96 Recognizing cryptic species: investigations into the molecular relatedness and morphological diversity of the species of the red algal genus plocamium (plocamiales, florideophyceae)

Specimens of Codium fragile (Suringar) Hariot ssp. tomentosoides were collected from 9 sites in New England, and Long Island, New York at intervals throughout the years 1999-2003. Segments were removed from the thalli and chopped into fine fragments, mostly individual utricles and medullary filaments. Fragments were incubated in enriched seawater in dim light at 15C, 12:12 LD. Within 2–3 days, in almost all cases (more than 300) motile cells formed in many of the utricles and filaments. These were 10–15 micrometer elongated biflagellate heterokont cells. They appeared to consume the chloroplasts, and within 24 hours were reduced to colorless cells, about 5 micrometers long. These cells are unable to grow in Codium chloroplast suspensions. They appear to be always associated with Codium thalli, despite attempts to clean the thalli, and were never seen in utricles or filaments of intact plants. Their ultrastructure is under investigation and will be reported on here.     

Development of the siphonous green alga Penicillus and the Espera state*

The thallus of Penicillus is composed of two filament types: axial filaments of indeterminate growth and laterals of determinate growth. In vegetative reproduction new plants arise from horizontal rhizoids. Four stages can be distinguished in development. In the primordium stage the tip of a rhizoid swells and forms a primordium. In the germling stage ascending and descending axial filaments arise from the primordium, the former grow into a fascicle and give rise to lateral saccate branches, each of which forms an ascending and a descending arm and branches further into rhizoid-like branchlets. Together these structures constitute the foundation of the stipe. At the same time the descending axial filaments elongate and become main rhizoidal filaments with lateral rhizoidal branchlets. In the early juvenile stage the stipe is formed. The elongating ascending axial filaments form a medulla while their laterals produce a cortex. In the late juvenile stage the axial filaments form the capitulum. The Espera state of Penicillus lacks a stipe since the ascending axial filaments do not join in a fascicle. Espera has been grown in laboratory cultures from Penicillus plants collected in the Caribbean region and also been found uncommonly in nature in this area. This state may be a response to environmental stress. The flattening of thalli and their orientation perpendicular to the direction of waves are discussed. A comparison of Penicillus and Codium indicates that at least two types of development exist in multiaxial Eusiphoniidae.     

A SIMPLE METHOD FOR ISOLATING FILAMENTS AS “ALGAL SEED STOCK” FROM MONOSTROMA LATISSIMUM (CHLOROPHYTA) GERMLINGS,AND APPLICATION FOR MASS CULTIVATION1

Germlings were grown from Monostroma latissimum Wittr. reproductive cells on nylon ropes. Holdfast threads and some uniseriate filaments were observed to have penetrated the fibers of the dispersed ropes. The algal filaments were easily isolated and prepared for cultivation, in comparison to the methods of enzymatically isolated algal protoplasts. Under low light (60–100 μmol photons · m^?2 · s^?1), the algal filaments grew to form a filamentous mass. When cultivated under stronger light (300–600 μmol photons · m^?2 · s^?1), they grew to initially form tubular thalli and then, when cultivated under light intensities >700 μmol photons · m^?2 · s^?1, formed foliaceous thalli. Consequently, the filaments were homogenized into small sections and then sewed on the nylon rope for algal mass cultivation. Under high‐intensity natural light, they grew to form leafy thalli.     

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.     

Production and application of filaments of <Emphasis Type="BoldItalic">Grateloupia turuturu</Emphasis> (Halymeniaceae,Rhodophyta)

Filaments from Grateloupia turuturu were obtained through germination of spores, regeneration from fragments of discoid crusts and erect thalli. The rates of filament formation through the three ways were 5.3 ± 1.2%, 100%, and 62.3 ± 5.6%, respectively. Discoid crusts were the best materials for the production of filaments. The obtained filaments were cloned in stationary and aerated culture. The differentiations of filaments were observed. When attached to the substrata, filaments differentiated into discoid crusts from which erect thalli grew, whereas for filaments in suspension culture, some cells in the filaments differentiated into spherical structures that also formed new erect thalli. Moreover, fragments of filaments (< 100 μ m) were seeded onto nori-nets. The regenerated plantlets grew into adult thalli in field cultivation.     

LIFE HISTORY AND SYSTEMATIC POSITION OF PSEUDOCHORDA GRACILIS SP. NOV. (LAMINARIALES,PHAEOPHYCEAE)1

Pseudochorda gracilis sp. Nov. (Pseudochordacease, Laminariales) is described from the Japan Sea coast of Hokkaido/ the species is subtidal, epilithic and annual, appearing in spring and maturing in winter. Erect thalli grow solitary or in tufts on a small discoid holdfast. They are simple, cord-shaped and hollow, with inner hyphal filaments, cylindrical medullary cells and paraphyses consisting of 3–6 cells. Hair tufts are observed only in young thalli. Unilocular sporangia are sessile and narrowly ovate. In culture, P. gracilis shows a heteromorphic life history with oogamy, characteristic of the order Laminariales. Gametophytes are dioecious and dimorphic. Gametophytes mature under lower temperature conditions (usually below 10°C), and sporophytes mature under low temperature and short-day conditions (5°C, SD). The seasonal growth pattern of the species results from the photoperiod-temperature conditions controlling saprophyte maturation.     

INTER-THALLIC SEXUAL REACTIONS IN ACHLYA,A GENUS OF THE AQUATIC FUNGI

Barksdale , Alma Whiffen . (New York Botanical Garden, New York.) Inter-thallic sexual reactions in Achlya, a genus of the aquatic fungi . Amer. Jour. Bot. 47(1): 14—23. Illus. 1960.– Inter-thallic sexual reactions, involving the coöperative induction of sexual organs, have been found to occur between members of homothallic, as well as heterothallic species of the sub-genus Achlya. The induction of sexual organs, both on single thalli and on paired thalli, is temperature-dependent. Sexual organs were initiated at 15°—20°C. in 14 of 27 heterothallic strains that were usually sterile at 25°. The hermaphroditic sexual organs were like those of either A. ambisexualis or A. bisexualis in 19 strains; like those of A. prolifera in 3 strains; like those of A. inflata in 1 strain; and like those of A. americana in 1 strain. In some instances, the sexual reaction between paired thalli was complete at 20°C. but incomplete (i.e., oospores were not formed) at 25°C. The heterothallic and homothallic strains studied could be assigned to 1 of 12 (6 female and 6 male) inter-action types, on the basis of their sexual responses. These sexual types are characterized and their possible usefulness in indicating kinship among species of Achlya is assessed. The possible origin of heterothallic from homothallic forms of Achlya is discussed.     

Melanothamnus maniticola sp. nov. (Ceramiales,Rhodophyta): an epizoic species evolved for living on the West Indian Manatee

Over 35 macroalgae have been documented growing epizoically on sea turtles, and macroalgae are also known to grow on the West Indian Manatee, but the number and identity of these latter species have not been determined. Analysis of DNA sequences of 12 samples collected from different manatees captured in three areas of Florida indicated that they represented a single undescribed species within the Rhodomelaceae genus Melanothamnus. Morphological analysis revealed Melanothamnus characteristics but also a previously undescribed combination of character states. These include eight to nine, but as many as 11, pericentral cells; heavy cortication restricted to the base of thalli, and a sharp transition between the corticated and ecorticate sections of the thallus; cells surrounding the ostiole being similar in size to the outer pericarp cells immediately below, and robust rhizoids that have no terminal lobes and develop from central axial cell filaments instead of pericentral cells. The unique characteristics of the rhizoids may be evolutionary adaptations for anchoring the thalli to manatee epidermis. This species is described as M. maniticola sp. nov.     

Charophyte-rich microfacies in the Barremian of the Eastern Iberian Chain (Spain)

Non-marine carbonate microfacies based on charophytes are a useful palaeoenvironmental tool that has been poorly developed to date. In the Barremian of the Maestrat Basin (Eastern Iberian Chain), five such microfacies are described and interpreted in terms of sedimentology and palaeoecology: (1) Microfacies of clavatoracean remains are mudstones and wackestones rich in utricles and well-preserved charophyte thalli of the genus Clavatoraxis, representing in situ deposition of clavatoracean meadows. (2) Microfacies of filamentous algae and clavatoraceans are mudstones with a high proportion of calcified filaments attributed to cyanobacteria and green algae. This facies may correspond to the open lake benthos. (3) Microfacies of Intraclasts and Charophyte remains are packstones formed on the shores of freshwater lakes or brackish swamps that underwent subaerial desiccation and wave reworking. (4) Microfacies of Porocharacean remains are wackestones and packstones with abundant gyrogonite fragments of genus Porochara. They are related to the reworking of a porocharacean meadow in shallow brackish marshes. (5) Microfacies of Munieria grambasti Bystricky 1976 fragments are grainstones attributed to the reworking of Munieria-dominated meadows. In consequence to these results, a palaeoecological model based on charophyte remains is proposed as a useful tool in carbonate facies analysis.     

Morphological observation,life history and rbcL gene sequence analysis of a new species,Grateloupia variata sp. nov. (Halymeniaceae,Rhodophyta) from Qingdao,China

Grateloupia variata sp. nov. H. W. Wang (Halymeniaceae, Rhodophyta) is newly described from Qingdao, China. The results indicated that: (1) thalli are cartilaginous or gelatinous in texture; (2) thalli are constricted at the base with frequent proliferations; (3) the end portion of branches are irregular, wide, flat or ungulate, split; (4) the cortex is 50–80?μm thick, consisting of four to six layers of cells; the medulla is 10–15?μm long and 1–3?μm wide, consisting of densely and irregularly intertwined filaments; (5) six-celled carpogonial ampullae branched and five-celled auxiliary cell ampullae branched; they are of the typical Grateloupia-type; (6) tetraspores and carpospores grow directly into discoid crusts through initial cell division and are of the immediate discal type; (7) Grateloupia variata sp. nov. shows a typical triphasic life history with homotypic gametophytes, carposporophytes and tetrasporophytes, and a typical homotypic alternation of generations; and (8) the rbcL sequences show no pairwise divergence and the species form a small single monophyletic subclade within the Grateloupia clade. Morphological observations, life history and molecular analysis support G. variata as a new species.     

Epi-endophytic symbiosis between Laminariocolax aecidioides (Ectocarpales, Phaeophyceae) and Undaria pinnatifida (Laminariales, Phaeophyceae) growing on Argentinian coasts

The present is the first study on epi-endophytic algae on thalli of Undaria pinnatifida growing along Argentinian coasts. The main goal is to describe the nature and the morphology of this symbiosis. Individuals of Laminariocolax aecidioides were detected in both June and December 2004, growing on U. pinnatifida sporophytes. In nature, the epi-endophyte were macroscopically observed as dark zones that partially covered the hosts’ fronds. L. aecidioides vegetative thalli were irregularly branched uniseriate filaments. The life cycle is described from laboratory cultures started from Patagonian populations. Caryology revealed that the sporophytic diploid phase presented 16 chromosomes whereas the gametophytic haploid phase presented 8 chromosomes. Isolates made from thalli growing in the interior of infected hosts developed into filamentous, branched sporophytes that reproduced by both unispores and plurispores that were produced in unilocular and plurilocular sporangia, respectively. The results of this paper also allowed us to conclude that L. aecidioides uses the thalli of U. pinnatifida as a proper substrate. The penetration of endophitic filaments among the host′s cortical cells produced a lateral compression and, finally, their thalli development generated perforations in the host tissues. The effects of the epi-endophytic infection of L. aecidioides on U. pinnatifida are neither severe nor deleterious.     

High morpho‐anatomical variability in Halimeda macroloba (Bryopsidales,Chlorophyta) in Thai waters

Halimeda macroloba is a common widespread and morphologically variable species in the Indo‐Pacific Ocean. A series of morphometric data (16 morphological and 46 anatomical variables) and the chloroplast‐encoded tuf A gene were examined to explore the morphological and genetic variations within the species and to better determine species boundaries in H. macroloba in Thai waters. Moreover, the environmental conditions, which may affect morphological variations in H. macroloba, especially of Haplotype I were examined. Our results showed that H. macroloba has more morpho‐anatomical variability and broader species boundaries than previously thought in several characters: holdfast type, segment shape and size, node height with differences in pore size, the presence of two additional patterns of peripheral utricle as seen in surface view, and additional layers of utricles (up to seven layers). Additionally, two new variable characteristics have been reported for the first time, i.e. shape and margin of peripheral utricles. Water depth and light correlated with thallus morphology, larger thalli and segments with a greater number of utricle layers were associated with subtidal plants, while smaller thalli and segments were found in the intertidal or high light intensity areas. The high plasticity of this species may explain why in the Indo‐Pacific Ocean this species is common and dominant.     

Photosynthetic Reactions in the Marine Alga Codium vermilara: I. CO(2) Fixation and Hill Reaction in Isolated Chloroplasts

Chloroplasts were isolated from the marine alga Codium vermilara (Siphonales). The isolated chloroplasts were active in CO₂ fixation in the light at a rate comparable to the rates obtained by fragments of thalli. Maximal rates of CO₂ fixation by isolated chloroplasts from Codium were obtained in the presence of salt or sorbitol isoosmotic with sea water. The conditions of isolation of Codium chloroplasts are much less stringent than those required for active chloroplasts from higher plants. The isolated chloroplasts comprise a homogeneous population of the intact “class I” type, as based on microscopic observations and on their inability to reduce ferricyanide unless osmotically shocked. The intact chloroplasts are able to reduce p-benzoquinone at a high rate.     

The influence of light quality on the development of the brown algae Petalonia and Scytosiphon

Petalonia fascia (O. F. Müll.) Kuntze, P. zosterifolia (Reinke) Kuntze and Scytosiphon lomentaria (Lyngb.) J. Ag. have been cultured in white, blue and red light of equal quantum irradiance at 15°C. Hairs, knot-filaments and Ralfsia-like crusts were formed only in blue light, whereas the prostrate system of red-grown plants consisted entirely of sparingly-branched and mostly uniseriate filaments. The production of erect thalli from the prostrate system was controlled or stimulated by red light, but these erect thalli became fertile only in the presence of blue light. All three species exhibited all of these types of response, although specific differences in the degree of certain types of response were observed.     

CYTOLOGICAL AND ULTRASTRUCTURAL STUDIES ON MUSCLE DIFFERENTIATION IN THE ASCIDIAN,PEROPHORA ORIENTALIS*,**

Muscle cell differentiation in the tail of the ascidian, Perophora orientalis, from early tail-bud embryos to swimming larvae, were studied cytologically and ultrastructurally. Myogenic cells did not form multinucleated myotubes, but remained as mononucleated cells. Nucleolar component increased prior to a marked increase in cytoplasmic RNA. Cytoplasmic RNA appeared first around nucleus and later concentrated in the peripheral cytoplasm. The fine filaments measuring 20–30 Å in their thin parts and 30–45 Å in their thick parts in diameter appeared initially, forming loose networks, in the peripheral cytoplasm where ribosome clusters had been concentrated. These filaments were tightly attached by particles of various size and density. These filaments tended to be arranged in parallel as they increased in their size. They seemed to be precursors of both actin and myosin filaments of formed myofibrils. Z band precursors were found as dense patches in association with loosely arranged myofilaments and consisted of particulate and filamentous materials. The myofibrils seemed to grow further by organizing free filaments into bundles and further by aligning bundles of myofilaments at both ends.     

Cultivation of the green alga, Codium fragile (Suringar) Hariot, by artificial seed production in Korea

Codium fragile (Suringar) Hariot is an edible green alga farmed in Korea using seed stock produced from regeneration of isolated utricles and medullary filaments. Experiments were conducted to reveal the optimal conditions for nursery culture and out-growing of C. fragile. Sampling and measurement of underwater irradiance were carried out at farms cultivating C. fragile at Wando, on the southwestern coast of Korea, from October 2004 to August 2005. Growth of erect thalli and underwater irradiance were measured over a range of depths for three culture stages. During the nursery cultivation stage (Stage I), growth rate was greatest at 0.5 m depth (0.055 ± 0.032 mm day⁻¹), where the average midday irradiance over 60 days was 924 ± 32 μmol photons m⁻² s⁻¹. During the pre-main cultivation stage (Stage II), the greatest growth rate occurred at a depth of 2 m (0.113 ± 0.003 mm day⁻¹) with an average irradiance of 248 ± 116 μmol photons m⁻² s⁻¹. For the main cultivation stage (Stage III) of the alga, thalli achieved the greatest increase in biomass at 1 m depth (7.2 ± 1.0 kg fresh wt m⁻¹). These results suggest that optimal growth at each cultivation stages of C. fragile could be controlled by depth of cultivation rope.     

Temperature,Herbivory and Epibiont Acquisition as Factors Controlling the Distribution and Ecological Role of an Invasive Seaweed

The invasive canopy alga, Codium fragile ssp. tomentosoides, first observed at the Isles of Shoals in 1983, has become the dominant canopy species to 8 m throughout the islands. Codium populations are replacing themselves at most sites in what appears to be a new, climax, canopy species. However, Codium densities have declined in protected Gosport Harbor areas where it first became established. Codium has only slowly expanded its presence in adjacent nearshore subtidal habitats. Recent studies suggest a combination of factors that may be influencing the relative success of populations between habitats. The herbivorous sea slug, Placida dendritica, may be reducing populations in protected areas in spite of predators such as the green crab, Carcinus maenas, while surge may inhibit herbivore buildup in exposed habitats. Temperature instability due to localized, wind-driven upwelling may be slowing the buildup of subtidal Codium populations in nearshore sites. The combination of Codium dominance and the acquisition of increasing epibiont diversity are producing a new, potentially more complex community state than the previous kelp-dominated climax typical of the Gulf of Maine.     

Porphyra lilliputiana sp. nov. (Bangiales, Rhodophyta): A diminutive New Zealand endemic with novel reproductive biology

A new species of Porphyra, Porphyra lilliputiana, is described for the New Zealand region. This species is very small ([5] 10–20 [35] mm) and is found growing epiphytically, epilithically and epizoically on upper inter-tidal shores of moderate exposure. Field-collected material of P. lilliputiana possessed archeosporangia, endosporangia, spermatangia and zygotosporangia. In culture, archeospores vi/ere released and germinated to form thalli. Endosporangia either developed directly into thalli or released endospores which individually formed thalli. Zygotospores developed into the concho-celis phase, which formed conchosporangia. Released conchospores formed thalli. This species is distinguished by its small size, arrangement of reproductive cells, occurrence of endosporangia, dentate margin and habitat.     

Cytological damage to the red alga Griffithsia pacifica from ultraviolet radiation

Continuous exposure for 7–10 days to 60% of ambient levels (sea level at mid-day in December) of UV-A and UV-B radiation caused cytological damage to regenerating fragments of Griffithsia pacifica under laboratory conditions. There was high mortality of individual cells and entire fragments in UV treated filaments. Rhizoid initiation was slower and rhizoids grew more slowly following UV treatment. After 7 days, UV radiated thalli showed chloroplast and nuclear degeneration. In addition, filaments tended to disarticulate so that single or groups of apparently healthy cells were common in the medium. These data suggest that the subtidal habitat of G. pacifica is based in part on lack of tolerance to UV radiation, and that UV protection mechanisms are not inducible or insufficient to prevent the accumulation of damage in this species.     

Nuclear number in cells of some species of Delesseriaceae (Rhodophyta)

The number of nuclei in multinucleate blade cells of 12 species of red algae in the family Delesseriaceae (Rhodophyta) is primarily restricted to a range that is characteristic of the species; in some species larger or older cells in other parts of thalli contain more nuclei than cells in blades. The numbers were similar in laboratory-grown and field-collected specimens and in gametangial and tetrasporangial plants. Most blade cells of Anisocladella pacifica, Sorella spp., and Phycodrys profunda contain 3–5 nuclei. Cells throughout thalli of Phrix gregarium most frequently have one or two nuclei. Other species showed larger ranges. Cells within stipes, midribs, or basal regions of blades of Anisocladella pacifica, Branchioglossum undulatum, Nienburgia andersoniana, Nitophyllum hollenbergii, Platysiphonia clevelandii, and two species of Cryptopleura can have 20+ nuclei, more than in distal blade cells. Chromosome numbers of n = 8–10 for Nienburgia andersoniana, n = 20–26 for Nitophyllum hollenbergii, and n = 7–8 for Phrix gregarium are reported.