NORTHERN AND SOUTHERN HEMISPHERE HYBRIDS OF MACROCYSTIS (PHAEOPHYCEAE)1

All combinations of individuals of Macrocystis pyrifera (L.) C. Agardh, M. integrifolia Bory, and M. angustifolia Bory hybridized. Gametophyte isolates obtained from 18 individuals were used, including M. pyrifera and M. integrifolia from the extremes of their Northern Hemisphere ranges along the Pacific Coast of North America and M. pyrifera and M. angustifolia from Tasmania, Australia. All combinations of gametophytes produced sporophytes of normal morphology, with the exception of crosses involving three gametophyte isolates. One female (M. integrifolia) and two male (M. pyrifera and M. angustifolia)gametophyte isolates were unable to produce normal sporophytes in combination with gametophytes of the opposite sex. Some cultures of female gametophytes produced abnormally shaped parthenogenetic sporophytes. Gametophytes and sporangia of M. pyrifera had n= 16 chromosomes. The M. integrifolia female gametophyte that was unable to produce normal sporophytes had n = ca. 32 chromosomes. These results show that these species of Macrocystis have not become reproductively isolated. Although these species may be considered conspecific according to the biological species concept, we recommend that they continue to be recognized as separate species based on morphological differences.     

EFFECT OF HIGH IRRADIANCE ON RECRUITMENT OF THE GIANT KELP MACROCYSTIS (PHAEOPHYTA) IN SHALLOW WATER1

Laboratory and field experiments were done hi Still-water Cove, Carmel Bay, California, and Monterey Harbor, California, to determine the effect of photosynthetically active radiation (PAR) on the shallow (upper) limit of giant kelp, Macrocystis pyrifera (L.) C. Agardh. At shallow depths, M. pyrifera did not recruit or grow to macroscopic size from gametophytes or embryonic sporophytes transplanted to vertical buoy lines; sharp decreases in PAR with depth coincided with observed recruitment and sporophyte distributions. Shade manipulations indicated that settlement of M. pyrifera zoospores was decreased, but not prohibited, by high PAR. Postsettlement stages (gametophytes and embryonic sporophytes), however, survived only under shade. These results suggest that high PAR can inhibit the recruitment of M. pyrifera to shallow water by killing its postsettlement stages; whether or not ultraviolet (UV) radiation also inhibits recruitment was not tested. In either case, however, it appears that high irradiance (PAR and/or UV) regulates the shallow limit of M. pyrifera prior to temperature and desiccation stresses inherent to intertidal regions. In an additional experiment, recruitment or growth of transplanted gametophytes or embryonic sporophytes of Macrocystis integrifolia Bory also did not occur at shallow depths, suggesting that this shallow water species accesses high irradiance regions via a method other than sexual reproduction.     

Effects of chelated iron on oogenesis and vegetative growth of kelp gametophytes (Phaeophyceae)

The supply of iron has been reported to affect gametogenesis in the gametophytes of some species of kelps (order Laminariales). Spores of the kelps Alaria marginata Postels & Ruprecht, Dictyoneurum californicum Ruprecht, Egregia menziesii (Turner) Areschoug, Laminaria setchellii Silva, and Macrocystis pyrifera (L.) C. Agardh were cultured in enriched seawater with and without added chelated iron (Fe‐ethylenediaminetetraacetate) to determine the effects of iron on oogenesis. All species showed a decrease in oogenesis without added Fe‐ethylenediaminetetraacetic acid (EDTA). Gametophytes of E. menziesii showed predominant gametogenesis with or without supplied iron, resulting in all cells being converted to gametes so that vegetative growth did not continue. Vegetative gametophytes were obtained in the other species used. Gametophytes of M. pyrifera did not show any oogenesis without added Fe‐EDTA, while those of L. setchellii, A. marginata and D. californicum were intermediate in their response, showing some gametogenesis without added Fe‐EDTA. When Fe‐EDTA supply was delayed by 6, 13 and 20 days with spores of M. pyrifera, the gametophytes produced fewer eggs, with a greater decrease as the delay grew longer. A range of Fe‐EDTA concentrations was investigated using isolated female gametophytes of two strains of M. pyrifera and one of Macrocystis integrifolia Bory. None of these three strains produced gametes without the addition of Fe‐EDTA. Gametophytes of M. integrifolia required the least amount of added Fe‐EDTA to achieve gametogenesis while gametophytes of M. pyrifera required higher amounts, with the two strains showing somewhat different responses. Iron nutrition appears to be an essential factor for gametogenesis in several species of kelps.     

Nuclear DNA level and life cycle of kelps: Evidence for sex‐specific polyteny in Macrocystis (Laminariales,Phaeophyceae)

Giant kelp, Macrocystis pyrifera (Linnaeus) C. Agardh, is the subject of intense breeding studies for marine biomass production and conservation of natural resources. In this context, six gametophyte pairs and a sporophyte offspring of Macrocystis from South America were analyzed by flow cytometry. Minimum relative DNA content per cell (1C) was found in five males. Unexpectedly, nuclei of all female gametophytes contained approximately double the DNA content (2C) of males; the male gametophyte from one locality also contained 2C, likely a spontaneous natural diploid variant. The results illustrate a sex‐specific difference in nuclear DNA content among Macrocystis gametophytes, with the chromosomes of the females in a polytenic condition. This correlates with significantly larger cell sizes in female gametophytes compared to males and resource allocation in oogamous reproduction. The results provide key information for the interpretation of DNA measurements in kelp life cycle stages and prompt further research on the regulation of the cell cycle, metabolic activity, sex determination, and sporophyte development.     

DEFINING TAXON BOUNDARIES IN MEMBERS OF THE MORPHOLOGICALLY AND GENETICALLY PLASTIC GENES CAULERPA (CAULERPALES,CHLOROPHYTA)1

Numerous attempts to capture the morphological variability of the genus Caulerpa have resulted in an unstable classification of numerous varieties and formae. In the present study we attempted to test taxon boundaries by investigating morphological and genetic variation within and between seven taxa of Caulerpa, supposedly belonging to four species, sampled at different sites in a Philippine reef system. Using both field and culture observations, we described the relation between the variability of a set of morphological characters and ecological parameters, such as wave exposure, light intensity, and substrate type. Statistical analyses showed that the limits between two (out of three) ecads of the C. racemosa (Forsskål) J. Agardh complex were obscured by the presence of morphological plasticity. Other studied taxa of Caulerpa (i.e. C. cupressoides [Vahl] C. Agardh, C. serrulata [Forsskål] J. Agardh, and two formae of C. sertularioides [S. Gmelin] Howe) could be grouped based on morphology despite the presence of morphological plasticity. Our results indicated a strong association between light intensity and several quantitative morphological variables. Genetic diversity of these taxa was assessed by partial sequencing chloroplast rbcL and tufA genes and the ycf10‐chlB chloroplast spacer. In all phylogenetic analyses, C. serrulata, C. cupressoides, C. sertularioides, and the three ecads of C. racemosa emerged as distinct genetic units. Despite the presence of morphological plasticity and morphological convergence, a subset of morphological characters traditionally used in taxonomic delimitation still had sufficient discriminative power to recognize the terminal phylogenetic clades.     

PURIFICATION OF SPECIES SPECIFIC ANTIBODIES TO CARBOHYDRATE COMPONENTS OF MACROCYSTIS PYRIFERA (PHAEOPHYTA)1

Polyclonal rabbit antibodies to cell wall components were produced against gametophytes of the giant kelp Macrocystis pyrifera (Linnaeus) C. Agardh. These antibodies were found to react with carbohydrates extracted from M. pyrifera and Pterygophora californica Ruprecht by carbohydrate based enzyme immunoassay (EIA). The antibodies reacted with carbohydrates from both species. After affinity purification on a column with M. pyrifera carbohydrate coupled to AH-Sepharose, the eluted antibody was specific for M. pyrifera carbohydrate with little cross reactivity to P. californica carbohydrate in the EIA test. In experiments carried out to characterize the antigenic specificity of unfractionated antibody using commercially prepared carbohydrates in the EIA, the antibodies were shown to react primarily with fucoidan and to a lesser degree, alginate. The unfractionated antibody was also shown to bind to proteins from both M. pyrifera and P. californica. These results indicate that species specific carbohydrate determinants may be present in the kelp cell wall.     

Molecular phylogeny and timing of radiation in Lessonia (Phaeophyceae,Laminariales)

The genus Lessonia Bory de Saint‐Vincent is distributed solely in the Southern Hemisphere, with four species found in South America and five in Australasia. Our goal was to determine the evolutionary relationships between the Lessonia species of the two disjunct areas and hypothesize dispersal patterns and timing of radiation. We combined mitochondria, plastid and nuclear markers in a comprehensive dataset from multiple individuals per known species. Furthermore, for some species we added samples from multiple populations to take account of their widespread distribution over known bioregions. In all analyses the Australasian Lessonia formed a monophyletic group and in most analyses the South American species form a paraphyletic grade. Delimitations of the accepted species are highly supported except for L. variegata J. Agardh and L. nigrescens Bory de Saint‐Vincent. We showed for the first time four lineages for the New Zealand endemic Lessonia variegata with an unexpected high level of genetic differentiation. Our analysis cannot resolve whether the dispersal of the common ancestor of this genus was from the Americas to Australasia or vice versa. Molecular clock analysis suggested that a sudden radiation took place in Australasia c. 3.5 Mya when almost all Australasian species diverged within a time frame of only 35 000 years.     

Interspecific interference among species of Gelidium from Central Chile

In the rocky intertidal of Central Chile, Gelidium lingulatum (C. Agardh) J. Agardh and G. fílicinum Bory occur at the same tidal level with no evidence of temporal or spatial segregation. This study documents the effects of this biotic interaction on temporal standing stock changes, phenology, reproductive potential, and morphological variation of these species.     

REPRODUCTIVE LONGEVITY OF DRIFTING KELP MACROCYSTIS PYRIFERA (PHAEOPHYCEAE) IN MONTEREY BAY,USA1

Drifting Macrocystis pyrifera (L.) C. Agardh sporophytes have long been viewed as the primary long‐distance dispersal vector; yet, few data exist that support the ability of reproductive viable sporophytes to actually travel the presumed hundreds to thousands of kilometers. This study addressed the reproductive longevity of experimental and naturally occurring M. pyrifera drifters. Temporal variability in sporophyte size and reproduction was estimated for experimental drifting sporophytes that were tethered to surface buoys and compared with attached plants (controls). Reproductive viability was also studied for beach‐cast drifters (BCD), and naturally drifting sporophytes observed during field surveys in Monterey Bay. Detached drifting sporophytes were tracked with radio transmitters to follow drifter trajectories and to measure drifting speed. Experimental drifters (ED) experienced a 74% reduction in frond length after 35 days, a 76% reduction in average frond number after 70 days, and a reduction in average sorus area by 83% after 28 days. Although zoospore production was reduced following detachment, sporophytes remained fertile with high zoospore germination success as long as sori were present (125 days). Zoospore production and germination success for natural and BCD was similar to ED. The average displacement of radio‐tagged drifters was 7.12 km·day^?1, suggesting that a sporophyte adrift for 125 days disperses viable propagules (zoospores) over 890 km (±363). Dispersal of propagules is important for population restoration, distribution, and genetic diversity. Such dispersal distances are long enough to connect potentially all Northern Hemisphere Macrocystis populations across a generational timescale and may facilitate inter‐hemispheric gene flow.     

PHOTOSYNTHETIC PERFORMANCE,LIGHT ABSORPTION,AND PIGMENT COMPOSITION OF MACROCYSTIS PYRIFERA (LAMINARIALES,PHAEOPHYCEAE) BLADES FROM DIFFERENT DEPTHS1

Macrocystis pyrifera (L.) C. Agardh is a canopy‐forming species that occupies the entire water column. The photosynthetic tissue of this alga is exposed to a broad range of environmental factors, particularly related to light quantity and quality. In the present work, photosynthetic performance, light absorption, pigment composition, and thermal dissipation were measured in blades collected from different depths to characterize the photoacclimation and photoprotection responses of M. pyrifera according to the position of its photosynthetic tissue in the water column. The most important response of M. pyrifera was the enhancement of photoprotection in surface and near‐surface blades. The size of the xanthophyll cycle pigment pool (XC) was correlated to the nonphotochemical quenching (NPQ) of chl a fluorescence capacity of the blades. In surface blades, we detected the highest accumulation of UV‐absorbing compounds, photoprotective carotenoids, ΣXC, and NPQ. These characteristics were important responses that allowed surface blades to present the highest maximum photosynthetic rate and the highest PSII electron transport rate. Therefore, surface blades made the highest contribution to algae production. In contrast, basal blades presented the opposite trend. These blades do not to contribute significantly to photosynthetate production of the whole organism, but they might be important for other functions, like nutrient uptake.     

A RIBOSOMAL DNA PHYLOGENY SUPPORTS THE CLOSE EVOLUTIONARY RELATIONSHIPS AMONG THE SPOROCHNALES,DESMARESTIALES, AND LAMINARIALES (PHAEOPHYCEAE)1

We report six complete 18S ribosomal DNA (rDNA) sequences representing five brown algal orders: Sporochnus comosus C. A. Agardh (Sporochnales), Chorda tomentosa Lyngbye (Chordaceae, Laminariales), Saccorhiza polyschides (Lightfoot) Batters (Phyllariaceae, Laminariales), Desmarestia ligulata (Lightfoot) Lamouroux (Desmarestiales), Ectocarpus siliculosus (Dillwyn) Lyngbye (Ectocarpales), and Scytosiphon lomentaria (Lyngbye) J. G. Agardh (Scytosiphonales). These sequences were compared with published laminarialean (Alaria marginata Postel et Ruprecht [Alariaceae] and Macrocystis integrifolia Bory [Lessoniaceae]) and fucalean (Fucus gardneri Silva) rDNA sequences for phylogeny inference using both the distance-matrix and parsimony methods. The inferred 18S phylogenies clustered Sporochnus, Desmarestia, Chorda, Saccorhiza, Alaria, and Macrocystis in an assemblage. This Sporochnales–Desmarestiales–Laminariales (S-D-L) complex was consistently separated from the Ectocarpales, Scytosiphonales, and Fucales by bootstrap analyses. The inferred phylogenies are consistent with several possible evolutionary processes leading to this S-D-L complex. Members in this assemblage lack eyespots in their sperm, and their sperm have the atypical brown algal flagellation: shorter anterior and longer posterior flagella. In addition, they are oogamous with a heteromorphic alternation of generations between a microscopic gametophyte and a macroscopic sporophyte. Members of the S-D-L complex can be separated into different phylogenetic lines based on the presence/absence of eyespots in their meiospores. Our findings support the contention that the Sporochnales, Desmarestiales, and Laminariales are closely related. In addition, our rDNA tree suggests that the Laminariales is paraphyletic.     

GRAZING ON GIANT KELP MICROSCOPIC PHASES AND THE RECRUITMENT SUCCESS OF ANNUAL POPULATIONS OF MACROCYSTIS PYRIFERA (LAMINARIALES,PHAEOPHYTA) IN SOUTHERN CHILE1

The giant kelp Macrocystis pyrifera (L.) C. Agardh is widely distributed in the Northern Hemisphere and Southern Hemisphere, yet it exhibits distinct population dynamics at local to regional spatial scales. Giant kelp populations are typically perennial with the potential for year‐round reproduction and recruitment. In southern Chile, however, annual giant kelp populations exist and often persist entirely on secondary substrata (e.g., shells of the slipper limpet Crepipatella fecunda [Gastropoda, Calyptraeidae]) that can cover up to 90% of the rocky bottom. In these populations, the macroscopic sporophyte phase disappears annually during winter and early spring, leaving a 3–4 month period in which a persistent microscopic phase remains to support the subsequent year’s recruitment. We tested the effects of a suite of grazers on the recruitment success of this critical microscopic phase at two sites in southern Chile. Field experiments indicated that the snail Tegula atra negatively impacted M. pyrifera sporophyte recruitment, but that recruitment was highest in the presence of sessile female limpets, C. fecunda. Conversely, small male C. fecunda (biofilm grazers) did not regulate kelp recruitment. Laboratory observations showed that C. fecunda males only grazed on microscopic kelp gametophytes and small (<250 μm) sporophytes, rejecting larger sporophytes, whereas T. atra grazed on all the kelp stages. Recruitment to the C. fecunda treatments far exceeded that to bare rock in the absence of grazers but was not due to the physical presence of C. fecunda shells. We concluded that the key to M. pyrifera recruitment success in southern Chile is its capacity to colonize secondary substrates provided by the slipper limpet C. fecunda.     

Die Entwirrung desBotrytella(Sorocarpus)-Komplexes (Ectocarpaceae,Phaeophyta)

Sorocarpus Pringsh. is a later synonym forBotrytella Bory. Four species of the genus are known from northern European coasts. Three of them are found at Helgoland (North Sea), type locality of bothBotrytella uvaeformis (Pringsh.) andB. reinboldii (Reinke). A third species, most frequently seen, still remains unnamed. Plants of similar habit, however, have been described from Japan. The type species for the genus,Botrytella micromora Bory, was originally published asEctocarpus siliculosus β.uvaeformis by Lyngbye. A sample from Danish waters proved to be identical with it. In a previous paper (Kornmann & Sahling, 1984), the life history of two species was studied and the investigation is now extended to include the other two, demonstrating the conformity of the four species with the genus characters as to morphological and developmental features.      

TAXONOMIC REVISION OF SARGASSUM SPECIES (FUCALES,PHAEOPHYCEAE) FROM NEW CALEDONIA BASED ON MORPHOLOGICAL AND MOLECULAR ANALYSES1

Sargassum C. Agardh (1820) is a taxonomically difficult genus distributed worldwide and reported as the most species‐rich genus of the Fucales. It is especially abundant in the Pacific where decreasing species richness is reported to occur from west to east. New Caledonia has been recognized as one of the hotspots of Sargassum diversity; however, species lists available for this region are old and incomplete and have not yet been updated with regard to the latest taxonomic revisions published. This study aimed at revising Sargassum diversity in New Caledonia and to assess its geographic affinities with neighboring Pacific regions. We used combined morphological and DNA analyses on new collections and examined numerous type specimens. Although 45 taxa have been listed in the literature, most of them have been either transferred to synonymy since or misidentified, and in this study, only 12 taxa were recognized as occurring in New Caledonia. They belong to the subgenus Sargassum sect. Binderianae (Grunow) Mattio et Payri (2), sect. Ilicifoliae (J. Agardh) Mattio et Payri (2), sect. Polycystae Mattio et Payri. (1), sect. Sargassum (4), sect. Zygocarpicae (J. Agardh) Setch. (2), and subgenus Phyllotrichia (Aresh.) J. Agardh (1). New Caledonian Sargassum flora appeared as the second richest in the region after the Pacific coast of Australia, with which it has shown high similarity, and shared species with all neighboring regions. One species, S. turbinarioides Grunow, is considered as endemic to New Caledonia. The low genetic diversity detected among several polymorphic species belonging to sect. Sargassum is also discussed.     

Sexual compatibility and hybrid formation between the giant kelp species <Emphasis Type="Italic">Macrocystis pyrifera</Emphasis> and <Emphasis Type="Italic">M. integrifolia</Emphasis> (Laminariales,Phaeophyceae) in Chile

Two species of giant kelp inhabit the coast of Chile: Macrocystis integrifolia and M. pyrifera, representing important economic resources. As part of our efforts to domesticate these kelps for mariculture, and to obtain superior cultivars, we studied their biological relationship. Hybridization experiments with clonal gametophyte cultures showed reciprocal cross-fertility and produced fertile hybrid sporophytes with intermediate morphological characters. This hybridization potential in the laboratory contrasts with the persistence of two morphologically well-defined sister taxa in natural habitats on the Pacific coast of South America. We conclude that M. integrifolia and M. pyrifera are conspecific and speculate that unknown mechanisms support the co-existence of two morphologically distinct taxa on the subspecific level.     

Further confirmation of the presence of DNA in the pyrenoid core of the siphonous green algal genus Caulerpa (Caulerpales, Ulvophyceae, Chlorophyta)

We re-examined the distribution of chloroplast DNA (ct-DNA) in the pyrenoid core of Caulerpa okamurae Weber van Bosse and C. lentillifera J. Agardh by fluorescence microscopy after staining the squashes and Technovit sections with DNA fluorochromes such as 4′6-diamidino-2-phenylmdole (DAPI), ethidium bromide, Hoechst 33258 and chromomycin A3. All fluorochromes stained specifically the pyrenoid core on the squashes and Technovit sections. In addition, we present new data on the localization of ct-DNA in the pyrenoid core of two other species of the genus Caulerpa: C. cactoides (Turner) Agardh and C. geminata Harvey.     

ITS1 sequences of type specimens of Gigartina and Sarcothalia and their significance for the classification of South African Gigartinaceae (Gigartinales,Rhodophyta)

Uncertainties about the identity of type specimens of red algae have frequently led to taxonomic and nomenclatural confusion. A procedure for extracting PCR-amplifiable DNA from formalin-fixed material and herbarium specimens was used to investigate the taxonomic status of several South African Gigartinaceae. We compared nucleotide sequences in the internal transcribed spacer 1 (ITS1) region in type specimens and other historically important collections presently referred to Gigartina bracteata, G. radula and G. stiriata. The following opinions are supported: (1) Fucus bracteatus S.G. Gmelin, Chondrodictyon capense Kützing and Iridaea clathrata Decaisne represent a single species of South African Gigartina (G. bracteata (S.G. Gmelin) Setchell & Gardner) in which the disintegration of tetrasporangial sori results in a reticulate thallus. (2) Mastocarpus polycarpus Kützing, M. incrassatus Kützing and Iridaea lapathifolia Kützing represent a single species of South African Gigartina (G. polycarpa (Kützing) Setchell & Gardner) that has often, but erroneously, been called G. radula (Esper) J. Agardh. (3) Mastocarpus verrucosus Kützing is a later heterotypic synonym of Iridaea papillosa Bory (Sarcothalia papillosa (Bory) Leister) and was based on material that probably came from southern South America rather than from South Africa, the provenance given by Kützing. (4) Fucus stiriatus Turner and Sphaerococcus burmannii C. Agardh represent a single species of South African Sarcothalia (S. stiriata (Turner) Leister).     

Colony Structure and Seasonal Differences in Light and Nitrogen Modify the Impact of Sessile Epifauna on the Giant Kelp Macrocystis pyrifera (L.) C Agardh

The presence of the stoloniferous hydroid Obelia geniculata (L.) had no effect on the pigment concentration or nitrogen status of underlying blade tissue of the giant kelp Macrocystis pyrifera (L.) C. Agardh. The sheet-like colonies of the bryozoan Membranipora membranacea (L.) markedly reduced the pigment concentration of colonized blade tissue, but only during winter. Reductions in pigment concentration are most likely a result of damage to underlying tissue due to some factor related to the presence of bryozoan colonies on blade surfaces. Blade tissue colonized by M. membranacea also had higher δ¹⁵N signatures than surrounding bryozoan-free tissue, possibly indicating the provision of nitrogen to M. pyrifera by bryozoan colonies. Results show that seasonal changes in nitrogen and colony size can strongly modify the effect of epifauna on macroalgae they colonize. Unlike bryozoans, hydroid colonies provided no barrier to nitrogen uptake by colonized M. pyrifera tissue and enhanced ammonium uptake was observed for tissue colonized by O. geniculata during nitrogen limitation. Epifauna with stoloniferous growth forms such as hydroids are more likely to have benign or even mutualistic relationships with macroalgae they colonize than the sheet-like colonies of bryozoans.