic signal in temperate lineages seemed to be due to either inadequate taxonomy or recent introductions. Grateloupia minima P. & H. Crouan, a taxon placed in synonymy under G. filicina, is reinstated as a separate species distributed in the northeast Atlantic Ocean. Grateloupia capensis sp. nov. is described to accommodate specimens from South Africa with a G. filicina‐type morphology, and G. filicina var. luxurians is elevated to species status. Morphological and anatomical characters were put forward that support the distinctiveness of these three distinct species.     

Taxonomic features of the red alga Meristotheca coacta (Solieriaceae, Gigartinales)

Vegetative and reproductive structures of the little‐known red alga, Meristotheca coacta Okamura (Solieriaceae, Gigartinales) were described on the basis of specimens collected from three localities in southern Japan, including an island close to the lectotype locality. Meristotheca coacta was shown to be a strongly supported monophyletic clade in our rbcL analyses. The taxonomic features of this species were reassessed. This species is characterized as follows: (i) the thalli consist of relatively thick (300–1400 μm from the uppermost to basal regions), elastic and rose‐red blades; (ii) the blades are irregularly lobed and branched into variously shaped segments with undulate and crispate margins; (iii) tetrasporangial initials are laterally attached to their parental cells and the lateral pit‐connections remain in mature tetrasporangia; (iv) gametophytes are dioecious; (v) carpogonial branches are three‐celled (occasionally two‐ or four‐celled); (vi) a darkly staining auxiliary cell complex is present and recognizable prior to diploidization; and (vii) cystocarps are produced along (or near) the margins of the blades and on the marginal proliferations and lack spinous outgrowths.     

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.     

A taxonomic revision of a Hypnea charoides-valentiae complex (Rhodophyta, Gigartinales) in Japan, with a description of Hypnea flexicaulis sp. nov.

Morphology of a Hypnea charoides‐valentiae complex (Rhodophyta, Gigartinales) in Japan is investigated on the basis of field‐collected and cultured plants. We can recognize two species, Hypnea charoides Lamouroux and H. flexicaulis Yamagishi et Masuda, sp. nov., which are distinguished by: (i) conspicuous percurrent main axes growing straight in the former versus less conspicuous percurrent main axes growing flexuosely in the latter; (ii) narrow branching angles (less than 90°) in the former versus wide branching angles (up to 150°) in the latter; (iii) the presence of branches and branchlets showing abrupt adaxial bending in the former versus those showing pronounced abaxial bending, some growing into hooked branchlets in the latter; and (iv) the absence (in the former) or presence (in the latter) of antler‐like branches. Nucleotide sequences of the plastid‐encoded large subunit of the ribulose‐1,5‐bisphosphate carboxylase/oxygenase gene (rbcL) support the differentiation of these two entities: the pairwise distance between them was 83 b.p. (6.2%).     

Ochtodes searlesii sp. nov. (Gigartinales,Rhodophyta), from the Pacific tropical coast of Mexico,based on morphological and molecular evidence

Ochtodes searlesii Mendoza‐González, Mateo‐Cid et Sentíes sp. nov. is described from Michoacán, tropical Mexican Pacific, on the basis of comparative morphology and rbcL sequence analysis. It is distinguished from other Ochtodes species by its erect axes arising from an encrusting base, its small terete fronds, regularly dichotomously branched axes, and obliquely divided zonate tetrasporangia. Phylogenetic analyses showed that three Pacific Mexican samples, from Caletilla, Zapote and La Majahuita (Michoacán), were identical and formed a distinctive and well supported Clade segregated from other species of Ochtodes from Brazil, Cuba, Ecuador, Guadeloupe and Mexico. The Mexican entity is morphologically distinct from other Ochtodes species as well. On this basis we propose a new Ochtodes species.     

New red alga Meristotheca imbricata (Solieriaceae, Gigartinales) from Japan

A new red alga Meristotheca imbricata Faye et Masuda (Solieriaceae, Gigartinales) was described on the basis of specimens collected from southern Japan. Although this species might have been for a long time included in the concept of M. coacta Okamura, until recently a very poorly known alga, it is distinguished from the latter by the following features: (i) each thallus consists of one primary blade and several secondary blades, the latter arising from the apical or subapical portion of terminal segments of the primary blade; (ii) the individual blades are repeatedly dichotomously divided into linear segments of which margins are neither undulate nor crispate, and are fleshy, imbricate, frequently anastomosing and often acervate; (iii) each tetrasporangial initial is attached by a basal pit-connection to the parental cell, and the position of the pit-connection changes from basal to lateral by inward growth of the tetrasporangium; and (iv) the carpogonial branches sometimes have a one-celled sterile lateral.     

Circumscription of species in the genus Sirodotia (Batrachospermales,Rhodophyta) based on molecular and morphological data

Species level taxonomy and phylogeographical distribution patterns in the freshwater rhodophyte Sirodotia are resolved through phylogenetic inferences based on rbcL and cox2–3 sequence data. Previous studies focused on the taxonomy of specific Sirodotia species or the distributions across a limited geographical region. Our molecular phylogenies included samples attributable to five recognized Sirodotia species and include collections from Australia, Brazil, Costa Rica, Canada, Finland, Mexico, New Zealand, South Africa and the United States. Both rbcL and cox2–3 phylogenies inferred S. suecica, S. tenuissima and S. goebelii as a monophyletic group with little sequence divergence. This result supports the synonymy of S. tenuissima and S. goebelii with S. suecica (the species name with priority). Within this clade, samples collected from Australia and New Zealand formed a monophyletic group with no other discernible phylogeographical patterns within S. suecica. This result seems to be somewhat unusual in the Batrachospermales, as other species have shown greater genetic variation among geographically distant locations. As in previous studies, S. huillensis and S. delicatula were inferred as a separate species based on the rbcL phylogeny, supporting the current taxonomy. A specimen of S. aff. huillensis from South Africa, may represent a new species but further research is necessary before it can be designated as such.     

Taxonomic and molecular studies of the family Sebdeniaceae (Sebdeniales,Rhodophyta): new species of Lesleigha gen. nov. and Crassitegula from Hawaii,east Asia and Lord Howe Island

The genus Lesleigha gen. nov., based on L. hawaiiensis sp. nov. from Oahu in the Hawaiian Islands, is characterized morphologically and anatomically and strongly supported as distinct from the other genera of the family Sebdeniaceae (Sebdenia and Crassitegula) primarily on the basis of mitochondrial COI-5P and nuclear SSU and LSU ribosomal DNA sequences. Characters distinguishing Lesleigha from Sebdenia include the repent, dorsiventral habit, nemathecial tetrasporangia, and irregularly cruciate tetrasporangial division pattern. Although these features are equally displayed by the recently described Crassitegula from Bermuda, that genus appears to differ from Lesleigha anatomically by its lack of a fusion cell subtending the gonimoblast, although in the absence of carposporophytes it is virtually necessary that DNA-sequence analyses be conducted before a distinction between Lesleigha and Crassitegula can be certain. Besides Lesleigha hawaiiensis, L. howensis sp. nov. is newly described from Lord Howe Island in the southern Pacific, and the Japanese and Korean Sebdenia yamadae is transferred to Lesleigha as L. yamadae, based on habit and anatomical conformity as well as DNA sequences. In addition, a new species of Crassitegula, C. imitans, is described from Lord Howe Island. The existence of further sebdenialean taxa is documented although not formally treated, indicating that substantial additional diversity most likely still awaits discovery in this reputedly species-poor order.