Morphology and taxonomy of Chondria tenuissima and Chondria dasyphylla (Rhodomelaceae,Rhodophyta) from European waters

Recent collections of fertile Chondria tenuissima, the type species of the genus, and Chondria dasyphylla (Woodward) C. Ag. from European waters have clarified details of their morphology and reproduction. This has allowed more detailed comparisons with southern Australian material hitherto placed under these species and has shown that neither occurs on these coasts. Records of these species from other countries may thus be open to doubt.

Ruthenium Red-positive cell wall thickenings are present in some populations of both species. In C. tenuissima the thickenings are thin and lenticular or band-like, occurring on both the upper ends and inner and radial walls of pericentral and subcortical cells. In C. dasyphylla the thickenings occur as band-like caps on upper ends of these cells. Older pericentral cells may also develop additional, separate thickenings on the inner and radial walls, and in cells near the base of the plant these become lobed.

The production of an auxiliary cell after fertilization of the procarp has been observed in both species. However its purported absence in material of C. tenuissima examined by Phillips (1896, p. 19) is not discounted as this situation has been observed in a number of southern Australian species of Chondria in which the division of the supporting cell is delayed until after the diploid nucleus has been transferred from the carpogonium. This variation appears to be more common in the Ceramiales than previously realized, however it does not appear to be sufficient to invalidate Kylin's differentiation of the order.     

Resolving the taxonomy of the Polysiphonia scopulorum complex and the Bryocladia lineage (Rhodomelaceae,Rhodophyta)

Cryptic diversity is common among marine macroalgae, with molecular tools leading to the discovery of many new species. To assign names to these morphologically similar species, the type and synonyms have to be examined, and if appropriate, new species must be described. The turf-forming red alga Polysiphonia scopulorum was originally described from Rottnest Island, Australia, and subsequently widely reported in tropical and temperate coasts based on morphological identifications. A recent study of molecular species delineation revealed a complex of 12 species in Australia, South Africa, and Europe. These species are placed in a taxonomically unresolved lineage of the tribe Polysiphonieae. The aim of this study was to resolve the genus- and species-level taxonomy of this complex and related species using molecular and morphological information. Three morphologically indistinguishable species of the complex were found at the type locality of P. scopulorum, preventing a straightforward assignment of the name to any of the molecular lineages. Therefore, we propose a molecularly characterized epitype. Polysiphonia caespitosa is reinstated for the only species found in its type locality in South Africa. We describe seven new species. Only one species of the complex can be morphologically recognized, with the other eight species indistinguishable based on morphometric analysis. The studied complex, together with another seven species currently placed in Polysiphonia and two Bryocladia species, formed a clade distinct from Polysiphonia sensu stricto. Based on observations of Bryocladia cervicornis (the generitype), we describe our seven new species in the genus Bryocladia and transfer another nine species from Polysiphonia to Bryocladia.     

Taxonomic notes on Laurencia brongniartii (Rhodomelaceae, Rhodophyta)

The red alga Laurencia brongniartii J. Agardh (Rhodomelaceae, Ceramiales) is characterized by: (i) the production of four periaxial cells from each vegetative axial segment; (ii) the presence of two or three corps en cerise per superficial cortical cell and one per trichoblast cell; (iii) the production of a single tetrasporangium-bearing periaxial (fourth) cell per fertile segment; (iv) a tetrasporangial arrangement that is intermediate between perpendicular and parallel types; (v) procarps produced from the last-formed (fifth) periaxial cell of the terminal segment of a two-celled female trichoblast; and (vi) distally positioned spermatangial nuclei, in addition to known features. The production of a single tet-rasporangium-bearing periaxial cell per fertile segment allies this species to Laurencia similis Nam et Saito.     

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.     

Molecular relationships within Bostrychia tenuissima (Rhodomelaceae, Rhodophyta)

Bostrychia tenuissima King and Puttock is restricted to southern Australia and New Zealand. Previous studies in Australia have revealed two distinct patterns in the presence of osmotically active polyols. Southern populations only have D-sorbitol whereas northern populations have both D-sorbitol and D-dulcitol. These polyol patterns lead to speculation on the ecotypic differentiation of these two population types. Using single-stranded confirmation polymorphism to rapidly score plastid haplotypes, plus DNA sequencing, a 100% congruence was found between polyol patterns and plastid haplotype. Analysis of 33 B. tenuissima isolates shows that the plastid relationships closely follow biogeo-graphic regions, with south-eastern Australia (southern NSW and Victoria) and South Australia and Tasmania having one haplotype and central and northern NSW having two alternate haplotypes. There is an overlap at the haplotype boundary just south of Sydney, New South Wales.     

Biogeography of sexual and asexual populations in Bostrychia moritziana (Rhodomelaceae, Rhodophyta)

Bostrychia moritziana (Sonder ex Kützing) J. Agardh is recorded from many regions around the world. Our laboratory culture investigations have verified a sexual life cycle in isolates from Australia, Venezuela, Colombia, South Africa, Fiji, New Zealand and Indonesia. By contrast, asexual isolates producing successive generations of tetrasporophytes in laboratory culture and, presumably, in the field, are known from Australia. New Caledonia and Japan. In Australia, asexual reproduction is absent only in Victoria. In Western Australia, Northern Territory and Queensland, 99% of the isolates have asexual reproduction. In New South Wales (NSW), asexual and sexual populations are often intermixed. Of the 176 worldwide field collections, 58% were vegetative, 39% were tetrasporic, 2% were female and 1% were male. After several years of observations on the asexual isolates in culture, at least 30 successive asexual tetrasporophytic generations have developed. Only two asexual isolates (3558 and 3575) from NSW have formed a single male and female gametophyte in culture. In a self-cross of 3568, the carpospores developed into tetrasporophytes that recycled asexually. All outcrosses done with normal sexual isolates produced normal carposporophytes and the carpospores developed into tetrasporophytes that also recycled asexually. Asexual populations may arise repeatedly by loss of meiosis in tetrasporangia of sexual populations. Asexual reproduction apparently does not diminish the overall dispersal and abundance in the field. Our present bio-geographic data show that sexually reproducing popuiations of B. moritziana occur worldwide, while asexuaily reproducing populations are confined to the western Pacific. Bostrychia bispora West et Zuccarello, initially described on the basis of its asexual reproduction to distinguish it from B, moritziana, is now reduced to synonymy with B. moritziana.     

Observations on Bostrychia radicosa comb. nov. (Rhodomelaceae, Rhodophyta)

Only two genera in the Rhodomelaceae share the morphological character of transverse division of periaxial cells into two or more tier cells in which the pit connection is retained between the lower cell and the axial cell: Bostrychia and Rhodolachne. One species, Rhodolachne radicosa Itono, has been reported from mangroves, a common habitat for Bostrychia. Many collections of an entity similar to Rhodolachne radicosa have been made from localities around the Indo‐Pacific. Culture observations show a Polysiphonia‐type sexual life history in Malaysia and New Caledonia isolates that produce self‐compatible bisexual gametophytes. The New Caledonia isolate also has unisexual gametophytes. An isolate from New South Wales (Australia) reproduces asexually through successive generations of tetrasporophytes. The Thailand isolate has successive generations of mixed‐phase tetrasporophytes. The tetrasporangial stichidia also bear male spermatangial sectors, but female structures are lacking. Western Australia and Madagascar isolates do not reproduce in culture. Molecular evidence, based on sequencing of the rbcL and the large subunit ribosomal RNA genes, shows that these isolates belong to the genus Bostrychia. Low molecular weight carbohydrate analysis reveals high levels of digeneaside in all isolates. The sugar hexitol sorbitol, an osmolyte characteristic of Bostrychia, occurs in all isolates, whereas the Madagascar and New Caledonia isolates have very low levels of dulcitol. Molecular, low molecular weight carbohydrate and morphological evidence show that Rhodolachne radicosa belongs within the genus Bostrychia. We transfer Rhodolachne radicosa to Bostrychia radicosa (Itono) West, Zuccarello and Hommersand.     

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.     

Transfer of Pterosiphonia pumila Yendo to the genus Symphyocladia (Rhodomelaceae, Rhodophyta)

The red alga Pterosiphonia pumila Yendo (Rhodomelaceae, Ceramiales) has flattened thalli in which there is complete lateral fusion between branches and their parental axes during early stages of development, the laterals only becoming distally free in reproduc-tively mature plants. This results in a leaf-like organization distinct from that displayed by all other species of the genus Pterosiphonia, the members of which have terete to compressed axes that are congenitally fused along only a few (<4.5) proximal segments of the laterals. The organization of P. pumila is, thus, similar to that of species of the genus Symphyocladia, to which it is here transferred as Symphyoctadia pumila (Yendo) Uwai et Masuda, comb. nov. Symphyoctadia pumila is restricted to Japan and Korea and is distinguished from the other three described species of Symphyocladia by its small (<3 cm long) ecorticate thalli and complete lack of vegetative trichoblasts. Symphyoctadia pennata Okamura has a similar suite of characteristics, has been successfully cross-bred with S. pumila in laboratory culture and is, thus, placed in synonymy with S. pumila.     

Phylogenomic analysis of pseudocryptic diversity reveals the new genus Deltalsia (Rhodomelaceae,Rhodophyta)

Molecular analyses, in combination with morphological studies, provide invaluable tools for delineating red algal taxa. However, molecular datasets are incomplete and taxonomic revisions are often required once additional species or populations are sequenced. The small red alga Conferva parasitica was described from the British Isles in 1762 and then reported from other parts of Europe. Conferva parasitica was traditionally included in the genus Pterosiphonia (type species P. cloiophylla in Schmitz and Falkenberg 1897), based on its morphological characters, and later transferred to Symphyocladia and finally to Symphyocladiella using molecular data from an Iberian specimen. However, although morphological differences have been observed between specimens of Symphyocladiella parasitica from northern and southern Europe they have yet to be investigated in a phylogenetic context. In this study, we collected specimens from both regions, studied their morphology and analyzed rbcL and cox1 DNA sequences. We determined the phylogenetic position of a British specimen using a phylogenomic approach based on mitochondrial and plastid genomes. Northern and southern European populations attributed to S. parasitica represent different species. Symphyocladiella arecina sp. nov. is proposed for specimens from southern Europe, but British specimens were resolved as a distant sister lineage to the morphologically distinctive Amplisiphonia, so we propose the new genus Deltalsia for this species. Our study highlights the relevance of using materials collected close to the type localities for taxonomic reassessments, and showcases the utility of genome-based phylogenies for resolving classification issues in the red algae.     

Unique distribution of epiphytic Neosiphonia harveyi (Rhodomelaceae,Rhodophyta) along sargassacean hosts

Although epiphytism is generally heaviest on older portions of the host thallus and both variation and abundance of epiphytes decrease with proximity to meristematic apices, Neosiphonia harveyi (Bailey) Kim, Choi, Guiry and Saunders was frequently found on upper parts of sargassacean hosts. This study compared density, thallus length and fertility of epiphytic N. harveyi among different regions of Sargassum patens C. Agardh thalli to reveal a unique distribution pattern of this epiphyte. The majority of epiphytic N. harveyi was observed on the upper part of host thalli and was seldom seen near the basal part. Over a 3‐month period, mean epiphyte density was greater on host apices while thallus length and fertility were greater on subapical portions of host thalli, suggesting the colonization by this epiphyte near the apical part of the host. Distribution patterns of this epiphyte were similar among S. patens growing at different depths. Apical portions of S. patens appear to be more suitable substrata for N. harveyi settlement and colonization compared with other portions of the host thallus, regardless of depth.     

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.     

Fertilization and the cytoskeleton in the red alga Bostrychia moritziana (Rhodomelaceae,Rhodophyta)

Time-lapse videomicroscopy was used to observe the effects of various cytoskeletal inhibitors on three important fertilization events in Bostrychia moritziana: spermatial mitosis, gamete fusion (formation of a fertilization pore) and nuclear migration along the trichogyne. The microtubule inhibitor oryzalin disrupted spermatial mitosis but had no other effect on fertilization. The actin inhibitors, jasplakinolide, cytochalasin B, latrunculin A and B and mycalolide B inhibited gamete fusion while BDM, a myosin-disrupting drug, inhibited all three major fertilization events. FL-Phallacidin was used to stain actin filaments in spermatia and trichogynes while microtubules were labelled with antibodies at appropriate stages of fertilization. Microtubules were only evident during spermatial nuclear division. Actin filaments were present in both trichogynes and spermatia throughout fertilization; they formed a discrete ring around the fertilization pore and ensheathed male nuclei as the latter migrated into and along the trichogyne. These results suggest that the actin/myosin system plays a role in the events of fertilization.     

Morphology and reproduction of Bostrychia pinnata (Rhodomelaceae, Ceramiales) in laboratory culture

In unialgal culture, isolates of vegetative plants of Bostrychia pinnata often developed tetrasporangial stichidia and released viable tetraspores. Most tetra‐sporelings developed normal branching before reproduction, however, some sporelings developed procarps, and later, spermatangia on juvenille unbranched stages (< 1 mm). Most normally branched gametophytes (> 2 mm) were initially female before becoming bisexual when older and larger (> 5 mm). Unisexual male gametophytes were not seen in culture. Carposporophytes developed slowly (40–70 days) and were sometimes abortive or produced reduced numbers of carpo‐sporangia (10–15 in cultures compared with 40–50 in field material). Carpospores germinated more slowly than tetraspores and the tetrasporophytes required up to 6 months to reach reproductive maturity. The Polysi‐phonia‐type life‐history was completed in 9–12 months in the laboratory. Peripherohaptera were absent in cultured plants. The isolates from Florida, Guatemala and Peru did not reproduce sexually, but regularly underwent lower branch abscission as a means of vegetative reproduction. After several years in culture, most individual isolates became self‐incompatible. In the female, 1 or 2 procarps formed on each axial segment and most were 4‐celled with a few having 3 cells. A single branched sterile group of 3–7 cells was also present. After fertilization, the diploid nucleus in the carpogonium divided twice, isolating the capping element and trichogyne and establishing the connecting element adjacent to the auxiliary cell. In mature normal cystocarps the sterile group persisted and secreted mucilage into the central cavity. The mature pericarp was four layers thick (one layer of axial filaments and three cortical cell layers). Pseudocystocarps were common, produced no carpo‐sporangia, contained elongate sterile cells, and were enclosed with a partially developed pericarp. Carpogonia in which 4 nuclei were usually seen may indicate developmental failure resulting in pseudocystocarps.     

Contrasting patterns of population structure and demographic history in cryptic species of Bostrychia intricata (Rhodomelaceae,Rhodophyta) from New Zealand

Spatial patterns of genetic diversity provide insight into the demography and history of species. Morphologically similar but genetically distinct “cryptic” species are increasingly being recognized in marine organisms through molecular analyses. Such species are, on closer inspection, often discovered to display contrasting life histories or occasionally minor morphological differences; molecular tools can thus be useful indicators of diversity. Bostrychia intricata, a marine red alga, is widely distributed throughout the Southern Hemisphere and comprises many cryptic species. We used mitochondrial cytochrome c oxidase I gene sequences to assess the genetic variation, population genetic structure, and demographic history of B. intricata in New Zealand. Our results supported the existence of three cryptic species of B. intricata (N2, N4, and N5) in New Zealand. Cryptic species N4, which was found throughout New Zealand, showed a higher genetic diversity and wider distribution than the other two species, which were only found in the North Island and northern South Island. Our analyses showed low to moderate genetic differentiation among eastern North Island populations for cryptic species N2, but high differentiation among North and South Island populations for N4, suggesting different population structure between these cryptic species. Data also indicated that N2 has recently undergone population expansion, probably since the Last Glacial Maximum (LGM), while the higher genetic diversity in N4 populations suggests persistence in situ through the LGM. The contrasting population structures and inferred demographic histories of these species highlight that life history can vary greatly even among morphologically indistinguishable taxa.