EVIDENCE FOR SEXUAL REPRODUCTION IN THE PRIMITIVE GREEN ALGA NEPHROSELMIS OLIVACEA (PRASINOPHYCEAE)1

Evidence for a specialized sexual process in Nephroselmis olivacea Stein is presented. This alga is a member of the Prasinophyceae, which is regarded by some as the most primitive Class of green plants. N. olivacea has a heterothallic type of mating system. Plus and minus gametes were morphologically similar but showed different behaviors during the mating process. The minus gamete settled to the substratum, attaching by its ventral side. The plus gamete attached to the dorsal side of the minus gamete by the region near the flagellar bases of the plus gamete. The mature zygotes were spherical and strongly adhered to the substratum. After zygote germination, two biflagellate daughter cells, each with two pyrenoids were liberated. These cells divided, resulting in four vegetative cells, each with a single pyrenoid.     

PHYLOGENETIC ANALYSIS OF PSEUDOCHLORODESMIS STRAINS REVEALS CRYPTIC DIVERSITY ABOVE THE FAMILY LEVEL IN THE SIPHONOUS GREEN ALGAE (BRYOPSIDALES,CHLOROPHYTA)1

The genus Pseudochlorodesmis (Bryopsidales) is composed of diminutive siphons of extreme morphological simplicity. The discovery of Pseudochlorodesmis‐like juveniles in more complex Bryopsidales (e.g., the Halimeda microthallus stage) jeopardized the recognition of this genus. Confronted with this uncertainty, taxonomists transferred many simple siphons into a new genus, Siphonogramen. In this study, we used a multimarker approach to clarify the phylogenetic and taxonomic affinities of the Pseudochlorodesmis‐Siphonogramen (PS) complex within the more morphologically complex bryopsidalean taxa. Our analyses reveal a new layer of diversity largely distinct from the lineages containing the structurally complex genera. The PS complex shows profound cryptic diversity exceeding the family level. We discuss a potential link between thallus complexity and the prevalence and profundity of cryptic diversity. For taxonomic simplicity and as a first step toward clarifying the taxonomy of these simple siphons, we propose to maintain Pseudochlorodesmis as a form genus and subsume Siphonogramen and Botryodesmis therein.     

CONTROL OF REPRODUCTION RHYTHMICITY BY ENVIRONMENTAL AND ENDOGENOUS SIGNALS IN ULVA PSEUDOCURVATA (CHLOROPHYTA)1

A field population of Ulva pseudocurvata Koeman et C. Hoek (hereafter termed Ulva) at Sylt Island (North Sea, Germany) exhibited biweekly peaks of gametophytic reproduction during the colder seasons and approximately weekly peaks during summer. The reproductive events lasted 1–5 d and were separated from each other by purely vegetative phases. Under constant conditions in the laboratory, a free‐running rhythm was observed with reproductive peaks occurring approximately every 7 d. When artificial moonlight was provided every 4 weeks, fewer reproductive events occurred, and the reproductive rhythm became synchronized to the environmental artificial moonlight rhythm. In the laboratory, apical disks were entirely converted into reproductive tissue after 8 d cultivation, while almost all basal disks stayed vegetative, which prevented the entire loss of the vegetative thallus during reproductive events. Seasonal size reduction of the thallus occurred from late autumn onward and was determined to be controlled by a genuine photoperiodic response, since size reduction could be induced from May onward by experimental short‐day (SD) treatment but was prevented in a long‐day (LD) or night‐break regime (NB). A daily fine‐tuning occurred with gamete release early in the morning at the first sign of daylight, following an obligatory dark (“night”) period of at least 1 h duration. No release took place if the overnight dark phase was replaced by continuous light. Blue, green, or red light all triggered gamete release after a dark phase at an irradiance of 0.1 μmol photons · m^?2 ·s^?1, while 0.001 μmol photons · m^?2 · s^?1 was equivalent to a dark control.     

OBSERVATIONS ON THE MORPHOLOGY AND SEXUAL REPRODUCTION OF COOLIA MONOTIS (DINOPHYCEAE)1

The surface morphology of the dinoflagellate Coolia monotis Meunier was compared with the surface morphology of Ostreopsis, The apical pore of C. monotis is similar in architecture to that of Ostreopsis but considerably longer (12 μm) than in O. heptagona (8–9 μm) and O. ovata (6–7 μm). A ventral pore in C. monotis is located on the right ventral margin between apical plate l′ and precingular plate 6″ and is similar in appearance and location to the ventral pore of O. ovata. The longitudinal flagellum (20 μm) in C. monotis is longer than in O. ovata (12 μ). Although Coolia and Ostreopsis appear to be distinctly different and should remain as two separate genera, they appear to be related. Cells of C. monotis divided by binary fission. Doubling time was 3–4 days in the logarithmic phase of growth at 23°C, 12:12 h L:D, 30–90 μE-m^?2·s^?1, and a salinity of 36%. Cultures reached cell densities of 2.5 × 10³ cells·L^?1 after 15 days of growth. The sexual process in C. monotis occurred in Erdschreiber's medium when Danish soil extract was substituted with mangrove sediment extract under the culture conditions described above. Gamete fusion produced large biflagellated planozygotes (70–75 μm diam). Planozygote maturation involved cytoplasmic reorganization, loss of motility, development of a spherical shape (80–90 μm diam), and two to three orange accumulation bodies. The cells at this stage appeared to be thin-walled cysts. Further development included reorganization of cyst contents, emergence of non-motile gametes, and development of chloroplasts, sulcus, and girdle. The nucleus of the newly formed cells occupied 50% or more of the total cell volume. Meiosis occurred in the cyst, but nuclear cyclosis was not observed. Four daughter cells were produced within 36–48 h, and motile gametes developed. The gametes exhibited sexuality for 2 months and completed the sexual life cycle by going through a thin-walled cyst stage.     

双角多甲藻有性生殖的初步观察

通过对双角多甲藻有性过程的初步研究,表明在这种间核生物的生活史中存在着典型的育性生殖。在双角多甲藻春季生长高峰的末期(3月底至4月初),有性生殖大量发生。形成的合子经过一系列变化后成为厚垣合子进入休眠期。     

ABIOTIC REGULATION OF INVESTMENT IN SEXUAL VERSUS VEGETATIVE REPRODUCTION IN THE CLONAL KELP LAMINARIA SINCLAIRII (LAMINARIALES,PHAEOPHYCEAE)1

Clonal kelp taxa may reproduce both sexually and vegetatively resulting in a potential trade‐off in the allocation of acquired carbon and nitrogen resources. Such trade‐offs may dictate a different response of clonal kelps to varying environmental conditions relative to aclonal kelp taxa. Laboratory temperature and nutrient manipulation experiments demonstrated that investment in sexual and vegetative reproduction in Laminaria sinclairii (Harv. ex Hook. f. et Harv.) Farl., C. L. Anderson et D. C. Eaton was regulated by different abiotic factors. Sorus production (investment in sexual reproduction) and blade growth were significantly higher at 12°C compared to 17°C, regardless of nutrient concentration. Net carbon storage and depletion in rhizomes were observed in the low‐ and high‐temperature treatments, respectively, suggesting that carbon stores were not responsible for increased growth. Rhizome elongation (investment in vegetative reproduction), on the other hand, was significantly higher in 12 μM NO₃^? than in 2 μM NO₃^?, irrespective of temperature. This increase in rhizome growth was concurrent with elevated rhizome percent tissue nitrogen levels also observed in treatments with higher nutrients, again indicating a growth response to treatment independent of previous nutrient stores. These results suggest that regulation of growth and investment in sexual reproduction in L. sinclairii is similar to that in aclonal kelps (i.e., warmer temperatures result in decreased reproductive output). Additionally, depletion of carbon and nitrogen from rhizomes in suboptimal conditions confirms the role of clonal kelp rhizomes in carbon and nutrient storage.     

SEASONAL CHANGE IN THE DISTRIBUTION OF CELL SIZE OF COCCONEIS SCUTELLUM VAR. ORNATA (BACILLARIOPHYCEAE) IN RELATION TO GROWTH AND SEXUAL REPRODUCTION1

Growth and sexual reproduction of the marine littoral diatom Cocconeis scutellum Ehrenb. var. ornata Grun. were investigated at 30 different combinations of temperature (5, 10, 14, 18, 22° C), irradiance (20, 60, 100 μE·m^?2·s^?1) and daylength (14:10 and 10:14 h LD cycle). Growth occurred at all combinations. The optimal growth was observed at 14–18° C, long daylength and highest-to-moderate irradiance, and at 18° C, short daylength and highest irradiance. Sexual reproduction on the other hand occurred between 5 and 18° C, and the optimal condition was 10–14° C and short daylength. Annual cyclic, and sesonal changes in the distribution of cell size (valve length) were observed in a field population. These changes were characterized by an annual minimum in mean cell size in autumn, an annual maximum in winter, a slight decrease from the mean in spring–middle summer, a rapid decrease from the mean in late summer–early autumn, and appearance of bimodal distribution of cell size in winter. These changes were caused by sexual reproduction in autumn, rapid growth in late summer–early autumn and slow growth in other seasons, and poor viability of small cells near the lower end of the size range.     

ASEXUAL AND SEXUAL REPRODUCTION IN GONIUM QUADRATUM (CHLOROPHYTA) WITH A DISCUSSION OF PHYLOGENETIC RELATIONSHIP WITHIN THE GONIACEAE1

Morphological details of asexual and sexual reproduction in Gonium Quadratum Pringsheim ex Nozaki (Goniaceae, Chlorophyta) were observed by light microscopy, based on clonal cultured materials originating from Nepal. In asexual reproduction, the alga exhibited two different patterns of cell cleavage during formation of 8-and 16-celled daughter colonies. Sexual reproduction was heterothallic and isogamous. The gametes bore a tubular mating structure (bilateral mating papilla) at the base of the flagella, and the papillae of the two gametes. The germinating zygote gave rise to four biflagellate gone cells joined in a colony (germ colony). Possible phylogenetic relationships within the Goniaceae at the species level are outlined, mainly on the basis of reproduction characteristics.     

THE ECOLOGY OF EPIPELIC ALGAE OF FIVE WELSH LAKES,WITH SPECIAL REFERENCE TO VOLVOCALEAN GREEN FLAGELLATES (CHLOROPHYCEAE)1

The ecology of epipelic algae on the marginal sediments of five Welsh lakes was studied over an annual cycle. The lakes, Llydaw, Cwellyn, Padarn, Maelog and Coron ranged from very oligotrophic to nutrient-rich. Attention was focussed on chlamydomonad flagellates, diatoms, blue-green algae and euglenoids and the different proportions of these in algae in the epipelon of lakes of contrasting water quality. A total of 75 algal taxa was found in the five lakes, 25 were species of volvocalean flagellates. Mean annual population density of these flagellates differed by an order of magnitude between the lakes. The greatest population density was recorded for Chlamydomonas anticontata Schiller in nutrient-rich Llyn Maelog. Twenty species of pennate diatoms were recorded frequently in the epipelon. In the nutrient-rich lakes, Maelog and Coron, pennate diatoms were dominant on the sediments, where they exhibited population maxima in spring and autumn. Increase in numbers of epipelic diatoms was recorded when silica concentrations were minimum in the overlying lake waters. Navicula hungarica Grun. achieved the maximum population density, 260 000 cells · cm^?2. Euglenoids formed large epipelic populations during late-summer and autumn in these nutrient-rich lakes. Blue-green algae were more important, proportionally, in the nutrient-poor mountain lakes, which had sediments of higher organic content. Chlamydomonads were the major algal component of the epipelon in the mountain lakes, Llydaw and Cwellyn, where the sediments were characterized by larger particle size, and higher organic content. In the nutrient-rich lakes, where the sediments had higher calcium content, chlamydomonads formed significant populations only during spring and summer, when nutrient levels were minimal in the overlying lakewaters.     

VARIABILITY IN THE ECOPHYSIOLOGY OF HALIMEDA SPP. (CHLOROPHYTA,BRYOPSIDALES) ON CONCH REEF,FLORIDA KEYS,USA1

The photosynthetic performance, pigmentation, and growth of a Halimeda community were studied over a depth gradient on Conch Reef, Florida Keys, USA during summer–fall periods of 5 consecutive years. The physiology and growth of H. tuna (Ellis & Solander) Lamouroux and H. opuntia (L.) Lamouroux on this algal dominated reef were highly variable. Maximum rate of net photosynthesis (P_max), respiration rate, and quantum efficiency (α) did not differ between populations of either species at 7 versus 21 m, even though the 21‐m site received a 66% lower photon flux density (PFD). Physiological parameters, as well as levels of photosynthetic pigments, varied temporally. P_max, saturation irradiance, compensation irradiance, and growth were greatest in summer months, whereas α, chl a, chl b, and carotenoid concentrations were elevated each fall. Halimeda tuna growth rates were higher at 7 m compared with 21 m for only two of five growth trials. This may have arisen from variability in light and nutrient availability. Individuals growing at 7 m received a 29% greater PFD in August 2001 than in 1999. In August 1999 and 2001 seawater temperatures were uniform over the 14‐m gradient, whereas in August 2000 cold water regularly intruded upon the 21‐m but not the 7‐m site. These results illustrate the potentially dynamic relationship between nutrients, irradiance, and algal productivity. This suggests the necessity of long‐term monitoring over spatial and temporal gradients to accurately characterize factors that impact productivity.     

EVIDENCE FOR SEXUAL REPRODUCTION IN THE MARINE DINOFLAGELLATES, PYROCYSTIS NOCTILUCA AND PYROCYSTIS LUNULA (DINOPHYTA)

Evidence for sexual reproduction was observed in two oceanic dinoflagellate species, Pyrocystis noctiluca Murray ex Haeckel and Pyrocystis lunula (Schütt) Schütt. Observations suggest that cells underwent fertilization as opposed to cell division because of the following: first, fusing cells had a conspicuous pore (fusion pore) connecting the two gametes; dividing cells lacked this feature. In culture, about 0.1% of P. noctiluca cells had a fusion pore, which serves as a possible site for gamete recognition on the cell wall. Second, we document a temporal progression of plasmogamy and karyogamy. Fusion events in both species were observed at the beginning of the day, whereas division stages were most apparent at the end of the day.     

REVISION OF THE MASTOPHOROIDEAE (CORALLINALES,RHODOPHYTA) AND POLYPHYLY IN NONGENICULATE SPECIES WIDELY DISTRIBUTED ON PACIFIC CORAL REEFS1

The subfamily Mastophoroideae (Corallinaceae, Rhodophyta) is characterized by species possessing nongeniculate, uniporate tetrasporangial conceptacles without apical plugs, the presence of cell fusions, and the absence of secondary pit connections. However, molecular phylogenetic studies not including the type genus Mastophora indicated that the Mastophoroideae was polyphyletic. Our molecular phylogenetic analysis of the subfamily including the type genus using DNA sequences of SSU rDNA and plastid‐encoded gene of PSII reaction center protein D1 (psbA) revealed that Mastophora formed a robust clade only with Metamastophora. The other mastophoroid genera were divided into six lineages within the family Corallinaceae. Five supported lineages—(i) Pneophyllum; (ii) Hydrolithon gardineri (Foslie) Verheij et Prud’homme, Hydrolithon onkodes (Heydr.) Penrose et Woelk., and Hydrolithon pachydermum (Foslie) J. C. Bailey, J. E. Gabel et Freshwater; (iii) Hydrolithon reinboldii (Weber Bosse et Foslie) Foslie; (iv) Spongites; and (v) Neogoniolithon—were clearly distinguished by the combination of characters including the presence or absence of palisade cells and trichocytes in large, tightly packed horizontal fields and features of tetrasporangial and spermatangial conceptacles. Therefore, we amend the Mastophoroideae to be limited to Mastophora and Metamastophora with a thin thallus with basal filaments comprised of palisade cells, tetrasporangial conceptacles formed by filaments peripheral to fertile areas, and spermatangia derived only from the floor of male conceptacles. This emendation supports Setchell’s (1943) original definition of the Mastophoroideae as having thin thalli. We also propose the establishment of three new subfamilies, Hydrolithoideae subfam. nov. including Hydrolithon, Porolithoideae subfam. nov. including the resurrected genus Porolithon, and Neogoniolithoideae subfam. nov. including Neogoniolithon. Taxonomic revisions of Pneophyllum and Spongites were not made because we did not examine their type species.     

GAMETE RELEASE IS INCREASED BY CALM CONDITIONS IN THE COENOCYTIC GREEN ALGA BRYOPSIS (CHLOROPHYTA)

Water motion did not inhibit gamete release in cultures of the coenocytic green alga Bryopsis plumosa (Hudson) C. Agardh; however, the number of gametangia that released gametes increased significantly under transiently calm conditions. This stimulatory effect of calm conditions in the laboratory was found in isolates from two different areas of the Maine coast. The isolates were all monoecious, but strong differences in levels of fertilization, numbers of male and female gametes remaining following fertilization, and levels of polygamy (=zygotes formed by fusions of more than two gametes) were observed among isolates. These data support the hypothesis that organisms with external fertilization are able to sense and respond to water motion in ways that favor reproductive success.     

MASS SEXUAL REPRODUCTION IN THE TOXIGENIC DIATOMS PSEUDO‐NITZSCHIA AUSTRALIS AND P. PUNGENS (BACILLARIOPHYCEAE) ON THE WASHINGTON COAST,USA1

Sexual reproduction is documented for the first time in field populations of the pennate diatoms Pseudo‐nitzschia australis Freng. and P. pungens (Grunow ex Cleve) Hasle (var. cingulata Villac and hybrids between var. cingulata and var. pungens). A bloom dominated by these species began on June 26, 2006, along Kalaloch Beach, Washington, USA, coincident with a drop in the Si(OH)₄:NO₃ ratio to below two. Multimodal size distributions were detected for both species, and synchronous auxosporulation occurred within the smallest size class during a 3‐week window. Auxospores and initial cells created a new class of large cells, and cells in the intermediate size classes increased in abundance during auxosporulation. Mating cells of both species were attached to colonies of surf‐zone diatoms. Paired gametangia, gametes, zygotes, auxospores, and large initial cells were found. Auxosporulation began first for P. pungens (June 30), apparently once a critical, high cell concentration was reached, followed by P. australis (July 5), when the total Pseudo‐nitzschia cell concentration reached 929,000 cells · L^?1. Low frequencies of auxosporulation occurred throughout the bloom but increased 4‐fold for P. australis and 3‐fold for P. pungens when macronutrients were reduced to low levels on July 11. A 2‐year life cycle was estimated for P. australis and 3 years for P. pungens, both with annual auxosporulation. Domoic acid (DA) in razor clams reached a maximum of 38 μg DA · g^?1 on July 18. A significant relationship existed between the percent of cells within the new size range and DA concentrations in razor clams on the same beach.     

A STUDY OF THE SEXUAL REPRODUCTION AND DETERMINATION OF MATING TYPE OF GYMNODINIUM NOLLERI (DINOPHYCEAE) IN CULTURE1

Sexual reproduction of Gymnodinium nolleri ( Ellegaard & Moestrup 1999 ) was studied by intercrossing experiments in all combinations of six clonal strains and backcrossing of five clonal F1 offspring. The results indicated that the conjugation of G. nolleri responded to the existence of more than two sexual types (complex heterothallism) and that compatibility between progeny of one cyst (inbreeding) was the rule. Sexual fusion, planozygote formation and development, cyst formation, and germination and planomeiocyte division were followed using time‐lapse photography. This study revealed many similarities between the sexual stages and life cycle pattern of G. nolleri and the related G. catenatum and the existence under culture conditions of an alternative cycle between vegetative cells and zygotes without a hypnozygote stage. The fate of zygotes, division or encystment, was influenced by the nutritional status of the external medium. The division of G. nolleri planozygotes was promoted by high levels of external nutrients, whereas the maximum percentage of encystment was recorded when phosphates were reduced in the isolation medium. The division of zygotes might be different from both vegetative and planomeiocyte division because it resulted in two‐cell chains with the cells not oriented in parallel.     

A PHYLOGENETIC PERSPECTIVE ON THE EVOLUTION OF SEXUAL DICHROMATISM IN TANAGERS (THRAUPIDAE): THE ROLE OF FEMALE VERSUS MALE PLUMAGE

The evolution of sexual dichromatism in tanagers (family Thraupidae) was studied from a phylogenetic perspective using a molecular-based phylogeny. Mapping patterns of sexual dimorphism in plumage onto the phylogeny reveals that changes in female plumage occur more frequently than changes in male plumage. Possible explanations for this pattern include sexual selection acting on female plumage and natural selection for background matching. The results of this study and other recent phylogenetic and comparative studies suggest that factors affecting female plumage are important in shaping patterns of sexual dimorphism.     

SEXUAL REPRODUCTIO IN CODIUM FRAGILE SSP. TOMENTOSOIDES (CHLOROPHYCEAE) FROM THE NORTHEAST COAST OF NORTH AMERICA1

Male and female gametes were formed within the same gametangiurn on reproductive plants of Codium fragile (Sur.) Hariot ssp. tomentosoides (Van Goor) Silva growing off Appledore Island in the Gulf of Maine, USA. The female gametes were approximately twice the diameter of male gametes and outnumbered male gametes in the same gametangium by approximately six times. Fusion appears to require gametes from different gametangia if not different plants.