POLYAMINES INFLUENCE MATURATION IN REPRODUCTIVE STRUCTURES OF GRACILARIA CORNEA (GRACILARIALES,RHODOPHYTA)1

Naturally occurring levels of putrescine, spermidine, and spermine were analyzed in female gametophyte (nonfertilized) and reproductive tissues (cystocarps) at two different stages of development in the marine red algae Gracilaria cornea J. Agardh. Endogenous polyamine levels changed at differential stages of cystocarp maturation. Highest polyamine values were found on tissue from the early post‐fertilization stage, decreasing as the cystocarp matured. Incubation experiments revealed that exogenous polyamines induced cystocarp maturation and promoted carpospore liberation, developing cell masses within 4 to 7 days in treatments with spermine. This is the first report on the effect of polyamines on cystocarp maturation in marine algae.     

The Seasonal Succession of Epiphytic Communities within an Equisetum fluviatile L. Stand in Lake Pääjärvi,Southern Finland

The seasonal succession of epiphytic communities on Equisetum fluviatile was controlled both directly and indirectly by the macrophyte. Decaying macrophytic material supported rich algal growth and biomass accumulation in spring and early summer. Emergence of the macrophytes severely reduced underwater light availability, and the epiphytic algal biomass declined rapidly as a result of both lower photosynthetic activity of the epiphytic algae, and more intensive grazing by invertebrate herbivores. Epiphytic N: P ratios were lower than those in the water around suggesting that either the water was not the only source of phosphorus for the epiphytic algae or the algae took phosphorus up selectively from the water. Low epiphytic C: N ratios suggested a high potential nutritional value for herbivores.     

Ceramialean epiphytism in an intertidal Gracilaria chilensis (Rhodophyta) bed in southern Chile

The cultivation of the agarophytic red alga Gracilaria has become an activity of major importance in several parts of the world. However, Gracilaria cultivation in Chile still faces problems such as epiphytism. We report ceramialean epiphytism abundance, recruitment patterns and the microscopic stages fixed on the Gracilaria thalli in an intertidal bed (Metri bay) of southern Chile. Using a factorial field experiment, we analyze the effect of intertidal elevation and the use of epiphyte free inoculum on the abundance of ceramialean epiphytic algae. This evidence is used to provide management and control recommendations for ceramialean epiphytes. The results show a summer recruitment and increase in abundance of ceramialean epiphytes. A seasonal and spatial pattern of abundance of microscopic stages of this epiphytic algae was also found, showing a significant increase from the apical (new tissues) to the central parts of the thalli (older tissues). Apical tissues are free of epiphytic propagules. The epiphyte-free tissue zone decreases significantly from winter to summer. Also, a significantly higher density of microscopic stages of ceramialean epiphytes was found on the thalli collected in the farming are a during the summer at low tidal levels. The experimental results indicated that the abundance of epiphytes was greater at the lower tidal levels than at the higher distribution limits of Gracilaria and it was not possible to control epiphytism by manual cleaning. This revised version was published online in September 2006 with corrections to the Cover Date.     

Sustained high yields of Gracilaria (Rhodophyta) grown in intensive large-scale culture

Gracilaria ferox J. Agardh was grown continuously in large, outdoor tanks under a pulse-fed nutrient regime for four years. Productivity ranged from 21.4 to 59.2 g d. wt m⁻² d⁻¹ with a mean of 39.7 g d. wt m⁻² d⁻¹ over the entire study period. Because the cultures were maintained under non-nutrient limiting conditions, productivity was regulated primarily by seasonal changes in light and temperature, which accounted for 75% of the variability of growth in algal yields. Salinity ranged from 31.0 to 36.5‰ and had insignificant effects on growth within this range. The original vegetative strain was maintained over the entire study without the need for additional supplementation from field-collected stock. Because of the pulse-fed nutrient supply, epiphytic growth on the target species was negligible (< 3% total biomass) throughout the study. The yields attained in this study rank among the highest reported for any intensively managed photosynthetic crop and demonstrate the feasibility of growing red macroalgae like Gracilaria at a sustained high yield in a large-scale, land-based culture system. This revised version was published online in July 2006 with corrections to the Cover Date.     

IDENTIFICATION OF GREEN ALGAL ENDOPHYTES AS THE ALTERNATE PHASE OF ACROSIPHONIA (CODIOLALES, CHLOROPHYTA) USING ITS1 AND ITS2 RIBOSOMAL DNA SEQUENCE DATA

The internal transcribed spacer regions of the nuclear ribosomal DNA cistron of green algal endophytes, previously identified as Chlorochytrium inclusum Kjellman and Codiolum petrocelidis Kuckuck, were sequenced. Culture studies have implicated marine Chlorochytrium and Codiolum -like cells in the life histories of a number of macrophytic green algae. The identity of the endophytes was resolved by comparing DNA sequences from unicells endophytic in the foliose red alga Mazzaella splendens (Setchell et Gardner) Hommersand and in the crustose phase of Mastocarpus papillatus (J. Agardh) Kylin, with sequences from species of Acrosiphonia, Urospora, Ulothrix, and Monostroma. All endophyte isolates were more closely related to species of Acrosiphonia than to those of Urospora, Ulothrix, or Monostroma. The results support previous culture studies, and conclusively identify the endophytes obtained in the field as alternate life history phases of one or more Acrosiphonia species in southern British Columbia, Canada.     

New additions to the marine algae of Mochima National Park, Sucre, Venezuela

The marine algal flora from the eastern coast of Mochima National Park, Sucre, Venezuela was studied with a total of 51 taxa identified, including eight new additions of red algae which are Erythrotrichia carnea (Dillwyn) J. Agardh, Sahlingia subintegra (Rosenvinge) Kornmann, Liagora ceranoides Lamouroux, Asparagopsis taxiformis (Delile) Trevisan, Dasya corymbifera J. Agardh, Chondria dasyphylla (Woodward) C. Agardh, Herposiphonia secunda f. tenella (C. Agardh) M.J. Wynne and Polysiphonia subtilissima Montagne. Morphological and anatomical aspects of the specimens are described and illustrated.     

Nitrogen and carbon isotope fractionation by two species of marine amphipods: Laboratory and field studies

The fractionation of the stable nitrogen and carbon isotopes between two species of marine amphipods and their food sources was investigated in both field and laboratory studies. The nitrogen isotope ratios showed a strong species effect. Amphithoe valida Smith grown in the laboratory averaged ?0.3‰ relative to several food sources. Parhyale hawaiensis (Dana) was +2.3‰ relative to its diets. Animals from the field showed similar differences from the macrophytic algae on which they were found. The carbon isotope composition of both species averaged ?0.7‰ relative to their diets. Laboratory animals fed fresh algae were slightly more negative than field animals or animals fed detrital algae.     

Ecological engineering in aquaculture: use of seaweeds for removing nutrients from intensive mariculture

Rapid scale growth of intensive mariculture systems can often lead to adverse impacts on the environment. Intensive fish and shrimp farming, being defined as throughput-based systems, have a continuous or pulse release of nutrients that adds to coastal eutrophication. As an alternative treatment solution, seaweeds can be used to clean the dissolved part of this effluent. Two examples of successfully using seaweeds as biofilters in intensive mariculture systems are discussed in this paper. The first example shows that Gracilaria co-cultivated with salmon in a tank system reached production rates as high as 48.9 kg m⁻² a⁻¹, and could remove 50% of the dissolved ammonium released by the fish in winter, increasing to 90–95% in spring. In the second example, Gracilaria cultivated on ropes near a 22-t fish cage farm, had up to 40% higher growth rate (specific growth rate of 7% d⁻¹) compared to controls. Extrapolation of the results showed that a 1 ha Gracilaria culture gave an annual harvest of 34 t (d. wt), and assimilated 6.5% of the released dissolved nitrogen. This production and assimilation was more than twice that of a Gracilaria monoculture. By integrating seaweeds with fish farming the nutrient assimilating capacity of an area increases. With increased carrying capacity it will be possible to increase salmon cage densities before risking negative environmental effects like eutrophication and toxic algal blooms sometimes associated with the release of dissolved nutrients. The potential for using mangroves and/or seaweeds as filters for wastes from intensive shrimp pond farming is also discussed. It is concluded that such techniques, based on ecological engineering, seems promising for mitigating environmental impacts from intensive mariculture; however, continued research on this type of solution is required. This revised version was published online in June 2006 with corrections to the Cover Date.     

Seasonal succession of planktonic and epiphytic algae in a canal in Southern England

The standing crop of phytoplankton in a canal in southern England remained low during 1973 and 1974, seldom exceeding 5 × 10⁴ cells/1. Since phosphorus, nitrogen and silicon occurred abundantly in the water, competition with higher plants for some other substance must have limited development. Although the standing crop of epiphytic algae associated withNasturtium officinale andGroenlandia densa seemed to be limited by the number of attachment sites, this factor was of little importance in the case of algae attached toCladophora glomerata. Achnanthes minutissima v.cryptocephala was always predominant in the epiphytic assemblages, representing 50–8o% by numbers of the flora. The limited pool of predominant epiphytic taxa may have restricted the communities' ability to adapt to fluctuations in environmental conditions. The grazing of isopods, amphipods and molluscs probably never limited algal densities.     

Induction of defenses and within-alga variation of palatability in two brown algae from the northern-central coast of Chile: Effects of mesograzers and UV radiation

Macroalgae possess different defense mechanisms in response to herbivory. Some species produce anti-herbivore secondary metabolites, but production of these substances can be costly. Therefore, algae may produce defensive metabolites only in response to herbivory (inducible defense) or defend particular parts of the alga differentially (within-alga variation). In the present study, we examined whether two species of brown algae from the SE-Pacific show evidence of inducible chemical defense (non-polar compounds) or within-alga variation of defense, which we estimated in form of palatability of differently treated algae to amphipod grazers (with live algae and agar-based food containing non-polar algal extracts). In Glossophora kunthii (C. Agardh) J. Agardh, we observed an increase in palatability after algae were acclimated for 12 days without grazers. Subsequent addition of grazers for 12 days then resulted in a reduction of palatability indicating the existence of inducible defense. After removal of grazers for 12 days, these induced effects again disappeared. The reaction of G. kunthii was triggered even by the mere presence of grazers, which suggests that this alga can respond to waterborne cues by reducing palatability. Effects were only found for agar-based food containing non-polar extracts, but not for live algae, suggesting that some parts of the algae are undefended. Our second experiment on within-alga variation confirmed that only apical (growth region) and basal parts (near the holdfast region) of G. kunthii are defended against herbivores. For the second species, Macrocystis integrifolia Bory, the first experiment revealed no induction of defense, while the second experiment on within-alga variation showed that amphipods avoided basal parts and in particular stipes of M. integrifolia but only in live algae. Although both studied algal species differed substantially in their defensive strategies, their reaction was independent of the presence or absence of UV radiation. Thus, it appears that UV effects play only a minor role in anti-herbivore defense, which is in accordance with most previous studies.     

MICROSATELLITE DEVELOPMENT IN RHODOPHYTA USING HIGH‐THROUGHPUT SEQUENCE DATA1

Shotgun genome sequencing is rapidly emerging as the method of choice for the identification of microsatellite loci in nonmodel organisms. However, to the best of our knowledge, this approach has not been applied to marine algae so far. Herein, we report the results of using the 454 next‐generation sequencing (NGS) platform to randomly sample 36.0 and 40.9 Mbp (139,786 and 139,795 reads, respectively) of the genome of two red algae from the northwest Iberian Peninsula [Grateloupia lanceola (J. Agardh) J. Agardh and a still undescribed new member of the family Cruoriaceae]. Using data mining tools, we identified 4,766 and 5,174 perfect microsatellite loci in 4,344 and 4,504 sequences/contigs from G. lanceola and the Cruoriaceae, respectively. After conservative removal of potentially problematic loci (redundant sequences, mobile elements), primer design was possible for 1,371 and 1,366 loci, respectively. A survey of the literature indicates that microsatellite density in our Rhodophyta is at the low end of the values reported for other organisms investigated with the same technology (land plants and animals). A limited number of loci were successfully tested for PCR amplification and polymorphism finding that they may be suitable for population genetic studies. This study demonstrates that random genome sequencing is a rapid, effective alternative to develop useful microsatellite loci in previously unstudied red algae.     

Snail communities increase submerged macrophyte growth by grazing epiphytic algae and phytoplankton in a mesocosm experiment

The relationships between producers (e.g., macrophytes, phytoplankton and epiphytic algae) and snails play an important role in maintaining the function and stability of shallow ecosystems. Complex relationships exist among macrophytes, epiphytic algae, phytoplankton, and snails. We studied the effects of snail communities (consisting of Radix swinhoei, Hippeutis cantori, Bellamya aeruginosa, and Parafossarulus striatulus) on the biomass of phytoplankton and epiphytic algae as well as on the growth of three species of submerged macrophytes (Hydrilla verticillata, Vallisneria natans, and one exotic submerged plant, Elodea nuttallii) in a 90‐day outdoor mesocosm experiment conducted on the shore of subtropical Lake Liangzihu, China. A structural equation model showed that the snail communities affected the submerged macrophytes by grazing phytoplankton and epiphytic algae (reduction in phytoplankton Chl‐a and epiphytic algal abundance), enhancing the biomass of submerged macrophytes. Highly branched macrophytes with high surfaces and morphologies and many microhabitats supported the most snails and epiphytic algae (the biomass of the snail communities and epiphytic algae on H. verticillata was greater than that on V. natans), and snails preferred to feed on native plants. Competition drove the snails to change their grazing preferences to achieve coexistence.     

INDUCTION AND REDUCTION OF ANTI‐HERBIVORE DEFENSES IN BROWN AND RED MACROALGAE OFF THE KENYAN COAST1

Herbivory is particularly intense in tropical benthic communities, suggesting preference of constitutive, rather than inducible, anti‐herbivory defense. The objective of the study was to examine whether anti‐herbivore defenses in the red alga Hypnea pannosa J. Agardh and the brown algae Sargassum asperifolium Hering and G. Martens ex J. Agardh and Cystoseira myrica (S.G. Gmelin) C. Agardh could be induced and subsequently reduced in response to grazing by the amphipod Cymadusa filosa Savigny. During a 14‐day treatment phase, algae were exposed to amphipod grazing or were left ungrazed (control). Subsequently, one subset of algae was used in feeding assays, whereas another was cultivated for additional 14 days without consumers (recovery phase). At the end of each phase, bioassays were conducted to detect defensive traits in terms of differences in consumption rates of grazed and control pieces of live algae and agar‐based food containing nonpolar algal extracts. Consumption of grazed live S. asperifolium and H. pannosa specimens was lower than of control algae. Furthermore, nonpolar extracts of grazed S. asperifolium and C. myrica were less preferred than those from control algae. Defensive responses were exclusively detected after the treatment phase, although strong preference of ungrazed H. pannosa and C. myrica over grazed conspecifics continued throughout the recovery phase. These findings suggest that phenotypic plasticity in anti‐herbivory defense of marine macroalgae 1) might be more common than previously shown, 2) could be switched on and off within 2 weeks, and 3) can be found in nonpolar algal extracts.     

MOLECULAR PHYLOGENY OF GRACILARIA SPECIES INFERRED FROM MOLECULAR MARKERS BELONGING TO THREE DIFFERENT GENOMES1

The nucleotide sequence data of molecular markers 18S rRNA, RUBISCO spacer, and cox2‐3 intergenic spacer were integrated to infer the phylogeny of Gracilaria species, collected from the western coast of India, reducing the possibility of misidentification and providing greater phylogenetic resolution. A phylogenetic tree was constructed using cox2‐3 and RUBISCO spacer sequences, exhibiting the same clustering but differing slightly from that of the rRNA‐based phylogenetic tree. The phylogeny inferred from the combined data set confers an analogous pattern of clustering, compared with those of trees constructed from individual data sets. The combined data set resulted in a phylogeny with better resolution, which supported the clade with higher consistency index, retention index, and bootstrap values. It was observed that Gracilaria foliifera (Forssk.) Børgesen is closer to G. corticata (J. Agardh) J. Agardh varieties, while G. salicornia (C. Agardh) E. Y. Dawson and G. fergusonii J. Agardh both originated from the same clade. The position of G. textorii (Suringar) De Toni faltered and toppled between G. salicornia and G. dura (C. Agardh) J. Agardh; however, G. gracilis (Stackh.) M. Steentoft, L. M. Irvine et W. F. Farnham was evidently distant from the rest of the species.     

山西省辛安泉泉域三种泉溪大型藻类植物附生藻类的研究

对三种泉溪大型藻类(脆弱刚毛藻, 弧形串珠藻和普生轮藻)藻体上附生藻类的种类、分布及环境因子与寄主本身对其的影响进行了研究。2004 年7 月至2005 年4 月在山西省辛安泉泉域采集三种大型藻类植物(每个季节采集一次)。研究结果表明, 三种大型藻类藻体上的附生藻类种类数不同, 普生轮藻上的附生藻类有49 种, 脆弱刚毛藻上37 种, 弧形串珠藻上6 种, 其中硅藻门种类数最多。这说明寄主本身结构的复杂程度在一定意义上会影响附生藻类的种类数及多样性。就季节变化而言, 三种大型藻类藻体上的附生藻类也表现出了明显的季节性,大致趋势为: 春季>秋季>夏季>冬季, 同时, 灰关联分析结果也表明, 环境因子在一定程度上对附生藻类也有影响, 最关键的影响因子是流速、水深和电导。这说明影响大型藻类植物藻体上附生藻类的影响因素是多方面的, 不仅有寄主本身, 同时流速、水深和电导这三种环境因子也起到了很关键的作用。       

Impact of power plant heated effluent on the abundance of sedentary organisms,off Kalpakkam,East coast of India

The impact of heated effluents from the condenser outfall of the Madras Atomic Power Station (MAPS), on the flora and fauna settled on an iron pile in the discharge area was studied. At temperatures ranging between 27.2 to 31.0 °C, the sedentary community was composed of epiphytic algae, sea anemones, tubeworms, polychaetes, crabs, amphipods, barnacles, periwinkles, mussels and ascidians. During January–February, ambient temperature ranged between 37.0 to 37.6 °C, and almost all the macroepifauna and flora, except for periwinkles and Cthamalid barnacles perished.     

IMPACTS OF MESOGRAZERS ON EPIPHYTE AND ENDOPHYTE GROWTH ASSOCIATED WITH CHEMICALLY DEFENDED MACROALGE FROM THE WESTERN ANTARCTIC PENINSULA: A MESOCOSM EXPERIMENT1

It has been hypothesized that the extensive mesograzer community along the western Antarctic Peninsula regulates epiphytic algae as well as emergent filaments from endophytic species. Should grazing limit growth of fouling or potentially pathogenic microphytes, then Antarctic macrophytes may actually benefit from the remarkably high densities of mesograzer amphipods that occur in these waters. Although initially counterintuitive, the negative impacts of epi/endophyte fouling may outweigh stresses caused by limited amphipod grazing on chemically defended macrophytes by reducing stress from endo/epiphyte biomass. If so, then alleviating mesograzing stress should result in significant increases in endo/epiphytic biomass. To test this hypothesis, a mesocosm experiment was conducted. Individuals representing four common species of Antarctic macroalgae were placed in flow‐through seawater mesocosms. Amphipods were added to five mesocosms at simulated natural densities, while the other five remained herbivore free. At the end of 7 weeks, endo/epiphytic growth on individual macrophytes was quantified. Most species of macroalgae demonstrated noticeably higher instances of endophyte coverage, epiphytic diversity, and diatom colonization in consumer‐free mesocosms than in the presence of amphipods. These data suggest that macroalgae along the western Antarctic Peninsula rely on grazers to control populations of potentially harmful epiphytes. We hypothesize that the chemically defended macroalgal flora lives in mutualism with high densities of mesograzers, providing amphipods with shelter from predation while continually being cleaned of potentially harmful endo/epiphytes.     

SYSTEMATICS OF THE GRACILARIACEAE (GRACILARIALES,RHODOPHYTA): A CRITICAL ASSESSMENT BASED ON RBCL SEQUENCE ANALYSES1

Generic concepts in the economically important agarophyte red algal family Gracilariaceae were evaluated based on maximum parsimony, Bayesian likelihood, and minimum evolution analyses of the chloroplast‐encoded rbc L gene from 67 specimens worldwide. The results confirm the monophyly of the family and identify three large clades, one of which corresponds to the ancestral antiboreal genera Curdiea and Melanthalia, one to Gracilariopsis, and one to Gracilaria sensu lato, which contains nine distinct independent evolutionary lineages, including Hydropuntia. The species currently attributed to Hydropuntia comprise a single well‐supported clade composed of two distinct lineages. The two most basal clades within Gracilaria sensu lato deserve generic rank: a new genus centered around G. chilensis Bird, McLachlan et Oliveira and G. aff. tenuistipitata Chang et Xia and a resurrected Hydropuntia encompassing primarily Indo‐Pacific (G. urvillei [Montagne] Abbott, G. edulis [S. Gmelin] P. Silva, G. eucheumatoides Harvey, G. preissiana [Sonder] Womersley, and G. rangiferina [Kützing] Piccone) and western Atlantic species (G. cornea J. Agardh, G. crassissima P. et H. Crouan in Mazé et Schramm, G. usneoides [C. Agardh] J. Agardh, G. caudata J. Agardh, and G. secunda P. et H. Crouan in Mazé et Schramm). Cystocarpic features within the Gracilaria sensu lato clades appear to be more phylogenetically informative than male characters. The textorii‐type spermatangial configuration is represented in two distinct clusters of Gracilaria. The rbc L genetic divergence among the Gracilariaceae genera ranged between 8.46% and 16.41%, providing at least 2.5 times more genetic variation than does the 18S nuclear rDNA. rbc L also resolves intrageneric relationships, especially within Gracilaria sensu lato. The current number of gracilariacean species is underestimated in the western Atlantic because of convergence in habit and apparent homoplasy in vegetative and reproductive anatomy.     

The yield,physical and chemical properties of agar gel from Gracilaria species (Gracilariales,Rhodophyta) of the Kenya coast

Six species of Gracilaria, G. corticata J. Agardh, G. crassa Harvey, G. millardetii J. Agardh, G. salicornia (J. Ag.) Dawson, G. verrucosa (Huds.) Papenfuss and Gracilaria sp, collected from different stations along the Kenya coast were studied. The yield of hot water native agar extract ranges from 8.1–30% of dry weight, with G. verrucosa and G. salicornia having the greatest and the least yield, respectively. The gel-strength of 1.5% agar solution was highest in G. verrucosa (220 g cm^–2) and lowest in G. corticata (< 60 g cm – 2) whereas the highest gelling temperature was recorded for Gracilaria sp. (40.4 °C) and the lowest in G. verrucosa (28.9 °C). 3,6 anhydrogalactose content was the highest in G. verrucosa and the lowest in G. corticata while sulphate content was higher in the latter.