BIOGEOGRAPHY AND ECOLOGY OF THE KELP/RED ALGAL SYMBIOSIS

A kelp/red algal symbiosis is described from nature based on extensive collections from the San Juan Islands, Washington. Kelp gametophytes were found as endophytes in the cell walls of seventeen species of red algae in three different kelp communities. Host red algae were mostly filamentous (e.g., Pleonosporium vancouverianum) or polysiphonous (e.g. Polysiphonia paniculata). The kelp gametophytes completed vegetative and reproductive development in the hosts with gametangia formed at the host surface and with sporophytes up to several mm in height being produced while still attached to the host. To date, none of the kelp gametophytes from nature have been identified to genus or species, although the gametophyte of Nereocystis luetkeana is a potential candidate for the symbiosis. Preliminary observations from Nova Scotia and the Isle of Man have not found the association in the Atlantic Ocean. Laboratory studies in Korea successfully reconstructed the symbiosis in the red alga Aglaothamnion oosumiense using zoospores of Undaria pinnatifida but not Laminaria religiosa. Here we outline the development of the symbiosis and discuss the potential adaptive significance of the kelp/red algal interaction.     

PHYLOGENETIC AND BIOGEOGRAPHIC AFFINITIES OF THE HALYMENIALES AND RHODYMENIALES (RHODOPHYTA) FROM THE GULF OF MEXICO

A kelp/red algal symbiosis is described from nature based on extensive collections from the San Juan Islands, Washington. Kelp gametophytes were found as endophytes in the cell walls of seventeen species of red algae in three different kelp communities. Host red algae were mostly filamentous (e.g., Pleonosporium vancouverianum ) or polysiphonous (e.g. Polysiphonia paniculata ). The kelp gametophytes completed vegetative and reproductive development in the hosts with gametangia formed at the host surface and with sporophytes up to several mm in height being produced while still attached to the host. To date, none of the kelp gametophytes from nature have been identified to genus or species, although the gametophyte of Nereocystis luetkeana is a potential candidate for the symbiosis. Preliminary observations from Nova Scotia and the Isle of Man have not found the association in the Atlantic Ocean. Laboratory studies in Korea successfully reconstructed the symbiosis in the red alga Aglaothamnion oosumiense using zoospores of Undaria pinnatifida but not Laminaria religiosa. Here we outline the development of the symbiosis and discuss the potential adaptive significance of the kelp/red algal interaction.     

Comparative development of gametophytes ofOsmunda lancea andO. japonica (osmundaceae): Adaptation of rheophilous fern gametophyte

Development of the gametophytes of a rehophytic fernOsmunda lancea and a related dryland speciesO. japonica was examined, and adaptation ofO. lancea gametophytes to rheophytic habitat was discussed. The spores ofO. lancea were larger in size and contained larger oil droplets than those ofO. japonica. The germination rate was similar. In agar culture,O. lancea gametophytes grew more rapidly and reached reprudctive maturity earlier thanO. japonica, although there was no prominent gross-morphological difference in developing and mature gametophytes. Sporophyte formation ofO. lancea took place at roughly two times higher rate than that ofO. japonica. TheO. lancea gametophytes might be adapted to the periodically flooded habitat by shortening its life, which is a different strategy from that of the sporophytic generation.     

Cloning and characterization of two types of ubiquitin genes from Gracilariopsis lemaneiformis (Gracilariales, Rhodophyta)

Two types of ubiquitin genes were isolated from the marine red alga Gracilaria lemaneiformis: a ubiquitin-52 amino acid fusion protein gene, and a 6-unit polyubiquitin gene. Alignment of polyubiquitins among three red algae (Gracilaria lemaneiformis, Gracilaria verrucosa, Aglaothamnion neglectum) and other species revealed that there were six ubiquitin repeats in all three red algae polyubiquitins, and that glutamine was the final amino acid residue in the terminal repeat of the polyprotein in the two Gracilaria sequences. Southern blot analysis revealed that both genes were encoded by low-copy number genes. Semi-quantitative RT-PCR was performed to investigate the expression of these two genes in two phases of G. lemaneiformis. The result revealed that the monoubiquitin was phase-relative, and upregulated in tetrasporophytes compared with female gametophytes. The polyubiquitin gene was expressed at similar levels in both phases.     

Balliella,a new genus of ceramiaceae (Rhodophyta)

Detailed studies of various aspects of the morphology and reproduction ofAntithamnion subcorticatum Itono andAntithamnion crouanioides Itono show that these species are incorrectly referred to as the genusAntithamnion. To include these species, a new genus,Balliella, is proposed. In addition, some other algae possibly referable to this genus are briefly discussed. The genusBalliella is here recognized as a member of the new tribeDelesseriopsieae which is most nearly related to the tribe Antithamnieae in the Ceramiaceae. The tribe Delesseriopsieae is now recognized to include the two genera, i.e.,Delesseriopsis andBalliella.     

Effects of temperature and nutrients on microscopic stages of the bull kelp (Nereocystis luetkeana,Phaeophyceae)

Warming ocean temperatures have been linked to kelp forest declines worldwide, and elevated temperatures can act synergistically with other local stressors to exacerbate kelp loss. The bull kelp Nereocystis luetkeana is the primary canopy-forming kelp species in the Salish Sea, where it is declining in areas with elevated summer water temperatures and low nutrient concentrations. To determine the interactive effects of these two stressors on microscopic stages of N. luetkeana, we cultured gametophytes and microscopic sporophytes from seven different Salish Sea populations across seven different temperatures (10–22°C) and two nitrogen concentrations. The thermal tolerance of microscopic gametophytes and sporophytes was similar across populations, and high temperatures were more stressful than low nitrogen levels. Additional nitrogen did not improve gametophyte or sporophyte survival at high temperatures. Gametophyte densities were highest between 10 and 16°C and declined sharply at 18°C, and temperatures of 20 and 22°C were lethal. The window for successful sporophyte production was narrower, peaking at 10–14°C. Across all populations, the warmest temperature at which sporophytes were produced was 16 or 18°C, but sporophyte densities were 78% lower at 16°C and 95% lower at 18°C compared to cooler temperatures. In the field, bottom temperatures revealed that the thermal limits of gametophyte growth (18°C) and sporophyte production (16–18°C) were reached during the summer at multiple sites. Prolonged exposure of bull kelp gametophytes to temperatures of 16°C and above could limit reproduction, and therefore recruitment, of adult kelp sporophytes.     

CULTURE STUDIES ON THE RELATIONSHIP BETWEEN SCHIZYMENIA AND HAEMATOCELIS (GIGARTINALES,RHODOPHYCEAE) FROM THE PACIFIC COAST OF NORTH AMERICA1

Carpospores from Schizymenia pacifica (Kylin) Kylin (Gymnophlaeaceae) from California formed crusts anatomically identical to Haematocelis rubens J. Agardh (Cruoriaceae). Tetraspores of H. rubens from Monterey, California, and Baja California, Mexico, germinated to form basal discs from which arose upright multiaxial blades with a filamentous medulla and cortical gland cells. Pro-carps and spermatangia were present on the same blades; subsequently, cystocarps characteristic of Schizymenia pacifica developed. Re-examination of herbarium specimens suggests that the foliose tetrasporangial phases previously reported as S. pacifica are referable to Halymenia, Dilsea, Cryptonemia, or Turnerella. Schizymenia pacifica (type locality: San Juan Islands, Washington) thus is considered to be the gametophyte in the life history of Haematocelis rubens (type locality: Brest, France), which has also been reported to be the tetrasporophyte of S. dubyi (Chauvin ex Duby) J. Agardh (type locality: Cherbourg, France). Atlantic and Pacific gametophytes and tetrasporophytes are anatomically very similar, suggesting that only one species is involved, but critical studies must be made before a decision on this taxonomic question can be reached. Haematocelis zonalis Dawson et Neushul (type locality: Anacapa Island, California) is considered to be a growth form of the tetrasporangial phase of S. pacifica.     

Arbuscular mycorrhizal fungi on adjacent semi-natural grasslands with different vegetation in Japan

Arbuscular mycorrhizal (AM) fungi on Japanese semi-natural grasslands were investigated at three adjacent sites with different vegetation. The predominant grasses at the three sites were 1)Pleioblastus chino, 2)Miscanthus sinensis andArundinella hirta (M. sinensis/A. hirta), and 3)Zoysia japonica, respectively. The degree of colonization was higher inM. sinensis/A. hirta than inP. chino andZ. japonica. AM fungi were recovered by spore extraction and by pot cultures started from soil inoculum or from transplanting of field plants. Total spore number obtained by the spore extraction method was highest in the rhizosphere ofM. sinensis/A. hirta and lowest in that ofP. chino. AGlomus sp. resemblingG. geosporum predominated in association withM. sinensis/A. hirta andP. chino. FromZ. japonica, three species,Acaulospora gerdemannii, Glomus leptotichum, and a species resemblingG. clarum, were isolated by pot culture from soil and two species,A. longula andScutellospora cerradensis, by pot culture from transplanting ofZ. japonica. FromM. sinensis/A. hirta, one species,A. longula, was found by pot culture from soil. FromP. chino, no AM fungus was detected by either method. Single-spore culture confirmed thatG. leptotichum andA. gerdemannii are conspecific.     

The effects of diet mixing on consumer fitness: macroalgae, epiphytes, and animal matter as food for marine amphipods

Herbivores are thought to achieve adequate nutrition by consuming numerous species of plants or by occasionally consuming animal tissue. Although active selection of diverse foods is common in nature, the relationship between diet mixing and consumer fitness is poorly understood, especially in marine environments. We studied the fitness-based consequences of dietary mixing in the sympatric amphipods Ampithoe marcuzzii, A. valida, Cymadusa compta, and Gammarus mucronatus by measuring survivorship, growth, and fecundity of these amphipods when they were offered single species of algae, a single animal food, a mixture of algal species, or a combination of algae and animal matter. For the more sedentary, tube-building amphipods A. marcuzzii, A. valida, and C. compta, fitness on mixed algal diets was matched by fitness on at least one of the monospecific algal diets, suggesting that they could benefit from preferential feeding on those algae in the field. The more mobile amphipod, G. mucronatus, survived and grew similarly on the mixed diets and on the filamentous brown alga Ectocarpus siliculosus. However, its fecundity was significantly higher when feeding on the algal and animal mixture than on Ectocarpus alone. Additionally, for G. mucronatus, fitness on mixed algae, mixed algae plus animal matter, and animal matter alone was equivalent, although female growth (but not gonad production) was slightly lower on animal matter alone than on the mixed algae combined with animal food. Thus the more mobile amphipod, G. mucronatus, was the only species able to perform well on animal food alone. In contrast, A. valida and C. compta experienced large negative effects when limited to consuming animal matter alone. For these two species, combining algae and animal matter did not enhance fitness over combining only algae. Fitness effects of specific algal diets showed some general similarities, but also considerable variance among the amphipods. For example, E. siliculosus was generally better food than other algae for all four amphipods, whereas Sargassum filipendula was generally poor. However, A. marcuzzii did not suffer negative effects of consuming only Sargassum. The red alga Polysiphonia sp. and the green alga Enteromorpha flexuosa decreased fitness in A. marcuzzii, C. compta, and G. mucronatus, but not A. valida, and the negative effects of Polysiphonia were considerably larger for A. marcuzzii than for the other amphipods. Our data show that nutritional requirements, even among related species (e.g., A. marcuzzii and A. valida), can be dramatically different. Diet mixing may benefit more mobile consumers like Gammarus that are better able to search for different foods, and may be less important for more sedentary herbivores like Ampithoe and Cymadusa that consume, and live in close association with, individual host plants. Received: 14 September 1998 / Accepted: 22 October 1999     

UNUSUAL PHENOMENA IN THE LIFE HISTORIES OF FLORIDEAE IN CULTURE1

Several Florideae grown in natural seawater media under defined laboratory conditions have interesting and unusual life histories. Antithamnion occidentale males of one generation produced tetraspores that gave rise to nonsporangiate males. The functional females of A. pygmaeum developed spermatangia and tetrasporangia; the tetraspores formed new females. Antithamnion defectum tetrasporophytes of one generation bore spermatangia in addition to tetrasporangia; the tetraspores gave rise to typical gametophytes. Tetraspores from successive generations of Callitham-nion sp. developed into tetrasporophytes and males but no females were produced. Functional female gametophytes of Platythamnion sp. bore abortive tetrasporangia. Field-collected plants of two species of Fauchea produced tetraspores that yielded additional sporangiate plants: those of F. pygmaea being bispo-rangiate and tetrasporangiate, and those of F. lacini-ata being strictly tetrasporangiate. Male plants of Pleonosporium vancouverianum from a running seawater table bore spermatangia and polysporangia when collected. The same plants in unialgal culture produced only spermatangia.     

Mycothallic/mycorrhizal symbiosis of chlorophyllous gametophytes and sporophytes of a fern, Pellaea viridis (Forsk.) Prantl (Pellaeaceae, Pteridales)

Gametophytes of Pellaea viridis that appeared spontaneously on the surface of substratum originating from an ultramafic area were found to form mycothallic symbiosis with arbuscular mycorrhizal fungi (AMF) under laboratory conditions. In gametophytes and sporophytes grown with Glomus tenue, abundant arbuscule formation was observed at both stages. In gametophytes, the fungus was found in the region where the rhizoids are initiated. If G. intraradices was added to the soil, the gametophytes were colonised mostly by G. tenue, and roots of sporophytes were colonised by G. intraradices. The presence of AM fungi in both gametophytes and sporophytes of P. viridis resulted in the development of larger leaf area and root length of the sporophyte. The analysis of gametophytes from the Botanical Garden in Krakow (Poland) showed that cordate gametophytes of Pteridales, namely Pellaea viridis (Pellaeaceae), Adiantum raddianum and A. formosum (Adiantaceae), were also mycothallic.     

A SCAR MOLECULAR MARKER SPECIFICALLY RELATED TO THE FEMALE GAMETOPHYTES OF SACCHARINA (LAMINARIA) JAPONICA (PHAEOPHYTA)1

PCR amplification was employed to identify female or male gametophyte associated markers in Saccharina japonica (Aresch.) C. E. Lane, C. Mayes et G. W. Saunders (=Laminaria japonica Aresch.). One pair of the primers, P5, was screened from five pairs designed based on a specific sequence (GenBank accession no. AB069714 ) of Marchantia polymorpha Y chromosome, resulting in a differential band ～500 bp in size between female and male gametophytes of Rongfu strain of S. japonica. According to the SCAR (sequence‐characterized amplified regions) strategies, one pair of primers, P51, was designed on the basis of the sequence of this band that was only present in female gametophytes. A SCAR marker, designated FRML‐494 (494‐bp Female‐Related Marker of S. japonica, GenBank accession no. EU931619 ), was developed successfully by PCR amplification using the designed P51 primer pair. The SCAR marker was verified to be present only in female gametophytes of another variety 901 of this kelp that was a hybrid between S. japonica as paternal and S. longissima (Miyabe) C. E. Lane, C. Mayes, Druehl et G. W. Saunders (=Laminaria longissima Miyabe) as maternal, suggesting that the FRML‐494 marker was specifically related to female gametophytes of the genus. This marker is the first molecular tool reported for sex identification in kelps. This study was beneficial for identifying gametophyte gender during vegetative growth and for judging whether the monogenetic sporophytes came from exclusive male or female gametophytes, as well as for further research on sex determination at the molecular level in kelps.     

Redescription ofRhinochimaera pacifica (mitsukuri) and first record ofR. africana compagno, stehmann & ebert from Japan (Chimaeriformes: Rhinochimaeridae)

Rhinochimaera pacifica is redescribed and a specimen from Tokyo Bay is designated as the neotype. A second species of the genus,R. africana, is newly recognized from Japan. A new scheme of measurements of the lateral line canals has been developed, and a comparison of proportional measurements of the snout width, eye length, and canal measurements clearly distinguishesR. pacifica andR. africana. Intraspecific variation ofR. pacifica and comparison ofR. pacifica andR. africana is discussed.Rhinochimaera pacifica has an irregular distribution in the north and south Pacific Ocean and Indian Ocean. This study also confirms a much wider range forR. africana than previously known and extends the range to the northern Pacific Ocean around the Japanese Archipelago.     

Calcium oxalate crystals in the red algaAntithamnion kylinii (Ceramiales): cytoplasmic and limited to indeterminate axes

Summary The red alga,Antithamnion kylinii Gardner, was found to have needle-shaped inclusions about 10 m long and less than 0.4 m thick. They ranged in abundance from one or a few in young cells to hundreds in fully enlarged cells. Under polarized light, the inclusions were birefringent, indicating crystallinity. Solubility tests suggested that the inclusions were composed of calcium oxalate: they dissolved in 1 N hydrochloric acid and in a saturated solution of aqueous cupric acetate, but they were not soluble in 10 N acetic acid or 5.25% sodium hypochlorite. X-ray microanalysis confirmed the presence of calcium. Calcium oxalate crystals were present in cells of indeterminate axes, but cells of determinate lateral filaments lacked them. Light and electron microscopic study demonstrated that the crystals were associated with the parietal cytoplasm. Calcium oxalate crystals were also present inA. defectum Kylin, but they were not found in ten more distantly related taxa.     

Porphyra nereocystis: A dual-daylength seaweed

Conchospores from the perennial conchocelis phase of the annual, epiphytic, marine red alga Porphyra nereocystis Anderson, which in nature lives on the large annual kelp Nereocystis luetkeana (Mertens) Postels et Ruprecht, are released in culture only in response to a short-day photoperiod treatment followed by a long-day treatment. Each treatment requires a minimum of three to four weeks and is enhanced by lower temperature during the second photoperiod treatment. To our knowledge P. nereocystis is the first known dualdaylength seaweed and requires a short-day-longday treatment for completion of its life cycle. This stringent environmental control of its reproduction appears to be an adaptation to coordinate conchospore production with the seasonal availability of its host kelp Nereocystis.Abbreviations LD long day - SD short day - SLD shortlong day     

Life and death beneath macrophyte canopies: effects of understory kelps on growth rates and survival of marine,benthic suspension feeders

Summary Experiments conducted on rocky bottoms at 7–11 m depth in the San Juan Archipelago, Washington assessed effects of canopies of understory kelps on growth of benthic suspension feeders, determined the mechanisms responsible for effects, and assessed the influence of kelp canopies on survivorship of benthic fauna. Kelp canopics influenced growth rates of diverse suspension feeders. At several sites the musselMytilus edulis, the barnacleBalanus glandula, and the serpulid polychaetePseudochitinopoma occidentalis grew faster on the bottom beneath kelp canopies than on nearby exposed substrata. The cheilostome bryozoanMembranipora membranacea showed a mixed response to kelp canopies, growing faster in exposed regions at one site, but faster beneath canopies at another. There were no differences in growth of 2 other species (the cheilostome bryozoanCheilopora praelonga and the spongeMyxilla incrustans) between kelp and no-kelp treatments; however, some processes influenced by plant canopies affected their growth. Specific mechanisms responsible for kelp effects on growth were assessed in a series of field experiments usingPseudochitinopoma, Membranipora, Cheilopora andMyxilla. Particulate deposition on the bottom, which is more intense beneath canopies, negatively affected growth of all 4 species. Kelps also reduced rates of flow and prevented devented development of microalgal turfs beneath the canopy.Pseudochitinopoma grew faster in the weaker flows below canopies and bothCheilopora andMyxilla grew faster where there were no microalgal turfs. These other effects of kelp canopies were at least as important to growth (in the cases ofCheilopora andMyxilla) or more important to growth (in the case ofPseudochitinopoma) than were the general, deleterious effects of higher sedimentation beneath canopies. The lower growth rates caused by higher sedimentation beneath kelp canopies did not result in higher rates of animal mortality. Surprisingly, kelp canopies typically did not influence mortality due to predation. For 7 of 12 taxa, mortality rates did not differ between kelp-covered and exposed treatments. Significantly higher mortality occurred outside canopies for only 4 of 12 taxa, and for at least 2 of these 4 differences probably were not related to predation.Mytilus, a species rare at these depths, exhibited higher mortality beneath kelp canopies due to predation by crabs. Other macrophytes in fresh and salt water, as well as some benthic animals that create complex, 3-dimensional habitats, should influence benthic organisms and assemblages in ways analogous to the kelps acting through their effects on flow, particle transport, and shading.     

Settlement behavior of cyphonautes larvae of the bryozoan Membranipora membranacea in response to two algal substrata

Abstract. Settlement is an important process in the biphasic life histories of many marine invertebrates. Little is known regarding the fine-scale behavioral mechanisms for finding and attaching to a suitable substratum, particularly under conditions that may impose a potential challenge, such as flow. In this study, we examined the settlement behavior of cyphonautes larvae of the bryozoan, Membranipora membranacea, in response to two different algal substrata. Larvae showed a strong preference for settling on the kelp Nereocystis luetkeana over the red alga Mazzaella splendens. We then tested whether the behavioral mechanisms used by larvae to attach to these algae differed when presented with the challenge of flowing water during settlement. We found that larvae exhibited different behaviors on the two species of algae in flowing water. Larvae were more often observed in direct contact with the preferred alga (N. luetkeana) exhibiting fine-scale active search behaviors. On the less preferred alga (M. splendens), larvae were less frequently observed in direct contact with the alga, and appeared to be exhibiting broad-scale passive search behaviors along the surface of the blade. Our results suggest that cyphonautes larvae alter their behavior in response to their preferred settlement habitat.     

STUDIES IN SOME FOLIOSE RED ALGAE OF THE PACIFIC COAST. I. CRYPTONEMIACEXE1

Pacific coast species of the red algae Halymenia and Cryptonemia are described Of the 8 previously reported species of Halymenia 3 (H. abyssicola Dawson, H. megaspore Dawson, H refugiensis Dawson) hive been placed in synonymy with other species. Halymenia hollenbergii from southern California is described as new; H cocinea is transferred from Schizymenia (and includes Aeodes gardneri Kylin) and H. templetonii is a transfer from Weeksia. Five species of Cryptonemia are described; one of them, C. taylorii from the Revillagigedo Archipelago, as new. Wider distribution ranges are given for the remaining species, C. ovalifolia, C. obovata, C. bore alis and C. angustata. Keys to genera of the west coast Cryptonemiaceae and to the species of Halymenia and Crypronemia are provided.     

ONTOGENY,HISTOCHEMISTRY AND FINE STRUCTURE OF CELLULAR INCLUSIONS IN VEGETATIVE CELLS OF ANTITHAMNION DEFECTUM (CERAMIACEAE,RHODOPHYTA)1

The ultrastructure and histochemistry of developing and mature cell inclusions in vegetative cells of Antithamnion defectum Kylin were examined. Those studied were chloroplast inclusions, cytoplasmic crystals and spherical bodies within the vacuole. Chloroplasts of mature vegetative cells contain an interthylakoidal, apparently noncrystalline deposit of undetermined chemical identity. The bodies are parallel to the long axis of the plastid, are square (0.13 μm) in cross-section, and up to 3 μm long. Spherical vacuolar bodies (0.5–1.5 μum diam) are formed during early stages of vacuole formation by accumulation of protein deposits in swelling endoplasmic reticulum (ER) cisternae. Swelling of smooth ER contiguous to the ER containing the deposits results in the vacuole enclosing the spherical bodies. In mature cells, vesicles appear to be secreted into the preformed vacuole. Cytoplasmic proteinaceous crystalloids develop without a bounding membrane and may serve as protein reserves.