TAXONOMIC REVISION OF SARGASSUM SPECIES (FUCALES,PHAEOPHYCEAE) FROM NEW CALEDONIA BASED ON MORPHOLOGICAL AND MOLECULAR ANALYSES1

Sargassum C. Agardh (1820) is a taxonomically difficult genus distributed worldwide and reported as the most species‐rich genus of the Fucales. It is especially abundant in the Pacific where decreasing species richness is reported to occur from west to east. New Caledonia has been recognized as one of the hotspots of Sargassum diversity; however, species lists available for this region are old and incomplete and have not yet been updated with regard to the latest taxonomic revisions published. This study aimed at revising Sargassum diversity in New Caledonia and to assess its geographic affinities with neighboring Pacific regions. We used combined morphological and DNA analyses on new collections and examined numerous type specimens. Although 45 taxa have been listed in the literature, most of them have been either transferred to synonymy since or misidentified, and in this study, only 12 taxa were recognized as occurring in New Caledonia. They belong to the subgenus Sargassum sect. Binderianae (Grunow) Mattio et Payri (2), sect. Ilicifoliae (J. Agardh) Mattio et Payri (2), sect. Polycystae Mattio et Payri. (1), sect. Sargassum (4), sect. Zygocarpicae (J. Agardh) Setch. (2), and subgenus Phyllotrichia (Aresh.) J. Agardh (1). New Caledonian Sargassum flora appeared as the second richest in the region after the Pacific coast of Australia, with which it has shown high similarity, and shared species with all neighboring regions. One species, S. turbinarioides Grunow, is considered as endemic to New Caledonia. The low genetic diversity detected among several polymorphic species belonging to sect. Sargassum is also discussed.     

Historical biogeography of the widespread macroalga Sargassum (Fucales,Phaeophyceae)

Sargassum is a cosmopolitan brown algal genus spanning the three ocean basins of the Atlantic, Pacific and Indian Oceans, inhabiting temperate, subtropical and tropical habitats. Sargassum has been postulated to have originated in the Oligocene epoch approximately 30 mya according to a broad phylogenetic analysis of brown macroalgae, but its diversification to become one of the most widespread and speciose macroalgal genera remains unclear. Here, we present a Bayesian molecular clock study, which analyzed data from the order Fucales of the brown algal crown radiation (BACR) group to reconstruct a time-calibrated phylogeny of the Sargassum clade. Our phylogeny included a total of 120 taxa with 99 Sargassum species sampled for three molecular markers – ITS-2, cox3 and rbcLS – calibrated with an unambiguous Sargassaceae fossil from between the lower and middle Miocene. The analysis revealed a much later origin of Sargassum than expected at about 6.7 mya, with the genus diversifying since approximately 4.3 mya. Current geographic distributions of Sargassum species were then analyzed in conjunction with the time-calibrated phylogeny using the dispersal-extinction-cladogenesis (DEC) model to estimate ancestral ranges of clades in the genus. Results strongly support origination of Sargassum in the Central Indo-Pacific (CIP) region with subsequent independent dispersal events into other marine realms. The longer history of diversification in the ancestral CIP range could explain the much greater diversity there relative to other marine areas today. Analyses of these dynamic processes, when fine-tuned to a higher spatial resolution, enable the identification of evolutionary hotspots and provide insights into long-term dispersal patterns.     

Biogeography of Plagiochila (Hepaticae): natural species groups span several floristic kingdoms

Aim This paper presents a synthesis of our recent results regarding the biogeography of Plagiochila using a molecular approach, and documents intercontinental ranges within this largest genus of the hepatics. Methods A maximum likelihood analysis of sixty‐one nrITS sequences of Plagiochila was performed and the molecular topology obtained was compared with morphological, phytochemical and geographical data. Results Our molecular data set allowed the identification of eleven Plagiochila sections, the majority of which cover at least two floristic kingdoms. Seven sections have species in Europe (sect. Arrectae, Carringtoniae, Fuscoluteae, Glaucescentes, Plagiochila, Rutilantes, Vagae). Plagiochila species from Atlantic Europe are usually close to or conspecific with neotropical taxa, whereas species widespread in Europe are closely related to Asian ones and not to those in the Neotropics. Plagiochila sect. Arrectae represents a neotropical – Atlantic European clade. The section is not closely related – as has often been suggested – to the morphologically similar sect. Zonatae from Asia and western North America. Sequence data show that the African P. integerrima and the neotropical P. subplana are members of the Asian sect. Cucullatae (sect. Ciliatae, syn. nov.), which becomes pantropical in distribution. An ITS sequence of P. boryana from Uganda confirms the Afro‐American range of the primarily neotropical sect. Hylacoetes. Similarities in sporophyte morphology between the sect. Cucullatae and sect. Hylacoetes are the result of parallel evolution. Main conclusions Our results indicate that intercontinental ranges at section and species level are common in Plagiochila. Carl's (1931) subdivision of Plagiochila into sections restricted to one floristic kingdom is outdated. Biogeographical patterns in Plagiochila are not dissimilar to those of other groups of bryophytes but elucidation of the geographical ranges of the taxa requires a molecular approach. Contrary to earlier belief, most Plagiochila species from Atlantic Europe do not have close relatives in Asia but are conspecific with or closely related to species from tropical America.     

GLOBAL SYSTEMATIC AND PHYLOGENETIC ANALYSIS OF SARGASSUM IN THE GULF OF MEXICO,CARIBBEAN AND PACIFIC BASIN

Sargassum is one of the most species‐rich genera in the brown algae with over 400 described species worldwide. The bulk of these species occurs in Pacific‐Indian ocean waters with only a small portion found on the Atlantic side of the Isthmus of Panama. Sargassum also has one of the most subdivided and complex taxonomic systems used within the algae. Systematic distinctions within the genus are further complicated by high rates of phenotypic variability in several key morphological characters. Molecular analyses in such systems should allow testing of systematic concepts while providing insights into speciation and evolutionary patterns. Global molecular phylogenetic analyses using both conserved and variable regions of the Rubisco operon (rbcL and rbcL‐IGS‐rbcS) were performed with species from the Gulf of Mexico, Caribbean, and Pacific basin. Results confirm earlier analyses based on rbcL‐IGS‐rbcS from Pacific species at the subgeneric and sectional level while providing additional insights into the systematics and phylogenetics on a global scale. For example, species east of the Isthmus of Panama form a distinct well‐resolved clade within the tropical subgenus. This result in sharp contrast to traditional systematic treatments but provides a window into the evolutionary history of this genus in the Pacific and Atlantic Ocean basins and a possible means to time speciation events.     

The genera Melyvonnea gen. nov. and Mesophyllum s.s. (Melobesioideae,Corallinales, Rhodophyta) particularly from the central Atlantic Ocean

We propose the new genus Melyvonnea to accommodate species previously included in Mesophyllum having: a) perithallial protuberances that may branch and dominate over the encrusting base, b) monoecious gametophytes with gametangial conceptacles occasionally developed in superimposition, c) spheroid carposporangial chambers (lacking a central pedestal), and d) filaments lining canals of multiporate roofs composed of 3 to 5 cells with distinctively elongate basal cells. The new genus shares with Mesophyllum the development of a predominantly coaxial hypothallium. Melyvonnea presently accommodates three species in the Central Atlantic, viz. the generitype Melyvonnea canariensis (Foslie) comb. nov. from the Canary Islands, Melyvonnea erubescens (Foslie) comb. nov. ( = Mesophyllum incertum; type locality: Bermuda) from the western Atlantic, Melyvonnea aemulans (Foslie & Howe) comb. nov. from Puerto Rico, and one Indo‐Pacific species, Melyvonnea madagascariensis (Foslie) comb. nov. We also emend Mesophyllum Lemoine to encompass Northern Hemisphere species that lack the above apomorphies of Melyvonnea and in addition develop a central pedestal in carposporangial conceptacles (via dissolution of the surrounding cells) with gonimoblasts bending down to fill the empty space. Mesophyllum sensu stricto currently includes six species in the northeast Pacific (M. aleuticum, M. conchatum, M. crassiusculum, M. lamellatum, M. megagastri, M. vancouveriense), two species in the western Atlantic (M. mesomorphum and M. syntrophicum), and three species in the northeast Atlantic and the Mediterranean Sea (M. expansum, M. lichenoides, M. philippii). Gametophytic species of each genus show a mainly disjunct distribution being restricted to the tropics–subtropics (Melyvonnea) and the temperate waters of the Northern Hemisphere (Mesophyllum s.s.). This classification is supported by a consensus of studies of all well‐known species of Mesophyllum sensu Adey (1970), and is based on a phylogenetic analysis of morphological and anatomical characters in addition to molecular evidence.     

Phylogenetic relationships within the genus Sargassum (Fucales, Phaeophyceae), inferred from ITS-2 nrDNA, with an emphasis on the taxonomic subdivision of the genus

Sequences from the ribosomal DNA internal transcribed spacer‐2 (ITS‐2) were compared among species of Sargassaceae including the genera Sargassum and Hizikia. Species of different subgenera and sections of Sargassum were used to assess the taxonomic relationships within the genus, especially the subdivisions of the subgenus Bactrophycus. Sequences were aligned in accordance with their common secondary structure. Phylogenetic trees were constructed using neighbor‐joining, maximum likelihood and maximum parsimony methods with three species of Turbinaria as outgroups. The resulting phylogenetic trees showed that the genus Sargassum is divided into three clades corresponding to the subgenera Phyllotrichia, Sargassum and Bactrophycus. This last subgenus is further divided into four distinct groups: a Spongocarpus clade, a Teretia clade, a Hizikia clade, and a Halochloa/ Repentia clade. The position of the section Phyllo‐cystae, excluded from the subgenus Bactrophycus and included within the subgenus Sargassum is once again confirmed by the present study. Current results strongly support the assignation of Hizikia fusiformis to the genus Sargassum. Based on morphological differences and a distinct position in the molecular trees, Hizikia should be recognized as a section in the subgenus Bactrophycus so that Hizikia (Okamura) Yoshida, stat. nov. is proposed. A remarkably low divergence of ITS‐2 sequences was observed for the species in the sections Repentia and Halochloa, suggesting very recent radiation of these species. The subgenus Sargassum is divided into three clades corresponding to the three known sections: Acanthocarpicae, Malacocarpicae and Zygocarpicae, previously recognized by the morphology of receptacles. The position of Sargassum duplicatum, S. carpophyllum, S.yendoi, S. piluliferum and S. patens within the subgenus Sargassum is discussed.     

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.     

Bullera anomala sp. nov. and Bullera pseudovariabilis sp. nov., two new ballistoconidium-forming yeast species from Yunnan,China

Two ballistoconidium-forming yeast strains, CH 2.504 and CH 2.509, were isolated from the wilting leaves of Parthenocissus sp. collected in Yunnan, China in 1996. These two strains were assigned to the genus Bullera Derx by the characteristics of forming rotationally symmetric ballistoconidia, containing xylose in the cell hydrolysates and having Q-10 as the major ubiquinone. Molecular phylogenetic analysis based on small subunit ribosomal RNA gene (18S rDNA) and internal transcribed spacer (ITS) region sequences revealed that CH 2.504 is closely related to B. variabilis Nakase & Suzuki, while CH 2.509 is located in a clearly separate clade among the described Bullera species and related taxa. The results indicated that CH 2.509 represents a new species, for which Bullera anomala sp. nov. is the proposed name (type strain AS 2.2094^T). Further DNA-DNA reassociation data showed that CH 2.504 also represents a distinct species, for which the name Bullera pseudovariabilis sp. nov. is given (type strain AS 2.2092^T). This revised version was published online in June 2006 with corrections to the Cover Date.     

Etheliaceae fam. nov. (Gigartinales,Rhodophyta), with a clarification of the generitype of Ethelia and the addition of six novel species from warm waters

Based upon COI‐5P, LSU rDNA, and rbcL sequence data and morphological characteristics, six new members of the noncalcified crustose genus of red algae Ethelia are described in a new family, Etheliaceae (Gigartinales), sister to the recently described Ptilocladiopsidaceae. The novel species are described from subtropical to tropical Atlantic and Indo‐Pacific Ocean basins; E. mucronata sp. nov. and E. denizotii sp. nov. from southern and northern Western Australia respectively, E. wilcei sp. nov. from the Cocos (Keeling) Islands of Australia, E. suluensis sp. nov. from the Philippines, E. umbricola sp. nov. from Bermuda and E. kraftii sp. nov. from Lord Howe Island, Australia. The generitype, Ethelia biradiata, originally reported from the Seychelles, Indian Ocean, is added to the Western Australian flora.     

Application of polymerase chain reaction (PCR) using random amplified polymorphic DNA (RAPD) primers in the molecular identification of selected Sargassum species (Phaeophyta,Fucales)

The random amplified polymorphic DNA-polymerase chain reaction (RAPD-PCR) was used for the molecular characterisation and identification of Sargassum spp. A total of 17 samples of Sargassum (Sargassaceae, Fucales) was obtained from various localities around Peninsular Malaysia and Singapore. On the basis of morphological characteristics, the samples were tentatively grouped into five species: Sargassum baccularia, S. glaucescens, S. oligocystum, S. polycystum and S. siliquosum. By RAPD-PCR, five of 31 random primers tested generated reproducible amplification products, and polymorphic loci were detected by four of them (OPA02, OPA03, OPA04, OPA13). The RAPD-PCR profiles did not correlate with the morphological grouping into five species and extensive variation was detected between different isolates of the same species. A 450 base pair fragment generated using OPA13 was detected in 12 of 17 samples of Sargassum. This fragment was also present in profiles from Turbinaria (Sargassaceae). This study suggests that RAPD-PCR is useful in discriminating individual samples of the genus Sargassum and in developing fingerprints for them.     

Research note: Amorphochlora amoebiformis gen. et comb. nov. (Chlorarachniophyceae)

A chlorarachniophycean alga, Lotharella amoebiformis, which has been classified in the genus Lotharella is placed into a new genus Amorphochlora gen. nov., based on its phylogenetic position, which has been clarified by the recently accumulated molecular phylogenetic information, and the morphological difference between the vegetative cells of the Lotharella species. Following this taxonomic treatment, a new combination Amorphochlora amoebiformis comb. nov., is proposed.     

Phenetic studies in Nuphar Sm. (Nymphaeaceae):variation in sect. Nuphar

Previous taxonomic assessments of the hydrophytic genus Nuphar Sm. (Nymphaeaceae) have inadequately considered the range of morphological variation existing in the genus. A comprehensive statistical analysis of morphological features among the recognized taxa of Nuphar is presented here, first using 10 fruit characters to delimit the separation of two infrageneric groups of taxa, and secondly using 31 characters to quantify the morphological variation found within sect. Nuphar. Results from combined phenetic analyses provided empirical support for recognition of two infrageneric groups of species in the genus (largely by fruit, stigmatic disk margin, neck, and stigmatic disk size) and for separation of taxa within sect. Nuphar (by leaf size, and fruit disk and neck size). Statistical analyses demonstrate that extensive variation in ranges of character states exists among sect. Nuphar taxa. However, a suite of qualitative and quantitative characters can be used to recognize five species and three subspecies within this predominantly Old World assemblage.     

Testing systematic concepts of Sargassum (Fucales, Phaeophyceae) using portions of the rbcLS operon

The taxonomic and phylogenetic concepts within the Sargassum C. Agardh (Sargassaceae) species complex were evaluated through molecular phylogenetic analyses using portions of the chloroplast encoded rbcLS Operon. According to more conservative sequences (rbcL), Turbinaria (Turner) J. Agardh is a close and well‐supported sister lineage to the Sargassum species complex and an appropriate external outgroup for analyses of subgenera and subsections within Sargassum. Both rbcL and more rapidly evolving rbcLS spacer sequences indicated that the East Asiatic genus Myagropsis (Mertens et Turner) Fensholt, along with Sargassum sinicola Setchell et Gardner, represent the closest lineage to Sargassum and form appropriate internal outgroups. The rbcLS spacer region supported three of four subgeneric designations by J. Agardh and sectional levels within the subgenus Sargassum. However, some aspects of Agardh's system were not supported: many of the subsectional ranks or the phyletic concepts; Phyllotrichia was not monophyletic as a subgenus, and its species were also not the most ancestral of Sargassum; and subgenus Sargassum was not the most derived subgenus within the genus. This modern phylogeny suggests a deep evolutionary history for subgenus Sargassum with rapid speciation in closely related subsections and series, and a sister relationship between subgenera Arthrophycus and Bactrophycus.