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.     

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.     

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。     

COMPARATIVE ULTRASTRUCTURE OF VEGETATIVE AND SEXUAL THECAE OF PERIDINIUM LIMBATUM AND PERIDINIUM CINCTUM (DINOPHYCEAE)

Peridinium limbatum (Stokes) Lemmerman reproduces sexually in nitrogen deficient medium. Sexual reproduction is homothallic and similar to that reported for other Peridinium species. Thecate planozygotes of P. limbatum and P. cinctum enlarge greatly. Suture bands connecting thecal plates enlarge unequally in P. cinctum producing a “warty” zygote and equally in P. limbatum maintaining the regular but enlarged appearance of the cell. Zygotes can be recognized by greatly enlarged striated suture bands.     

双角多甲藻有性生殖及厚垣合子形成条件的研究

双角多甲藻在MCV合成培养基中的最短加倍时间约6天,其有性生殖可用缺氮培养基进行诱导。批量培养的迟滞期和指数生长期末细胞的诱导效率最高,合子产率可达70％。厚垣合子形成的最适温度为19℃、pH7.4、光强5200lx。在鉴定的70个克隆藻株中,有2株为同宗配合;48株为异宗配合,其中“+”株23个,“-”株25个;其余20株未能诱导出有性生殖。合子开始萌发时,在中层壁和内层壁之间的空间出现1至多个“管状结构”,协助原生质体将合子壁顶破,外包一层薄壁的原生质体放出。     

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.     

EFFECTS OF PARENTAL FACTORS AND MEIOSIS ON SEXUAL OFFSPRING OF GYMNODINIUM NOLLERI (DINOPHYCEAE)1

Clonal strains of the dinoflagellate Gymnodinium nolleri Ellegaard and Moestrup were intercrossed to determine if cyst‐related traits are genetically regulated and to clarify unknown aspects in the sexuality of this species. The objectives were to determine whether the parental identity influenced the physiological and morphological aspects of the cyst offspring, and to describe and compare nuclear development and cell division of encysting and non‐encysting zygotes. Variables characteristic of each parental cross (difference in growth rates among parents, cyst production (CP), and genetic distance (GD) among parents assessed via an amplified fragment length analysis analysis) were studied to seek for possible relationships of the parental crosses with some characteristics of the cyst offspring (cyst size, length of dormancy period, germination success, and germling viability (V)). A principal component analysis using these variables showed three main results: (1) the dormancy period of cysts responded to a simple pattern of inheritance, (2) the larger the GD between parents, the smaller the CP, and progeny V, and (3) the size of cysts was influenced by both CP and the parental strain identity. A stable inheritance of the short dormancy period (14.6±5.5 days), dominant over medium (31.0±8.5 days) and long periods (52.7±9.2 days), was confirmed through two subsequent generations of cysts. The regulation of the sexual processes by a multiple loci system is discussed based on the pattern of inheritance of the dormancy period and the number of sexual recombination events recorded within cultures with self‐CP capability. Fusion of the gamete nuclei happened 0–48 h after the total cytoplasmic fusion. The nucleus of the zygote was bilobed and had thick and distinct chromosomes. Similar processes of nuclear and cell division occurred in the non‐encysting or encysting planozygote, and were characterized by the loss of the chromosomal structure, an apparent increase of the DNA content, and the formation of thinner chromosomes.     

SEXUAL REPRODUCTION OF PERIDINUM CINCTUM F. OVOPLANUM (DINOPHYCEAE)1,2

Sexual reproduction is induced in the dinoflagelate Peridinium cinctum f. ovoplanum Lindemann when exponentially growing cells are inoculated into nitrogen deficient medium. Small, naked vegetative cells, produced by division of the thecate cells, then act as gametes. The zygote remains motile for 12–13 days during which time it enlarges and the theca which it forms becomes warty. Thirteen to 14 day s following plasmogamy the zygote is nonmotile, the protoplast contracts, a large red oil droplet appears, the wall thickens and becomes chitinized. This hypnozygote germinates within 7–8 weeks at 20 c producing 1 post-zygotic cell retaining the large red oil droplet. The presence of 4 nuclei in these post-zygotic cells may be demonstrated by staining them with acetocarmine. Two of these nuclei are smaller than the other two and probably abort. One may infer that meiosis occurs immediately prior to or at the germinartion of the hypnozygote. This post-zygotic cell divides within 24 h into 2 daughter cells each with a promment red oil droplet. These daughter cells divide after 2–3 days into ordinary vegetative cells. Attempts to induce sexual reproduction by changes in temperature or light and by inoculation of cells into media deficient in a number of basic elements were unsuccessful.     

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.     

VEGETATIVE AND SEXUAL ASPECTS OF DINOPHYSIS PAVILLARDI (DINOPHYCEAE)

Phases in the life history of the dinoflagellate Dinophysis pavillardi Schroeder from cultured phytoplankton assemblages are described. Under stressful conditions, induced in the laboratory through substantial thermic and nutritive changes, vegetative cells divided repeatedly. Scanning electron and light microscopy of dividing specimens showed that thecal fission began with the separation of the sulcal and ventral epithecal plates and the simultaneous dislocation of the pore plates from the right cell half. The posterior progression of the division led to pairs of cells attached antapically, which produced a new wall of reduced size. This phase of the life cycle coincided with the appearance and development of small forms of D. pavillardi, which displayed cytological features and behavior typical of male gametes, suggesting a process of gametogenesis through depauperating mitotic divisions. Anisogamy occurred at the time of the maximum production of small cells and involved the shedding of thecal components by the smaller gamete and subsequent cytoplasmic fusion and formation of planozygotes. Although the dormancy aspects of this species remain unknown, these observations provide the first evidence of sexuality .     

SEXUAL REPRODUCTION IN STEPHANODISCUS NIAGARAE (BACILLARIOPHYTA)1

We observed sexual reproduction in a clonal culture of Stephanodiscus niagarae Ehrenb. and used light and scanning electron microscopy to absent flagellated male cells, auxospore growth, initial valve structure and production, and subsequent daughter cell division. Free auxospores were spherical and nonsiliceous throughout growth, producing hemispherical initial valves devoid of spines and with nonfasciculate striae. Pregametangial cells averaged 43% of the diameter of the daughter cell population and were 1/9 the biovolume of initial, cells. This paper is the first confirmed report of sexual reproduction in S. niagarae, although it appears that specimens of Actinocyclus niagarae H. L. Smith, described from Lake Erie in 1878, are actually initial valves of S. niagarae.     

SEXUAL REPRODUCTION IN NAVICULA CRYPTOCEPHALA (BACILLARIOPHYCEAE)1

Homothallic sexual reproduction and auxosporulation were studied in monoclonal cultures and seminatural populations of the freshwater epipelic diatom Navicula cryptocephala Kütz. Gametangia paired via the girdle, one gamete was formed per gametangium (and hence one zygote per pair of gametangia), and gamete fusion took place without the formation of any copulation envelope or copulation canal. Superfluous nuclei from meiosis survived unusually long, so that gametes and young zygotes were probably functionally polyploid; later, all but two haploid nuclei degenerated. Expanded auxospores had a swollen center, but during formation of the initial valves, the auxospore contracted away from the perizonium to produce linear‐lanceolate valves. The pattern of reproductive behavior found in N. cryptocephala can be classified as type IIA2a auxosporulation in Geitler's system. The same type of zygote and auxospore formation seen in clonal cultures was observed in seminatural material from four lakes in Scotland and the Czech Republic. Variation in nuclear structure and auxosporulation in the N. cryptocephala species complex is discussed, as is the evolution of type II auxosporulation (one zygote per pair of gametangia) from type I auxosporulation (two zygotes per pair). The penalty of smaller numbers of zygote produced in type II may be outweighed by formation of larger auxospores (prolonging the vegetative phase) or more vigorous auxospores. The variation present among members of the N. cryptocephala complex indicates that biogeographical analyses based on use of the name N. cryptocephala, as performed recently to support the ubiquity hypothesis of protist distributions, are almost meaningless.     

SEXUAL AND APOMICTIC REPRODUCTION IN CALAMAGROSTIS (GRAMINEAE) FROM EASTERN NORTH AMERICA

Reproductive mode and chromosome numbers were determined for populations of several species of Calamagrostis from eastern North America. Calamagrostis pickeringii (2n = 28), C. perplexa (2n = 70), C. porteri subsp. porteri (2n = ca. 88–100) and C. porteri subsp. insperata (2n = 56) all have a sexual pattern of megagametophyte formation; the basal megaspore of a tetrad develops into a Polygonum-type embryo sac with proliferating antipodal cells characteristic of the Gramineae. In these four taxa self-incompatability, population structure and infrequent flowering limit seed production; they persist primarily by rhizomes and occupy relatively stable, late-successional habitats. Calamagrostis stricta subsp. inexpansa includes apomictic variants (2n = ca. 104–123)that produce megagametophytes by diplospory; the single archesporial cell divides mitotically to produce an embryo sac appearing identical to those formed by sexual species. The embryo and endosperm develop autonomously from egg cell and polar nuclei, respectively. Some apomictic individuals occasionally produce some pollen and may have the potential for reproducing sexually. Their seed set insured by apomixis, variants of subsp. inexpansa colonize disturbed, open habitats and have achieved wide distributions in glaciated regions of North America. Reinterpretation of relationships among taxa I examined necessitates the following new nomenclatural combinations; C. porteri subsp. insperata (Swallen) comb. nov. is based on C. insperata Swallen; C. stricta subsp. inexpansa (A. Gray) comb. nov. is based on C. inexpansa A. Gray and includes C. lacustris (Kearney) Nash and C. fernaldii Louis-Marie.     

SEXUAL REPRODUCTION AND INTERCROSSING IN VOLVULINA STEINW1

Gametes of Volvulina steinii bear near-apical mating papillae. Zygospore germination yields a single biflagellate cell that develops into a colony xuhose asexual progeny are all of the same mating type. Backcrossing of clones of progeny indicated a 1 :1 ratio of mating types among the progeny. Of 20 clones from a number of localities, none was homothallic and 3 showed no matins: reaction. Mating reactions of clones crossed in all possible combinations indicated the presence of 2 sexually isolated groups of clones producing smooth-walled zygospores and 1 group that produced spiny-walled zygospores. In the latter group weak and nonreciprocal mating reactions occurred in some clone combinations. Failure of germination of spiny-walled zygospores from certain crosses suggests further subdivision into genetically isolated groups.     

UTEX 1336: PERIDINIUM CINCTUM OR PERIDINIUM VOLZII (DINOPHYCEAE)?1

Plate pattern variation in UTEX clone 1336 and published photographs using this clone indicate the presence of two taxa: Peridinium cinctum and Peridinium volzii. All subclones established from UTEX 1336 contain the same two cell types. The identification of UTEX 1336 and its future use by researchers are discussed.     

THE SEXUAL AND ASEXUAL LIFE CYCLE OF GLENODINIOPSIS STEINII (DINOPHYCEAE)

The sexual and asexual portions of the life cycle of Glenodiniopsis steinii were examined at both the light and scanning electron microscopic levels. Asexual reproduction by cell division occurs every 2 to 3 d under optimal conditions. Gamete formation and sexual reproduction can be induced by either nitrogen deprivation or aging of cultures. Fusion of gametes peaks about 10 hr into the light cycle and requires about 10 hr for completion. Cells remain motile during fusion but the zygote loses motility. The cell then undergoes changes that give it a thick-walled, warty appearance.     

SEXUAL REPRODUCTION IN CLOSTERIUM MONILIFERUM AND CLOSTERIUM EHRENBERGII1

Sexual reproduction in C. moniliferum is described for the first time. The morphology of conjugation is quite like that of C. ehrenbergii. Homothallic strains of both species usually produce single zygospores between daughter cells that have just divided. However, 2 homothallic clones of C. moniliferum form twin zygospores between conjugants which have paired before division and conjugation. This has not been observed in C. ehrenbergii. Heterothallic strains of both species form twin zygospores in the same manner. Heterotfiallisrn seems a well-established feature in both species. Germination and the survival of 2 products of meiosis arc typical of other desmids which have been investigated.     

NUTRITION AND SEXUAL REPRODUCTION IN THE WATER MOLD ACHLYA

The entire sexual reproductive progression in the heterothallic water mold Achlya ambisexualis Raper occurs in the absence of exogenous nutrients. Mated male and female mycelia, previously grown on a chemically defined medium, can utilize endogenous materials for the formation and maturation of sexual reproductive structures. Under these conditions the initiating sexual hormone antheridiol, produced by the female, exerts its morphogenetic effect on male mycelium in the absence of exogenous nutrients.     

PERIDINIUM BALTICUM (LEVANDER) LEMMERMANN,AN UNUSUAL DINOFLAGELLATE WITH A MESOCARYOTIC AND AN EUCARYOTIC NUCLEUS123

Light and electron microscopy indicate that Peridinium balticum possesses 2 Feulgen-positive, membrane-bound nuclei which divide synchronously. One nucleus has the typical structure of dinocaryotic dinoflagellates, while the other nucleus has a structure typical of eucaryotic organisms. Connections between each nucleus and the endoplasmic reticulum are common. Membrane-bound vesicles are intimately associated with the nuclear envelope of the eucaryotic nucleus.