Generationswechsel,Kernphasenwechsel und Sexualität der Braunalge Ectocarpus siliculosus im Kulturversuch

Zusammenfassung Der Entwicklungscyclus von Ectocarpus siliculosus wurde unter Kulturbedingungen untersucht. Gametophyten und Sporophyten können morphologisch und funktionell unterschieden werden. Die Gametophyten sind diöcisch, die Geschlechtsbestimmung erfolgt genotypisch. Sporophyten können in der haploiden, diploiden und tetraploiden Phase vorliegen. Sporophyten aller Kernphasen können unilokuläre Sporangien ausbilden, tetraploide und diploide Sporophyten führen dabei eine Reduktionsteilung durch. Schwärmer haploider Gametophyten und Sporophyten können sich spontan zu diploiden oder tetraploiden homozygoten Sporophyten entwickleln. Gametophyten können nur zusammen mit Sporophyten aus Schwärmern unilokulärer Sporangien entstehen (Heteroblastie). Aus der Reduktionsteilung tetraploider Sporophyten gingen diploide männliche Gametophyten hervor, deren Gameten mit normalen weiblichen Gameten kopulierten. Die verschiedenen Kernphasen und Wuchsformen stehen durch Reduktionsteilungen, Heteroblastie, Kopulation und spontane Aufregulierung der Chromosomenzahl miteinander in Verbindung.

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Culture experiments on life cycle, nuclear phases, and sexuality of the brown alga Ectocarpus siliculosus

Summary The life cycle of Ectocarpus siliculosus from Naples (Italy) was investigated, using well defined cultured material. Gametophytes and sporophytes differ morphologically and functionally. The gametophytes are dioecious, with genotypic determination of their sex. Sporophytes exist in the haploid, diploid and tetraploid phase. All sporophytes can form unilocular sporangia. In tetraploid and diploid sporophytes the formation of unilocular sporangia is connected with meiosis. Certain motile cells of haploid plants may spontaneously give rise to diploid or tetraploid sporophytes which are homozygous. Gametophytes can only be formed together with sporophytes form the swarmers of unilocular sporangia (heteroblasty). Meiosis in tetraploid sporophytes resulted in diploid gametophytes, the gametes of which fused with haploid female gametes. All observed nuclear phases and growth forms are connected with each other by meiosis, heteroblasty, fusion of gametes and spontaneous increase in chromosome number.

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Mit Unterstützung durch die Deutsche Forschungsgemeinschaft.     

Hybridization between two cryptic filamentous brown seaweeds along the shore: analysing pre‐ and postzygotic barriers in populations of individuals with varying ploidy levels

We aimed to study the importance of hybridization between two cryptic species of the genus Ectocarpus, a group of filamentous algae with haploid–diploid life cycles that include the principal genetic model organism for the brown algae. In haploid–diploid species, the genetic structure of the two phases of the life cycle can be analysed separately in natural populations. Such life cycles provide a unique opportunity to estimate the frequency of hybrid genotypes in diploid sporophytes and meiotic recombinant genotypes in haploid gametophytes allowing the effects of reproductive barriers preventing fertilization or preventing meiosis to be untangle. The level of hybridization between E. siliculosus and E. crouaniorum was quantified along the European coast. Clonal cultures (568 diploid, 336 haploid) isolated from field samples were genotyped using cytoplasmic and nuclear markers to estimate the frequency of hybrid genotypes in diploids and recombinant haploids. We identified admixed individuals using microsatellite loci, classical assignment methods and a newly developed Bayesian method (XPloidAssignment), which allows the analysis of populations that exhibit variations in ploidy level. Over all populations, the level of hybridization was estimated at 8.7%. Hybrids were exclusively observed in sympatric populations. More than 98% of hybrids were diploids (40% of which showed signs of aneuploidy) with a high frequency of rare alleles. The near absence of haploid recombinant hybrids demonstrates that the reproductive barriers are mostly postzygotic and suggests that abnormal chromosome segregation during meiosis following hybridization of species with different genome sizes could be a major cause of interspecific incompatibility in this system.     

Maintenance of Complex Life Cycles Via Cryptic Differences In The Ecophysiology Of Haploid And Diploid Spores Of An Isomorphic Red Alga1

Recognition of the wide diversity of organisms that maintain complex haploid–diploid life cycles has generated interest in understanding the evolution and persistence of such life cycles. We empirically tested the model where complex haploid–diploid life cycles may be maintained by subtle/cryptic differences in the vital rates of isomorphic haploid–diploids, by examining the ecophysiology of haploid tetraspores and diploid carpospores of the isomorphic red alga Chondrus verrucosus. While tetraspores and carpospores of this species did not differ in size or autofluorescence, concentrations of phycobiliproteins of carpospores were greater than that of tetraspores. However, tetraspores were more photosynthetically competent than carpospores over a broader range of photosynthetic photon flux densities (PPFDs) and at PPFDs found at both the depth that C. verrucosus is found at high tide and in surface waters in which planktonic propagules might disperse. These results suggest potential differences in dispersal potential and reproductive success of haploid and diploid spores. Moreover, these cryptic differences in ecological niche partitioning of haploid and diploid spores contribute to our understanding of some of the differences between these ploidy stages that may ultimately lead to the maintenance of the complex haploid–diploid life cycle in this isomorphic red alga.     

ADDITIONAL EVIDENCE FOR ECOLOGICAL DIFFERENCES AMONG ISOMORPHIC REPRODUCTIVE PHASES OF IRIDAEA LAMINARIOIDES (RHODOPHYTA: GIGARTINALES)1

Gametophytes are more abundant thou sporophytes in wave exposed rocky intertidal populations of Iridaea laminarioides Bory in Central Chile. In this study we experimental tested the differential effects of selected ecological factors on karyologically different life history phases. In the field, gametophytes dominated at higher elevation and during summer; tetrasporophytes were most abundant low in the intertidal and during the fall. Laboratory responses correlated with these patterns. Gametophytes exhibited greater desiccation tolerance than tetrasporophytes. Optimum growth of gametophytes occurred at higher temperatures (20°C) and longer photoperiods (16:8 h LD) than sporophytes (15°C and 12:12 h LD). Grazing preferences changed with the developmental stage of the alga, but all herbivores tested had increased preference for diploid tissues as compared to haploid. Number of spores produced with respect to total plant surface, or total rocky surface, or settlement of spores and their germination rate did not show significant differences between phases but showed great variability in space and time. Spontaneous spore release, however, was always higher in cystocarpic than in tetrasporangial thalli. Such a combination of results suggests that some real ecological differences exist between the two life history phases of I. laminarioides. Such ecological differences permit a prediction of vertical and temporal patterns of distribution for both phases. Horizontal patterns of distribution cannot be explained because the several selection factors probably interact differently in various habitats.     

Ploidy Distribution of the Harmful Bloom Forming Macroalgae Ulva spp. in Narragansett Bay,Rhode Island,USA, Using Flow Cytometry Methods

Macroalgal blooms occur worldwide and have the potential to cause severe ecological and economic damage. Narragansett Bay, RI is a eutrophic system that experiences summer macroalgal blooms composed mostly of Ulva compressa and Ulva rigida, which have biphasic life cycles with separate haploid and diploid phases. In this study, we used flow cytometry to assess ploidy levels of U. compressa and U. rigida populations from five sites in Narragansett Bay, RI, USA, to assess the relative contribution of both phases to bloom formation. Both haploid gametophytes and diploid sporophytes were present for both species. Sites ranged from a relative overabundance of gametophytes to a relative overabundance of sporophytes, compared to the null model prediction of √2 gametophytes: 1 sporophyte. We found significant differences in cell area between ploidy levels for each species, with sporophyte cells significantly larger than gametophyte cells in U. compressa and U. rigida. We found no differences in relative growth rate between ploidy levels for each species. Our results indicate the presence of both phases of each of the two dominant bloom forming species throughout the bloom season, and represent one of the first studies of in situ Ulva life cycle dynamics.     

Alternate life history phases of a common seaweed have distinct microbial surface communities

Macroalgal life histories are complex, often involving the alternation of distinct free‐living life history phases that differ in morphology, longevity and ploidy. The surfaces of marine macroalgae support diverse microbial biofilms, yet the degree of microbial variation between alternate phases is unknown. We quantified bacterial (16S rRNA gene) and microeukaryote (18S rRNA gene) communities on the surface of the common intertidal seaweed, Mastocarpus spp., which alternates between gametophyte (foliose, haploid) and sporophyte (encrusting, diploid) life history phases. A large portion (97%) of bacterial taxa on the surface Mastocarpus was also present in samples from the environment, indicating that macroalgal surface communities are largely assembled from the surrounding seawater. Still, changes in the relative abundance of bacterial taxa result in significantly different communities on alternate Mastocarpus life history phases, rocky substrate and seawater at all intertidal elevations. For microeukaryote assemblages, only high intertidal samples had significant differences between life history phases although sporophytes were not different from the rocky substrate at this elevation; gametophytes and sporophytes did not differ in microeukaryote communities in the mid and low zones. By sequencing three host genes, we identified three cryptic species of Mastocarpus in our data set, which co‐occur in the mid‐to‐low intertidal zone. In these samples, M. alaskensis sporophytes harboured distinct bacterial communities compared to M. agardhii and M. intermedius sporophytes, which were not distinguishable. Conversely, microeukaryote communities did not differ among species.     

Untersuchungen zur Entwicklungsgeschichte der Braunalge Ectocarpus siliculosus Aus Neapel

Zusammenfassung Kulturversuche und cytologische Untersuchungen an Ectocarpus siliculosus aus dem Mittelmeer (Neapel) führten zu folgenden Ergebnissen: die haploide Chromosomenzahl liegt zwischen 18 und 31. Es sind zwei morphologisch verschiedene Wuchsformen vorhanden, die dem Freilandmaterial gleichenden haploiden Gametophyten und die nur in Kultur beobachteten Sporophyten. Letztere können sowohl in der Haplophase als auch in der Diplophase vorliegen und bilden in jedem Fall neben pluriloculären auch uniloculäre Fortpflanzungsorgane aus. In den uniloculären Sporangien der diploiden Sporophyten findet die Reduktionsteilung statt, während sie in denselben Sporangien auf haploiden Pflanzen unterbleibt. Unter den Schwärmern aus den Gametangien der Geschlechtspflanzen finden sich viergeißlige bewegliche Zygoten. Unter den Nachkommen eines Gemisches von unkopulierten Schwärmern und Zygoten wurden haploide und diploide Sporophyten gefunden. Unter den Nachkommen aus uniloculären Sporangien haploider und diploider Pflanzen sind fast stets Vertreter beider Wuchsformen zu finden. Einige der hier mitgeteilten Beobachtungen stimmen nicht mit den Angaben früherer Autoren überein.

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Studies on the life cycle of the brown alga Ectocarpus siliculosus from Naples, Italy

Summary Culture experiments and cytological studies were carried out under well-defined conditions. The haploid chromosome number is between 18 and 31. There are two morphological types of plants: the well-branched gametophyte resembling the plants found in nature, and the unbranched sporophytic form which may be haploid or diploid, found only in cultures. Meiosis takes place in the unilocular sporangia on diploid sporophytes. Haploid sporophytes form unilocular sporangia without reduction of the chromosome number. The spores from unilocular sporangia both on diploid and haploid sporophytes give rise to plants with either sporophytic or gamethophytic growth. The gametophytes are homothallic, the gametes forming motile zygotes with four flagella. Parthenogenetic development of gametes exclusively results in the formation of haploid sporophytes. Both the haploid and diploid sporophytes can propagate by means of zoids from plurilocular sporangia. Several observations reported here disagree with the findings of other workers.

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Mit Unterstützung durch die Deutsche Forschungsgemeinschaft.     

ON THE LIFE CYCLE,CYTOLOGY AND TAXONOMY OF CLADOPHORA CALLICOMA FROM INDIA

The present investigation deals with the cultural observations on the morphology, reproduction, life cycle, cytology and taxonomy of a freshwater Cladophora, C. callicoma (L.) Kütz, from India. There is a regular isomorphic alternation of generations between quadriflagellate zoospore-producing diploid plants and biflagellate isogamete-producing haploid plants, coupled with heterothallism in the latter. Chromosome numbers of n = 12 and 2n = 24 were determined respectively for gametophytes and sporophytes. Twelve bivalents were counted in meiosis where chiasmata varied from one to three. In light of above observations, the affinity of C. callicoma with C. glomerata, to which the former has often been assigned, has been discussed. It is proposed that the C. glomerata complex, which consists of intraspecific polyploid races, might have two distinct lines of evolution with regard to life cycle; one with forms having higher ploidy levels and lacking an alternation of generations, and the other with forms having low ploidy levels and alternation of generations.     

Ploidy chimeras induced in haploid sporophytes of <Emphasis Type="Italic">Osmunda claytoniana</Emphasis> and <Emphasis Type="Italic">Osmunda japonica</Emphasis>

Haploid sporophytes of Osmunda claytoniana (2n = x = 22) were apogamously produced from calli when cultivated on a hormone-free medium. Flow cytometric analysis showed that ploidy chimeras were spontaneously produced in a haploid sporophyte of O. claytoniana and those of O. japonica that were obtained in the previous study. In the haploid sporophyte of O. claytoniana, a diploid pinnule and a partially diploid terminal segment were produced in a haploid pinna. In O. japonica, a haploid sporophyte yielded a diploid pinna in a haploid frond, and another haploid sporophyte yielded a diploid pinnule in a haploid pinna. Diploid chimeras were large in size and could be readily distinguished from other haploid parts of the fronds. It is likely that the chimeras were produced clonally from a single diploid cell that established chromosome doubling.     

Nuclear behavior and roles indicate that Codiolum phase is a sporophyte in Monostroma angicava (Ulotrichales,Ulvophyceae)

The life‐cycle system of Ulotrichales, a major order of Ulvophyceae, remains controversial because it is unclear whether the Codiolum phase, a characteristic unicellular diploid generation in ulotrichalean algae, is a zygote or a sporophyte. This controversy inhibits the understanding of the diversified life cycles in Ulvophyceae. To distinguish between zygotes and sporophytes, we have to examine not only whether diploid generations function as sporophytes, but also whether mitosis occurs before meiosis in diploid generations. However, the nuclear behavior in the Codiolum phases is largely unknown, probably because no suitable methods are available. Using fluorescent microscopy with ethidium bromide and transmission electron microscopy of cell‐wall‐dissected specimens, we report the nuclear behavior in the Codiolum phases of an ulotrichalean alga with a representative life cycle, Monostroma angicava. Each vegetative Codiolum phase had a single polyploid nucleus due to endoreduplication, a type of mitosis without nuclear division. During zoosporogenesis, the nucleus had a structure that would be a meiosis‐specific complex. We quantitatively showed that Codiolum phases grew extremely large and produced numerous zoospores. Our results suggest that an event comparable to mitosis occurs before meiosis in the Codiolum phase of M. angicava. This nuclear behavior and the functions (growth and zoospore production abilities) correspond to those of sporophytes. Therefore, the life‐cycle system of M. angicava is a heteromorphic haplo‐diplontic cycle. This system appears to be widely adopted among other ulotrichalean algae.     

Salt Tolerance of Ectocarpus siliculosus (Dillw.) Lyngb.: Comparison of Gametophytes,Sporophytes and Isolates of Different Geographic Origin

Forty Ectocarpus siliculosus isolates from a wide geographical range, including gametophyte and sporophyte plants, have all been acclimated to the same salinity for several years. Their salinity tolerances in respect of cell viability, photosynthesis and dark respiration were evaluated over the salinity range: 8 to 96 ‰. Significant differences in the physiological tolerances to salt stress compared with viability measurements were evident. Genotypic differences in salt tolerances between groupings of the isolates, and also differences in responses of gametophyte and sporophyte generations were found. However, diploid and haploid sporophyte material had similar tolerances. Triploid and tetraploid sporophytes did not have improved tolerances over those of diploid plants. Culture plants originating from low salinities in the Baltic Sea had broader tolerances than field material collected from Baltic waters of similar salinity.     

THE OCCURRENCE AND PHYLOGENETIC SIGNIFICANCE OF PUTATIVE PLACENTAL TRANSFER CELLS IN THE GREEN ALGA COLEOCHAETE

Following fertilization, zygotes of the green alga Coleochaete orbicularis, which are retained on the haploid thallus, first enlarge, then become covered with a layer of vegetative cells. Light microscopy and high-voltage electron microscopy revealed the presence of localized wall ingrowths in vegetative cells adjacent to zygotes. These covering cells resemble the gametophytic placental transfer cells of embryophytes in their morphology, location, and time of development. If Coleochaete cells with wall protuberances function as do placental transfer cells of embryophytes, their presence is evidence that photosynthates may be transported between haploid thallus cells and zygotes. Thus, a nutritional relationship between different phases of the life cycle, similar to that which occurs in embryophytes, may also have evolved in green algae. This first report of putative placental transfer cells in a green alga supports Bower's (1908) ideas concerning the origin of land plant sporophytes and alternation of generations. The presence or absence of cells with wall ingrowths in several species of Coleochaete was correlated with estimates of zygote-plant area ratios.     

The evolution of life cycles with haploid and diploid phases

Sexual eukaryotic organisms are characterized by an alternation between haploid and diploid phases. In vascular plants and animals, somatic growth and development occur primarily in the diploid phase, with the haploid phase reduced to the gametic cells. In many other eukaryotes, however, growth and development occur in both phases, with substantial variability among organisms in the length of each phase of the life cycle. A number of theoretical models and experimental studies have shed light on factors that may influence life cycle evolution, yet we remain far from a complete understanding of the diversity of life cycles observed in nature. In this paper we review the current state of knowledge in this field, and touch upon the many questions that remain unanswered. BioEssays 20 :453–462, 1998. © 1998 John Wiley & Sons, Inc.     

The evolution of the life cycle of brown seaweeds

The brown seaweeds (Phaeophyta) are well-suited for testing theories of the evolution of the sexual alternation of haploid and diploid generations because of the great diversity of life cycles within the phylum. Three theories are investigated in this paper. (1) Diploid growth evolves because it has the effect of complementing deleterious recessive mutations. This is rejected because (a) ancestral haplonty is not a parsimonious inference from current phylogenies; (b) the exaggeration of diploid growth does not evolve in a comb-like fashion; (c) forms with predominantly haploid growth have evolved from smaller isomorphic ancestors; and (d) there is no correlation between haploid growth and monoecy. (2) Diploid growth evolves when gamete dimorphism leads to intense sexual selection, favouring the production of genetically diverse gametes through meiosis. This is rejected because diere is no correlation between the dominance of the diploid generation and the degree of gamete dimorphism. It is possible to show that gamete dimorphism itself has evolved in the Phaeophyta through the increase in size of the macrogamete in forms that have evolved larger sporophytes. (3) Microthalli become specialized as gametophytes because fusion is promoted by releasing gametes into the boundary layer; macrothalli become specialized as sporophytes because dispersal is promoted by releasing zoospores into the water column. This is consistent with the sexual and reproductive biology of Phaeophyta. The classic sexual cycle can then be interpreted as evolving from an asexual alternation of microthallus and macrothallus, governed largely by environmental factors, through selection for the appropriate association of ploidy with vegetative size. The exceptions to this general rule are forms in which gametes are released from macrothalli, where a different suite of sexual characters has evolved.     

Attachment strength differs amongst life‐history stages of an intertidal,isomorphic red alga

Complex haploid‐diploid life cycles amongst marine organisms may be maintained by ecological differences in life‐history phases. For red algal species within the Gigartinaceae, such differences may be driven, in part, by different cell wall composition and resultant biomechanical strengths of haploid and diploid phases. A field experiment tested the attachment strengths of gametophytes and tetrasporophytes of the isomorphic red alga, Chondrus verrucosus (with comparisons of fertile and vegetative fronds, with and without natural tissue damage across three wave‐exposed sites). Seventy‐nine percent of all fronds broke at the stipe‐holdfast junction. There were significant differences in attachment strength (break force and break stress), but not gross morphology (frond length, number of branch axes, wet weight and cross‐sectional area of fronds that dislodged at the stipe‐holdfast junction) of life‐history phases, with tetrasporophytes exhibiting weaker tissue strength and attachment, and therefore greater susceptibility to dislodgement by waves. However, fertility and tissue damage did not consistently influence dislodgement in pull‐to‐break tests simulating the effects of single waves. The ecological and evolutionary consequences of greater susceptibility to dislodgement of tetrasporophytes (relative to gametophytes) warrant further investigation.     

Taxonomic value of a betaine lipid in Ectocarpus (Phaeophyceae) and the first record of a sexual population of E. fasciculatus for the Pacific Ocean

The betaine lipid DGTA differentiates between two species of Ectocarpus: it is present in E. fasciculatus Harvey, and lacking in E. siliculosus (Dillwyn) Lyngbye, Two ectocarpoid isolates from the coast of Chile, which could not be identified to species level, were found to belong to opposite DGTA types. Culture experiments showed that these plants were sporophytes. Their meiospores produced gametophytes of the species predicted by the lipid analysis. Promoted by this new approach, a sexual population of Ectocarpus fasciculatus has been detected for the first time in the Pacific Ocean.     

Absence of major differences between soluble proteins from haploid gametophytes and diploid sporophytes in the green alga Ulva mutabilis Føyn

Soluble proteins from haploid gametophytes and diploid sporophytes of the marine green alga Ulva mutabilis Føyn have been reexamined, using polyacrylamide gel electrophoresis and isoelectric focusing. A two-dimensional system resolved about 150 protein spots. In contrast to an earlier report (Hoxmark (1976) Planta 130, 327–332), no major differences could be detected between soluble proteins from the two generation types by any of the methods used.To whom correspondence should be addressed     

Gametogenesis and chromosome number in Postelsia palmaeformis (Laminariales, Phaeophyceae)

Gametophytes of the ‘sea palm’, the kelp Postelsia palmaeformis Ruprecht, produced gametes whether or not chelated iron was supplied in the culture medium, in contrast to the inhibition of gametogenesis seen with the absence of iron in many other kelps. As gametogenesis proceeded, every cell of the gametophytes was converted into a gamete so that the gametophytes did not continue to grow vegetatively. The portion of the life history from spore release through germination, gametophyte growth, gametogenesis, fertilization and growth of the young sporophyte was completed in 9–10 days under laboratory conditions. Chromosome counts showed that sporophytes had a diploid number of 26–34 chromosomes while sporangia and gametophytes had a haploid number of 14–17 chromosomes, indicating a typical haplodiplophasic life history as seen in other Laminariales.     

Evolution of alternation of haploid and diploid phases in life cycles

Eukaryotic sex leads to an alternation of haploid and diploid nuclear phases. Because all multicellular animals are diploid, diploidy is often considered a 'biological success' and many arguments have been advanced to explain the evolution of a prolonged diploid phase. Nevertheless, among eukaryotes three basic situations are encountered, where the vegetative individuals are diploid or haploid or both. These three basic life cycles are widely distributed among kingdoms and in some taxa the occurrence of different life cycles within the same species has been reported. This article briefly summarizes the different hypotheses on the evolution of reproductive life cycles and underlines how possibilities of variation for this trait may open new perspectives for research.     

Culture studies on the life history of Haplospora globosa and Tilopteris mertensii (tilopteridales,phaeophyceae)

The complete life histories of Tilopteris mertensii and haplospora globosa were followed in culture. Tilopteris shows a succession of identical plants through uninucleate “eggs” which develop parthenogenetically. In Haplospora, sporophytes alternate with gametophytes without sexuality and nuclear alternation. However, evidence for meiotic stages is found in sporangium initials. Gametophytes produce oogonia and antheridia, and eggs develop parthenogenetically. The chromosome number of Tilopteris is n = 62 (60–65). In both phases of Haplospora numbers are n = 50 (43–54). Haplospora from Heligoland perpetuates the sporophyte only at chromosome numbers of n = 25 (22–28).