95 Development of macroalgal (seaweed) taxonomic keys utilizing digital & media technology

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.     

Growth of a dissociated, filamentous stage of Codium species in laboratory culture

Unialgal cultures were established from isolated utricles for all five Codium species occurring in the British Isles and growth studies performed under different conditions of temperature (10-30 °C), photon irradiance (0-175 μmol m-2 s-1) and salinity (0-100% seawater growth medium). The species studied were C. fragile ssp. tomentosoides, C. fragile ssp. atlanticum, C. adhaerens, C. bursa, C. tomentosum and C. vermilara. All cultures developed into growth forms comprised of branched, dissociated, coenocytic filaments. All grew rapidly in culture, although the conditions for optimum growth sometimes differed for the individual entities. The development of typical Codium plants, with utricles, similar to the parental material, occurred only occasionally. The rapid growth of the dissociated, filamentous forms in laboratory culture led to the provision of sufficient quantities of plant material to enable detailed studies to be made of the lectins present in several of the species. This revised version was published online in August 2006 with corrections to the Cover Date.     

DEVELOPMENT OF PROTOPLASTS FROM HOLDFASTS AND VEGETATIVE THALLI OF MONOSTROMA LATISSIMUM (CHLOROPHYTA, MONOSTROMATACAE) FOR ALGAL SEED STOCK

The aim of this study was to isolate and cultivate the protoplasts of the green alga Monostroma latissimum Wittrock and subsequently induce them to form algal filaments to act as an algal "seed" stock. Protoplasts of the alga were isolated enzymatically with 4% cellulase Onozuka R-10 and 2% Macerozyme R-10. The highest number of protoplasts was obtained on a 50-rpm shaker with 1.2 M of sorbitol after 6 h of incubation, with a yield of 9 × 106 protoplasts·g−1 of fresh thallus (including holdfast). Protoplasts from both holdfasts and erect thalli usually began to form new cell walls within 5 h after isolation and began to divide from day 6 to day 9 in PES medium; cell clusters, filaments, and/or tubular thalli were formed from day 14 to day 18. For algae collected in March, about 60% of protoplasts isolated from vegetative thalli regenerated to form tubular thalli, and about 45% of protoplasts isolated from holdfasts regenerated to form filaments. However, for algae collected in May, about 1% of protoplasts isolated from vegetative thalli developed directly to form tubular thalli, and 59% of protoplasts regenerated to form cell clusters without the ability to differentiate, whereas protoplasts isolated from holdfasts failed to develop. Regenerated filaments were kept in an incubator for more than 3 years at 24° C under the low irradiance of 66μmol photons·m−2·s−1. After this time, they retained the ability to develop to form tubular thalli under irradiance of 166 and 300 μmol photons·m−2·s−1 at 18°–30° C. Subsequently, these tubular thalli can develop to form leafy thalli after being cultivated at high irradiance of 300 μmol photons·m−2·s−1 and at 18°–22° C. Therefore, the filaments could serve as"seed" stock for algal mass culture.     

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.     

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.     

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.     

Photosynthetic Reactions in the Marine Alga Codium vermilara: II. Structural Studies

Chloroplasts from Codium vermilara, isolated by relatively crude methods, are able to fix CO₂ at rates comparable to the rates of intact plants. Sections in thalli of Codium vermilara show that the chloroplasts are surrounded by a thin layer of cytoplasm. This surrounding layer of cytoplasm, is retained also in isolated chloroplasts, and presumably preserves the intactness of the chloroplast envelope.     

Dispersal potential of the invasive green alga Codium fragile ssp. fragile

Since its introduction in 1989, the invasive green alga Codium fragile ssp. fragile (formerly Codium fragile ssp. tomentosoides) has spread rapidly in Atlantic Canada. Although its spread has likely been facilitated by human transport, C. fragile possesses diverse modes of natural dispersal. In addition to parthenogenetically developing swarmer cells, it can propagate through the release of vegetative buds, thallus fragments, and entire, dislodged thalli. We examined the natural dispersal potential of these propagules using a combination of field and laboratory experiments. Vegetative buds were most abundant on thalli in late August, coinciding with increasing late summer wave activity. At two locations within two field sites, we examined the effects of tidal state, wave action and topography on the dispersal of fragments and intact thalli. Over a 4-hour period, retention rate was generally lower and dispersal distance of retained propagules higher: during flood than ebb tide; at the more wave-exposed locations; and for fragments than thalli. Dispersal direction corresponded with topography or flood tide currents at certain locations, but was often bi-directional or random, suggesting an added role of wave action. Over periods of weeks, the retention rate of marked detrital thalli was negatively related to the magnitude of wave activity. In the laboratory, settling rate was negatively related to propagule length, and large buds had higher critical shear erosional velocities than either small buds or fragments. Our results indicate that large propagules of C. fragile, such as intact adult thalli, have a generally low dispersal potential (meters), but may be transported longer distances (kilometers) during storms or when positively buoyant. Fragments and buds are regularly transported 10's of meters at a time at average flow velocities. However, they also become resuspended in average flows, and probably disperse longer distances over multiple resuspension events, or when positively buoyant. Once settled, the smallest propagules may be less easily resuspended or transported along the bottom as they become entrapped by small-scale topographic features or turf algae. The wide variety of propagules produced by C. fragile and the variable distances over which different propagule types may be transported give C. fragile the advantage of both short- and long-distance dispersal, and have likely played a role in the invasive success of this alga.     

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.     

Differential Location of Different Types of Intermediate-Sized Filaments in Various Tissues of the Chicken Embryo

The location of constitutive proteins of different types of intermediate-sized (about 10 mm) filaments (cytokeratin, vimentin, desmin, brain filament protein) was examined in various tissues of 11–20 day chick embryos, using specific antibodies against the isolated proteins and immunofluorescence microscopy on frozen sections and on isolated serous membrane. The tissues studied which contained epithelia were small intestine, gizzard, esophagus, crop, liver, kidney, thymus, mesenteries, and epidermis. The results show that the different intermediate filament proteins, as seen in the same organ, are characteristic of specific lines of differentiation: Cytokeratin filaments are restricted to – and specific for – epithelial cells; vimentin filaments are seen – at this stage of embryogenesis – only in mesenchymal cells, including connective tissue, endothelial and blood cells, and chondrocytes; filaments containing protein(s) related to the subunit protein prepared from gizzard 10 nm filaments (i.e., desmin) are significant only in muscle cells; and intermediate filament protein of brain, most probably neurofilament protein, is present only in nerve cells. We conclude that for most tissues the expression of filaments of cytokeratin, vimentin, desmin, and neurofilament protein is mutually exclusive, and that these protein structures provide useful markers for histochemical and cytochemical differentiation of cells of epithelial, mesenchymal, myogenic, and neurogenic differentiation.     

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.     

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.     

DIFFERENTIATION OF THE GREEN ALGA CODIUM FRAGILE

Clonal cultures of Codium fragile were established from both swimming cells and vegetative filaments. In the laboratory axis primordia differentiate from heterotrichous juveniles only when cultures are agitated on a reciprocating shaker. The shear forces created by mechanical agitation are essential both for initiation and maintenance of primordia. Contact guidance of growing coenocytic filaments indicates mutual adhesion of filaments as the basis for the differentiation process.     

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.     

Phylogeny recapitulates geography, or why New Zealand has so many species of skinks

The evolutionary history of 25 New Zealand scincid lizards in the endemic genera Oligosoma and Cyclodina was examined using 12S rRNA sequence data. Phylogenetic resolution was poor, despite there being up to 9% sequence divergence between taxa. Lack of resolution was not attributable to biases in the data, such as site saturation or differences in sites free to vary, so we infer that New Zealand skinks underwent two relatively rapid phases of divergence. The rate of substitution for the skink sequences appears to be similar to some bird and mammal groups for which times of divergence have been estimated. Using fhese calibrations diversification of Oligosoma skinks probably began at least 23 million years ago (Mya). The pattern of relationships and the timing of this diversification are interpreted as resulting from rapid allopatric speciation during the Oligocene (25–35 Mya) when New Zealand was fragmented into many low lying islands. A second major phase of speciation involving the Cyclodina seems to have occurred during the Miocene (15–24 Mya), probably as a consequence of increasing land area and habitat diversity. This pattern of skink evolution contrasts with the Oligocene 'environmental crisis' hypodiesis of Cooper & Cooper (Proc. R. Soc. Land. B. 261, 293–302), but can be attributed to differences in the ecology of different taxa. This can be tested by examination of other groups, such as land snails and geckos. The large number of lizard species in New Zealand can be considered a legacy both of past geography as well as the absence of small mammals which would have been both competitors and predators.     

In situ development of the foliicolous lichen Phyllophiale (Trichotheliaceae) from propagule germination to propagule production

The vegetative cycle of the foliicolous lichen Phyllophiale, from propagule germination to propagule production, was studied by light microscope observation of thalli colonizing plastic cover slips placed within a lowland tropical forest. Discoid propagules germinated by growth of radially arranged fungal cells and developed directly into lichen thalli. The young lichen comprised a single disc of closely branched, radiating filaments of the algal symbiont Phycopeltis, covered by a network of fungal hyphae extending onto the substrate as a prothallus. The prothallic hyphae incorporated additional Phycopeltis thalli encountered on the substrate. The phycobiont formed a single layer, with individual algal thalli clearly distinguishable within the lichen. Radial growth ceased at points of contact between adjacent phycobiont thalli. The visible shape of the crustose lichen thallus corresponded to the perimeter of the phycobiont thalli within. Propagules were initiated at points corresponding to the margins of the phycobiont thalli, by vertical reorientation of horizontal algal filaments surrounded by fungal hyphae. The lichenized alga produced intercalary gametangia. Degeneration of propagules unsuccessful in lichen establishment sometimes resulted in free growth of the phycobiont. The alga generally maintained its shape, growth pattern, and reproductive independence within the lichen, while also participating in the formation of unique symbiotic propagules.     

Seasonal variation of phlorotannin in sargassacean species from the coast of the Sea of Japan

Variations in phlorotannin concentrations among the developmental stages of brown algae have been reported; however, the phlorotannin concentration plasticity associated with fluctuations in environmental factors make it difficult to determine the essential ontogenetic variation. The phlorotannin concentrations in five perennial sargassacean species where newly sprouted branches appear in summer and become fertile the following spring were examined every month during a year; and correlation with the developmental or seasonal environmental factors was determined. Although the phlorotannin fluctuated greatly throughout the year, the fluctuation patterns were relatively similar among the five species: phlorotannin showed a peak during July and August; gradually decreased in the winter; and increased in April. Performing a multiple regression analysis, the phlorotannin concentration did not correlate with thallus size in all species; and phlorotannin amounts were significantly affected by ambient abiotic factors in some species. The phlorotannin contents in newly sprouted branches were always higher than those in the long main branches during all seasons. When the phlorotannin contents were determined monthly for S. fulvellum (Turner) C. Agardh where the thalli were cultured from embryos in outdoor tanks, the phlorotannin concentrations were 3–4% of the dry matter (DM) in the juveniles and decreased to less than 1% of the DM in thalli >7.5 cm in length. However, the phlorotannin in these cultured thalli suddenly increased to 5.3% DM after being transplanted to the inshore coast; and then the concentration gradually decreased. The data show higher phlorotannin concentrations in younger sargassacean algae thalli and fluctuation of the phlorotannin amounts with extrinsic environmental factors.     

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.     

New data on multicellular thallophytes and fragments of cellular tissues from Late Proterozoic phosphate rocks, South China

Zhung Yun & Yuan Xun-lai 1992 01 15: New data on multiccllular thallophytes and fragments of cellular tissues from Late Proterozoic phosphate rocks. South China, Lethaia . Vol. 25. pp. 1–18. Oslo. ISSN 0024–1164.
A fossil assemblage consisting of multicellular thallophytes. acritarchs. cyanophytes. bacteria and some forms of uncertain affinity were found in black phosphorites of the Late Protcrozoic Doushantuo Formation (upper Sinian) in Wengan Phosphate Mine. central Guizhou Province. South China. This paper reports new forms of multiecllular thallophytes and cellular tissue. Four new genera and seven new species are described. Cytomorphological-histomorphological study of these lossil tissues and comparison with modern rhodophyle Corallina sp. suggest that (I) most fossils described here are thalloid plants with a certain degree of tissue differentiation. Some of them may be hisiomorphologically comparable to modern rhodophytcs. but all are still of uncertain systematic position Some exhibit histomorphological features of mctazoan tissues. (2) The fossil cells appear to have undergone plasmolysis that might be due to osmosis caused by the high phosphate content in the sea water. (3) Most of the thallophytes were sessile benthos with recognizable basal and upper parts to the thallus. Forms with leafshaped or nodular thalli may have moved passively or ncuslonically. New fossil forms discovered from the local rock bed demonstrate the possible existence of a community with relatively high diversity in Wengan shallow sea. This confirms that an evolutionary radiation of mctaphylcs look place during Ediacaran time, shortly after the Nantuo glacial epoch. Multicellular tlhallophytes. mctaphytes. phosphate rocks, phosphorites. Late Proterozoic .     

Effects of irradiance and water flow on formation and growth of spongy and filamentous thalli of Codium fragile

Effects of irradiance and water flow on formation and growth of filamentous and spongy thalli of Codium fragile (Suringar) Hariot growing on vinylon threads were investigated at the laboratory culture. They showed clear differences in their irradiance and water flow requirements for their formation and growth. Spongy thalli were formed from the cultured filamentous thalli only at the high water flow velocity (10 cm s⁻¹). Number of the spongy thalli remarkably increased with increasing irradiance because those at 10, 50 and 100 μmol m⁻² s⁻¹ reached 0, 2 and 76 thalli m⁻¹, respectively, by 10 weeks of culture. In contrast, filamentous thalli were formed from the cultured spongy thalli at 0 and 3 cm s⁻¹, and difference in irradiance had no effect on their formation. Growth of the spongy thalli greatly accelerated under the combination of the high irradiance and high water velocity (200 μmol m⁻² s⁻¹ and 10 cm s⁻¹) because their relative growth rate in wet weight under the condition was two–four times higher than those at the other examined irradiances and water velocities. On the other hand, difference in water velocity had no effect on growth of the filamentous thalli under flowing water, and their growth decelerated at the high irradiance (200 μmol m⁻² s⁻¹). This demonstrates that water flow is a major factor controlling the formation of the spongy and filamentous thalli. The formation and growth of the spongy thalli surely occur under the combination of the high irradiance and fast flowing water. In contrast, the formation of the filamentous thalli occurs in the calm water, and their growth is inhibited under the high irradiance.