The effect of phosphate concentration on growth and agar content of Gelidium robustum (Gelidiaceae,Rhodophyta) in culture

Phosphate concentration of the growth medium was found to affect the growth rate and agar yield of a clone of Gelidium robustum grown in the laboratory. To study differences in growth we used phosphate concentrations from 0 to 200 µM. To determine the effect of phosphate on agar yield and its properties we used concentrations from 0 to 20 µM. Growth rates generally increased with increasing phosphate concentration, with the highest growth rate (21% d^–1) obtained at 150 µM. Agar yield as percentage of fresh weight was highest (10%) in the algae grown with low phosphate concentrations, but agar yield as percentage of dry weight was highest(43%) at 20 µM of phosphate. Gel strength increased with phosphate concentration with a maximum of 160 g m^–2 for 0.75% gels for the cultures at 20 µM. Melting and gelling temperatures of the gels were also affected by phosphate concentration of the growth medium. Starch yield was highest in algae grown in low phosphate concentrations.     

Protoplasts of Gelidium robustum (Rhodophyta)

Viable protoplasts were isolated from apices of the agarophyte Gelidium robustum (Gardn.) Hollenb. & Abb. using a combination of commercial cell-wall degrading enzymes and extracellular wall-degrading enzymes isolated from a marine bacterium. The protoplasts were approximately 8–15 µm in diameter, liberated mainly from the surface cell layers and from cells at the distal ends of medullary filaments. The bacterial enzyme alone was not sufficient to liberate significant numbers of protoplasts. Maximum yield was 9 × 10⁵ protoplasts/g tissue (wet wt.). Optimum osmolality occurred between 1750–1950 mOs kg^–1; yield and viability were severely diminished at osmolalities less than 1350 mOs kg^–1. Viability, as determined by flurorescein diacetate staining and Evans Blue exclusion 1 hr after removal from the enzyme solution, was approximately 80–95%. Roughly 80% of the cells did not show Calcofluor fluorescence, while 40% stained positively for the presence of sulfated polysaccharides. Cell wall regeneration was observed with inconsistent reproducibility, and no cell division was observed when the protoplasts were placed in culture medium.Dedicated to the memory of Professor Michael Neushul.     

Gelidium cultivation in the sea

Gelidium fronds were grown in the sea under a variety of experimental conditions: on rigid, damped and tensioned test farms of various designs, in calmer and more turbulent habitats, at various depths, with and without commercial fertilizer supply. Initially, the effectiveness of a given cultivation strategy was based on the survival and growth of the fronds, here termed bio-assay mariculture. Ambient seawater temperature, nutrient availability, hydrodynamics and other environmental parameters were measured periodically. In-the-sea irrigation of test plants with commercial fertilizers was apparently effective, at least with some farm designs, and when ambient nutrient levels were low. Under optimal conditions, achieved through experimental manipulation of farm design, specific growth rates of over 2% per day were recorded. However, considerable variation in growth rates and in plant performances was observed. It was not always possible to correlate these variations with design modifications or other experimental parameters. In view of these findings, we have reviewed our initial bio-assay approach, namely the assumptions about the design and operation of farm structures and their interactions with the water and the fronds. Methods were developed to quantify these interactions. We advocate a quantitative, hydrodynamic approach in developing an effective cultivation strategy for gelidioid algae and are optimistic about progressing from test-to-commercial scale farms in the near future.     

Spore Adhesion and Cell Wall Formation in Gelidium Floridanum (Rhodophyta, Gelidiales)

The attachment of spores to a substratum is essential for their germination and, therefore, to the completion of the life cycle of the red algae. In most red algae, spores are liberated without a cell wall, within a sheath of mucilage which is responsible for their primary attachment. Utilizing fluorescent-labeled lectins, we identified carbohydrate residues and their locations in the mucilage and cell walls of spores of Gelidium floridanum. Cell wall formation and mucilage composition were studied with calcofluor, toluidine blue (AT-O), alcian blue (AB) and periodic acid-Schiff (PAS). In the mucilage we identified α-D mannose, α-D glucose, β-D-galactose, N-acetyl-glucosamine and N-acetyl-galactosamine. The first two sugar residues were not found in the cell wall of the germ tube but they were present on the rhizoid’s cell wall indicating their importance to substrate adhesion. A cell wall is produced soon after the spore’s attachment, beginning with a polar deposition of cellulose and its gradual spread around the spore as indicated by calcofluor. The cell wall matrix was positive to AB and metachromatic to AT-O, indicating acidic polysaccharides, while cellulose microfibrills were positive to PAS. A polar disorganization of the cell wall triggers the process of germination. As spores are the natural form of propagation of Gelidium, the understanding of the mechanisms of spore attachment may contribute to the cultivation of this valuable seaweed.     

Production of plantlets of the red alga of Gelidium genus (Rhodophyta) from thallus fragments

Experimental research on production of plantlents from fragments of thallus of the alga Gelidium were carried out. The best results on development of plantlets without rhizoids were achieved with cultivation of fragments of subapical parts of the plant. Thus, the average growth rate of plantlets in the last for 4 weeks of rearing was 6.7 ± 1.2% a day. Gelidium plantlets with rhizoids were grown from the top branches of plants in an intensively stirred culture with shading of the lower parts of the branches. It is suggested to use the same method for obtaining planting material during bottom rearing of Gelidium in the sea as that for intensive culture in tanks.     

Production ecology of Gelidium

Available data on determinants of production in species of Gelidium suggest several general patterns. Species diversity is higher in tropical latitudes, whereas in temperate latitudes the size of the fronds is larger, the species are ecologically dominant and commercially viable. Typically, the species occur on rocky substrate, often on coralline crusts, associated with rapid water movement and arranged in successive belts that can extend down to 25 m depth. Yields vary among species, to a maximum of 2.0 kg m^–2 y^–1. Growth and production in many species can best be explained by complex interactions between irradiance and nutrients. Temperature can interact synergistically with irradiance, while water movement compensates for nutrient limitations. Increased water movement or the addition of nutrients can prevent, to an extent, bleaching by high light and high temperature. Available data suggest the existence of at least eight biological factors affecting predicted productivity of Gelidium crops: morphology, age of the frond, thallus part, reproductive state, seasonality, crop density, life history phase and geographic and ecological origin of the species. At least four events can remove or destroy Gelidium crops: extreme low tides, storms, grazing and careless harvesting. Only the last-named factor has been analyzed over more extensive experimental periods.     

Intrageneric differences in cystocarp structure in Gelidium and Pterocladia

Unilocular cystocarps, with ostioles opening to one frond surface, have traditionally distinguished Pterocladia from Gelidium, described as having bilocular cystocarps, with ostioles opening to both surfaces; however, unequally developed locules have been described in Pterocladia and differences in cystocarpic architecture between Pterocladia capillacea and the type species of the genus, P. lucida, have been recently found. As heterogeneity in cystocarp architecture raises questions of basic intergeneric distinction, a survey of reproductive morphology of species in both genera is presented in this study. Six morphologically-different types of cystocarps are distinguished among the five species of Pterocladia and the seven species of Gelidium examined.     

Annual variations of the biochemical composition of Gelidium latifolium (Greville) Thuret et Bornet

Agar, floridean starch, protein, ash and water content in Gelidium latifolium from nature were followed monthly over one year. Comparison of variations in these contents, algal growth and cytological observations enable us to establish a cycle for agar synthesis. In autumn, after reproduction of alga, there is an active algal growth period with agar synthesis and utilization of floridean starch. In winter, synthesis is shifted toward protein accumulation while there is a partial agar hydrolysis. In spring another active growth period of the alga occurs with accumulation of floridean starch and synthesis of agar. In summer and during reproduction, there is a depletion of thalli nitrogen content while the agar synthesis pauses.     

Distribution and Recent Reduction of Gelidium Beds in Toyama Bay, Japan

The distribution and recent reduction of Gelidium beds, i.e. mat-like beds dominated by the agarophyte G. elegans Kützing in Toyama Bay (Sea of Japan), in which 95% of the coastline is protected artificially, are reported. Gelidium beds were common in shallow waters (usually < 10 m deep); most of the large beds (> 1 ha) were restricted to the inner coasts of the bay. In calm and eutrophic areas, however, G. elegans was heavily colonized by epiphytes. In the last decade, two beds were buried in situ and beds in their vicinity were damaged by the stagnation of coastal water and/or sedimentation by silts which accompanied land reclamation. At the other two beds monitored since 1988, Gelidium declined a few times but most prominently in 1998, when episodic long summer rain was recorded. This is the first report, not only on the current status of Gelidium beds other than for the central Pacific Coast of Honshu in Japan, but also concerning reduction of the beds caused by both anthropogenic and natural events.     

Physiological responses of Pterocladia and Gelidium (Gelidiales,Rhodophyta) from the Azores,Portugal

Manometric studies were conducted on Pterocladia capillacea, Gelidium latifolium and Gelidium spinulosum from the Azores, Portugal to determine optimal values of temperature, light and salinity for growth. Physiological responses were considered in relation to vertical distribution patterns of these species commonly observed throughout the Azores. Optimal parameters for the growth of Pterocladia capillacea, Gelidium latifolium and G. spinulosum were 17 to 25 °C, a photon flux density between 200 and 300 µmol m^–2 s^–1 and salinities of 25 to 35.     

Chemical composition of agars from a newly reported Japanese agarophyte,Gracilariopsis lemaneiformis

The chemical composition of agars from Gracilariopsis lemaneiformis, newly reported from Japan, was investigated. Native agars were isolated by a sequential extraction of plants in water at 22 °C and 100 °C, and in boiling 20, 40 and 60° ethanol. Agars in each extract were analyzed by chemical methods, ¹H, ¹³C NMR; and IR spectroscopy. The highest yield of agar (total carbohydrate) was obtained from the 40° ethanol extract (55°). Highest sulfate content was attained in non-alkali treated agars extracted with hot water (4.81°, DS 0.2). The 3,6-anhydrogalactose content was highest in the 40° ethanol extract (36.1° in non-alkali treatment, 40.3° in alkali treatment). The highest methoxyl content (6.51°, DS 0.66) was obtained in the 60° ethanol extract. The G. lemaneiformis agar is composed of the biological precursor to agarobiose repeating units and agarobiose containing 6-O-methyl agarobiose and a small amount of 2-O-methyl-α-l-galactopyranose residues. Alkali treatment improved the chemical quality of the agar fractions, which was comparable with Japanese commercial agar and agarose.     

Biological criteria for the exploitation of the commercially important species of Gelidium in Spain

Exploitation of the commercially-important species of Gelidium in Spain (G. sesquipedale and G. latifolium) and development of the industry occurred after World War II, as a consequence of their use as resources for the extraction of agar. This resulted in the implementation of several harvesting methods, the most important of which is the gathering of cast seaweeds, both from the shore and the sea. From the very beginning, direct exploitation of these species (i.e. plucking) was controversial because of possible adverse ecological effects. Consequently, several biological and ecological studies of both species of Gelidium were begun. This included such aspects as growth, biomass production, productivity, reproduction, regeneration capacity and agar yield. Recently, the growing interest in using and conserving this resource has led to increased knowledge of the biology and ecology of these species. In this paper, we provide an overview of the research carried out by different Spanish groups and suggest guidelines for rational management of these resources.     

Effect of climatic variation on the relative abundance of the red alga Gelidium robustum in Baja California Sur, Mexico

The red alga Gelidium robustum is important,because of its commercial exploitation in Mexico as araw material for the agar industry, providing 10% ofthe world production of agarophytes. In recent years,its annual harvest in Mexico has shownobvious,variations partly because of an increasedharvesting effort, but also because of environmentalchanges. An analysis is presented of the effect thatinterannual variability of the sea surfacetemperature, wind speed, and upwelling index had onthe relative abundance of this alga from 1980 to 1990.The results indicate a close relation betweenenvironmental fluctuations and the relative abundanceof this species. The response of G. robustum tothe different environmental conditions has not alwaysbeen equal. During El Niño 1982–84, the seasurface temperature was the most important factor andthere was a high negative correlation with therelative abundance. A lag period of three monthsshowed a positive correlation with upwelling index andwind speed. Under normal conditions and during LaNiña, the relative abundance of the alga showed apositive correlation with the sea surface temperature. For the upwelling index and wind speed, therelationship was similar to that during El Niño.     

Field and culture studies of species of Gelidium (Gelidiales,Rhodophyta) from their northern limit of distribution in Europe

Two species of Gelidium, provisionally referred to as G. pusillum and G. latifolium, are included in the Scandinavian flora and reach their northern limit of distribution on the Norwegian west coast. Small frond sizes due to adverse growth conditions, extreme phenotypic variability and lack of sexual reproduction make identification of specimens very difficult. Both species were isolated into unialgal culture and were compared with cultured strains referable to G. pusillum from Ireland and France. Temperature and salinity requirements and tolerance ranges were determined and discussed in view of distribution along the Norwegian coast. Little or no growth occurs below 9 °C, corresponding to a northern summer growth limit. On the Norwegian Skagerrak coast, low winter temperature rather than reduced salinity is the limiting factor.     

Fecundity,spore recruitment and size in Gelidium sesquipedale (Gelidiales,Rhodophyta)

Gelidium sesquipedale fecundity was quantified by counting tetrasporangial sori and cystocarps per meter squared and by estimating the number of spores contained inside them. These were obtained by regression on a size metric of reproductive structures. Tetrasporangial sori length and cystocarp thickness were the best estimators of spore number. To assess spore recruitment, 12 pottery tiles were fixed to the bottom, and the appearance of small fronds was monitored.No clear seasonal pattern of reproduction was found. Tetraspore production peaked in March 1990 with 10.4 × 10⁶ spores m^–2, whereas the carpospore peak was lower, 4.9 × 10⁵ spores m^–2 in July 1989. Recruitment followed tetraspore peaks. The probability of a G. sesquipedale tetraspore making the transition to a recruit was 4.7 × 10^–5. Frond length was significantly related to tetrasporangial sori number, while cystocarp number was only related to frond branching order. Minimum size for reproduction was 6.9 cm for gametophytes and 5.4 cm for tetrasporophytes; very rarely were cystocarpic fronds smaller than 9 cm, while tetrasporic fronds were often longer than 15 cm. Cystocarpic fronds were significantly shorter and had more branches than tetrasporic fronds.     

Worldwide distribution of commercial resources of seaweeds including Gelidium

The commercial exploitation of seaweeds for use as food and for the production of agar, alginate and carrageenan is outlined. The quantities of seaweed harvested for each purpose are tabulated and discussed. Seaweeds for food are derived chiefly from China, Japan and Korea, with almost 94% obtained by cultivation. Alginophytes are collected in 15 countries but six of these account for more than 80% of the total harvest; all are from natural stocks except for a large quantity of Laminaria cultivated in China. Natural carrageenophytes, from 12 countries, now account for only 20% of the total harvest; the remainder is cultivated Eucheuma species, 99% of which is produced in only two countries, the Philippines and Indonesia. Of the four categories of commercial resources of seaweeds considered, agarophytes are spread more evenly over a greater number of countries; they come from 20 countries and only five of these are minor contributors to the total. Gelidium species are particularly important because of the high quality agar they yield; their distribution and location are discussed.     

Deviations in the life-history of Gracilaria sp. (Rhodophyta,Gigartinales), from Coquimbo,Chile, under different culture conditions

Cystocarpic branches of a species of Gracilaria from Coquimbo, Chile, were cultured in vitro. A Polysiphonia-like life history was completed in about 6 months, but some abnormalities were observed: i. carpospores gave rise to plants producing either tetrasporangia and spermatangia, or tetrasporangia only; ii. tetraspores cultured without aeration developed into plants bearing spermatangia only; tetraspores cultured with aeration developed into 1:1 female and male gametophytes; iii. plant originated from tetraspore produced spermatangia and tetrasporangia; one of these tetraspores developed into a male gametophyte; iv. some tetraspores gave rise to spherical bodies instead of the ordinary cylindrical branches; one of them bore spermatangia after three months. The results show that environmental factors seem to be interfering with the mechanism of sex determination and induce the development of spermatangia on putative female gametophytes, or on putative tetrasporophytes. Noted added in proof: The Gracilaria sp. studied here was recently described as a new species, G. chilensis by Bird C. J., McLachlan, J & Oliveira, E. C. de, 1986. Can. J. Bot. 64: 2928–2934. Noted added in proof: The Gracilaria sp. studied here was recently described as a new species, G. chilensis by Bird C. J., McLachlan, J & Oliveira, E. C. de, 1986. Can. J. Bot. 64: 2928–2934.     

Life history, aspects of reproductive biology and production of Corophium orientale (Crustacea: Amphipoda) in Monolimni lagoon (Evros Delta, North Aegean Sea)

Monthly samples of Corophium orientale were collected during February 1998–February 1999 in both parts of Monolimni lagoon (0.3 < Sal. < 6 psu, 2 < Temp. < 28.5 °C). Corophium orientaleshowed a semiannual life cycle. In the southern part, breeding occurred from early spring to mid autumn having three peaks, in early spring, early summer and early autumn, and three cohorts were produced, a spring cohort, a summer cohort and an overwintering one. In the shallower innermost northern part of Monolimni lagoon, breeding peaked in mid-spring and, also, in early autumn, and two discrete cohorts, a spring and an overwintering one, were produced. A lack of large individuals, attributed to a temporary size – selective predation by migrating shorebirds, was observed during summer. That lack possibly contributed to the hiatus in reproduction. Photoperiod seemed to be a more important cue than temperature in the initiation of the reproductive cycle, while extremely low salinities (<1 psu) did not deter breeding. Females attained maturity at a smaller size in late spring and, especially, in summer than in early spring at lower temperatures. Brood size was a function of females body size. Females, matured during summer at high temperatures, showed the lowest brood size, body length ratio. Mean brood size was small (12–13 early embryos) and embryo loss during development high (54%), possibly due to a negative effect of low salinities. A 1:1 sex ratio existed in the small body length classes, but females preponderated in the large ones. The spring cohort in the southern part, which developed at moderate temperatures, showed the highest growth rate (40 m day^–1). Secondary production of C. orientalecalculated by Hyness method gave a mean annual density of 4562.5 and 9327.6 ind. m^–2, a mean annual crop (B) of 1.03 and 2.67 g DW m^–2, an annual production (P) of 6.91 and 22.54 g DW m^–2 and a P/B ratio of 6.7 and 8.4 in the northern and southern part of the lagoon, respectively.