A Molecular Phylogenetic Study of the Tribe Corallineae (Corallinales,Rhodophyta) with an Assessment of Genus‐Level Taxonomic Features and Descriptions of Novel Genera

A multigene phylogeny using COI‐5P (mitochondrial cytochrome c oxidase subunit 1), psbA (PSII reaction center protein D1), and EF2 (elongation factor 2) sequence data for members of the tribe Corallineae was constructed to assess generic boundaries. We determined that traditional reliance on conceptacle position as an indicator of generic affinities in the Corallineae is not supported and taxonomic changes are required. We found that species currently assigned to Pseudolithophyllum muricatum resolved within the Corallineae in all analyses. This is the first record of crustose members in the subfamily Corallinoideae. Further‐more, the genus Serraticardia was polyphyletic; we propose to synonomize Serraticardia with Corallina, transfer the type species S. maxima to Corallina (C. maxima (Yendo) comb. nov.), and describe the new genus Johansenia for S. macmillanii (J. macmillanii (Yendo) comb. nov.). Our molecular data also indicate that species in the genus Marginisporum have evolutionary affinities among species of Corallina and these genera should also be synonymized. This necessitates the combinations C. aberrans (Yendo) comb. nov. for M. aberrans (Yendo) Johansen & Chihara, C. crassissima (Yendo) comb. nov. for M. crassissimum (Yendo) Ganesan, and C. declinata (Yendo) comb. nov. for M. declinata (Yendo) Ganesan. Corallina elongata was divergent from all other members of Corallina and is transferred to a new genus, Ellisolandia (E. elongata (J. Ellis & Solander) comb. nov). In addition, COI‐5P and internal transcribed spacer (ITS) data combined with morphological characters were used to establish that rather than the four Corallina species recognized in Canada, there are nine.     

Activities of Corallina (Corallinales) and other Rhodophyta polymers in the modulation of calcification

The effects of a range of fractions from calcareous and non-calcareous marine red algae on the crystal growth of calcite-seeded supersaturated calcium carbonate solutions were monitored by measuring the change in conductivity. An effective inhibitor of crystallization was a macromolecular fraction derived from the coralline red alga, Corallina officinalis, by a series of acid and alkali treatments. A preliminary characterization of the extract suggests the presence of a protein-polysaccharide complex. The possibility that this participates in the modulation of calcification in vivo is suggested.     

Genetic and morphological variation in an ecosystem engineer,Lithophyllum byssoides (Corallinales,Rhodophyta)

Lithophyllum byssoides is a common coralline alga in the intertidal zone of Mediterranean coasts, where it produces biogenic concretions housing a high algal and invertebrate biodiversity. This species is an ecosystem engineer and is considered a target for conservation efforts, but designing effective conservation strategies currently is impossible due to lack of information about its population structure. The morphological and molecular variation of L. byssoides was investigated using morphoanatomy and DNA sequences (psbA and cox2,3) obtained from populations at 15 localities on the Italian and Croatian coasts. Lithophyllum byssoides exhibited a high number of haplotypes (31 psbA haplotypes and 24 cox2,3 haplotypes) in the central Mediterranean. The psbA and cox2,3 phylogenies were congruent and showed seven lineages. For most of these clades, the distribution was limited to one or a few localities, but one of them (clade 7) was widespread across the central Mediterranean, spanning the main biogeographic boundaries recognized in this area. The central Mediterranean populations formed a lineage separate from Atlantic samples; psbA pair‐wise divergences suggested that recognition of Atlantic and Mediterranean L. byssoides as different species may be appropriate. The central Mediterranean haplotype patterns of L. byssoides were interpreted as resulting from past climatic events in the hydrogeological history of the Mediterranean Sea. The high haplotype diversity and the restricted spatial distribution of the seven lineages suggest that individual populations should be managed as independent units.     

Different speciation for bromine in brown and red algae,revealed by in vivo X‐ray absorption spectroscopic studies

Members of various algal lineages are known to be strong producers of atmospherically relevant halogen emissions, that is a consequence of their capability to store and metabolize halogens. This study uses a noninvasive, synchrotron‐based technique, X‐ray absorption spectroscopy, for addressing in vivo bromine speciation in the brown algae Ectocarpus siliculosus, Ascophyllum nodosum, and Fucus serratus, the red algae Gracilaria dura, G. gracilis, Chondrus crispus, Osmundea pinnatifida, Asparagopsis armata, Polysiphonia elongata, and Corallina officinalis, the diatom Thalassiosira rotula, the dinoflagellate Lingulodinium polyedrum and a natural phytoplankton sample. The results highlight a diversity of fundamentally different bromine storage modes: while most of the stramenopile representatives and the dinoflagellate store mostly bromide, there is evidence for Br incorporated in nonaromatic hydrocarbons in Thalassiosira. Red algae operate various organic bromine stores – including a possible precursor (by the haloform reaction) for bromoform in Asparagopsis and aromatically bound Br in Polysiphonia and Corallina. Large fractions of the bromine in the red algae G. dura and C. crispus and the brown alga F. serratus are present as Br^? defects in solid KCl, similar to what was reported earlier for Laminaria parts. These results are discussed according to different defensive strategies that are used within algal taxa to cope with biotic or abiotic stresses.     

The dodecameric vanadium-dependent haloperoxidase from the marine algae Corallina officinalis: cloning, expression, and refolding of the recombinant enzyme

The dodecameric vanadium-dependent bromoperoxidase from Corallina officinalis has been cloned and over-expressed in Escherichia coli. However, the enzyme was found to be predominantly in the form of inclusion bodies. This protein presents a challenging target for refolding, both due to the size (768 kDa) and quaternary structure (12 × 64 kDa). Successful refolding conditions have been established which result in an increase in the final yield of active bromoperoxidase from 0.5 mg to 40 mg per litre of culture. The refolded protein has been characterised and compared to the native enzyme and was shown to be stable at temperatures of 80 °C, over a pH range 5.5–10 and in organic solvents such as ethanol, acetonitrile, methanol, and acetone. The novel refolding approach reported in this paper opens up the full potential of this versatile enzyme for use in large scale biotransformation studies.     

Systematische Stellung und Vorkommen einer Corallinoidee (Corallinaceae,Cryptonemiales, Rhodophyceae) aus der Karibischen See: Corallina panizzoi nom. nov. et stat. nov.

Corallina officinalis L. ß fastigata Kütz. aus der Karibischen See wird mit Thalli von Corallina officinalis L. (einschließlich C. mediterranea Aresch.) verschiedener Herkünfte verglichen. Wegen anatomischer (Länge und Verhältnis Länge: Breite der Markzellen der Glieder) und morphologischer Unterschiede (Stellung der Konzeptakeln, Durchmesser der Thallusglieder, Ausbildung von Hörnchen an endständigen Konzeptakeln) werden die karibischen Pflanzen als zu einer eigenständigen Art gehörig angesehen, für die der Name Corallina panizzoi vorgeschlagen wird. C. panizzoi tritt an der venezolanischen und kolumbianischen Küste nur in Gebieten auf, in denen es wenigstens während der Trockenzeit zur Auftriebwasserbildung kommt. Weitere Funde liegen von Costa Rica und Puerto Rico vor. Die nördliche Verbreitungsgrenze von C. panizzoi erreicht offenbar nicht das Verbreitungsgebiet von C. officinalis. In Kolumbien wächst C. panizzoi im Eulitoral und im oberen Sublitoral bis in 3 m Tiefe.     

DISTRIBUTION AND RECRUITMENT OF SUBTIDAL GENICULATE CORALLINE ALAGE1

Geniculate coralline algae are common members of kelp forest communities. The structure provided by their stiff branches greatly influences the abundance and species composition of benthic animals and can affect associated algae by inhibiting recruitment, but the branches are themselves substrate for a large number of other taxa. However, other than qualitative observations, little is known about the within-site distribution, recruitment, and growth of these algae. We examined the distribution of the dominant corallines at a subtidal site in central California. Abundances of Calliarthron tuberculosum (Post. & Rupr.) Dawson, Bossiella californica ssp. schmittii (Manza) Johans., Calliarthron cheilosporioides Manza, Corallina vancouveriensis Yendo, and unidentifiable juveniles were determined at depths of 10, 15, and 20 m and on horizontal rock, vertical rock, and cobble. Calliarthron tuberculosum was most abundant (≤39% cover) at all depths, growing primarily on horizontal surfaces. Vertical surfaces and cobbles were dominated by B. californica ssp. schmittii (40 and 15% cover, respectively). These two most abundant species had the highest cover at 15 m. Calliarthron cheilosporioides and C. vancouveriensis were relatively rare (<1% cover) and generally grew on horizontal rocks and at shallower depths. Unidentified juveniles were also rare and occurred mainly on horizontal rocks and cobbles at 20 m. The settlement and growth rates of coralline crusts and the initiation and growth rates of young erect fronds from these crusts were determined in clearings made in the spring and fall at the three depths. Crust densities and diameters were highest at 10 m and in spring clearings. Settlement and growth tended to decrease with increasing depth. Trends were similar in fall clearings, but initial settlement was lower. Initiation and growth of fronds decreased with depth and were also higher in fall clearings. These variations in depth and substrata distribution, as well as settlement and growth, suggest there is considerable variation in the population biology between species in this group of subtidal plants.     

A comparison of the north-and southe european associations of Corallina officinalis L.

A comparison betweenCorallina officinalis associations from the northern Adriatic and the Icelandic coastal area is given regarding their floristic composition, zonal position and physionomy, as related to the environmental parameters and the different origins of the respective floras.     

Use of serially coupled capillary columns with different polarity of stationary phases for the separation of the natural complex volatile mixture of the marine red alga Corallina elongata

Separation of a complex of natural volatile compounds using serially coupled capillary columns with different polarity of stationary phases by gas chromatography–mass spectrometry from the medicinal marine red alga Corallina elongata is reported. Nearly 200 hydrocarbons, halogen compounds, fatty acids, and other metabolites were found. Using this gas chromatography procedure we demonstrate the successful separation of different volatile organic compounds.     

Phylogenetic relationships within the tribe Janieae (Corallinales,Rhodophyta) based on molecular and morphological data: a reappraisal of Jania1

Generic boundaries among the genera Cheilosporum, Haliptilon, and Jania—currently referred to the tribe Janieae (Corallinaceae, Corallinales, Rhodophyta)—were reassessed. Phylogenetic relationships among 42 corallinoidean taxa were determined based on 26 anatomical characters and nuclear SSU rDNA sequence data for 11 species (with two duplicate plants) referred to the tribe Corallineae and 15 species referred to the tribe Janieae (two species of Cheilosporum, seven of Haliptilon, and six of Jania, with five duplicate plants). Results from our approach were consistent with the hypothesis that the tribe Janieae is monophyletic. Our data indicate, however, that Jania and Haliptilon as currently delimited are not monophyletic, and that Cheilosporum should not be recognized as an independent genus within the Janieae. Our data resolved two well‐supported biogeographic clades for the included Janieae, an Indian‐Pacific clade and a temperate North Atlantic clade. Among anatomical characters, reproductive structures reflected the evolution of the Janieae. Based on our results, three genera, Cheilosporum, Haliptilon, and Jania, should be merged into a single genus, with Jania having nomenclatural priority. We therefore propose new combinations where necessary of some species previously included in Cheilosporum and Haliptilon.     

Molecular systematics,species limits,and diversification of the genus Dendrocolaptes (Aves: Furnariidae): Insights on biotic exchanges between dry and humid forest types in the Neotropics

The true diversity and interspecific limits in the Neotropical endemic avian genus Dendrocolaptes (Furnariidae) remain a highly controversial subject, with previous genus‐wide assessments, based mostly on morphological characters, producing poorly resolved phylogenies. The lack of well‐resolved, robust, and taxonomically densely sampled phylogenies for Dendrocolaptes prevents reliable inferences on the genus’ actual species diversity and evolutionary history. Here, we analyzed 2,741 base pairs of mitochondrial and nuclear genes from 43 specimens belonging to all species and the majority of subspecies described for Dendrocolaptes to evaluate species limits and reconstruct its diversification through time. Our phylogenies recovered a monophyletic Dendrocolaptes, with two main highly supported internal clades corresponding to the D. certhia and D. picumnus species complexes. Also, our analyses supported the monophyly of most Dendrocolaptes species recognized today, except D. picumnus, which was consistently recovered as paraphyletic with respect to D. hoffmannsi. A coalescent‐based test supported a total of 15 different lineages in Dendrocolaptes and indicated that the number of currently accepted species within the genus may be greatly underestimated. Particularly relevant, when combined with previous analyses based on plumage characters, comparative high levels of genetic differentiation and coalescent analyses support the recognition of D. picumnus transfasciatus as a full species that is already under threat. Ancestral area reconstructions suggest that diversification in Dendrocolaptes was centered in lowland Amazonia, with several independent dispersal events leading to differentiation into different adjacent dry and high elevation forest types throughout the Neotropics, mainly during the Middle and Late Pleistocene.     

Investigating seed bank potential of crustose coralline algae using DNA metabarcoding

To examine the potential for the autogenic ecosystem engineers, crustose coralline algae (CCA), to serve as seed banks or refugia for life stages of other species, it is critical to develop sampling protocols that reflect the diversity of life present. In this pilot study on two shallow water species of CCA collected from Raoul Island (Kermadec Islands; Rangitāhua) New Zealand, we investigated two preservation methods (ethanol vs. silica gel), sampled inner and outer regions of the crusts, and used DNA metabarcoding and seven genes/gene regions (16S rRNA, 18S rRNA, 23S rRNA, cox1, rbcL, and tufA genes and the ITS rRNA region) to develop a protocol for taxa identification. The results revealed immense diversity, with typically more taxa identified within the inner layers than the outer layers. As highlighted in other metabarcoding studies and in earlier work on rhodoliths (nodose coralline algae), reference databases are incomplete, and to some extent, the use of multiple markers mitigates this issue. Specifically, the 23S rRNA and rbcL genes are currently more suitable for identifying algae, while the cox1 gene fares better at capturing the diversity present inclusive of algae. Further investigation of these autogenic ecosystem engineers that likely act as marine seed banks is needed.     

Phylogeny and morphology of Nemalionopsis shawii (Thoreales,Rhodophyta), a new generic record from mainland China

The freshwater red alga Nemalionopsis shawii Skuja is first reported for mainland China from specimens collected in Guangdong and Yunnan Province. Morphological observations and molecular sequences of rbcL and cox1 genes were used to identify and analyze the phylogenetic position of the samples. Samples from China formed a monophyletic clade with other N. shawii samples from Japan, Indonesia, and Nepal with robust support values. The pairwise genetic distances for N. shawii between the samples from China and other samples were 0.2–1.5% and 1.0–2.4% for rbcL and cox1, respectively. Both male and female reproductive structures were observed in the specimens from Guangdong, but only monosporangia in the specimen from Yunnan. The samples from China increase the diversity of morphological measurements for N. shawii. The discovery of this genus in mainland China results in a new record of a freshwater red alga for this country.     

SCANNING ELECTRON MICROSCOPY OF CORALLINA AND HALIPTILON (CORALLINACEAE,RHODOPHYTA): SURFACE FEATURES AND THEIR TAXONOMIC IMPLICATIONS1

Scanning electron microscopy of intergenicula in members of the subfamily Corallinoideae reveals two distinctive surface morphologies: a Corallina-type (C-type) with round to irregular cell outlines and round trichocyte bases, and a Jania-type (J-type) with elongate, polygonal cell outlines and elongate trichocyte bases with excentric pores. The surface results from the calcified lateral walls of the epithallial cells projecting up from around collapsed protoplasts. Since J-type surfaces and trichocytes only occur in unequivocal members of the tribe Janieae—especially the genera Jania and Haliptilon, the presence of J-type surfaces in questionable members of Corallina reveals that they in fact belong to Haliptilon. Thus the two surface types clarify previously difficult taxonomic distinctions between Haliptilon and Corallina and allow identification to genus from purely vegetative material. Seventeen new combinations in Haliptilon are proposed. These results have considerable biogeographic implications with tropical species found to belong to Haliptilon, and Corallina sensu stricto being recognized primarily as a temperate and cold water genus.     

Exploring levels of egg parasitism and variation in egg cuticular chemistry in different Clavigralla spp.

Clavigralla spp. (Hemiptera: Coreidae) are major pests of cowpea (Vigna unguiculata (L.) Walp, Fabacae), common bean (Phaseolus vulgaris L., Fabacae) and pigeon pea (Cajanus cajan L., Fabacae) in Africa. Clavigralla spp. egg parasitoids, Gryon spp. (Hymenoptera: Scelionidae), have previously been reported as potential biological control candidates. Little is known about the parasitism levels and their potential relationship with cuticular chemistry of Clavigralla spp. The aims of this study were to determine parasitism levels of Clavigralla tomentosicollis Stål (Hemiptera: Coreidae) and C. elongata Signoret (Hemiptera: Coreidae) eggs, and to explore the relationship between egg parasitism and egg cuticular chemistry. High parasitism levels were determined for C. tomentosicollis by collecting eggs from plants in mono‐cropping and multi‐cropping systems in farmers’ fields in Bénin and Kenya between April and June 2016. Three species of Clavigralla were recorded: C. tomentosicollis, C. shadabi and C. elongata. Clavigralla tomentosicollis was the most common in both countries, while C. shadabi and C. elongata were only collected in Bénin and Kenya, respectively. An egg parasitoid (Gryon sp.) was recovered from egg batches collected from both countries. In parasitism assays using Gryon sp., the incidence of parasitism was higher in C. tomentosicollis eggs than that of C. elongata. Chemical analysis by coupled gas chromatography/mass spectrometry (GC/MS) of cuticular extracts obtained from C. tomentosicollis and C. elongata eggs identified fifteen compounds including ten alkanes of which the amounts varied between the two species. We speculate that Clavigralla spp. cuticular chemistry may serve as potential host location cues for Gryon sp.     

Screening and evaluation of different algal extracts and prospects for controlling the disease vector mosquito Culex pipiens L.

Continual application of synthetic insecticides in controlling mosquito larvae has resulted in several problems as build-up of mosquito resistance beside to negative impacts on human health and environment. Discovering new and affordable bio-insecticidal agents with high efficiency, cost effective and target specific become a crucial need. The current study assessed the larvicidal activity of eight methanolic algal extracts belong to three different algal divisions against the 3^rd larval instar of Culex pipiens L. (Diptera: Culicidae). Comparative studies showed that four species of red and green algal extracts exhibited good larvicidal activity. Galaxaura elongata and Jania rubens (Rhodophyta), Codium tomentosum and Ulva intestinales (Chlorophyta) showed higher larvicidal potencies than Padina boryana, Dictyota dichotoma, and Sargassum dentifolium (Phaeophyta) and Gelidium latifolium (Rhodophyta). The maximum level of toxicity was achieved by exposure to G. elongata extract with LC₅₀ (31.13 ppm), followed by C. tomentosum (69.85 ppm) then J. rubens (84.82 ppm) and U. intestinalis (97.54 ppm), while the lowest toxicity exhibited by G. latifolium (297.38 ppm) at 72 h post- treatment. The application of LC₅₀ values of G. elongate, J. rubens, C. tomentosum, and U. intestinalis extracts affected the activities of antioxidant enzymes viz. superoxide dismutase, catalase and glutathione peroxidase as oxidative stress markers. An increase of antioxidant enzymes activities was recorded. Therefore, a significant elimination of free radicals, causing toxic effects. Overall, this study casts light on the insecticidal activity of some algal extracts, suggesting the possibility of application of these bio- agents as novel and cost- effective larvicides.     

Patterns of macroalgal diversity, community composition and long-term changes along the Swedish west coast

This study analyses the complicated patterns of vertical distribution of the macroalgal vegetation in an area where brackish and marine waters meet and mix. Variables used to record vegetation characteristics are algal cover, species composition and diversity. The data set includes 64 diving profiles, all from sites exposed to wave action, along a ca. 260 km long coastline. The profiles belong to four categories: coastal sites in the Skagerrak (more marine), coastal sites in the Kattegat (more brackish), coastal sites in the Kattegat after a toxic phytoplankton bloom, and submerged offshore stone reefs in the Kattegat. The highest species diversity was found at the reefs, which are not affected directly by land runoff. At the reefs the 18 most common perennial species penetrate 2–11 m (on average 5.5 m) deeper than at the coastal sites. The virtual absence of sedimentation, and thus the availability of substratum, at the reefs may explain the differences so that the lower limit for the algae is determined by light penetration or by recruitment problems caused by strong currents at the reefs, whereas sedimentation limits the settlement of algae in coastal sites. Ordination analysis based on species composition reveals that the major environmental gradients structuring the algal vegetation in the Kattegat and the Skagerrak are salinity and water depth. The large data set of this study made it possible to quantify the downward dislocation of Atlantic intertidal species to the sublittoral along the Swedish west coast. For example, the mean upper limit of Corallina officinalis is 2 m in the Skagerrak but 12.5 m in the Kattegat and the mean occurrence interval of Fucus serratus is 0.9–2.7 m in the Skagerrak, but 1.1–6.3 m in the Kattegat. This downward dislocation is suggested to be the result of decreased competition when species successively disappear with lower salinity. Comparisons of the present study's results with those of previous investigations show that eight common red algal species have moved upwards compared to the situation before the large-scale eutrophication started in the 1960s, e.g. Cystoclonium purpureum and Polysiphonia elongata by ca. 8 m, Phycodrys rubens and Delesseria sanguinea by ca. 5 m. A toxic phytoplankton bloom affected macroalgal community composition on the whole only slightly, but it had a negative effect on algal cover and species richness below a water depth of ca. 5 m, the algae were visibly damaged and the lower vegetation limit temporarily moved upwards.     

Design of hybrid PVA–CA–Jania rubens biomatrix for removal of lead

Polyvinyl alcohol–sodium alginate (PVA–SA) matrix was fabricated and red algae Jania rubens was embedded for removal of lead from aqueous solutions. The Pb(II) uptake rate was rapid primarily at 1 h and equilibrium was achieved within 2 h. The optimum pH was 5, the data were well fitted by Langmuir and Freundlich models, and R_L values are in the range of 0.1–0.38. The sorption capacity (q_e) of PVA–calcium alginate (CA)–J. rubens matrix increased from 10.77 to 37.195 mg g^?1 with increasing Pb(II) concentration from 24.86 to 98.75 mg L^?1 at the temperature of 30°C and pH 5. The sorption capacity (q_e) and maximum biosorption (q_m) were noted as 37.179 ± 0.32 and 71.43 mg/g, respectively. The adsorption process was well described by pseudo-second-order model. The reaction is endothermic, is spontaneous, and increases in randomness. The functional groups present on matrix, i.e., –OH, –C–N, –C–O,–CO–NH, –NH₂, –SH, and –C–OH, were intensely involved in the process. Scanning electron microscopy results revealed the morphological changes due to adsorption of Pb(II) on and inside of PVA–CA–J. rubens matrix. Desorption study indicates the efficient regeneration of PVA–CA–J. rubens biomass matrix for three cycles and is a promising matrix for removal of Pb(II) and can be used in continuous systems.     

THE INFLUENCE OF SEA ANEMONES ON THE MORPHOLOGY AND PRODUCTIVITY OF TWO INTERTIDAL SEAWEEDS1

The intertidal seaweeds Corallina vancouveriensis Yendo and Gelidium coulteri Harv. respond to the experimental removal of associated mats of the clonal anemone Anthopleura elegantissima Brandt with morphological changes that affect their photosynthetic performances. With the removal of anemones, these seaweeds develop a tight compaction of thalli which aids in the retention of moisture during aerial exposure. The morphological response, however, results in greatly reduced net photosynthesis in C. vancouveriensis (–98%) and G. coulteri (–73%) on an areal basis. Association with the clonal anemones allows these seaweeds to maintain a morphology that is more highly productive yet inconsistent with the exposure stresses in many microhabitats at similar tidal levels.