Determination of the nutritional value of proteins obtained from Ulva armoricana

The in vitro digestibility of Ulva armoricana proteins by trypsin, chymotrypsin and human intestinal juice was determined to evaluate their nutritional value. The amino acid composition of the protein fraction and its changes during a sampling period from October to February were also studied. Some differences in the protein pattern shown by SDS PAGE were found in different months, such as the presence of a 54 kDa protein in February. The protein fraction is composed mainly of aspartic and glutamic acids (24–35% of protein fraction, according to season) and the essential amino acids constitute 27–36% of the total fraction. The efficiencies of trypsin and chymotrypsin in Ulva protein digestion are comparable. Only four proteins with apparent molecular weights of 86, 68, 40, and 29 KDa are digested by these proteolytic systems. The proteins from the October sample were more sensitive to chymotrypsin than those from the February sample. For instance, two proteins with apparent molecular weights of 100 and 67 kDa were weakly digested by chymotrypsin in the February extract, were fully digested in the October sample. The February sample differed from two others in the presence of glycosylated proteins, most of which have apparent molecular weights higher than 43 KDa. With the October sample, the activity of human intestinal juice was more effective than two other proteolytic systems. This is especially evident with a 27 kDa protein, which was only partially digested by the intestinal liquid and not digested by chymotrypsin or trypsin. However, human intestinal juice in the February apparently did not attack the 27 kDa protein. These data suggest a change in protein structure making it less sensitive to human intestinal juice. The glycosylation of protein extract, which was especially marked in February, could explain the differences in behaviour of U. armoricana proteins in response to the digestive action of human enzymes. This revised version was published online in July 2006 with corrections to the Cover Date.     

Surface sensing and stress-signalling in Ulva and fouling diatoms – potential targets for antifouling: a review

Understanding the underlying signalling pathways that enable fouling algae to sense and respond to surfaces is essential in the design of environmentally friendly coatings. Both the green alga Ulva and diverse diatoms are important ecologically and economically as they are persistent biofoulers. Ulva spores exhibit rapid secretion, allowing them to adhere quickly and permanently to a ship, whilst diatoms secrete an abundance of extracellular polymeric substances (EPS), which are highly adaptable to different environmental conditions. There is evidence, now supported by molecular data, for complex calcium and nitric oxide (NO) signalling pathways in both Ulva and diatoms being involved in surface sensing and/or adhesion. Moreover, adaptation to stress has profound effects on the biofouling capability of both types of organism. Targets for future antifouling coatings based on surface sensing are discussed, with an emphasis on pursuing NO-releasing coatings as a potentially universal antifouling strategy.     

Seasonal changes of fatty acid composition and thermotropic behavior of polar lipids from marine macrophytes

Major glyco- and phospholipids as well as betaine lipid 1,2-diacylglycero-O-4'-(N,N,N-tri-methyl)-homoserine (DGTS) were isolated from five species of marine macrophytes harvested in the Sea of Japan in summer and winter at seawater temperatures of 20-23 and 3 degrees C, respectively. GC and DSC analysis of lipids revealed a common increase of ratio between n-3 and n-6 polyunsaturated fatty acids (PUFAs) of polar lipids from summer to winter despite their chemotaxonomically different fatty acid (FA) composition. Especially, high level of different n-3 PUFAs was observed in galactolipids in winter. However, the rise in FA unsaturation did not result in the lowering of peak maximum temperature of phase transition of photosynthetic lipids (glycolipids and phosphatidylglycerol (PG)) in contrast to non-photosynthetic ones [phosphatidylcholine (PC) and phosphatidylethanolamine (PE)]. Different thermotropic behavior of these lipid groups was accompanied by higher content of n-6 PUFAs from the sum of n-6 and n-3 PUFAs in PC and PE compared with glycolipids and PG in both seasons. Seasonal changes of DSC transitions and FA composition of DGTS studied for the first time were similar to PC and PE. Thermograms of all polar lipids were characterized by complex profiles and located in a wide temperature range between -130 and 80 degrees C, while the most evident phase separation occurred in PGs in both seasons. Polarizing microscopy combined with DSC has shown that the liquid crystal - isotropic melt transitions of polar lipids from marine macrophytes began from 10 to 30 degrees C mostly, which can cause the thermal sensitivity of plants to superoptimal temperatures in their environment.     

Effect of Temperature and Illumination on Growth and Reproduction of the Green Alga Ulva fenestrata

The combined effect of temperature (5, 10, 15, and 20°C) and illumination (40 and 60 mE/(m² s)) on growth and reproduction of the green marine alga Ulva fenestrata P. et R. from the sublittoral zone of Amursky Bay, Sea of Japan, was studied in the laboratory environment in the months April–July, 2000. It was demonstrated that the temperature of 5°C and illumination of 40 mE/(m² s) are the most favorable for maintaining the vegetative mass of the algae. A water temperature of 10°C and illumination of 40 mE/(m² s) are the optimum conditions for vegetative growth of U. fenestrata thalli. A temperature decrease and increase by 5°C reduces the growth rate on average by 30%. Sporo- and gametogenesis in U. fenestrata are the most regular (every 10 days) and occupy the greatest disk area at a water temperature of 15°C and illumination of 40 mE/(m² s). Vegetative growth of thalli is sharply inhibited at the stage of cell preparation to gametogenesis a day before the beginning of gamete formation.     

Asexual life history by quadriflagellate swarmers of Ulva spinulosa (Ulvales, Ulvophyceae)

This is the first report of a Ulva species reproducing asexually solely by quadriflagellate swarmers. Ulva spinulosa Okamura et Segawa specimens were collected from Ukibuchi on the Pacific coast of Kochi Prefecture, southern Japan. Quadriflagellate swarmers were released from these specimens. The swarmers showed negative phototaxis before settlement. Thalli cultured from these swarmers also released quadriflagellate swarmers in culture. Microspectrophotometric studies demonstrated equivalent DNA in nuclei of vegetative cells in thalli of U. spinulosa and in sporo‐phytes of the other Ulva species with sexual life history (U. fasciata Delile). Furthermore, the quadriflagellate swarmers of U. spinulosa had the same DNA value, demonstrating that the quadriflagellate swarmers are produced without meiosis.     

Improved isolation of glucuronan from algae and the production of glucuronic acid oligosaccharides using a glucuronan lyase

A new source for the production of bioactive glucuronic acid oligosaccharides (GlcUAOs) from the depolymerization of green seaweed Ulva lactuca glucuronan (Algal glucuronan) has been investigated. Algal glucuronan purification was optimized by the acidic precipitation method which allowed us to separate the polysaccharide mixture extracted from the cell wall of Ulva lactuca using hot water containing sodium oxalate. A series of the GlcUAOs were obtained by enzyme degradation of algal glucuronan with a glucuronan lyase (GL) isolated from Trichoderma strain. The putative bioactive GlcUAOs generated were then purified by size-exclusion chromatography in gram quantity and characterized by ¹H/¹³C NMR spectroscopy and ESI-Q/TOF-mass spectrometry.     

New species of freshwater Ulva, Ulva limnetica (Ulvales, Ulvophyceae) from the Ryukyu Islands, Japan

Ulva limnetica Ichihara et Shimada, sp. nov. (Ulvales, Ulvophyceae) is described from the Ryukyu Islands, Japan, and is characterized by thalli that are: (i) branched, tubular, fragile and wrinkled; (ii) up to 80 cm in height and up to 2 cm in diameter; (iii) light to yellowish green in color; and (iv) having an asexual reproduction by means of quadriflagellate swarmers. Rhizoidal cells bear tubular extensions on the outside of the cell layer in the stipe. Ulva limnetica is distinguished from species with similar thalli by chloroplast disposition, branching pattern, number of pyrenoids per cell and gross morphology. It is also distinguished by sequences of the nuclear-encoded 18S ribosomal RNA gene, internal transcribed spacer 2 region and the plastid-encoded large subunit of ribulose-1,5-bisphosphate carboxylase/oxgenase gene ( rbc L). Ulva limnetica was clustered with other Ulva species in an early diverging lineage within the genus.     

THE HORMESIS OF THE GREEN MACROALGA ULVA FASCIATA WITH LOW‐DOSE 60COBALT GAMMA RADIATION1

Homogenous germlings of the marine macroalga Ulva fasciata D. (synonym, Ulva lactuca L.) were used to study hormesis effects in macroalgae grown under a low dose of ⁶⁰Co γ‐ray radiation. The results of this study are the first to confirm the effects of macroalgal hormesis. Here it was demonstrated that growth of U. fasciata germlings was promoted substantially under 15 Gy of ⁶⁰Co γ‐ray radiation, with an average increase of algal biomass of 47.43%. The levels of polysaccharides and lipids varied among the tested material and showed no effects from the ⁶⁰Co γ‐ray radiation. However, the amount of protein was higher in the irradiated algae than in the control; the highest protein content of the irradiated algae was 3.958% (dry weight), in contrast to 2.318% in nonirradiated samples. This technique was applied to a field algal mass culture, which decreased the harvest time from 90 to 60 d. The mass culture approach may facilitate the production of macroalgae under unstable weather conditions such as typhoons in the summer or strong waves in the winter. The mass‐cultured macroalgae could be used as a source of bioenergy through the fermentation of algal simple sugars that derived from polysaccharides to produce ethanol.     

TRUE IDENTITY OF THE EUROPEAN FRESHWATER ULVA (CHLOROPHYTA,ULVOPHYCEAE) REVEALED BY A COMBINED MOLECULAR AND MORPHOLOGICAL APPROACH1

A set of 18 freshwater and morphologically similar marine samples of Ulva were collected from inland and coastal waters throughout Europe to assess their taxonomic identity and invasive potential. An additional 11 specimens were obtained from herbaria. The material was studied using a combination of classical morphological methods and molecular techniques; the latter included sequencing of the nuclear internal transcribed spacer (ITS) region (ITS1‐5.8S‐ITS2) and the chloroplast RUBISCO LSU (rbcL) gene and comparison of the ITS2 secondary structure predictions. Based on classical methods, all the specimens could be determined as U. flexuosa Wulfen and could be further divided into three groups matching three infraspecific taxa. This pattern was generally well supported by molecular phylogenetic analyses. All sequenced samples formed a monophyletic lineage within Ulva, showing a putative synapomorphy in the ITS2 secondary structure. The individual subspecies corresponded to phylogenetic clusters within this lineage. In freshwater habitats, the dominant taxon was U. flexuosa subsp. pilifera, but subsp. paradoxa was also occasionally recorded. In marine habitats, only U. flexuosa subsp. flexuosa and subsp. paradoxa were located. These findings support the view that U. flexuosa subsp. pilifera is primarily a freshwater alga that probably dominates in Europe. As confirmed by the study of herbarium specimens, U. flexuosa should be regarded as indigenous, although it has a tendency to form blooms under certain conditions. Besides clarifying the identity of prevailing European freshwater Ulva, the study provides novel data concerning the distribution and morphological plasticity within the U. flexuosa complex.     

Ulva flexuosa (Ulvaceae,Chlorophyta) inhabiting inland aquatic ecosystems: molecular,morphological and ecological discrimination of subspecies

Species of Ulva (Ulvaceae, Chlorophyta) are relatively common macroalgae in most coastal areas. In this paper, we examine Ulva in inland aquatic ecosystems to determine relationships between the chemical parameters of the water and the Ulva taxa present. Two groups of samples of freshwater Ulva were selected from: (i) habitats with high salinity levels, and (ii) habitats with fresh or slightly saline waters. We showed that these two groups of Ulva, which occupy distinct habitats, represent two separate taxa. The first group represents Ulva flexuosa subsp. pilifera (Kützing) M.J. Wynne, which is very common and regularly encountered in fresh waters, but is equally well represented in slightly saline waters. The second is U. flexuosa subsp. paradoxa (C. Agardh) M.J. Wynne, which is less common, preferring mainly salt-wedge saline waters. We did not find both Ulva flexuosa subspecies together at any of the research sites.

There were significant morphological differences between the Ulva flexuosa subspecies. Ulva flexuosa subsp. pilifera had longer and wider thalli and wider cells than those of subsp. paradoxa. Ulva flexuosa subsp. paradoxa cells were longer and of greater circumference than those of subsp. pilifera. Analyses of the rRNA ITS region and partial rbcL gene sequences demonstrated minor sequence variation within the two subspecies. The results of this study provide detailed information on the characteristics of limnic Ulva habitats and indicate the niches where such populations are likely to occur.