Biogeography of sexual and asexual populations in Bostrychia moritziana (Rhodomelaceae, Rhodophyta)

Bostrychia moritziana (Sonder ex Kützing) J. Agardh is recorded from many regions around the world. Our laboratory culture investigations have verified a sexual life cycle in isolates from Australia, Venezuela, Colombia, South Africa, Fiji, New Zealand and Indonesia. By contrast, asexual isolates producing successive generations of tetrasporophytes in laboratory culture and, presumably, in the field, are known from Australia. New Caledonia and Japan. In Australia, asexual reproduction is absent only in Victoria. In Western Australia, Northern Territory and Queensland, 99% of the isolates have asexual reproduction. In New South Wales (NSW), asexual and sexual populations are often intermixed. Of the 176 worldwide field collections, 58% were vegetative, 39% were tetrasporic, 2% were female and 1% were male. After several years of observations on the asexual isolates in culture, at least 30 successive asexual tetrasporophytic generations have developed. Only two asexual isolates (3558 and 3575) from NSW have formed a single male and female gametophyte in culture. In a self-cross of 3568, the carpospores developed into tetrasporophytes that recycled asexually. All outcrosses done with normal sexual isolates produced normal carposporophytes and the carpospores developed into tetrasporophytes that also recycled asexually. Asexual populations may arise repeatedly by loss of meiosis in tetrasporangia of sexual populations. Asexual reproduction apparently does not diminish the overall dispersal and abundance in the field. Our present bio-geographic data show that sexually reproducing popuiations of B. moritziana occur worldwide, while asexuaily reproducing populations are confined to the western Pacific. Bostrychia bispora West et Zuccarello, initially described on the basis of its asexual reproduction to distinguish it from B, moritziana, is now reduced to synonymy with B. moritziana.     

A polysaccharide fraction from the red seaweed Champia novae-zealandiae Rhodymeniales,Rhodophyta

The polysaccharide recovered after extraction of Champia novae-zealandiae is a galactan with alternating 3-linked d-galactopyranosyl units sulfated at the 2-position, and 4-linked galactopyranosyl units sulfated at both the 2- and 3-positions that are predominantly of the l- and partly of the d-configuration. Other minor substitution includes 6-O-methyl ether or 4,6-pyruvate acetal on the 3-linked residues. Techniques used in determining the structure include infrared and ¹³C-NMR spectroscopy, and GC-MS analysis of alditol acetate derivatives produced by reductive hydrolysis/acetylation of native, methylated, and/or desulfated samples. These results are of particular interest because 4-linked 2,3-desulfated galactosyl residues have not been encountered as major constituents of red algal polysaccharides.     

Carbohydrate regulation of attachment, encystment, and appressorium formation by Pythium porphyrae (Oomycota) zoospores on Porphyra yezoensis (Rhodophyta)

Pythium porphyrae Takahashi et Sasaki, a facultative parasite of Porphyra spp., is the common microbial agent responsible for red rot disease of this red alga in Japan. Host infection by this species and other plant parasitic members of the Pythiaceae is initiated by motile biflagellate zoospores. Factors regulating host specificity and the initial steps involved in the infection process, consisting of attachment, encystment and appressorium formation, are not known. Zoospore encystment and appressorium formation of P. porphyrae were monitored by staining of the fungal cell walls using calcofluor. The zoospores infected only Porphyra spp. and Bangia atropurpurea (Roth) C. Agardh thalli, although they attached to, and encysted on, many other members of the Rhodophyceae (Stylonema alsidii[Zanardini] Drew, Gelidium elegans Kützing, Pterocladiella capillacea[Gmelin] Santelices et Hommersand, Carpopeltis affinis[Harvey] Okamura, Gloiosiphonia capillaris[Hudson] Carmichael in Berkeley, Grateloupia turuturu Yamada, Callophyllis adhaerens Yamada, Gracilaria spp., Lomentaria hakodatensis Yendo, Rhodymenia intricata[Okamura] Okamura, Griffithsia subcylindrica Okamura, Wrangelia tanegana Harvey, and Polysiphonia morrowii Harvey). No attachment or encystment was observed on the red alga Kappaphycus striatum (Schmitz) Doty ex Silva in Silva et al., the brown algae Undaria pinnatifida (Harvey) Suringar, Scytosiphon sp., and Sargassum thunbergii (Mertens ex Roth) Kuntze as well as members of the Ulvaceae (green algae). Sequential extraction of carbohydrates from Porphyra yezoensis Ueda thalli and the addition of diverse monosaccharides, polysaccharides, and amino acids to zoospore suspensions indicated that encystment and appressorium formation were induced only by sulfated galactans (porphyran, commercial agar, agarose, and carrageenans). Zoospore attachment and encystment on thalli of P. yezoensis was abolished by periodate oxidation of the thallus surface and was reduced by 80–90% after enzymatic removal of sulfated galactan (porphyran). It appears that the interaction of zoospore surface receptors with sulfated galactan (porphyran) determinants on the thallus surface induced specific attachment and encystment on Porphyra spp. thalli. Zoospores encysted, germinated, and formed appressoria on sulfated galactan films and in suspensions of this carbohydrate. Attachment and encystment were induced on commercial agar and agarose films, but appressoria were not induced on agarose films. Supplementation of agarose media with both cold and hot water fractions and with porphyran from P. yezoensis–induced appressoria implicated sulfated galactans (porphyran) in appressorium formation.     

Olpidiopsis sp., an oomycete from Madagascar that infects Bostrychia and other red algae: Host species susceptibility

Olpidiopsis sp. (Oomycota) was cultured with its original host Bostrychia moritziana (Sonder ex Kützing) J. Agardh from Madagascar. Bean‐shaped zoospores with two heterokont flagella attached to the host cell wall surface and in 2 days host cells began collapsing and one or more syncytia developed in each infected cell. Zoospores were cleaved and an exit tube with a small plug was formed. Complete development and zoospore discharge occurred in 3 days. Infection occurred in cells of polysiphonous branches, monosiphonous branches, rhizoids and reproductive stichidia. Dead cells of plants treated with microwave were not infected. Susceptibility was variable in other Bostrychia species from different countries. Bostrychia moritziana (Sonder ex Kützing) J. Agardh, and Bostrychia radicans (Montagne) Montagne from Madagascar were susceptible but one Bostrychia tenella (J. V. Lamouroux) J. Agardh isolate from Madagascar was susceptible and two were not. B. radicosa (Itono) J. A. West, G. C. Zuccarello et M. Hommersand isolates from Madagascar, Thailand, Australia and New Caledonia were susceptible but an isolate from Malaysia was not. B. radicans isolates from Mexico and Brazil were non‐susceptible as were Bostrychia flagellifera Post, Bostrychia harveyi Montagne, Bostrychia montagnei Harvey, Bostrychia simpliciuscula Harvey ex J. Agardh, Bostrychia tenuissima R. J. King et Puttock, Stictosiphonia intricata(Bory de Saint‐Vincent) P. C. Silva, Stictosiphonia kelanensis (Grunow) R. J. King et Puttock and Stictosiphonia tangatensis (Post) R. J. King et Puttock, Lophosiphonia sp., Neosiphonia sp. and Polysiphonia spp. isolates were also non‐susceptible. Many non‐susceptible strains showed initial cell‐collapse followed by rapid wound‐repair cell formation without syncytia or sporangia developing. Caloglossa leprieurii (Montagne) G. Martens from Madagascar showed cell‐collapse and wound‐repair in periaxial cells, but wing cells died and became purple without wound‐repair. Caloglossa ogasawaraensis Okamura and Caloglossa postiae M. Kamiya et R. J. King had no symptoms of infection. Dasysiphonia chejuensis I. K. Lee et J. A. West was not infected. Surprisingly, the conchocelis phase but not the blade phase of Porphyra pulchella J. A.West, G. C. Zuccarello and Porphyra suborbiculata Kjellman was infected. The conchocelis of Porphyra tenera Kjellman and Porphyra linearis Greville were infected but no blade stages were tested. Porphyra miniata (C. Agardh) C. Agardh and Porphyra dentata Kjellman conchocelis were not infected. Bangia atropurpurea (Roth) C. Agardh gametophyte filaments were not infected. Other red, brown and green algae were not infected. Time lapse videomicroscopy of development and spore release was done.     

Hybridization studies in Bostrychia. 1: B. radicans (Rhodomelaceae, Rhodophyta) from Pacific and Atlantic North America

Bostrychia radicans(Montagne) Montagne is a pantropical/temperate red alga associated with mangroves and saltmarsh plants. Collections were made from a similar north-south geographic distribution along both the Pacific and Atlantic coasts of North America. Hybridization studies were performed with cultured isolates to assess the extent of interfertility and reproductive isolation along these two coastlines. All male and female gametophytes derived from single tetrasporophytes were intercompatible. Almost all isolates extending over 1500 km of coast line from northern Pacific Mexico are compatible, forming cystocarps that released viable carpospores. Even isolates which morphologically would be placed in two species [B. radicans and B. moritziana(Sender ex Kützing) J. Agardh], based on the presence or absence of monosiphonous branches, were capable of hybridizing. Crosses of isolates from the Atlantic USA showed a greater amount of incompatibility. Certain isolates were not compatible with any other isolates including isolates collected in close proximity (North Carolina isolates), while other isolates from the same locality were compatible (South Carolina). An isolate from South Carolina formed tetrasporophytes with isolates from Pacific Mexico but tetraspores were not viable. Certain incompatible crosses formed ‘pseudocystocarps’ but viable carposporophytes did not develop. Generalizations about reproductive isolation within a species must also consider differences between populations from different biogeographic regions that may reflect different paleoclimatological histories, founder effects and unique dispersal events.     

The Galactan Sulphates of the Conchocelis Phases of Porphyra leucostricta and Bangia atropurpurea (Rhodophyta)

Galactan sulphates have been extracted from mechanically isolatedcell walls of laboratory cultured conchocelis phases of Porphyraleucostricta and Bangia atropurpurea and their structure establishedby chemical methods and ¹³C-n.m.r. spectroscopy. The polysaccharidesrepresent an extreme structure for porphyran-type polymers inthat they consist almost entirely of 3-linked ß-D-galactosylresidues alternating with 4-linked -L-galactose-6-sulphate residues,the molar ratio of D to L units being approximately 1: 1. ¹³C-n.m.r.and gas-liquid and paper chromatography gave no indication of6-O-methyl-D-galactose in these polymers. Conchocelis galactansare chemically distinct from porphyrans of generic phases ofPorphyra and Bangia. Key words: Rhodophyta, Galactans, Cell walls     

HOST SPECIFICITY IN THE RED ALGAL PARASITES BOSTRYCHIOCOLAX AUSTRALIS AND DAWSONIOCOLAX BOSTRYCHIAE (CHOREOCOLACACEAE,RHODOPHYTA)1

The determinants of host specificity, which are poorly understood in red algal parasites, were studied in the red algal parasites Bostrychiocolax australis Zuccarello et West and Dawsoniocolax bostrychiae (Joly et Yamaguishi-Tomita) Joly et Yamaguishi-Tomita. Culture studies were performed to determine host range, sites of host resistance, and genetics of transmission of resistance. Both species parasitize Bostrychia radicans (Montagne) Montagne, whereas Bostrychiocolax australis also parasitizes Bostrychia moritziana (Sonder ex Kützing) J. Agardh and Stictosiphonia kelanensis (Grunow ex Post) R. J. King et Puttock. Isolates of B. radicans resistant to both parasites were found worldwide, often within the same population as susceptible isolates. On resistant Bostrychia species and isolates, specificity was manifested at three stages: 1) host penetration, in which the spore germ peg failed to penetrate the host cuticle/wall; 2) parasite–host cell fusion, in which the fusion cell died and the parasite died; and 3) growth, in which parasites grew but soon died; parasites rarely reproduced and infections did not continue in culture. Resistance to parasite infection was usually transmitted as a dominant trait and did not segregate as a single locus during meiosis. In certain crosses, transmission of resistance was non-mendelian.     

Effects of initial enrichment of nitrogen and phosphorus on Bostrychia and Caloglossa (Ceramiales, Rhodophyta) growth using digital imaging

Growth experiments measured the effects of an initial starvation period followed by a single nutrient pulse. Nutrient pulses were conducted at four different N-levels (65, 1514, 2900, and 6080 μg N L^-1) and four different P-levels (84, 281, 639, and 849 μg P L^-1) using isolates of Bostrychia moritziana (Sonder ex Kutzing) J.Agardh and Caloglossa leprieurii (Mon-tagne) G.Martens. Specific growth rates (% day^-1) in primary axis length (6. moritziana only) and surface area were measured using digital imaging. Results showed that the specific growth rates of both algae were highly dependent on the N-levels of the seawater (P < 0.001–0.05). Specific growth rates for the B. moritziana were approximately 3.5% day^-1 at t = 7 days declining to approximately 1.0–1.5% day^-1 at t = 49 days for the primary axis length, and approximately 8–9% day^-1 declining to approximately 2–3% day^-1 for surface area. The specific growth rate declined more rapidly with decreasing N initially added to the medium. In C. leprieurii, the specific growth rates for all four conditions at t = 8 days were approximately 11–13% day^-1, but N/O declined to approximately 3% day-¹ whereas the others declined to only approximately 7–8% day^-1 by t = 49 days. The effects of initial P-levels on growth varied, but generally indicated a direct relationship with specific growth rate. In C. leprieurii, the number of nodes and blades per plant was also measured via digital imaging and were found to increase with increasing N-levels, whereas P-levels had no influence. It was concluded that B. moritziana and C. leprieurii are prone to N-limitation, but P-limitation is less prone. Using digital imaging to measure the specific growth rate in total surface area and primary axis length provided a significantly more accurate depiction of the rate of growth than some of the more conventional means of measuring growth rate.     

The xylogalactan sulfate from Chondria macrocarpa (Ceramiales,Rhodophyta)

A structure is proposed for the complex xylogalactan sulfate from Chondria macrocarpa. The hot-water extract of C. macrocarpa was desulfated or alkali-treated and Smith degraded. Constituent sugars and their substitution patterns were identified using a modified Hakamori methylation procedure suited to sulfated polysaccharides and a double hydrolysis-reduction protocol that yielded derivatives from all of the sugar residues, including the labile 3,6-anhydrogalactosyl residues. The polymer has an agar-type backbone of alternating 3-linked \-d- and 4-linked -L-galactopyranosyl units. The d-residues are partially sulfated on O-2 (50%) and O-6 (20–30%). About 40% of the l-residues are present as the 3,6-anhydride and 25% as its precursor l-galactose 6-sulfate. A significant proportion of the remaining l-galactosyl residues have both a d-xylopyranosyl substituent on O-3 and a sulfate ester on O-6 and are stable to alkali.     

Cell Wall Regeneration in Bangia atropurpurea (Rhodophyta) Protoplasts Observed Using a Mannan-Specific Carbohydrate-Binding Module

The cell wall of the red alga Bangia atropurpurea is composed of three unique polysaccharides (β-1,4-mannan, β-1,3-xylan, and porphyran), similar to that in Porphyra. In this study, we visualized β-mannan in the regenerating cell walls of B. atropurpurea protoplasts by using a fusion protein of a carbohydrate-binding module (CBM) and green fluorescent protein (GFP). A mannan-binding family 27 CBM (CBM27) of β-1,4-mannanase (Man5C) from Vibrio sp. strain MA-138 was fused to GFP, and the resultant fusion protein (GFP–CBM27) was expressed in Escherichia coli. Native affinity gel electrophoresis revealed that GFP–CBM27 maintained its binding ability to soluble β-mannans, while normal GFP could not bind to β-mannans. Protoplasts were isolated from the fronds of B. atropurpurea by using three kinds of bacterial enzymes. The GFP–CBM27 was mixed with protoplasts from different growth stages, and the process of cell wall regeneration was observed by fluorescence microscopy. Some protoplasts began to excrete β-mannan at certain areas of their cell surface after 12 h of culture. As the protoplast culture progressed, β-mannans were spread on their entire cell surfaces. The percentages of protoplasts bound to GFP–CBM27 were 3%, 12%, 17%, 29%, and 25% after 12, 24, 36, 48, and 60 h of culture, respectively. Although GFP–CBM27 bound to cells at the initial growth stages, its binding to the mature fronds was not confirmed definitely. This is the first report on the visualization of β-mannan in regenerating algal cell walls by using a fluorescence-labeled CBM.     

Chemical structure and quality of agars from Gracilaria

Agar polymers synthesized by species of the genus Gracilaria constitute a complex mixture of molecules, containing several extremes in structure. Sulphate hemi-esters, methyl ethers and pyruvic ketals can alter in a number of ways the structural regularity of agar based on strictly 3-O-linked β-l-galactopyranose and 4-O-linked α-l-galactopyranose residues. In comparison with agars from Gelidium and Pterocladia, agars from Gracilaria can have higher degrees of sulphation, methoxylation and pyruvylation. The gelling ability of agars from most of Gracilaria species is considerably improved by adopting, before extraction, an alkali pretreatment which converts α-l-galactose 6 sulphate into 3,6-anhydro-α-l-galactose. Native agars obtained from Gracilaria cannot be classified, with few exceptions, as bacteriological grade agar as they have a high content of methoxyls and consequently high gelling temperatures. On the contrary, the genus Gracilaria is considered the most important source of food and sugar-reactive grade agars. Among techniques which can be used to study algal polysaccharides, combined ¹H and ¹³C nuclear magnetic resonance spectroscopy represent the most effective and powerful method for the investigation of the chemical structure of agarocolloids.     

149 Distribution and Invasion of Bangia Atropurpurea (Rhodophyta) in the Laurentian Great Lakes

Müller et al. (1998) noted that freshwater collections of the genus Bangia formed a distinct group separate from marine entities in gene sequence analyses. Recently, the species epithet B. atropurpurea has been resurrected to represent this freshwater lineage. This taxon is one of many invasive species within the Laurentian Great Lakes. B. atropurpurea was first observed in Lake Erie in 1964 and by 1982 was observed in all of the Great lakes except Lake Superior. The present study was initiated to examine the further spread of B. atropurpurea and determine the origin of these populations. Hence, a survey of all the Great Lakes was conducted in 1995 (86 sites) and again in 2002 (104 sites). Bangia was observed at 43 sites in 1995 and 39 sites in 2002. For the first time, this alga has been observed to be present in the St. Lawrence River (1995), Georgian Bay on Lake Huron (2002) and Lake Simcoe (eastern shore, 2002) and hence this alga appears to be spreading into new locations. Cluster analyses of morphological data reveal three distinct groupings that do not separate according to location or lake basin. Preliminary analyses of ITS 1 and 2 sequences show differences among samples within Lake Ontario and among all Lakes; however, collections from Lake Simcoe are very similar in sequence. We are continuing to examine the relationship of Great Lakes populations with freshwater collections from Europe.     

THE UTILITY OF PROTEOMICS IN ALGAL TAXONOMY: BOSTRYCHIA RADICANS/B. MORITZIANA (RHODOMELACEAE,RHODOPHYTA) AS A MODEL STUDY1

A comparison of the proteome of eight genetically well‐characterized isolates of the Bostrychia radicans (Mont.) Mont./B. moritziana (Sond. ex Kütz.) J. Agardh species complex was undertaken to establish if genetic relationships among them can be determined using proteome data. Genetic distances were calculated on the basis of common and distinct spots in two‐dimensional gel electrophoresis (2‐DE). Proteomes of the male and female plants of each population were compared to analyze the range of genetic difference within an isolate. Haploid male and female plants of the same species had 3.7%–7.1% sex‐specific proteins. The degree of similarity of the proteome was consistent with previous DNA sequence data and sexual compatibility studies between the isolates. Two sexually compatible isolates from Venezuela showed a pair‐wise distance ranging from 0.14 to 0.21. The isolates from Mexico and Venezuela, which were partially compatible, showed a maximum pair‐wise distance of 0.26. A high level of genetic difference was found among isolates that were sexually incompatible. The isolate from Brazil was reproductively isolated from the Mexico and Venezuela isolates and showed a maximum pair‐wise distance of 0.65 and 0.58, respectively. Comparative proteomics may be helpful for studying genetic distances among algal samples, if intraisolate variation (gene expression) can be minimized or tested.     

Structure and properties of carrageenan-like polysaccharide from the red alga Tichocarpus crinitus (Gmel.) Rupr. (Rhodophyta, Tichocarpaceae)

Sulfated polysaccharides occurring in the red algae Tichocarpus crinitus cell wall were fractionated and purified. NMR and FT-IR spectroscopy analyses revealed that the non-gelling fraction contained a sulfated galactans having a new carrageenan-like structure. It is built with alternatively linked 1,3-linked β-D-galactopyranosyl-2,4-disulphates and 1,4-linked 3,6-anhydro-α-D-galactopyranosyl residues. Minor amounts of its biosynthetic precursor were detected in a water-extracted specimen. Brief analysis of rheological and biological properties of the non-gelling fraction was carried out. The carrageenan-like polysaccharide from T. crinitus displayed the properties of “random coil” polymer at high temperature, and possesses high anticoagulant activity at low concentration.     

CHEMICAL COMPOSITION OF THE CELL WALLS OF THE FRESHWATER RED ALGA LEMANEA ANNULATA (BATRACHOSPERMALES)1

Cell walls of the generic phase of the freshwater red alga Lemanea annulata Kütz were mechanically isolated and chemically characterized. Walls consisted mainly of polysaccharide with lesser quantities of associated protein and lipid. The major wall component was alkali-soluble xylan, comprised mainly of 4-linked β-xylopyranosyl residues and small amounts of 3-O-substituted β-xylopyranosyl residues. Hot water extracts yielded non-sulfated polymers, with 3- or 3,4-linked β-galactosyl residues alternating with 4-linked α-glucuronosyl residues as the predominant structural features. This acid polysaccharide shares many characteristics of the mucilage previously described from the freshwater genus Batrachospermum. Isolated cell walls of L. annulata contained approximately 6% cellulose. Cellobiohydrolase/colloidal gold labelling of cell walls revealed β-4-glycan throughout the fibrillar portion of the wall. Wall protein consisted of at least 17 amino acids, of which threonine and alanine were the most abundant. Polysaccharides of the cell walls of L. annulata differ from those of marine red algae and are similar to those described for other Batrachospermales.     

The galactan sulfate from the edible,red alga Porphyra columbina

A galactan sulfate has been isolated from the seaweed Porphyra columbina, and its structure established by a combination of methylation, methanolysis, treatment with alkali followed by methylation, and ¹³C-n.m.r. spectroscopy. The polysaccharide belongs to the porphyran class, and consists of 3-linked β-d-galactosyl residues and 4-linked α-l-galactosyl residues. 3,6-Anhydro-l-galactose and l-galactose 6-sulfate residues total approximately half of the sugar units, the other half being made up of d-galactose and 6-O-methyl-d-galactose residues. Some evidence is presented that suggests that the galactan sulfate does not have a completely alternating structure.     

Physiological responses of Caloglossa leprieurii (Ceramiales, Rhodophyta) to salinity stress

Marine, estuarine and freshwater isolates of Caloglossa leprieurii (Montagne) J. Agardh exhibit a high salinity tolerance, reflected by broad cell viability and growth rate. Osmotic adjustment is shown to rely to a large extent on ion-transport systems, with K⁺ and Cl- accumulated in osmotically- significant quantities and active Na⁺ extrusion. The ion concentrations contribute a large proportion (67–94%) to internal osmotic pressure. The concentration of the organic osmolyte mannitol in all populations was strongly salinity dependent. Mannitol made a lower contribution to the internal osmotic pressure, when compared to ion concentrations, but nonetheless represented an important proportion of the internal osmolality. The response of the three isolates is discussed in relation to the salinity of their respective environments.     

Fertilization and the cytoskeleton in the red alga Bostrychia moritziana (Rhodomelaceae,Rhodophyta)

Time-lapse videomicroscopy was used to observe the effects of various cytoskeletal inhibitors on three important fertilization events in Bostrychia moritziana: spermatial mitosis, gamete fusion (formation of a fertilization pore) and nuclear migration along the trichogyne. The microtubule inhibitor oryzalin disrupted spermatial mitosis but had no other effect on fertilization. The actin inhibitors, jasplakinolide, cytochalasin B, latrunculin A and B and mycalolide B inhibited gamete fusion while BDM, a myosin-disrupting drug, inhibited all three major fertilization events. FL-Phallacidin was used to stain actin filaments in spermatia and trichogynes while microtubules were labelled with antibodies at appropriate stages of fertilization. Microtubules were only evident during spermatial nuclear division. Actin filaments were present in both trichogynes and spermatia throughout fertilization; they formed a discrete ring around the fertilization pore and ensheathed male nuclei as the latter migrated into and along the trichogyne. These results suggest that the actin/myosin system plays a role in the events of fertilization.     

Random amplified polymorphic DNA (RAPD) identification of genetic variation in three species ofPorphyra (Bangiales,Rhodophyta)

The random amplified polymorphic DNA (RAPD) technique was used to characterize three species ofPorphyra from the western North Atlantic and adjacent Gulf of Mexico. Twenty 10-mer primers were screened for DNA amplification usingPorphyra template DNA. Nine of these oligonucleotide primers, all (G+C)-rich, were positive or band-producing, but yielded poor or variable band resolution. Subsequent use of the universal 20-mer M 13 primer resulted in both clear band resolution with a minimum of secondary bands and a high degree of reproducibility. Amplification products for DNA from six regional isolates ofPorphyra carolinensis Coll et Cox,P. leucosticta Thuret in Le Jolis andP. rosengurttii Coll et Cox were compared to each other and toBangia atropurpurea (Roth) C. Agardh. Results provide evidence of both genetically hetero- and homogeneous populations. Use of the RAPD method with the M 13 primer yields amplification products which can be used to fingerprint specific genotypes. This procedure could be used to discriminate between hetero- and homokaryotic fusion products from previously characterized donor strains.     

Fine‐Scale Spatial Patterns of Mature and Immature Filaments and Bioorganic Compounds of the Rhodophyte Bangia atropurpurea in the Supra‐ and Eulittoral Zones of a Large Lake1

Bangia atropurpurea (Mertens ex Roth) C. Agardh is a freshwater red alga species that is distributed worldwide. B. atropurpurea is highly adaptable due to its stress‐tolerance, which ensures survival under desiccation periods and under radiation extremes typical of the supra‐ and upper eulittoral zones. Whereas a number of previous investigations addressed some of the physiological and biochemical traits involved in stress‐tolerance, we studied the spatial arrangement of the mature (multiseriate) and immature (uniseriate) filaments and of selected bioorganic compounds along a gradient defined by distance from the waterline. Substantial physiological and biochemical differences were previously observed among phenological stages in the marine environment. In this study, we showed a nonrandom spatial structure of both phenological stages and photosynthetic pigments and photoprotective compounds, R‐phycocyanin and R‐phycoerythrin along the supralittoral‐eulittoral gradient. This observed pattern strongly suggests a complex interplay between physio‐morphological regulation and spatial arrangement of mature and immature filaments in conferring the typical stress tolerance of B. atropurpurea.