SIMILAR UNSTABLE MUTATIONS IN THREE SPECIES OF GRACILARIA (RHODOPHYTA)1

Unstable mutants with similar variegated pigmentation were genetically characterized in the red algae. Gracilaria tikvahiae (McLachlan), G. foliifera (Forsk.) Børg. and. G. sjoestedtii (Kylin). All three mutants were green plants with flecks of red tissue where cells had reverted to wild type. The mutant green phenotypes were all recessive, and their genetic behavior in crosses indicated that each was the result of a single, unstable, nuclear gene. Wild-type revertant tissue was stable one it arose. Revertant plants obtained from spores and revertant fronds taken from variegated plants could not be distinguished from the normal wild type, either phenotypically or genetically. Reversion to wild type occurred during all phases of the life cycle. In crosses between the mutants and wild type, most of the F₁ tetrasporophytes were heterozygous wild-type plants, an observation consistent with the recessive nature of the mutations; however, a low frequency of homozygous unstable-green F₁ tetrasporophytes was also obttained from these crosses. The molecular basis of neither the mutant instability, i.e. the reversion to wild type, nor of the process producing the unstable green F₁ tetrasporophytes can yet be deduced, but the phenotype of the plants and genetic results suggest the involvement of transposable genetic elements.     

FATTY ACID DISTRIBUTION AMONG SOME RED ALGAL MACROPHYTES1

Macrophytic members of the Rhodophyta, which were grown under controlled conditions, were analyzed for their fatty acid distribution. Significant differences were found between some species of the Gelidiales and Gracilariales. Two dominant polyunsaturated fatty acids of the 20 carbon chain group, 20:4ω6 and 20:5ω3, were found in all the Gelidiales species, whereas only 20:4ω6 was found in the Gracilariales species. An inverse relationship between the content of these two polyunsaturated fatty acids in Gelidiales species and among the two groups of species are reported and discussed. All other fatty acid characteristics were more or less similar in these two orders. We cannot draw any solid conclusion about the relationship between Gracilariopsis cf. lemaneiformis (formerly Gracilaria lemaneiformis) and the genus Gracilaria, since our results (the existence of 20:4ω6 and 20:5ω3 in Gracilariopsis) contradict results found by other researchers. The diverse growth conditions of photon flux density and temperature caused some differences in the distribution of the fatty acids in each species. These differences could not explain the different results in similar species reported in the literature.     

Assessment of allozyme variation among New Zealand populations of Gracilaria chilensis (Gracialiares,Rhodophyta) using starch-gel electrophoresis

New Zealand populations of Gracilaria chilensis are uniform in anatomical reproductive characteristics but vary morphologically and have been found to separate into two distinct groups with respect to agar methylation level, namely low (24–30%) and high (43–47%). To investigate the genetic variation within New Zealand populations of this species, 14 isozyme loci detected by starch-gel electrophoresis were examined in 17 wild populations from a wide range of localities, and in cultures derived from these populations. Five of these loci were polymorphic, but the genetic variation within populations was low: of the 17 populations examined, 15 were fixed at all loci (heterozygosity 0.000) and in the remaining two populations the observed heterozygosity was still low (0.004 and 0.011). The genetic distances between the populations ranged from 0.00 to 0.43. UPGMA cluster analysis separated the populations into two groups, a northern group and a group found throughout the country. Although these two groups do not correlate with the two groups based on agar methylation level at every locality, the correlation is sufficiently striking to merit further investigation.     

Antiviral compounds in extracts of Korean seaweeds: Evidence for multiple activities

Extracts of 13 Korean seaweeds, previously shown to contain antiviral activity, were investigated in more detail in order to learn the nature of the antiviral compounds and their mechanisms of action. One extract, from Codium fragile, was active against all three test viruses (herpes simplex, HSV; Sindbis, SINV; polio), whereas the others were more selective. Thus four species, Enteromorpha linza, Colpomenia bullosa, Scytosiphon lomentaria, and Undaria pinnatifida, were active against HSV and SINV, but not poliovirus. The other eight were active against either HSV or SINV. In all cases there was evidence for photosensitizers, since the antiviral activities required or were enhanced substantially by light. In general UVA (long wave ultraviolet) was much more effective than visible light in promoting activity, although the extract of Sargassum sagamianum could be activated equally by either. In experiments to determine the site of action of these antiviral extracts, the predominant activity was virucidal (i.e. direct inactivation of virus particles), rather than inhibition of virus replication, although Sargassum sagamianum also could protect cells against subsequent virus infection. These results imply that different antiviral compounds are present among the extracts, and furthermore the activities cannot be explained in terms of common ingredients such as polysaccharides or tannins. We suggest that seaweeds may be a source of potentially useful and interesting antiviral compounds. This revised version was published online in June 2006 with corrections to the Cover Date.     

Affinities of the marine flora of the Revillagigedo Islands,Mexico

The benthic algal flora reported for the Revillagigedo Islands comprises 205 specific infraspecific taxa: 42 Chlorophyta, 29 Phaeophyta and 134 Rhodophyta. This insular flora shares 131 taxa (54%) with other regions of the Mexican Pacific and 74 (36%) are restricted apparently to the islands. One hundred three taxa (50%) are shared with areas of the Mexican tropical Pacific, 69 (34%) with warm temperate Pacific Mexico and 66 (32%) with La Paz, the transitional zone between tropical and warm temperate Pacific Mexico. Considering more general regions, the Revillagigedo Islands flora includes apparently restricted distribution (34 spp., 16.6%), exclusively tropical (51 spp., 25%) and widely distributed eastern Pacific (33 spp., 16%) taxa. Even though we consider that the inventory of the Revillagigedo Islands and to a lesser degree the eastern tropical Pacific flora is still incomplete and in need of further taxonomic study, the floristic comparison shows a greater affinity of the Revillagigedo Islands flora with the Mexican tropical Pacific than with any other part of Mexico.     

Systematic studies of the antarctic species of the phyllophoraceae (Gigartinales,Rhodophyta) based on rbcL sequence analysis

The taxonomic placement of four antarctic species of the marine red algal family Phyllophoraceae (Gigartinales) is assessed within a preliminary molecular phylogeny of the family based on direct sequence analysis of the chloroplast gene rbcL. Parsimony analysis of rbcL sequences indicates that Gymnogongrus antarcticus and Gymnogongrus turquetii cluster in a clade consisting predominantly of southern hemisphere species currently placed in Gymnogongrus and Ahnfeltiopsis, whereas Phyllophora ahnfeltioides and Phyllophora antarctica cluster in a separate clade that is widely divergent from the northern hemisphere Phyllophora clade. Results from molecular and morphological data challenge the current taxonomic concept that type of life history is a phylogenetically valid criterion for recognition of genera in the Phyllophoraceae.     

Amino acid composition, protein content and calculation of nitrogen-to-protein conversion factors for 19 tropical seaweeds

The use of nitrogen‐to‐protein conversion factors (N‐Prot factors) is the most practical way of determining protein content. The accuracy of protein determination by this method depends on the establishment of N‐Prot factors specific to individual species. Experimental data are needed to allow the use of this methodology with seaweeds. The present study was designed to characterize the amino acid composition and to establish specific N‐Prot factors for six green, four brown and nine red marine algae. Mean values for individual amino acids tended to be similar among the three groups, but some differences were found. Green algae tended to show lower percentages of both aspartic acid and glutamic acid than the other two groups of algae. The percentages of both lysine and arginine were higher in red algae, while brown algae tended to show more methionine than green and red algae. The actual protein content of the species, based on the sum of amino acid residues, varied from 10.8% (Chnoospora minima, brown algae) to 23.1% (Aglaothamnion uru‐guayense, red algae) of the dry weight. Nitrogen‐to‐protein conversion factors were established for the species studied, based on the ratio of amino acid residues to total nitrogen, with values ranging from 3.75 (Cryptonemia seminervis, red algae) to 5.72 (Padina gymnospora, brown algae). The relative importance of non‐protein nitrogen is greater in red algae, and consequently lower N‐Prot factors were calculated for these species (average value 4.59). Conversely, protein nitrogen content in both green and brown algae tends to be higher, and average N‐Prot factors were 5.13 and 5.38, respectively. An overall average N‐Prot factor for all species studied of 4.92 ± 0.59 (n = 57) was established. This study confirms that the use of the traditional factor 6.25 is unsuitable for seaweeds, and the use of the N‐Prot factors proposed here is recommended.     

An evaluation of methods for extraction and quantification of protein from marine macro- and microalgae

Comparison of data of protein content in algae is very difficult, primarily due to differences in the analytical methods employed. The different extraction procedures (exposure to water, grinding, etc.), protein precipitation using different amounts of 25% trichloroacetic acid and quantification of protein by two different methods and using two protein standards were evaluated. All procedures were tested using freeze-dried samples of three macroalgae: Porphyra acanthophora var. acanthophora, Sargassum vulgare and Ulva fasciata. Based on these results, a protocol for protein extraction was developed, involving the immersion of samples in 4.0 mL ultra-pure water for 12 h, followed by complete grinding of the samples with a Potter homogeniser. The precipitation of protein should be done with 2.5:1 25% TCA:homogenate (v/v). The protocol for extraction and precipitation of protein developed in this study was tested with other macroalgae (Aglaothamnion uruguayense, Caulerpa fastigiata, Chnoospora minima, Codium decorticatum, Dictyota menstrualis, Padina gymnospora and Pterocladiella capillacea) and microalgae (Amphidinium carterae, Dunaliella tertiolecta, Hillea sp., Isochrysis galbana and Skeletonema costatum). Comparison with the actual protein content determined from the sum of amino acid residues, suggests that Lowry's method should be used instead of Bradford's using bovine serum albumin (BSA) as protein standard instead of casein. This may be related to the reactivity of the protein standards and the greater similarity in the amino acid composition of BSA and algae. The current results should contribute to more accurate protein determinations in marine algae.     

The arrival of propagules of marine macroalgae in the intertidal zone

The composition and abundance of macroalgal propagules contained in sea water arriving at intertidal rocky shores was estimated monthly at Pelancura, central Chile, from June to December, 1984. Samples from surface water and from water running off rocky platforms with mixed algal vegetation were cultured in laboratory conditions and examined for development of sporelings. Thirty eight macroalgal entities grew in the cultures, 75% of them with opportunistic life-styles. The total number of sporelings was similar in the surface water of sites separated by 300 m. Marked variability in composition and number of sporelings was, however, observed between samples taken simultaneously at one site and between different months, suggesting patchiness in the dispersal of propagules. Marked differences were established in the ratio run-off water/surface water in the number of sporelings, which could be related to the dispersal shadows of the various taxa. Sporelings of late successional algae showed marked spatial and temporal variations in number. Some taxa of opportunistic algae were characterized by the development of several thousand sporelings per litre of sea water while in others the number of sporelings was two or three orders of magnitude less, suggesting differences in reproductive effort.     

Chemical defenses of seaweeds against microbial colonization

Any living or non-living surface immersed in seawaterrapidly acquires a bacterial biofilm. For living marineorganisms, biofilm formation can result in the death ofthe host, and thus there is strong evolutionary pressure formarine eukaryotes to evolve mechanisms which inhibit orcontrol the development of biofilms on their surfaces.Some marine eukaryotes are indeed successful incontrolling biofilms on their surfaces, and in manyinstances this control is achieved by the production ofinhibitory chemicals which act at or near the surface ofthe organism. In some cases these natural inhibitors aresimply toxic to bacteria. However, increasingly it appearsthat at least some of these compounds act by interferingspecifically with bacterial characteristics which effect theability of bacteria to colonize their hosts, such asattachment, surface spreading, or the production ofextracellular macromolecules. As an example, theAustralian seaweed Delisea pulchra appears tocontrol bacterial colonization by interfering with abacterial regulatory system (the acylated homoserinelactone system) that regulates several colonizationrelevant bacterial traits. Understanding how marineorganisms control specific bacterial colonization traitsshould provide us with insights into new technologies forthe control of biofilms on artificial surfaces.