POPULATION STRUCTURE AND PHYSIOLOGICAL DIFFERENTIATION OF HAPLOTYPES OF CALOGLOSSA LEPRIEURII (RHODOPHYTA) IN A MANGROVE INTERTIDAL ZONE

Zonation of macroalgae in the intertidal zone has been well documented. However, studies of zonation of macroalgae have predominantly examined the distribution of different species rather than the distribution of variants within a species. This study investigated the spatial variation of plastid haplotypes of the mangrove red alga Caloglossa leprieurii (Montagne) J. Agardh at a site in eastern Australia and tests for physiological differences (growth, photosynthesis) between those haplotypes. RUBISCO spacer plastid haplotypes were scored using single-stranded comformational polymorphism, and the population structure at two sites was examined using a nested sampling design comparing between sites, among transects within sites, and among quadrats within transects. Growth rates at various salinities and light intensities and the photosynthesis–irradiance curves of the three main haplotypes were compared. The two sites showed a high degree of genetic differentiation across a short distance, suggesting limited gene flow. The distribution of haplotypes was patchy and did not reflect a zonation pattern along the intertidal gradient. The three haplotypes were physiologically differentiated with haplotype A, with a lower growth rate and a lower photosynthetic efficiency at higher light intensities. There is some evidence of physiological differentiation between life history phases in C. leprieurii with sporophytes having a higher growth rate than females under most conditions. Our results suggest a correlation between our culture results and our population data. Haplotypes (haplotype A) and life history phases (gametophytes) with lower performance (growth and photosynthetic efficiency) under our culture conditions were correlated with a minor representation in the field. This is the first study to integrate population-level data with physiological parameters toward an understanding of the distribution and relative abundance of red algal genetic variants.     

A morphological study and hybridization analysis of Caloglossa leprieurii (Ceramiales, Rhodophyta) from Japan, Singapore and Australia

Morphological and hybridization experiments were performed on Caloglossa leprieurii (Montagne) J. Agardh collected from Japan, Singapore and Australia in order to evaluate taxonomic characters of this species. Within C. leprieurii at least four mating groups were recognized from the Indo-Pacific region. These mating groups could be characterized by the blade width at the inter-node and the cell-row numbers on the opposite side derived from the first axial cell at the main axis, though these properties showed a certain variability even in the same plant under both field and culture conditions. The phylogenetic relationship and geographic distribution of the four mating groups are discussed.     

Caloglossa intermedia, sp. nov. (Rhodophyta) from the western atlantic coast: molecular and morphological analyses with special reference to C. leprieurii and C. monosticha

Caloglossa intermedia , sp. nov. is described from estuaries and salt marshes of Atlantic North America. This species is related to C. leprieurii (Montagne) G. Martens based on the position of rhizoids, whereas it is more similar to C. monosticha Kamiya in the number of cell rows at the nodes. Rhizoidal development of this alga also shows an intermediate characteristic between C. leprieurii and C. monosticha. Crosses were successful between the strains of C. intermedia from Georgia and South Carolina, but neither strain was interfertile with morphologically related species. C. intermedia from Georgia, New Jersey, and South Carolina had identical DNA sequences in the Rubisco spacer and flanking regions of rbc L and rbc S. This alga was more closely related to C. leprieurii in a molecular phylogenetic analysis. The evolution of each morphological character is discussed.     

Studies on the mechanism of light-dependent germination of akinetes of blue-green algae

This paper deals with the role of light in the germination of akinetes of Anabaena azollae. The two maxima action spectra are situated at 385 and 615 nm and the stimulation of the germination process by photosynthate was confirmed. The photoreceptor absorbing at 385 nm was identified as a flavin and that at 615 nm as a phytochrome. A model is suggested for the mode of action of light in the germination of akinetes of blue-green algae.C. Tsui     

The morphology and taxonomy of Womersleya monanthos, an endemic species and genus of Delesseriaceae (Ceramiales, Rhodophyta) from southeastern Australia

Womersleya monanthos (J. Agardh) Papenfuss is typically an epiphyte of larger brown and red algae that are common in drift along the southeastern coasts of Australia. A hitherto little-known member of the Phycodrys group of the Nitophylloideae, its reproductive features have been studied in detail and its taxonomic position clarified. Blades are polystromatic throughout and lack veins or nerves, with blades originating from apical cells of primary and second-order cell rows. Intercalary cell divisions take place in primary cell rows and all other branch orders, with third-order laterals arising both abaxially and adaxially on cells of second-order rows. Fertile central cells bear procarps on pericentral cells on both sides of the blade, the procarps consisting of two 4-celled carpogonial branches and a single central group of sterile cells that enlarge and persist at the distal end of a bicampanulate fusion cell at maturity. Spermatangia and tetrasporangia form in circular subapical sori on both sides of the blade or in marginal lobes or proliferations. After comparing it to other members of the Phycodrys group, we conclude that Womersleya is a monotypic genus well distinguished from other genera and with probable closest affinities to the Northern Hemisphere Polyneura, Erythroglossum and Sorella, as well as the Australian endemic, Crassilingua.     

The influence of daylength,light intensity and temperature on the growth rates of planktonic blue-green algae

The in vitro growth rates under continuous light of the four dominant blue-green algae in Lough Neagh, Anabaena flos-aquae Bréb., Aphanizomenon flos-aquae Ralfs fa. gracile Lemm., Oscillatoria agardhii Gom. and Oscillatoria redekei van Goor were slower than in situ rates from Lough Neagh that had been corrected for hours of light received by the algae. However, by culturing on a 6: 18 light-dark cycle in vitro growth rates were obtained that were similar to the in situ rates. Under continuous light small species showed the fastest growth with Oscillatoria redekei the dominant species. However, this pattern was almost completely reversed under the light-dark cycle with Oscillatoria redekei only exhibiting the fastest growth rate under low light conditions. This observation showed agreement with Lough Neagh field data which showed that Oscillatoria redekei reached its maximum crop in April while the other three species were dominant during the summer months. Compared to the generally assumed high thermal tendency of blue-green algae the temperature maxima of the four species were low. No growth was observed at 35°C for any species while Anabaena flos-aquae was severely inhibited at 25°C.     

Erythroglossum latum sp. nov. (Delesseriaceae, Rhodophyta) from the east coast of Honshu, Japan

A new species, Erythroglossum latum, is described based on specimens from Fukushima Prefecture along the east coast of Honshu, Japan. This species differs from Erythroglossum pinnatum Okamura in having a broader main blade, a clear midrib and lateral branches that are smaller in size and not clearly constricted at the base.     

Polymorphic molecular markers from anonymous nuclear DNA for genetic analysis of populations

A simple method is presented for developing polymorphic, anonymous DNA markers suitable for population genetic studies. Anonymous DNA fragments are screened for sequence variability using a common mutation detection technique (single strand conformation polymorphism analysis; SSCP) and locus-specific PCR primers are designed for polymorphic DNA fragments. Detection of the markers by SSCP analysis coupled with sequence analysis of SSCP variants allows rapid screening while retaining information about the genealogical relationship among alleles. Variability detected for six markers was assessed in rainbow trout Oncorhynchus mykiss and was compared with variability detected by similar analysis of intron loci. Between three and 12 distinct alleles were observed at each marker locus, and average within-population heterozygosity ranged from 0.12 to 0.44. Advantages and limitations of the methodology for population genetic analysis are discussed.     

The occurrence of the hydrogenase in some blue-green algae

Several blue-green algae were surveyed for the occurrence of the hydrogenase which was assayed by the oxyhydrogen or Knallgas reaction in the intact organisms. In aerobically grown cultures, the reaction was detectable in Anabaena cylindrica, Nostoc muscorum and in two Anabaena variabilis species, whereas virtually no activity was observed in Anacystis nidulans and Cyanophora paradoxa. In these latter two algae, the reaction was, however, found after growth under molecular hydrogen for several days, which drastically increased the activity levels with all the algae tested. In the nitrogen fixing species, the activity of the Knallgas reaction was enhanced when all combined nitrogen was omitted from the media. H₂ and hydrogenase could not significantly support the CO₂-fixation in photoreduction experiments with all blue-green algae investigated here. Hydrogenase was assayed by the dithionite and methyl viologen dependent evolution of hydrogen and was found to be present with essentially the same specific activity levels in preparations of both heterocysts and vegetative cells from Anabaena cylindrica. Na₂S₂O₄ as well as H₂ supported the C₂H₂-reduction of the isolated heterocysts. The H₂-dependent C₂H₂-reduction did not require the presence of oxygen but was strictly light-dependent where H₂ served as an electron donor to photosystem I of these cells. It is concluded that hydrogen can be utilized by two different pathways in blue-green algae.Abbreviations Chl chlrophyll - CP creatine phosphate - CP kinase creatine phosphokinase - DCMU N-(3,4-dichlorophenyl)N,N-dimethylurea     

Sorellocolax stellaris gen. et sp. nov., a hemiparasitic alga (Delesseriaceae, Rhodophyta) from the east coast of Honshu, Japan

A hemiparasitic alga, Sorellocotax stellaris sp. nov. is described growing on plants of Sorella repens collected from Onagawa, Miyagi Prefecture, east coast of Honshu, Japan. The thallus is small, up to 2 mm high, once or twice branched from the margin. The growing apex has a transversely dividing apical cell, and intercalary cell divisions occur in the cells of first-order rows. Tetrasporangia are cut off from the cells of the inner cortex, The procarp is composed of a supporting cell, one group of sterile cells and two carpogonial branches. Carposporangia are borne in short chains.     

Screening of DNA polymorphisms in samples of archived scales from New Zealand snapper

Primers and protocols were developed to screen effciently DNA sequence polymorphism of nuclear and mitochondrial loci in samples of dried archived scales collected over the last 50 years from the New Zealand snapper Pagrus auratus (Sparidae).     

The evolutionary history of grey wolf Y chromosomes

Analyses of Y chromosome haplotypes uniquely provide a paternal picture of evolutionary histories and offer a very useful contrast to studies based on maternally inherited mitochondrial DNA (mtDNA). Here we used a bioinformatic approach based on comparison of male and female sequence coverage to identify 4.7 Mb from the grey wolf (Canis lupis) Y chromosome, probably representing most of the male‐specific, nonampliconic sequence from the euchromatic part of the chromosome. We characterized this sequence and then identified ≈1,500 Y‐linked single nucleotide polymorphisms in a sample of 145 resequenced male wolves, including 75 Finnish wolf genomes newly sequenced in this study, and in 24 dogs and eight other canids. We found 53 Y chromosome haplotypes, of which 26 were seen in grey wolves, that clustered in four major haplogroups. All four haplogroups were represented in samples of Finnish wolves, showing that haplogroup lineages were not partitioned on a continental scale. However, regional population structure was indicated because individual haplotypes were never shared between geographically distant areas, and genetically similar haplotypes were only found within the same geographical region. The deepest split between grey wolf haplogroups was estimated to have occurred 125,000 years ago, which is considerably older than recent estimates of the time of divergence of wolf populations. The distribution of dogs in a phylogenetic tree of Y chromosome haplotypes supports multiple domestication events, or wolf paternal introgression, starting 29,000 years ago. We also addressed the disputed origin of a recently founded population of Scandinavian wolves and observed that founding as well as most recent immigrant haplotypes were present in the neighbouring Finnish population, but not in sequenced wolves from elsewhere in the world, or in dogs.     

Genetic structure of brown trout (Salmo trutta, L.) populations from south-western France: data from mitochondrial control region variability

The genetic relationships between several French Atlantic populations of brown trout were analysed using polymerase chain reaction-single-strand conformation polymorphism and sequencing of two parts of the mitochondrial control region. Ten mitochondrial haplotypes were identified, separated by a small number of mutations. The distribution of these haplotypes confirmed our hypothesis of the existence of two genetically well-differentiated groups of populations in this area, which has already been suggested using nuclear markers. Nuclear and mitochondrial markers, however, show different patterns of genetic differentiation: some populations which belong to one group according to allozyme results appear to be part of the other when using mitochondrial DNA. The origin of these different forms is discussed; they seem to belong to the Atlantic grouping. Nevertheless, the hypothesis of an ancestral and a modern form suggested for northern populations does not appear valid in this particular case.     

SSCP is not so difficult: the application and utility of single-stranded conformation polymorphism in evolutionary biology and molecular ecology

All genetic markers are estimators of DNA nucleotide sequence variation. Rather than obtaining DNA sequence data, it is cheaper and faster to use techniques that estimate sequence variation, although this usually results in the loss of some information. SSCP (single-stranded conformation polymorphism) offers a sensitive but inexpensive, rapid, and convenient method for determining which DNA samples in a set differ in sequence, so that only an informative subset need be sequenced. In short, most DNA sequence variation can be detected with relatively little sequencing. SSCP has been widely applied in medical diagnosis, yet few studies have been published in population genetics. The utility and convenience of SSCP is far from fully appreciated by molecular population biologists. We hope to help redress this by illustrating the application of a single simple SSCP protocol to mitochondrial genes, nuclear introns, microsatellites, and anonymous nuclear sequences, in a range of vertebrates and invertebrates.     

Phylogeography of the endangered Cathaya argyrophylla (Pinaceae) inferred from sequence variation of mitochondrial and nuclear DNA

Cathaya argyrophylla is an endangered conifer restricted to subtropical mountains of China. To study phylogeographical pattern and demographic history of C. argyrophylla, species-wide genetic variation was investigated using sequences of maternally inherited mtDNA and biparentally inherited nuclear DNA. Of 15 populations sampled from all four distinct regions, only three mitotypes were detected at two loci, without single region having a mixed composition (G(ST) = 1). Average nucleotide diversity (theta(ws) = 0.0024; pi(s) = 0.0029) across eight nuclear loci is significantly lower than those found for other conifers (theta(ws) = 0.003 approximately 0.015; pi(s) = 0.002 approximately 0.012) based on estimates of multiple loci. Because of its highest diversity among the eight nuclear loci and evolving neutrally, one locus (2009) was further used for phylogeographical studies and eight haplotypes resulting from 12 polymorphic sites were obtained from 98 individuals. All the four distinct regions had at least four haplotypes, with the Dalou region (DL) having the highest diversity and the Bamian region (BM) the lowest, paralleling the result of the eight nuclear loci. An AMOVA revealed significant proportion of diversity attributable to differences among regions (13.4%) and among populations within regions (8.9%). F(ST) analysis also indicated significantly high differentiation among populations (F(ST) = 0.22) and between regions (F(ST) = 0.12-0.38). Non-overlapping distribution of mitotypes and high genetic differentiation among the distinct geographical groups suggest the existence of at least four separate glacial refugia. Based on network and mismatch distribution analyses, we do not find evidence of long distance dispersal and population expansion in C. argyrophylla. Ex situ conservation and artificial crossing are recommended for the management of this endangered species.     

Phycodrys valentinae sp. nov. (Delesseriaceae, Rhodophyta) with Discussion of Other Species of the Phycodrys Genus from the North Pacific

A new species Phycodrys valentinae Seliv. et Zhigad. from the northwest of the Bering Sea, distinct from other species of the genus Phycodrys by the presence of a distromatic lamina and by the arrangement of basal generative prolifications, was described. Based on the analysis of the morphological and anatomic traits of species of the genus Phycodrys, P. serratiloba (Rupr.) A. Zin., which was considered as synonym of P. riggii Gardn. to the present [27, 28, 30], was reinstated as an independent species. The taxonomic status of other representatives of the genus Phycodrys of the studied region is considered. In our opinion, five species of the genus Phycodrys (P. serratiloba, P. riggii, P. amchitkensis, P. vinogradovae, and P. valentinae) grow in the seas of the Russian Far East.     

Genetic variation within and among North Atlantic and Baltic populations of the benthic alga Phycodrys rubens (Rhodophyta)

Genetic variation was examined within and among North Atlantic, North Sea and Baltic populations of the benthic red alga Phycodrys rubens using allozymes and random amplified polymorphic DNA (RAPD) markers. On western and eastern North Atlantic coasts distinct allozyme types were found, with the exception of western Newfoundland where East and West Atlantic types co-occur. Along the European coasts, two genetic groups were distinguished by fixed allelic differences: an outer oceanic group and a North Sea/Baltic group. The two genetic types co-occur in the Skagerrak and Kattegat region. Reproductive isolation between the two types is suggested by the lack of hybrids in the overlap zones, and they may therefore represent sibling species. Unexpectedly, an analysis of RAPD variation was unable to recover the two cryptic species identified using allozymes. Within-population RAPD variation was similar to or greater than between-population variation. The lack of structure in the RAPD data cannot be attributed solely to technical artefacts of the method but appears to reflect real biological variability. Within-population genomic polymorphisms caused by frequent mutational events are discussed, as are high amounts of genetic drift and possible disruptive selection brought about by stressed habitats. Finally, Baltic and extra-Baltic salinity ecotypes are known to exist in P. rubens. However, no correlation between ecotypic variation and allozyme groups was detected.     

The electrophoretic isolation and partial characterization of three chlorophyll-protein complexes from blue-green algae

Three chlorophyll-protein complexes have been resolved from blue-green algae using an improved procedure for membrane solubilization and electrophoretic fractionation. One complex has a red absorbance maximum of 676 nm and a molecular weight equivalency of 255 000 ± 15 000. A second complex has an absorbance maximum of 676 nm, a molecular weight equivalency of 118 000 ± 8000, and resembles the previously described P-700-chlorophylla-protein (CPI) of higher plants and algae. The third chlorophyll-protein has a red absorbance maximum of 671 nm and a molecular weight equivalency of 58 000 ± 5000. Blue-green algal membrane fractions enriched in Photosystem I and heterocyst cells do not contain this third chlorophyll-protein, whereas Photosystem II-enriched membrane fractions and vegetative cells do. A component of the same spectral characteristics and molecular weight equivalency was also observed in chlorophyll b-deficient mutants of barley and maize. It is hypothesized that this third complex is involved in some manner with Photosystem II.