LIFE HISTORY AND IN SITU GROWTH RATES OF ALEXANDRIUM TAYLORI (DINOPHYCEAE, PYRROPHYTA)

Alexandrium taylori Balech is a phototrophic marine dinoflagellate. It produced recurrent blooms during the summer months (July and August) of 1994 to 1997 in La Fosca beach (NW Mediterranean). In addition to a motile vegetative form, A. taylori had two benthic forms: temporary cysts and resting cysts. Temporary cysts were a temporally quiescent stage produced from the ecdysis of the vegetative cell in both natural populations and laboratory cultures. Temporary cysts may divide to form motile cells. Resting cysts had a thicker wall than the temporary cysts and had a red accumulation body. Gametes and planozygotes were also observed in laboratory cultures. Alexandrium taylori showed in situ diurnal vertical migration with an increase of vegetative cells in the water column in the morning through midday, with concentrations peaking in the afternoon followed by lower levels at night. Most vegetative cells lost their thecae and flagella, and with them their motility, turning into temporary cysts that settled in the early evening. The number of temporary cysts in the water column rose in the evening and at night. The temporary cysts gave rise to motile cells the following morning. Synthesis of DNA occurred in vegetative cells at night, and a preferential period of cell division occurred at sunrise. The estimated division rate in the field was 0.4–0.5 vegetative cells·day⁻¹. Temporary cysts had twice the DNA of a G₁ vegetative cell. The minimum in situ division rate of the temporary cysts was 0.14 day⁻¹. The role of the resting and temporary cyst population in the annual recurrence and maintenance of the A. taylori bloom is discussed.     

Light is a crucial signal for zoosporogenesis and gametogenesis in some green microalgae

Patterns of reproduction were investigated in some microalgal species of Chlorophyceae (Botryosphaerella sudetica, Neochloris aquatica, Neochloris vigensis, Bracteacoccus minor). Under continuous light, the microalgae reproduced asexually producing autospores. However, appropriate manipulation of external conditions led to a change in the reproduction pattern towards production of zoospores or gametes. Production of zoospores and gametes was inhibited by light; motile cells emerged when microalgae were cultivated in darkness. The period of dark treatment necessary for zoosporogenesis or gametogenesis differed substantially among species that were tested. Sexual reproduction was observed in Neochloris vigensis and Bracteacoccus minor, whose generative life cycle had not been previously reported. The morphology of motile cells, the mode of sexual reproduction, and the efficiency of both the production of motile stages and mating events, were investigated. In order to gain detailed insights into patterns of reproduction, Botryosphaerella sudetica was selected for investigation under different light treatments. Non-actinic red light applied in the early phase of dark cultivation (up to 2 h) suppressed both zoosporogenesis and gametogenesis. However, after a 3-h dark pre-treatment, red light treatment had no effect on zoosporogenesis or gametogenesis. In contrast, non-actinic blue light did not block zoosporogenesis or gametogenesis, regardless of the time of treatment. The possible role of a red-light photoreceptor in zoosporogenesis and gametogenesis is discussed.     

THE LIFE CYCLE OF PEDIASTRUM SIMPLEX1

A sexual cycle and a newly discovered mode of asexual reproduction by azygotes arc described for Pediastrum simplex. In the sexual cycle isogametes from colony cells fuse in pairs, develop into zygotes, which grow and divide, producing motile cells which become angular polyeders. Polyeders may divide internally to form 4-celled colonies, or they enlarge, cleave, and produce zoospores or nonmotile cells. Zoospores are released in vesicles, swarm, adhere to each other, and form, flat colonies I cell thick. The nonmotile cells adhere to each other, enlarge, break out of the polyeder, and become spheroidal colonies. In the asexual cycle, nonfusing gametes, and motile cells derived from colony cells, become 4-spined azygotes which enlarge, divide internally, and release motile cells developing into polyeders. These polyeders develop further like those from zygotes.     

ENCYSTMENT OF THE DINOFLAGELLATE GYRODINIUM UNCATENUM: TEMPERATURE AND NUTRIENT EFFECTS1

Sexual reproduction and encystment of the marine dinoflagellate Gyrodinium uncatenum Hulburt were induced in nitrogen and phosphorus-limited batch cultures. Sexuality did not occur under nutrient-replete conditions even when growth rate was reduced by non-optimal temperatures. Growth was optimal over a broader temperature range than encystment and virtually no cysts were produced at some low and high temperatures where growth occurred. Most cells initiated sexuality as intracellular pools of each limiting nutrient reached minimum or subsistence levels as much as four days after extracellular nutrients were exhausted. High nitrogen cell quotas during the phosphorus experiment indicate that sexuality was induced by a shortage of phosphorus and not by an indirect effect on nitrogen uptake. Total cyst yield corresponded to successful encystment of 9–13% of the motile populations, yet 60–85% of the plateau-phase motile cells were planozygotes (swimming zygotes formed from fusing gametes). Batch culture studies monitoring total cyst yield may thus seriously underestimate the extent of sexuality. More importantly, the number of cysts produced in a dinoflagellate population may be significantly reduced by environmental factors acting on the cells after sexual induction and fusion.     

Observation on the Sexual Reproduction of Peridinium bipes Stein (Dinophyceae)

Sexual reproduction of the dinoflagellate Peridinium bipes Stein was observed. At the late growth season (from late March to early April), the small, unarmored motile cells i.e. gametes are produced by division of the cate cell. Isogamy occurs in this species. Two gametes in fusion are morphologically indistinguishable, but their behavior are different. Before fusion, two gametes are connected by a transparent granular structure and move quickly for about ten minutes. When the plasmogamy almost completes, the fusing cells stop moving for a while and the transverse flagellum of one gamete is cast off. By staining with modified carbol fuchsin, it was proved that the karyogamy takes place soon after plasmogamy and the change of chromosomes in this period was also observed. The zygote keeps motile for about 14 days before casts off its two flagella and becomes aplanozygote. During this period i.e. planozygote stage, the zygote enlarges from 55 × 50 μm to 75 ×70 μm, intercolary bands connecting thecai plates widen, lots of oil droplets are produced as storage granules. After sinking to the bottom of flask, aplano- zygote continues changing: exospore wall is cast off, mesospore wall and endospore wall are thickened, oil droplets turn to starch grains, protoplast contracts and becomes spherical, a large red lipid granule, perhaps eye-spot, appears. Afterward, aplanozygote has become hypnozygote i.e. resting cyst. The type of sexual reproduction, the amphiesma of the zygote, the resistance of hypnozygotic wall to acid and alkali, the relationship between fossil dinoflagellate remain and the wall of hypnozygote were discussed. It was also considered that the three formas of P. bipes named by Huber-Pestalozzi were the different stages of zygote development.     

SEXUAL REPRODUCTION OF PERIDINIUM WILLEI (DINOPHYCEAE)1

Sexual reproduction was induced in the dinoflagellate Peridinium willei Huitfeld-Kass when exponentially growing cells were inoculated into nitrogen deficient medium. Small, naked vegetative cells produced by division of thecate cells acted as gametes. The zygote remained motile 13–14 days, during which time it enlarged and the theca formed became warty. Fourteen to 15 days following plasmogamy the zygote was nonmotile with the protoplast contracted. A large red oil droplet appeared and the wall thickened, becoming chitinized. Hypnozygotes with 4 nuclei were observed 7–8 wk following formation. Meiosis was inferred. The hypnozygote germinated, within 8 wk producing one post-zygotic cell retaining the red oil droplet. This cell divided within 24 h into 2 daughter cells each with a prominent red oil droplet. These daughter cells divided after 2 to 3 days into ordinary vegetative cells. Attempts to induce sexual reproduction by inoculation of cells into media deficient in a number of basic elements were unsuccessful.     

Life history stages of Pyramimonas tychotreta (Prasinophyceae, Chlorophyta), a marine flagellate from the Ross Sea, Antarctica

During a summer cruise to the Ross Sea (Antarctica) areas of snow‐covered sea ice were red‐coloured due to high concentrations of the recently described Pyramimonas tychotreta Daugbjerg. Light microscopy of living material revealed that the population was comprised of quadriflagellate motile cells and thick‐walled cysts. The red colour was due to large numbers of secondary carotenoid‐containing granules, positioned in the periphery of motile cells and cysts. Mature cysts also contained numerous starch grains and lipid droplets. Cells from a red‐coloured field sample turned green overnight as the secondary carotenoids disappeared when cells were placed in low light conditions. The sample then exhibited the typical grass‐green colour of motile cells observed in water samples from the area. Under reduced light motile cells showed strong positive phototaxis. The encystment process involved the asexual transformation of quadriflagellate cells into cysts. A single type of square cyst scale, with perforated floors and walls, replaced the body scales of motile cells. A marked extension, often ending in a hook was at each corner of the cyst scales. Germinating cysts produced four motile cells. Electron microscopy showed the cyst wall to be tri‐layered, with a thin, electron‐dense inner layer, a thick middle layer and a thin outer layer. Sea ice samples with dense populations of motile cells and cyst stages also contained elongate uniflagel‐late cells. These cells were covered with box scales, foot‐print scales, an underlayer of pentagonal scales, limuloid scales and flagellar hair scales identical to those present on the quadriflagellate stage. We tentatively suggest that the uniflagellate stage represents a gamete and its presence implies the occurrence of sexual reproduction. Although, fusion of gametes was not observed, a biflagellate cell with a larger volume was seen which may have been a zygote. How this stage fits into of the life history remains to be explained.     

SEXUAL REPRODUCTION OF PERIDINIUM GATUNENSE (DINOPHYCEAE)1

Sexual reproduction was induced in the dinoflagellate Peridinium gatunense Nygaard when exponentially growing cells were inoculated into nitrogen deficient medium. Small thecate cells produced by division of vegetative cells then acted as gametes. Thecae of fusing gametes broke in the girdle region and were lost. Zygotes thus formed remained motile 3–5 days during which time they enlarged slightly with the newly formed theca becoming warty. Three to 5 days following plasmogamy the zygote became nonmotile, the protoplast contracted, and the cell wall thickened. Hypnozygotes with 4 nuclei were observed ca. 10–12 h following formation. Meiosis was inferred. Hypnozygotes germinated within 12 h of formation producing 2 vegetative cells which divided within a 24 h period. Attempts to induce sexual reproduction by inoculation of cells into media deficient in a number of basic elements other than N were unsuccessful.     

Novel distribution pattern between coexisting sexual and obligate asexual variants of the true estuarine macroalga Ulva prolifera

Where sexual and asexual forms coexist within a species, the asexuals are often found to be prevalent in marginal habitats. This asexual distribution pattern has received evolutionary attention linked to the paradox of sex. In many marine species, there is a distributional trend of asexual modes being more common in lower salinity waters regarded as the ecogeographic marginal, being explained by negative effects of low salinities on sexual reproductive success. However, the distribution pattern of estuarine species recently adapted to low salinity waters has remained unknown. The brackish macroalga Ulva prolifera being a major benthic component of estuarine ecosystems includes a sexual variant and obligate asexual variants by means of motile spores. We examined the sexual–asexual distribution pattern of this alga along a salinity gradient in river estuaries. Surprisingly, opposite to the distributional trend of marine organisms, the results clearly showed the persistent predominance of sexuals in the lower salinity reaches than the asexuals. In addition, a frequent alternating of dioecious gametophytes and sporophytes in the sexual population was observed, suggesting the sexual reproductive process would be robustly performed in the low salinity waters. Considering U. prolifera had evolved from the ancestral marine species to become a true estuarine species of which the core habitat is the low salinity reaches, in a broad sense its sexual–asexual distribution pattern would be involved in asexual marginal occupations of the species range previously reported in other organisms. Based on the frozen niche variation model, we can give a concise explanation for the evolutionary process of this pattern; multiple asexuals with frozen genotypic variation had arisen from sexual ancestors undergoing low salinity adaptation and share the estuarine habitat with the sexuals at present.     

The life cycle of Gyrodinium instriatum (Dinophyceae) in culture*

The life cycle events of an unarmored dinoflagellate Gyrodinium instriatum Freudenthal et Lee has been investigated using clonal cultures. After the inoculation of vegetative cells into fresh medium, clumping of gametes was observed after a period of 10 days. In the clumps, a number of gametes were observed to be swimming in close contact with each other, pairing successively and then forming a plano-zygote. This clumping behavior is considered to be useful in the performance of sexual reproduction, particularly in the event of low cell density, because it increases the chance of fusion. Gametes of this species were most often isogarnous, although apparent amsogamous fusions were occasionally observed. When planozygotes were isolated and placed in fresh medium, they enlarged their size and finally divided into two cells. The daughter cells continued to multiply by binary fission and produced vegetative cells. Thus, G. instriatum has an alternative cycle between vegetative cells and zygotes without a hypnozygote stage. However, cysts of this species were transformed from large motile cells (pre-cyst cells) which are oblong and dorso-ventrally flattened in shape. These mottle pre-cyst cells have two longitudinal flagella, which may indicate that cysts of this species are of zygote origin. On the basis of these results, the relationship between zygotes and pre-cysts of G. instriatum is discussed. Excystment was enhanced by dark and cold treatments prior to the incubation for germination experiments with a germination success rate of 26–64%. Encystment was greatly inhibited by the lack of dark and cold treatments.     

蓖齿眼子菜繁殖多样性研究

 繁殖是影响水生植物生存竞争、分布以及群落结构动态变化的关键因素。通过野外观察和室内试验,较详细地介绍了蓖齿眼子菜(Potamogeton pectinatus)的繁殖策略。研究结果表明:蓖齿眼子菜有性繁殖中具有水表和水下气泡两种传粉方式;种子发生量较大,但种子发芽率低,不到6%,种子繁殖对种群恢复贡献不大,其主要作用在于远程传播和保持持久的种子库。蓖齿眼子菜具有广泛而高效的无性繁殖系统,可以通过断株、根状茎、地上块茎、地下块茎、地上茎节以及叶腋基部进行繁殖,其中最重要的是地下块茎和根状茎,次年种群恢复主要来自于地下块茎和根状茎。此外,对采用蓖齿眼子菜进行沉水植被恢复与重建提出了建议。     

ACROSYMPHYTON FIRMUM SP. NOV. (RHODOPHYTA,CRYPTONEMIALES), A NEW SUBTIDAL RED ALGA FROM NEW ZEALAND; DEVELOPMENTAL MORPHOLOGY AND DISTRIBUTION OF THE GAMETOPHYTE1

Acrosymphyton firmum sp. nov., is described from the northeastern coast of North 1., New Zealand. Gametophytes are spring–summer annuals which grow subtidally on cobbles. Thalli are uniaxial; each axial cell bears a whorl of four indeterminate and one determinate branchlets. Indeterminate branchlets are alternately arranged giving the thallus a distichous and feather-like appearance. Numerous corticating rhizoidal filaments are produced from the periaxial and lower whorl branchlet cells. These rhizoids entwine and obscure the main axis as the thallus develops until in the mature plant the axes have a firm consistency and lubricous texture. The carpogonial branch bearing short lateral filaments and auxiliary cell branch with terminal auxiliary cell place this new species in the genus Acrosymphyton Sjöstedt. Of the three described species in the genus, A. firmum is most similar to A. taylori. This is the first report of the genus Acrosymphyton and the only confirmed report of the family Dumontiaceae in New Zealand waters.     

Elimination of plastids during spermatogenesis and fertilization in the plant kingdom

Ultrastructural and genetic investigations involving diverse species of plants have demonstrated that plastids may be transmitted either biparentally or maternally during sexual reproduction. In species in which plastid transmission is maternal, elimination of plastids from the paternal parent may occur in a number of ways: exclusion from the male gamete during spermatogenesis, loss from the motile sperm, exclusion during fertilization, or degradation within the zygote. These diverse ways in which maternal inheritance of plastids is achieved suggest that this inheritance pattern may have evolved independently many times in response to different selective pressures in different phyletic lineages.     

What’s wrong with a little sex?

In many species, most (or all) offspring are produced by sexual means. However, theory suggests that selection should often favour the evolution of species in which a small fraction of offspring are produced sexually, and the rest are produced asexually. Here, we present the analysis of a model that may help to resolve this paradox. We show that, when heterozygote advantage is in force, members of species in which sex is rare will tend to produce poorly adapted offspring when they mate. This problem should be less severe in species where most offspring are produced by sexual means. As a consequence, once the rate of sexual reproduction becomes sufficiently rare, the benefits of sex may vanish, leading to the evolution of obligate asexuality. Substantial benefits of sexual reproduction may tend to accrue only if a large proportion of offspring are produced sexually. We suggest that similar findings are likely in the case of epistatic interactions between loci.     

Multiple direct transitions from sexual reproduction to apomictic parthenogenesis in Timema stick insects

Transitions from sexual reproduction to parthenogenesis may occur along multiple evolutionary pathways and involve various cytological mechanisms to produce diploid eggs. Here, we investigate routes to parthenogenesis in Timema stick insects, a genus comprising five obligate parthenogens. By combining information from microsatellites and karyotypes with a previously published mitochondrial phylogeny, we show that all five parthenogens likely evolved spontaneously from sexually reproducing species, and that the sexual ancestor of one of the five parthenogens was probably of hybrid origin. The complete maintenance of heterozygosity between generations in the five parthenogens strongly suggests that eggs are produced by apomixis. Virgin females of the sexual species were also able to produce parthenogenetic offspring, but these females produced eggs by automixis. High heterozygosity levels stemming from conserved ancestral alleles in the parthenogens suggest, however, that automixis has not generated the current parthenogenetic Timema lineages but that apomixis appeared abruptly in several sexual species. A direct transition from sexual reproduction to (at least functional) apomixis results in a relatively high level of allelic diversity and high efficiency for parthenogenesis. Because both of these traits should positively affect the demographic success of asexual lineages, spontaneous apomixis may have contributed to the origin and maintenance of asexuality in Timema .     

Studies on woloszynskioid dinoflagellates VI: description of Tovellia aveirensis sp. nov. (Dinophyceae), a new species of Tovelliaceae with spiny cysts

A new species of Tovellia, T. aveirensis, is described on the basis of light (LM) and scanning electron microscopy (SEM) of motile cells and resting cysts, complemented with transmission electron microscopy (TEM) of flagellate cells and phylogenetic analysis of partial sequences of the large subunit ribosomal rRNA gene. Both vegetative cells and several stages of a life cycle involving sexual reproduction and the production of resting cysts were examined in cultures established from a tank in the University of Aveiro campus. Vegetative cells were round and little compressed dorsoventrally; planozygotes were longer and had a proportionally larger epicone. Chloroplast lobes were shown by TEM to radiate from a central, branched pyrenoid, although this was difficult to ascertain in LM. The amphiesma of flagellate cells had mainly 5 or 6-sided vesicles with thin plates, arranged in 5–7 latitudinal series on the epicone, 3–5 on the hypocone. The cingulum had 2 rows of plates, the posterior row extending into the hypocone and crossed by a series of small projecting knobs along the lower edge of the cingulum. A line of narrow amphiesmal plates extended over the cell apex, from near the cingulum on the ventral side to the middle of the dorsal side of the epicone. Eight or 9 narrow amphiesmal plates lined each side of this apical line of plates (ALP). Resting cysts differed from any described before in having numerous long, tapering spines with branched tips distributed over most of the surface. Most mature cysts showed an equatorial constriction. Neither cysts nor motile cells were seen to accumulate red cytoplasmic bodies in any stage of the cultures. The phylogenetic analysis placed, with high statistical support, the new species within the genus Tovellia; it formed a clade, with moderate support, with T. sanguinea, a species notable for its reddening cells.     

Hybridization and sexual reproduction in the invasive alien Fallopia (Polygonaceae) complex in Belgium

BACKGROUND AND AIMS: The knotweed complex, Fallopia spp. (Polygonaceae), belongs to the most troublesome invasive species in Europe and North America. Vegetative regeneration is widely recognized as the main mode of reproduction in the adventive regions. However, the contribution of sexual reproduction to the success of these invasive species has only been detailed for the British Isles. An examination was made as to how hybridization may influence the sexual reproduction of the complex in Belgium and to determine how it may contribute to the dispersal of the species. METHODS: Studies were made of floral biology, reproductive success, seed rain, seed bank, germination capacity, seedling survival and dispersal capacity in order to characterize the reproductive biology of the species. Moreover, chromosome counts and flow cytometry were used to assess the hybrid status of seedlings produced by sexual reproduction. KEY RESULTS: In the area investigated, extensive sexual reproduction by hybridization within the complex, including one horticultural species, was demonstrated. A small percentage of seeds may be dispersed outside the maternal clone (>16 m) allowing the formation of genetically differentiated individuals. Seed germination was possible even after a winter cold period. CONCLUSIONS: The extensive sexual reproduction by hybridization could further contribute to the dramatic invasive success of knotweeds in Belgium and should not be underestimated when considering control and management measures.     

Scrippsiella hexapraecingula sp. nov. (Dinophyceae), a tida pool dinoflagellate from the Northwest Pacific

Specimens of dinoflagellate collected in tide pools along the Pacific coast of central and southern Japan are described as a new species,Scrippsiella hexapraecingula Horiguchi et Chihara, of the Peridiniaceae (Class Dinophyceae). The plate formula is pp, x, 4′, 3a, 6″, 6c, 5‴, 2″" and, 5s, the same as that of other species ofScrippsiella, except in lacking one precingular plate. The genus must be emended, therefore, as having either six or seven precingular plates. This dinoflagellate migrates diurnally. In the morning motile cells are released from non-motile cells attached to the substrate and in the evening the motile cells swim down to settle on the bottom of the tide pool. Attached non-motile cells form either motile mono- or bispores. Sexual reproduction was not observed.     

Sexual reproduction development in apomictic Eulaliopsis binata (Poaceae)

Apomixis is a particular mode of reproduction that allows progeny formation without meiosis and fertilization. Eulaliopsis binata, a tetraploid apomictic species, is widely used for making paper, rope and mats. There is great potential for fixation of heterosis in E. binata due to autonomous endosperm formation in this species. Although most of its embryo sac originates from nucellus cells, termed apospory, we observed sexual reproduction initiation in 86.8 to 96.8% of the ovules, evidenced by callose deposition on the walls of cells undergoing megasporogenesis. However, only 2-3% mature polygonum-type sexual embryo sacs were confirmed by embryological investigation. Callose was not detected on aposporous initial cell walls. The aposporous initial cells differentiated during pre- and post-meiosis of the megaspore mother cell, while the sexual embryo sac degenerated at the megaspore stage. DNA content ratio of embryo and endosperm in some individuals was 2C:3C, based on flow cytometry screening of seed, similar to that of normal sexual seed. These results confirm that apomictic E. binata has conserved sexual reproduction to a certain degree, which may contribute to maintaining genetic diversity. The finding of sexual reproduction in apomictic E. binata could be useful for research on genetic mechanism of apomixis in E. binata.     

夜光藻有性繁殖研究进展

夜光藻是全球最主要的赤潮生物之一,也是我国近海常见的浮游甲藻。根据营养方式分为异养的红色夜光藻和混合营养的绿色夜光藻,前者广泛分布于温带和亚热带近岸水域,后者仅分布于热带西太平洋、阿拉伯海、阿曼湾和红海。夜光藻的生活史包括无性繁殖和有性繁殖过程。少部分营养细胞自发转变为配子母细胞,启动了有性繁殖。每个配子母细胞可形成大量配子,具有横沟、纵沟和2根鞭毛,形态与裸甲藻接近。配子两两融合形成合子,合子不经过休眠孢囊阶段直接发育成新的营养细胞。目前,对配子母细胞形成的调控机制、合子发育的影响因素的认识还存在分歧。研究发现,营养细胞经过一定次数的二分裂后都会转变为配子母细胞,而配子的存在能够中止此过程,使营养细胞继续进行二分裂。因此,有性繁殖可能通过产生新个体对种群增长做出贡献,还可能通过释放配子维持无性繁殖,进而促进种群增长。配子在相模湾水域全年都有分布,其丰度峰值与营养细胞丰度峰值同步或提前出现,配子的大量出现可能是赤潮形成的必要条件。对有性繁殖的研究佐证了夜光藻在甲藻的系统进化中处于较为古老的地位。此外,还简单介绍了研究夜光藻有性繁殖的主要方法,回顾了国内的夜光藻研究,并对相关研究进行了展望。