SEX RATIO VARIATION IN THE LESSONIA NIGRESCENS COMPLEX (LAMINARIALES,PHAEOPHYCEAE): EFFECT OF LATITUDE,TEMPERATURE, AND MARGINALITY1

Little is known about variation of sex ratio, the proportion of males to females, in natural populations of seaweed, though it is a major determinant of the mating system. The observation of sexual chromosomes in kelps suggested that sex is partly genetically determined. However, it is probably not purely genetic since the sex ratio can be modified by environmental factors such as salinity or temperature. In this paper, sex ratio variation was studied in the kelp Lessonia nigrescens Bory complex, recently identified as two cryptic species occurring along the Chilean coast: one located north and the other south of the biogeographic boundary at latitude 29°–30° S. The life cycle of L. nigrescens is characterized by an alternation of microscopic haploid gametophytic individuals and large macroscopic fronds of diploid sporophytes. The sex ratio was recorded in progenies from 241 sporophytic individuals collected from 13 populations distributed along the Chilean coast in order (i) to examine the effect of an environmental gradient coupled with latitude, and (ii) to compare marginal populations to central populations of the two species. In addition, we tested the hypothesis that the sex ratios of the two cryptic species would be affected differently by temperature. First, our results demonstrate that sex ratio seems to be mainly genetically determined and temperature can significantly modify it. Populations of the northern species showed a lower frequency of males at 14°C than at 10°C, whereas populations of the southern species showed the opposite pattern. Second, both species displayed an increased variation in sex ratio at the range limits. This greater variation at the margins could be due either to differential mortality between sexes or to geographic parthenogenesis (asexual reproduction).     

LAMINARIOCOLAX SP. (PHAEOPHYCEAE) ASSOCIATED WITH GALL DEVELOPMENTS IN LESSONIA NIGRESCENS (PHAEOPHYCEAE)1

As part of a long‐term study on the biology and ecology of the intertidal kelp Lessonia nigrescens Bory, we report on the occurrence of gall development on this alga, identified the possible causal agent, and assessed the extent of the phenomenon in two wild stands of the host. Our results showed that galls affecting natural populations of L. nigrescens were associated with the infection by a filamentous brown algal endophyte of the genus Laminariocolax. Assignment to Laminariocolax of the endophytes isolated from cultured gall tissue was based on the (i) high internal transcribed spacer 1 (ITS1) sequence similarity and phylogenetic relationship between the Chilean isolates and several species of the genus Laminariocolax endophytic in other kelps, (ii) reproductive and vegetative features of the endophyte in culture, and (iii) anatomical agreement of fully developed galls of Lessonia with those described for other kelp galls caused by endophytic members of Laminariocolax. Unequivocal identification at the species level of the endophytes infecting Lessonia, however, awaits further studies.     

NICHE PARTITIONING AND THE COEXISTENCE OF TWO CRYPTIC DICTYOTA (DICTYOTALES,PHAEOPHYCEAE) SPECIES FROM THE CANARY ISLANDS1

Coexistence in a homogeneous environment requires species to specialize in distinct niches. Sympatry of cryptic species is of special interest to both ecologists and evolutionary biologists because the mechanisms that facilitate their persistent coexistence are obscure. In this study, we report on two sympatric Dictyota species, D. dichotoma (Huds.) J. V. Lamour. and the newly described species D. cymatophila sp. nov., from the Canary Islands. Gene sequence data (rbcL, psbA, nad1, cox1, cox3, and LSU rDNA) demonstrate that D. dichotoma and D. cymatophila do not represent sister species. Rather, D. cymatophila and D. dichotoma have converged on a nearly identical morphology, only to be distinguished with detailed morphometric observations. Both species co‐occur in eulittoral pools and the shallow subtidal in Tenerife. Even though D. cymatophila was more dominant in wave‐exposed places and D. dichotoma in less exposed areas, the spatial distribution of both species overlapped in intermediate habitats. The species display radically different phenologies. D. dichotoma reached its highest density in winter and early spring and disappeared nearly completely in autumn, while D. cymatophila dominated the study site from July until November. The timing of gamete release also differs between both species, D. dichotoma releasing gametes twice every lunar cycle, while the release of gametes in D. cymatophila occurred roughly every other day.     

PHYLOGEOGRAPHIC STUDIES IN THE TROPICAL SEAWEED CLADOPHOROPSIS MEMBRANACEA (CHLOROPHYTA,ULVOPHYCEAE) REVEAL A CRYPTIC SPECIES COMPLEX1

The seaweed Cladophoropsis membranacea (Hofman Bang ex. C. Agardh) Børgesen is a widely distributed species on coral reefs and along rocky coastlines throughout the tropics and subtropics. In a recent population‐level survey openface>1600 individuals with eight microsatellite loci, a number of isolates from biogeographically disjunct locations could not be amplified for any of the loci. Nonamplifiable and amplifiable isolates co‐occurred within the Canary Islands, Cape Verde Islands, and in the Caribbean. These unexpected results led to question whether or not C. membranacea is a single species. Phylogenetic relationships were evaluated using rDNA ITS1 and ITS2 sequence comparisons from 42 isolates sampled from a subset of 30 of the 66 locations. Four well‐supported clades were identified. Sequence divergence within clades was <1%, whereas between‐clade divergence was 2%–3%. Intraindividual variation was extremely low with no effects on the analysis. A strong, but imperfect, correspondence was found between ITS clades and amplifiable microsatellite loci. It is concluded that C. membranacea consists of three cryptic species. Using Pacific isolates as an outgroup, the most basal clade included the Central Canary Islands, Cape Verde, and Bonaire (Caribbean) isolates and thus spanned the widest latitude. Two derived sister clades consisted of a southern transtropical group stretching across the SE Caribbean to the Cape Verde Islands and African coast (but not the Canary Islands) and a NE‐Canary Island‐Mediterranean clade that also included the Red Sea. The detection of overlapping biogeographic distributions highlights the importance of ecotypic differentiation with respect to temperature and the importance of shifting sea surface isotherms that have driven periodic extinctions and recolonizations of the Canary Islands—a crossroads of marine floral exchange—since the last glacial maximum.     

GENETIC AND MORPHOLOGICAL ANALYSES OF THE SOUTHERN BULL KELP DURVILLAEA ANTARCTICA (PHAEOPHYCEAE: DURVILLAEALES) IN NEW ZEALAND REVEAL CRYPTIC SPECIES1

Many macroalgae exhibit considerable intraspecific morphological variation, but whether such variation reflects phenotypic plasticity or underlying genetic differences is often poorly understood. We quantified both morphological and genetic variation of 96 plants from seven field sites across eastern South Island, New Zealand, to assess genetic differences between morphotypes of the southern bull kelp Durvillaea antarctica (Cham.) Har. Consistent DNA sequence differentiation across mitochondrial, plastid, and nuclear loci was correlated with two broadly sympatric morphotypes: “cape” and “thonged.” These ecologically, morphologically, and genetically distinct bull‐kelp lineages were previously considered to be environmentally determined phenotypes with no underlying genetic basis. Interestingly, the sheltered “cape” lineage appears essentially genetically uniform across its South Island range, whereas the exposed “thonged” lineage exhibits marked phylogeographic structure across its range. Results suggest that D. antarctica in New Zealand comprises two reproductively isolated species.     

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.     

CRYPTIC DIVERSITY AND PHYLOGENETIC RELATIONSHIPS WITHIN THE MASTOCARPUS PAPILLATUS SPECIES COMPLEX (RHODOPHYTA,PHYLLOPHORACEAE)1

Mastocarpus papillatus (C. Agardh) Kütz. is a common intertidal red alga occurring along the west coast of North America from Baja California to Alaska. Sequencing of both the chloroplast‐encoded rbcL gene and the nuclear ribosomal internal transcribed spacer (ITS) regions of ～200 specimens from California to Alaska revealed that M. papillatus is actually a complex of at least five species. All five species have high bootstrap support in phylogenetic analyses of both genetic regions, and in the case of the ITS marker, the species also have distinctive patterns of indels. Three of the species are localized in the mid‐ to upper intertidal, whereas two of the species occur in the low intertidal. The species also have different geographic ranges that overlap in the Vancouver Island area of British Columbia. No distinctive, reliable morphological differences were observed among the species. Although a variety of names are available for species in the complex, it is not yet clear which name goes with which species. As part of the survey, I also sequenced other species of Mastocarpus in the northeast Pacific region, and I provide new distribution records for M. jardinii ( J. Agardh) J. A. West and for a nonpapillate and probably undescribed species of Mastocarpus.     

THE RECLASSIFICATION OF LESSONIA LAMINARIOIDES (LAMINARIALES,PHAEOPHYCEAE): PSEUDOLESSONIA GEN. NOV.1

A new genus, Pseudolessonia, is proposed for the kelp Lessonia laminarioides Postels et Ruprecht (Laminariales, Phaeophyceae), which occurs on the northwest side of the Sea of Okhotsk, in the northwest Pacific Ocean. Pseudolessonia is monotypic and differs from Lessonia in its short primary stipes and its corrugated, unilaterally arranged blades with entire margins. This species is transferred on the basis of morphology and plastid gene sequence comparisons. We determined psaA and rbcL gene sequences from 17 taxa of Pseudolessonia, Lessonia, and putative relatives. Analyses of individual and combined data sets resulted in congruent trees showing a clear separation of Pseudolessonia laminarioides from Lessonia, but suggesting its sister relationships with the clade of Nereocystis, Macrocystis, Pelagophycus, and Postelsia in the North Pacific Ocean. On the other hand, Lessonia species from the South Pacific Ocean formed a strongly supported clade. The results indicate that the basal splitting of the blade, which has been considered a diagnostic character for the family Lessoniaceae, is a result of convergent evolution.     

SOURCE-SINK RELATIONSHIPS IN THE BLADE OF ALARIA ESCULENTA (LAMINARIALES,PHAEOPHYCEAE)1

The translocation of ¹⁴C-labelled photoassimilate was studied in blades of the kelp, Alaria esculenta (L.) Grev., which had been surgically modified to produce source and sink regions of various sizes. Thirty cm above the blade base a 2.5 cm dia. section of the blade received a 1 h pulse of H¹⁴CO₃^?. Efflux of ¹⁴C-labelled photoassimilate from this “source disc” was monitored over the next 8 days using a Geiger-Müller detector probe. Accumulation of ¹⁴C-labelled solutes by a “sink disc” of similar diameter, 20 cm below the source disc, was also monitored. Rate of ^l4C efflux from the source disc was governed by two factors: (1) total sink size and (2) feed-back from competing sources. In the latter case, source export was depressed if the portion of the blade, just distal to the source disc, was present. While the initial ¹⁴C influx rate into the sink disc was greater when competing sinks were present, the total accumulation of ¹⁴C-photoassimilates was 2–3 times higher in the sink disc when competing sources were not present. Basipetal translocation velocity (1.3–1.7 cm h^?1) was unaffected by competing sources and sinks.     

THE RATE OF TRANSLOCATION IN ALARIA ESCULENTA (LAMINARIALES,PHAEOPHYCEAE)12

The rate of translocation of organic carbon in blades of Alaria esculenta (L.) Grev. (Laminariales) was calculated from blade growth data. The cross sectional area of sieve filaments in the midrib medulla was estimated from light microscopic examination of fresh material. Files of these filaments form a perimedullar ring occupying ca. 2/3 of the medulla. Values computed for specific mass transfer of carbon into the blade meristem ranged from 36.2 to 60.8 mg C.wk^?1.0.1 mm^?2 sieve filaments.     

云南省库态按蚊复合体的亲缘种型研究(双翅目,蚊科)(英文)

库态按蚊是1个由5种亲缘种型组成的按蚊复合体,这些种型分别被命名为A、B、C、D 和 E 型。在使用PCR 检测还没有条件的地区,鉴定库态按蚊亲缘种型的一个重要方法是作染色体组型分析。使用库态按蚊幼虫作染色体涂片,对云南地区的库态按蚊种型作了初步研究,并首次报道了该地区存在的亲缘种型成员,认为在云南省库态按蚊 A型和 B 型在同一个地区并存。同时,还展示了库态按蚊复合体的染色体组型、核型模式图及种型在中国毗邻国家的分布地图。     

ULTRASTRUCTURE OF SWARMERS IN THE LAMINARIALES (PHAEOPHYCEAE). I. ZOOSPORES1

Zoospores of 17 species in 14 genera of Laminariales, collected in the northeast Pacific Ocean, were studied by electron microscopy. These zoospores are unique in the brown algae in lacking both an eyespot in the single chloroplast and any associated swelling at the base of the shorter, posterior flagellum. Spores of all species examined possess a distal whiplash portion on the longer, mastigoneme-bearing anterior flagellum. This appendage may sometimes be as long as the mastigoneme-bearing portion of the flagellum, but it is only seldom preserved in the preparations for electron microscopy. A microtubular cytoskeleton is probably responsible for maintaining the shape of the spore. It consists of a short band of about 10 microtubules between the two basal bodies, scattered tubules converging at the anterior of the spore, a band of 7–9 tubules directed anteriorly from the anterior basal body, and a band directed posteriorly from the posterior basal body. These anterior and posterior bands may form one continuous band looping around the periphery of the spore. Variation with possible taxonomic significance was found in the ultrastructure of vesicles which apparently contain adhesive material, and which are extruded through the plasmalemma when the zoospores settle.     

ULTRASTRUCTURE OF SWARMERS IN THE LAMINARIALES (PHAEOPHYCEAE). II. SPERM1

The ultrastructure of sperm from 13 species in 11 genera of Laminariales collected in the northeast Pacific Ocean is unique in the brown algae. The sperm are elongate, and possess a nucleus, several mitochondria and two or three chloroplasts, but no eyespot. The anterior flagellum bears mastigonemes on the proximal half of its length; a distal “whiplash” portion lacks mastigonemes and is an extension of only the two central singlet microtubules of the axoneme. A peculiar feature of these sperm is the posterior flagellum, which is longer than the anterior flagellum and tapers distally as the doublet microtubules become singlets and decrease in number. This feature contrasts with the laminarialean zoospore, which possesses a short posterior flagellum with the usual “9 + 2” axoneme. The structure of these sperm differs from that reported for Chorda, the sperm of which resembles a primitive brown algal zoospore. The facts support the concept that Chorda is the most primitive member of the Laminariales.     

MORPHO‐FUNCTIONAL PATTERNS OF PHOTOSYNTHESIS IN THE SOUTH PACIFIC KELP LESSONIA NIGRESCENS: EFFECTS OF UV RADIATION ON 14C FIXATION AND PRIMARY PHOTOCHEMICAL REACTIONS1

The morpho‐functional patterns of photosynthesis, measured as ¹⁴C‐fixation and chl fluorescence of PSII, also as affected by different doses of UV radiation in the laboratory were examined in the South Pacific kelp Lessonia nigrescens Bory of the coast of Valdivia, Chile (40°S). The results indicated the existence of longitudinal thallus profiles in physiological performance. In general, blades exhibited higher rates of carbon fixation and pigmentation as compared with stipes and holdfasts. Light‐independent ¹⁴C fixation (LICF) was high in meristematic zones of the blades (3.5 μmol ¹⁴C·g^?1 fresh weight [FW]·h^?1), representing 2%–16% (percentage ratio) of the photosynthetic ¹⁴C fixation (20 μmol ¹⁴C·g^?1 FW·h^?1). Exposures to UV radiation indicated that biologically effective UV‐B doses (BED_{photoinhibition300}) of 200–400 kJ·m^?2 (corresponding to current daily doses measured in Valdivia on cloudless summer days) inhibit photosynthetic ¹⁴C fixation of blades by 90%, while LICF was reduced by 70%. The percentage ratio of LICF to photosynthetic ¹⁴C fixation increased under UV exposure to 45%. Primary light reactions measured as maximum quantum yield (F_v/F_m) and electron transport rate (ETR) indicated a higher UV susceptibility of blades as compared with stipes and holdfasts: after a 48 h exposure to UV‐B, the decrease in the blades was close to 30%, while in the stipes and holdfasts it was <20%. The existence of translocation of labeled carbon along the blades suggests that growth at the meristem may be powered by nonphotosynthetic processes. A possible functional role of LIFC, such as during reduction of photosynthetic carbon fixation due to enhanced UV radiation, is discussed. These results in general support the idea that the UV‐related responses in Lessonia are integrated in the suite of morpho‐functional adaptations of the alga.     

THE RELEASE,SETTLEMENT AND GERMINATION OF ZOOSPORES IN CHORDA TOMENTOSA (PHAEOPHYCEAE,LAMINARIALES)1,2

Details of zoospore germination in Chorda tomentosa Lyngb. are outlined. Uninucleate zoospores, when released are embedded in a mucilaginous mass of carbohydrate which dissolves and the biflagellate zoospores become motile. The long anterior flagellum is composed of a highly coiled terminal region and a rigid lower section bearing mastigonemes. The rigid, short posterior flagellum lacks mastigonemes. After initial contact by the tightly coiled region of the anterior flagellum, the zoospore draws itself to the substrate by flagellar resorbtion. After deposition of 3 wall layers the germling produces a germ tube. During this time the disc-shaped chloroplast enlarges undergoing changes in shape. As the germ tubes reach ca. 15 μm they cease forward growth and swell at their tips. The majority of cytoplasm of the original zoospore moves into the tube. Just before the nucleus enters the tube, centriole replication occurs. Mitosis is presumed to take place somewhere in the germ tube so that at 24 h, 2-celled gametophytes are produced.     

COORDINATNE NUCLEAR AND CHLOROPLAST DIVISION IN UNILOCULAR SPORANGIA OF LAMINARIA ANGUSTATA (LAMINARIALES,PHAEOPHYCEAE)1

Changes in the number of nuclei and chloroplasts were examined during the process of unispore formation in unilocular sporangia of Laminaria angustata. Just before meiosis, eight chloroplasts were always present in unilocular sporangial mother cells. The number of chloroplasts remained constant through meiosis. After the resulting four nuclei divided again (third nuclear division), a close association between a nucleus and a chloroplast developed among each of the eight nuclei and eight chloroplasts. The eight chloroplasts divided ahost synchronously before the synchronous division of the eight nuclei. Following the 16 nucleate stage with 16 chloroplasts and the final 32 nucleate stage with 32 chloroplasts, 32 unispores, each with a nucleus and a chloroplast, were fomd in unilocular sporangza of L. angustata. Immunofluorescence microscopy using an anti-centrin antibody showed that two anticentrin-stained structures (as future mitotic poles) occurred adjacent to each of the premitotic four nuclei, and each spot was located near a chloroplast. Therefore, after the third division, each of the eight nuclei established close contact with a chloroplast presumably mediated by the centrosomes.     

LIFE HISTORY AND SYSTEMATIC POSITION OF PSEUDOCHORDA GRACILIS SP. NOV. (LAMINARIALES,PHAEOPHYCEAE)1

Pseudochorda gracilis sp. Nov. (Pseudochordacease, Laminariales) is described from the Japan Sea coast of Hokkaido/ the species is subtidal, epilithic and annual, appearing in spring and maturing in winter. Erect thalli grow solitary or in tufts on a small discoid holdfast. They are simple, cord-shaped and hollow, with inner hyphal filaments, cylindrical medullary cells and paraphyses consisting of 3–6 cells. Hair tufts are observed only in young thalli. Unilocular sporangia are sessile and narrowly ovate. In culture, P. gracilis shows a heteromorphic life history with oogamy, characteristic of the order Laminariales. Gametophytes are dioecious and dimorphic. Gametophytes mature under lower temperature conditions (usually below 10°C), and sporophytes mature under low temperature and short-day conditions (5°C, SD). The seasonal growth pattern of the species results from the photoperiod-temperature conditions controlling saprophyte maturation.     

A REASSESSMENT OF SPECIES BOUNDARIES IN CYSTOSEIRA AND HALIDRYS (PHAEOPHYCEAE,FUCALES) ALONG THE NORTH AMERICAN WEST COAST1

Application of phylogenetic species recognition to morphologically recognized species in the genera Cystoseira Agardh and Halidrys Lyngbye on North American west coasts revealed little genetic variation despite a remarkable degree of morphological variation currently used to recognize and delineate species. Whereas morphological characteristics allow recognition of two genera, four morphological species and three informal forms, maximum genetic variation among them was similar to that characteristic of the intraspecific level in European congeners and other Fucales. Among morphological species and forms, nucleotide variation in a combined 26S (large subunit (LSU)) and internal transcribed spacer (ITS) ribosomal DNA analysis was below 3% while it was 1% or less for the RUBISCO spacer of the chloroplast DNA. Comparison of the LSU data to available data for European congeners showed that the genera Cystoseira and Halidrys are not monophyletic and that the previously recognized Cystoseiraceae should be included within the family Sargassaceae. These observations suggest that the current taxonomy for the Sargassaceae fails to reflect evolutionary history because Atlantic and Pacific Cystoseira and Halidrys appear to have arrived at similar morphologies independently. Our results indicate a comparatively recent establishment on the west coast of North America of a sargassacean progenitor whose descendant taxa have experienced limited genetic divergence and are characterized by a high capacity for phenotypic variation despite their overall genetic similarity.     

SPECIES ISOLATION,GENITAL MECHANICS,AND THE EVOLUTION OF SPECIES-SPECIFIC GENITALIA IN THREE SPECIES OF MACRODACTYLUS BEETLES (COLEOPTERA,SCARABEIDAE, MELOLONTHINAE)

The question asked was why male genitalic structures have diverged in three syntopic species of Macrodactylus beetles. Four hypotheses were evaluated: 1. The ways in which male genitalia mesh with internal female structures indicate that selection for species isolation via mechanical exclusion (“lock and key”) is unlikely to explain the genitalic differences. 2. The specific mate recognition hypothesis also clearly fails to explain genitalic differences due to the implausibility of postulated environmental effects on genitalia, and lack of postulated coevolution of male and female morphologies. 3. Selection for species isolation via differences in genitalic stimulation (sensory lock and key) is unlikely due to relatively infrequent cross-specific pair formation and intromission in the field, and “excessive” numbers of species-specific genitalic structures and male courtship behavior patterns which nevertheless occasionally fail. It also fails to explain the frequent failure of intraspecific copulations to result in sperm transfer. This hypothesis cannot, however, be rejected as confidently as the previous hypotheses. 4. Conditions under which sexual selection by cryptic female choice could take place are common. Females frequently exercise their ability to prevent sperm transfer by conspecific males even after intromission has occurred, and females generally mate repeatedly, probably with different males. Males behave as if cryptic female choice is occurring, courting assiduously while their genitalia are within the female. Sexual selection by female choice could thus contribute to the divergence in genitalic structures.