Two new species of Gelidium (Rhodophyta, Gelidiales), Gelidium tenuifolium and Gelidium koshikianum, from Japan

Two new marine red algae, Gelidium tenuifolium sp. nov. and Gelidium koshikianum sp. nov. (Gelidiales, Gelidiaceae) are described from Japan. Gelidium tenuifolium with large‐sized thalli (up to 30 cm tall) is distinguished from other species with such thalIi by the production of wide, flattened and thin branches (up to 2 mm wide and 60–80 urn thick), the presence of an apical depression and simple determinate branches. Gelidium koshikianum with middle‐sized thalli (5–8 cm tall) is distinguished from other species with such thalli by having wide axes (up to 2.5 mm wide) and short (2.0–3.2 mm), unbranched, second‐ and third‐order branches issuing at short intervals (0.8–1.4 mm). In phylogenetic analyses of rbcL sequences, four Gelidium species that are chiefly distributed in Japan including G. tenuifolium were clustered together with 99% bootstrap value (Japanese Gelidium‐complex clade). Gelidium linoides Kützing came to the position of the sister group to G. tenuifolium with 99% bootstrap value. There were four substitutions (0.3% divergence) between G. linoides and G. tenuifolium sequences. Gelidium koshikianum and Gelidium allanii Chapman were clustered together with 100% bootstrap value and they came to the position of the sister group to the Japanese GeIidium‐complex clade with 83% bootstrap value. There were six substitutions (0.4% divergence) between G. koshikianum and G. allanii sequences.     

A NEW AGAROPHYTE SPECIES,GELIDIUM EUCORNEUM SP. NOV. (GELIDIALES,RHODOPHYTA), BASED ON MOLECULAR AND MORPHOLOGICAL DATA1

Gelidium is an economically and ecologically important agar‐producing genus. Although the taxonomy of Gelidium has been the focus of many published studies, there is still a need to reevaluate species‐level diversity. Herein, we describe Gelidium eucorneum sp. nov. based on specimens collected off Geojedo on the southern coast of Korea. G. eucorneum is distinguished by cartilaginous thalli with brush‐like haptera, rhizoidal filaments concentrated in the medulla, and globose cytocarps that are horned with multiple determinate branchlets. The species occurs in wave‐exposed intertidal sites, sometimes in association with other mat‐forming algae. Phylogenetic analyses (rbcL, psaA, and cox1) reveal that G. eucorneum is unique and clearly distinct from other species of the genus. The clade containing Gelidium vagum and Acanthopeltis longiramulosa was resolved as a sister group to G. eucorneum. We suggest that the diverse morphologies of G. eucorneum, G. vagum, and Acanthopeltis developed from a common ancestor in East Asian waters. This study shows that even in well‐studied areas, more agarophyte species are to be added to the world inventory of red algae.     

NUCLEOTIDE SEQUENCES OF SMALL-SUBUNIT AND INTERNAL TRANSCRIBED SPACER REGIONS OF NUCLEAR rRNA GENES SUPPORT THE AUTONOMY OF SOME GENERA OF THE GELIDIALES (RHODOPHYTA)

The lack of homogeneity in all previously proposed, distinguishing characteristics has left the relationships of taxa within the Gelidiales as one of the most enduring taxonomic uncertainties in the Rhodophyta. Although a precise knowledge of the taxonomy of commercially harvested members of the Gelidiales would assist resource management, agronomic practices, and marketing, even the distinction between two major groups, Gelidium and Pterocladia, has long remained controversial. In this study, the 18S ribosomal RNA (rRNA), internal transcribed spacer 1 (ITS1), 5.8S rRNA, and ITS2 regions of Gelidium latifolium (Greville) Bornet et Thuret, G. sesquipedale (Clemente) Thuret in Bornet et Thuret, G. vagum Okamura, Pterocladia lucida (Brown ex Turner) J. Agardh, and a recent segregate from Pterocladia, Pterocladiella capillacea (Gmelin) Santelices et Hommersand, were sequenced and analyzed. The ITS1, 5.8S rRNA, and ITS2 regions of G. arbuscula (Montagne) B=orgesen, G. canariensis (Grunow) Seoane-Camba, G. capense (Gmelin) Silva, and G. pristoides (Turner) Kützing were also sequenced. Phylogenetic analyses based on the 18S rRNA genes from four Gelidium species, Pterocladia lucida, and Pterocladiella capillacea , compared with 18S rRNA genes from several other red algae confirmed the division between Gelidium and the Pterocladia/Pterocladiella isolates and were consistent with the recently proposed segregation of Pterocladiella from Pterocladia. Analyses based on the ITS regions of seven Gelidium species, Pterocladia lucida, and Pterocladiella capillacea were completely consistent with the conclusions drawn from the 18S rRNA data. There were extensive length and sequence differences between Gelidium and Pterocladia/Pterocladiella . In addition, there were larger sequence differences between Pterocladiella and Pterocladia than exist among Gelidium isolates, in keeping with the recently proposed separation of the former two taxa.     

Agars from Chilean Gelidiaceae

Cystocarpic, tetrasporic and vegetative thalli of Gelidium chilense were extracted with water at 95 °C. The contents of 3,6-anhydro-galactose and sulfate group of the hydrocolloids correspond to those of an agar-type polysaccharide. The percentages of 6-O-methyl-galactose and of pyruvic acid ranged between 5.7–6.2% and 0.42–0.54%, respectively. The gel melting and gelation temperatures of Gelidium chilense, G. rex and G. lingulatum agars were determined. A correlation between 6-O-methylgalactose content and gelation temperatures was not observed. It was found by anion-exchange chromatography that 19.8% of tetrasporic and 4.9% of vegetative G. chilense agars are unsulfated polymers. Structural studies on the neutral fraction from tetrasporic G. chilense agar by partial hydrolysis and ¹H NMR spectroscopy have shown that it is mainly composed of agarose. Methylation analysis, oxidative hydrolysis and partial hydrolysis, followed by ¹H NMR spectroscopy, have shown that the neutral fraction of the agar from tetrasporic Gelidium rex is agarose. The results obtained in this work are compared with previously reported data on studies of agars from Chilean Gelidium species.     

Characterisation of the enigmatic,endemic red algaGelidium allanii (Gelidiales) from northern New Zealand — morphology,distribution, agar chemistry

Gelidium allanii Chapman is endemic to northern New Zealand with a highly localised distribution. This species has remained little known, having been collected from only one locality for more than 40 years. We compare this species with other New Zealand members of the Gelidiaceae, in particularGelidium caulacantheum andPterocladia capillacea, presenting new data on morphology, distribution and agar chemistry to more fully characteriseG. allanii.G. allanii possesses a very distinctive, highly pyruvated agar. New Zealand records of two Australian speciesG. australe andG. asperum are examined and it is concluded that there is no evidence for their occurrence in New Zealand.Author for correspondence     

Genotyping of a Miso and Soy Sauce Fermentation Yeast,Zygosaccharomyces rouxii,Based on Sequence Analysis of the Partial 26S Ribosomal RNA Gene and Two Internal Transcribed Spacers

We analyzed sequences of the D1D2 domain of the 26S ribosomal RNA gene (26S rDNA sequence), the internal transcribed spacer 1, the 5.8S ribosomal RNA gene, and the internal transcribed spacer 2 (the ITS sequence) from 46 strains of miso and soy sauce fermentation yeast, Zygosaccharomyces rouxii and a closely related species, Z. mellis, for typing. Based on the 26S rDNA sequence analysis, the Z. rouxii strains were of two types, and the extent of sequence divergence between them was 2.6%. Based on the ITS sequence analysis, they were divided into seven types (I–VII). Between the type strain (type I) and type VI, in particular, a 12% difference was detected. The occurrence of these nine genotypes with a divergence of more than 1% in these two sequences suggests that Z. rouxii is a species complex including novel species and hybrids. Z. mellis strains were of two types (type α and type β) based on the ITS sequence. Z. rouxii could clearly be distinguished from Z. mellis by 26S rDNA and ITS sequence analyses, but not by the 16% NaCl tolerance, when used as the sole key characteristic for differentiation between the two species.     

POLYPHYLETIC ORIGIN OF THE DICTYOSPHAERIUM MORPHOTYPE WITHIN CHLORELLACEAE (TREBOUXIOPHYCEAE)1

The green algal Dictyosphaerium morphotype is characterized by spherical or oval cells connected by gelatinized strands to microscopic colonies, which are covered by prominent mucilaginous envelopes. Combined SSU and ITS rRNA gene sequence analyses revealed that this morphotype evolved independently both in the Chlorella and Parachlorella clades of the Chlorellaceae. It was shown that strains exhibiting the morphology of the type species Dictyosphaerium ehrenbergianum Nägeli established a sister lineage to Parachlorella. The strain D. ehrenbergianum CCAP 222/1A was designated as an authentic strain for establishing the epitype of the genus Dictyosphaerium. The comparison of this strain with the authentic strain of Parachlorella beijerinckii Krienitz, E. Hegewald, Hepperle, V. Huss, T. Rohr et M. Wolf (SAG 2046) showed considerable differences in the secondary structure of the ITS region. Within the whole ITS‐1 and ITS‐2 region, 27 compensatory base changes (CBCs) were recognized. In the conserved Helix III of the ITS‐2, five CBCs/HemiCBCs were detected. This is a conclusive argument for separation of these two species. The clear definition of Dictyosphaerium is intended to be the necessary starting point of taxonomic reevaluation of Dictyosphaerium‐like algae within different evolutionary lineages of the Chlorellaceae.     

Gelidium commercial exploitation: natural resources and cultivation

The state of the Gelidium agarophyte resource was last reviewed in 1990 during the First International Workshop on Gelidium. The main objective of the present study was to gather and analyse the new information made available since then, with emphasis on the G. sesquipedale resource from the Iberian Peninsula and Morocco. Other Gelidium-based resources world-wide are also reviewed, as well as new attempts to cultivate Gelidium species in the sea. In general, since the late 1980s, G. sesquipedale yields have decreased in Portugal, and increased in Spain and Morocco. Current reported values, in dry weights, are ca. 670 t yr-1 for Portugal (average 1995–1997), 5200 t yr-1 in Spain, and 6950 t yr-1 for Morocco (average 1994–1996). Efforts to develop cultivation of G. sesquipedale in the sea in Spain have yet to progress beyond the demonstration scale. Other important Gelidium-based agarophyte resources are located in: (i) South Africa where, in addition to G. pristoides, new species are being harvested, with total agarophyte landings of 140 t yr-1 in 1996; (ii) Mexico, where G. robustum harvest averaged 750 t yr-1 in 1987–1996; (iii) and Chile where harvest of Gelidium species yielded ca. 460 t yr-1 in 1990–1992. Preliminary research on new techniques for commercial cultivation of Gelidium species is also reported from these countries. This revised version was published online in June 2006 with corrections to the Cover Date.     

Genetic structure and distribution of Gelidium elegans (Gelidiales,Rhodophyta) in Korea based on mitochondrial cox1 sequence data

Gelidium elegans Kützing is commonly found in Korea, China, and Japan, and is the economically most important agarophyte in the northwest Pacific. To assess the genetic structure of the Korean species, we analyzed 1200 base pairs of the mitochondrial cox1 gene from 272 individuals collected from 36 locations. A total of 34 haplotypes were found, most of which were unique, including 27 (79%) ‘private’ haplotypes. The nucleotide and haplotype diversities of cox1 within G. elegans were 0.711 ± 0.028 (H) and 0.00736 ± 0.00038 (π), respectively. The distribution of cox1 haplotypes, pairwise F_ST values, results of neutrality tests, AMOVA, and mismatch distribution revealed the existence of a deep genetic break between central Pacific Japan and all the other locations, corresponding to the surface seawater current patterns as well as the genetic signature of potential demographic expansion.     

Taxonomy of Chrysotoxum festivum Linnaeus, 1758 (Diptera: Syrphidae) – an integrative approach

An integrative taxonomic approach revealed two taxa within Chrysotoxum festivum (Linnaeus, 1758) (Diptera, Syrphidae), C. festivum A and C. festivum B. In addition to morphological differences, results also showed significant distinction in geometric morphometrics of wings and surstyli, and in DNA sequence data (nuclear ITS2 sequences) between C. festivum A, C. festivum B, and the closely related species C. elegans Loew, 1841. From examination of type material, the name C. tomentosum Giglio‐Tos, 1890 is proposed for C. festivum B, and the taxon is redefined. © 2013 The Linnean Society of London     

Phylogenetic Diversity of New Zealand Gelidiales as Revealed by rbcL Sequence Data

Diversity and phylogenetic relationships of New Zealand representatives of the red algal order Gelidiales have been examined using rbcL sequence data. Extensive field collections have been made from throughout the New Zealand region. Six genera have been reported previously from New Zealand (Capreolia, Gelidium, Pterocladia, Pterocladiella, Pterocladiastrum, Ptilophora). This research has revealed species with very restricted local distributions, as well as the discovery of several undescribed, cryptic taxa. The common and widespread Gelidium caulacantheum is confirmed to be more closely related to Capreolia than to other species of Gelidium. The generic concept of Capreolia, based on life history characters, will need to be modified to accommodate additional species possessing “Gelidium” life histories. A species endemic to New Zealand, Gelidium ceramoides, has been found to differ significantly from all other members of the Gelidiales and requires reclassification in another genus and order. Examination of field collections and herbarium specimens in addition to molecular sequence data have led us to conclude that specimens previously placed in the genera Ptilophora and Pterocladiastrum belong within Pterocladia lucida.     

Ulva diversity in the Yellow Sea during the large‐scale green algal blooms in 2008–2009

In the Yellow Sea of China, large‐scale green tides have broken out for three consecutive years from 2007 to 2009. As part of the efforts to localize the algal source, two cruises were conducted in the early stage and the outbreak stage of the bloom in 2009. We analyzed the morphological and genetic diversity of drifting Ulva specimens and culture‐derived isolates from seawater sampled in different localities. For phylogenetic analyses, the nuclear encoded ribosomal DNA internal transcribed spacer region (ITS nrDNA) and the plastid encoded large subunit of ribulose‐1, 5‐bisphosphate carboxylase/oxgenase gene (rbcL) were used. Our molecular and morphological data indicate that the dominant free‐floating Ulva species in 2008 and 2009 possibly belonged to a single strain of the U. linza‐procera‐prolifera (LPP) clade. The ITS sequences from bloom‐forming algal samples with dense branches were identical to those from U. linza‐like specimens without branches derived from the Yellow Sea. Microscopic individuals of the dominant Ulva strain were detected in eight stations, revealing that spore dispersal in the water helped to enlarge biomass in the water during the outbreak stage of green tide in the Yellow Sea.     

Examination of prezygotic and postzygotic isolating barriers in tropical Ulva (Ulvophyceae,Chlorophyta): evidence for ongoing speciation

Phylogenetic clades based on DNA sequences such as the chloroplast rbcL gene and the nuclear ITS region are frequently used to delimit algal species. However, these molecular markers cannot accurately delimit boundaries among some Ulva species. Although Ulva reticulata and Ulva ohnoi occasionally bloom in tropical to warm‐temperate regions and are clearly distinguishable by their reticulate or plain blade morphology, they have few or no sequence divergences in these molecular markers and form a monophyletic clade. In this study, to clarify the speciation and species delimitation in the U. reticulata‐ohnoi complex clade, reproductive relationships among several sexual strains from the Philippines and Japan including offspring that originated from the type specimen of U. ohnoi were examined by culturing and hybridization in addition to the ITS‐based analysis. As a result, both prezygotic and postzygotic reproductive isolation were revealed to occur between genetically perforated U. reticulata and imperforate U. ohnoi. They were also separated on the basis of sequence analysis of the ITS region. That strongly supports that the two taxa are independent biological species. Although no prezygotic barrier among the Philippine and Japanese strains of U. reticulata was observed, unexpectedly zoospores produced by hybrid sporophytes in some of their combinations mostly failed to develop, indicating partial formation of a postzygotic barrier despite a 0.2% divergence in the ITS sequence. These findings suggest speciation is still ongoing in U. reticulata.     

INTER‐ AND INTRASPECIFIC COMMUNITY STRUCTURE WITHIN THE DIATOM GENUS PSEUDO‐NITZSCHIA (BACILLARIOPHYCEAE)1

Pseudo‐nitzschia‐specific PCR primers (PnAll F/R) were designed to amplify a polymorphic region of the internal transcribed spacer 1 (ITS1) from at least 11 Pseudo‐nitzschia species. The primers were used to generate environmental clone libraries from Puget Sound, Washington, and Vancouver Island, British Columbia, to confirm that the primers were specific for Pseudo‐nitzschia and to determine the extent of ITS1 sequence diversity within individual species. All environmental ITS1 sequences generated with PnAll primers displayed the greatest similarity to known Pseudo‐nitzschia ITS1 sequences. The length of cloned ITS1 fragments differed among species but was conserved within a species. Intraspecific genotypes exhibited <3% sequence divergence for seven of the 10 species detected in clone libraries. Several ITS1 genotypes unique to the Pacific Northwest were identified in environmental samples, and other genotypes were more broadly distributed. The Pseudo‐nitzschia primers were also used to develop an automated ribosomal intergenic spacer analysis (ARISA) to rapidly identify Pseudo‐nitzschia species in environmental samples based on species‐specific variation in the length of the targeted ITS1 region. The ARISA peaks were then associated with the environmental clone sequences for Pseudo‐nitzschia species. Surveying the genetic composition of communities at both the inter‐ and intraspecific levels will enhance our understanding of Pseudo‐nitzschia bloom dynamics.     

Callionymus omanensis,a new species of dragonet from Oman,north‐western Indian Ocean (Teleostei: Callionymidae)

A new species of deep‐living dragonet Callionymus omanensis from Oman is described on the basis of a single male specimen collected in a trawl from 500 m depth off the coast of Oman. The new species is characterized within the subgenus Bathycallionymus by having a small branchial opening; head short (3·9 in proportion to standard length); eye large (2·4 in proportion to head length); preopercular spine with a long, upcurved main tip, with a small antrorse barb and a larger antrorse spine, and with a strong antrorse spine laterally at the preopercular‐spine base, ventral margin smooth; first dorsal fin slightly higher than second dorsal fin (male); second dorsal fin distally straight; 17 pectoral fin rays; distal end of caudal fin slightly pointed, with two median unbranched rays bearing short filaments; first dorsal fin with basal black spot reaching from first to fourth membranes, third membrane with an ocellated distal black blotch; second dorsal fin with vertical dark grey bars; distal three‐fourths of anal fin black; upper half of caudal fin with oblique dark grey bars; pelvic fin dark grey, second ray basally with a black blotch. The new species is compared with similar species. Revised keys to callionymid species of the western Indian Ocean and the Red Sea, as well as species of the subgenus Bathycallionymus, are presented.     

Molecular Characterization of a Distinct Begomovirus Infecting Euphorbia pulcherrima in China

Virus isolate G35 was obtained from Euphorbia pulcherrima showing leaf curl and vein thickening symptoms in Tianyang, Guangxi Province, China. The virus was transmitted by whiteflies to Nicotiana tabacum, Lycopersicon esculentum, Datura stramonium and E. pulcherrima. DNA‐A contains 2746 nucleotides, with two open reading frames (ORFs) in the virion‐sense DNA and four ORFs in the complementary‐sense DNA. When compared with the DNA‐A sequence of other begomoviruses, the total DNA‐A of isolate G35 was most closely related to that of Ageratum enation virus (79.9% sequence identity). However, the deduced coat protein of G35 is most like that of Pepper leaf curl virus from Bangladesh (94.9% amino acid sequence identity), and the AC1 of G35 is most like that of Cotton leaf curl Multan virus‐Okra (87.2% amino acid sequence identity). The molecular data showed that G35 is a distinct Begomovirus species, for which the name Euphorbia leaf curl virus (ELCV) is proposed.     

Characterization of Prorocentrum elegans and Prorocentrum levis (Dinophyceae) from the southeastern Bay of Biscay by morphology and molecular phylogeny

Benthic Prorocentrum species can produce toxins that adversely affect animals and human health. They are known to co‐occur with other bloom‐forming, potentially toxic, benthic dinoflagellates of the genera Ostreopsis, Coolia, and Gambierdiscus. In this study, we report on the presence of P. elegans M.Faust and P. levis M.A.Faust, Kibler, Vandersea, P.A. Tester & Litaker from the southeastern Bay of Biscay. Sampling was carried out in the Summer‐Autumn 2010–2012 along the Atlantic coast of the Iberian Peninsula, but these two species were only found in the northeastern part of the Peninsula. Strains were isolated from macroalgae collected from rocky‐shore areas bordering accessible beaches. Morphological traits of isolated strains were analyzed by LM and SEM, whereas molecular analyses were performed using the LSU and internal transcribed spacer (ITS)1‐5.8S‐ITS2 regions of the rDNA. A bioassay with Artemia fransciscana and liquid chromatography–high‐resolution mass spectrometry analyses were used to check the toxicity of the species, whose results were negative. The strains mostly corresponded to their species original morphological characterization, which is supported by the phylogenetic analyses in the case of P. levis, whereas for P. elegans, this is the first known molecular characterization. This is also the second known report of P. elegans.     

Vegetative and reproductive morphology of Herpochondria corallinae (Martens) Falkenberg and H. elegans (Okamura) Itono (Ceramiaceae,Rhodophyta) from Korea

The vegetative and reproductive morphology of two epiphytic Herpochondria species, H. corallinae (Martens) Falkenberg (the type species) and H. elegans (Okamura) Itono from Korea were investigated. They are bilaterally compressed and alternate-distichously branched, and lateral branches are produced by oblique divisions of the apical cell. Procarps are restricted to the first periaxial cell of the branches. Spermatangial mother cells produce one or two spermatangia. H. corallinae forms tetrasporangia on six periaxial cells, whereas H. elegans has them on only the two lateral periaxial cells. The prostrate habit, the production of six periaxial cells in opposite pairs, and tetrasporangial stichidia are confirmed as diagnostic characters of the genus Herpochondria.