Peronosporomycetes (Oomycota) from a Middle Permian Permineralised Peat within the Bainmedart Coal Measures,Prince Charles Mountains,Antarctica

The fossil record of Peronosporomycetes (water moulds) is rather sparse, though their distinctive ornamentation means they are probably better reported than some true fungal groups. Here we describe a rare Palaeozoic occurrence of this group from a Guadalupian (Middle Permian) silicified peat deposit in the Bainmedart Coal Measures, Prince Charles Mountains, Antarctica. Specimens are numerous and comprise two morphologically distinct kinds of ornamented oogonia, of which some are attached to hyphae by a septum. Combresomyces caespitosus sp. nov. consists of spherical oogonia bearing densely spaced, long, hollow, slender, conical papillae with multiple sharply pointed, strongly divergent, apical branches that commonly form a pseudoreticulate pattern under optical microscopy. The oogonia are attached to a parental hypha by a short truncated stalk with a single septum. Combresomyces rarus sp. nov. consists of spherical oogonia bearing widely spaced, hollow, broad, conical papillae that terminate in a single bifurcation producing a pair of acutely divergent sharply pointed branches. The oogonium bears a short truncate extension where it attaches to the parental hypha. We propose that similarities in oogonium shape, size, spine morphology and hyphal attachment between the Permian forms from the Prince Charles Mountains and other reported Peronosporomycetes from Devonian to Triassic strata at widely separated localities elsewhere in the world delimit an extinct but once cosmopolitan Palaeozoic to early Mesozoic branch of the peronosporomycete clade. We name this order Combresomycetales and note that it played an important role in late Palaeozoic and early Mesozoic peatland ecosystems worldwide.     

Oogonial release in Turbinaria conoides (J. Agardh) Kützing (Fucales,Sargassaceae)

The oogonium in Turbinaria conoides (J. Agardh) Kürzing at the time of release is 8-nucleate and consists of 3 wall layers namely exochiton, mesochiton and endochiton. The exochiton contains alginic acid. The mesochiton a mixture of alginic acid and sulphated polysaccharides. The endochiton that is evident in the unreleased 8-nucleate oogonium, stains for sulphated polysaccharides, but is PAS negative. In a released oogonium, the endochiton is Alcian blue and PAS positive and consists of both alginic acid and sulphated polysaccharides.Polysaccharide granules are secreted near the perinuclear region and in thecytoplasmic vacuoles during the initiation of mesochiton. Furthermore, at the distal end the mesochiton froms a pad that fuses with the exochiton. The pad consists of alternate thick and thin bands. The exochiton opens at one side due to the swelling of the oogonium and accumulation of sulphated polysaccharides between exochiton and mesochiton followed by formation and elongation of a mesochiton stalk. The 8-nucleate oogonium is, thus, extruded from the conceptacle, but remains attached to the exochiton by means of a long mesochiton stalk. The secretory paraphyses, at the time of oogonial release, produce copious amounts of polysaccharides. The oogonium is, thus, extruded through the ostiole by the combined efforts of the 3 wall layers that are histochemically and histologically different, in combination with secretion produced by secretory paraphyses.     

Developmental patterns of the hermaphroditic gonad in dicyemid mesozoans (Phylum Dicyemida)

Abstract. The development of the hermaphroditic gonad, the infusorigen, was studied in fixed specimens of 16 species belonging to four genera ( Conocyema, Dicyema, Dicyemennea , and Pseudicyema ) of dicyemid mesozoans. In addition, the literature on the cell lineages of infusorigens of eight other species was reviewed. After an agamete (axoblast) undergoes an unequal first division, excluding a small cell (the paranucleus), the resulting large cell divides to form the infusorigen. Five infursorigen cell lineage patterns can be identified: type I, both the first oogonium and the first spermatogonium are produced at the third division; type II, the first spermatogonium is produced at the second division and the first oogonium is produced at the third division; type III, the first spermatogonium is produced at the second division and the first oogonium is produced at the fourth division; type IV, the first oogonium is produced at the second division and the first spermatogonium is produced at the third division; and type V, the first oogonium is produced at the second division and the first spermatogonium is produced at the fourth division. Later development is similar in members of all genera. Mature infusorigens are similarly organized in all species examined. Sizes of agametes and infusorigen axial cells, the number of infusorigens, and the type of reproductive strategy were not correlated with infusorigen developmental patterns or cell lineage patterns.     

Fine structure of antheridia and oogonia of Phytophthora macrospora,the downy mildew fungus of rice plants

In the present paper fine structure of antheridium and oogonium ofPhytophthora macrospora (Sacc.)S. Ito etI. Tanaka, the downy mildew fungus of rice plants was discussed.Before the fertilization some nuclei and a large number of mitochondria were scattered in the cytoplasm of the antheridium. Many lipid granules were observed in the peripheral region, but vacuoles did not appear at this stage of antheridium. Many mitochondria were associated in the neighborhood of the fertilization pore. The wall at the pore was very thin, but the wall surrounding the pore was slightly swollen towards the inside.In the oogonium, many nuclei, mitochondria and cytoplasmic matrix were observed at the peripheral part. A large number of lipid granules was found in the oogonium, but they were more numerous in the peripheral region. The vacuoles developed as the oogonium matured. They were enveloped by tonoplast and contained vacuolar matrix. Many electron dense granules were in contact with the tonoplast or free in the vacuoles, and they were larger in the central part. As stated above, wall at the fertilization pore was thin. However, the oogonial wall surrounding the pore swelled protruding into the oogonium. An electron-dense layer was recognized between the antheridial and oogonial wall, and the walls of both the organs were closely in contact with each other.Contribution No. 252.     

Gametogenesis and auxospore development in actinocyclus (bacillariophyta)

cGametogenesis and auxospore development have been studied in detail in surprisingly few centric diatoms. We studied the development of sperm, eggs and auxospores in Actinocyclus sp., a radially symmetrical freshwater diatom collected from Japan, using LM and electron microscopy of living cultures and thin sections. Actinocyclus represents a deep branch of the 'radial centric' diatoms and should therefore contribute useful insights into the evolution of sexual reproduction in diatoms. Spermatogenesis was examined by LM and SEM and involved the formation of two spermatogonia (sperm mother-cells) in each spermatogonangium through an equal mitotic division. The spermatogonia produced a reduced 'lid' valve, resembling a large flat scale with irregular radial thickenings. Sperm formation was merogenous, producing four sperm per spermatogonium, which were released by dehiscence of the 'lid' valve. The sperm were spindle-shaped with numerous surface globules and, as usual for diatoms, the single anterior flagellum bore mastigonemes. One egg cell was produced per oogonium. Immature eggs produced a thin layer of circular silica scales before fertilization, while the eggs were still contained within the oogonium. Sperm were attracted in large numbers to each egg and were apparently able to contact the egg surface via a gap formed between the long hypotheca and shorter epitheca of the oogonium and a small underlying hole in the scale-case. Auxospores expanded isodiametrically and many new scales were added to its envelope during expansion. Finally, new slightly-domed initial valves were produced at right angles to the oogonium axis, after a strong contraction of the cell away from the auxospore wall. At different stages, Golgi bodies were associated with chloroplasts or mitochondria, contrasting with the constancy of Golgi-ER-mitochondrion (G-ER-M) units in some other centric diatoms, which has been suggested to have phylogenetic significance. Electron-dense bodies in the vacuole of Actinocyclus are probably acidocalcisomes containing polyphosphate.     

Protein kinase C is likely to be involved in zoosporogenesis and maintenance of flagellar motility in the peronosporomycete zoospores

The motility of zoospores is critical in the disease cycles of Peronosporomycetes that cause devastating diseases in plants, fishes, vertebrates, and microbes. In the course of screening for secondary metabolites, we found that ethyl acetate extracts of a marine Streptomyces sp. strain B5136 rapidly impaired the motility of zoospores of the grapevine downy mildew pathogen Plasmopara viticola at 0.1 μg/ml. The active principle in the extracts was identified as staurosporine, a known broad-spectrum inhibitor of protein kinases, including protein kinase C (PKC). In the presence of staurosporine (2 nM), zoospores moved very slowly in their axis or spun in tight circles, instead of displaying straight swimming in a helical fashion. Compounds such as K-252a, K-252b, and K-252c structurally related to staurosporine also impaired the motility of zoospores in a similar manner but at varying doses. Among the 22 known kinase inhibitors tested, the PKC inhibitor chelerythrine was the most potent to arrest the motility of zoospores at concentrations starting from 5 nM. Inhibitors that targeted kinase pathways other than PKC pathways did not practically show any activity in impairing zoospore motility. Interestingly, both staurosporine (5 nM) and chelerythrine (10 nM) also inhibited the release of zoospores from the P. viticola sporangia in a dose-dependent manner. In addition, staurosporine completely suppressed downy mildew disease in grapevine leaves at 2 μM, suggesting the potential of small-molecule PKC inhibitors for the control of peronosporomycete phytopathogens. Taken together, these results suggest that PKC is likely to be a key signaling mediator associated with zoosporogenesis and the maintenance of flagellar motility in peronosporomycete zoospores.     

鳖性腺发育的研究

湖南地区鳖的性成熟年龄是四年。成熟雌鳖每年产卵3—5次(窝),一般每次产卵8—12个,最多可达20—26个,是典型的多次产卵类型。由8—10微米直径的卵原细胞,长足到17—20毫米的成熟卵子,其间可以分为卵原细胞期、初级卵泡期、生长卵泡期和成熟卵泡期。在生长卵泡期可以看到由卵表伸向放射膜内的微绒毛(microvilli),这种结构与卵黄形成所需的物质转运有关。曲细精管中精子的正常形成和存活,不受季节变化的影响。进入雌鳖输卵管中的精子经过冬季到来年的生殖季节仍保持受精能力。       

Comparison of reproductive development in triploid and diploid female rainbow trout Oncorhynchus mykiss

The diploid rainbow trout Oncorhynchus mykiss reached sexual maturity 3 years after hatching and its oogenesis underwent four stages, which were oogonia, primary oocyte, secondary oocyte and egg. Reproductive development and hormone changes of 4 to 35 month‐old female O. mykiss were investigated using histological and radioimmunoassay methods in order to provide a theoretical and practical basis for the use of triploid female O. mykiss. The oogonium of the triploid female could develop into the oocytes of the prophase with abortion occurring later; the oogonium was surrounded by stroma cells to form the oogonium cluster and the gonads showed a virilescent tendency when the oogonium clusters were gradually replaced by spermatogenic‐like cytocysts. After 13 months, amounts of gonadotropic hormone (GtH‐I, GtH‐II) and oestradiol (17β‐E₂) in triploid females were lower than in diploid fish at corresponding time periods, but the amounts of testosterone (T) increased consistently after 21 months and were more than in diploid fish in the corresponding time periods (P > 0·05). The infertility of triploid females resulted from meiosis failure, which caused developmental abortion of oocytes and oogonium formed cytocysts before the prophase oocytes. The cytocyst formation was due to the lack of the normal interaction of ovum and follicular cells, the development of follicular cells producing steroids were inhibited, the arylate path from T to 17β‐E₂ was interrupted, concentration of 17β‐E₂ decreased and concentration of T increased in the blood, the content of vitellogenin (Vg) decreased in the liver with a low 17β‐E₂ and high T caused to ovaries to show a tendency to be virilescent.     

Malathion Toxicity: Effect on the Ovary of the Zebra Fish Brachydanio rerio (Cyprinidae)

The nucleic acids (DNA and RNA), protein contents and the activity of alkaline phosphatase was significantly decreased in the ovary of Brachydanio rerio after exposure to different concentrations of malathion, an organophosphorus pesticide. Total free amino acids and acid phosphatase activity, however, were enhanced. The decrease in the measured parameters depended on both concentration and time. Histopathological studies revealed developmental arrest at all levels from oogonium to tertiary stage oocytes. After transfer to fresh water without the pesticide, there was a marked trend of recovery in al' the parameters studied.     

腐霉属的一个新种和二个新记录

本文报告作者1982—1983年在北京和广东一些地区土壤中用已知方法(余永年,1975)分离到的几种腐霉,其中1个新种(Pythium borealis sp.nov.)、2个国内未报道的种(P.oligandrum和P.myriotylum)和1个已知种的变异株(P.ultimum variant)。文中对以上各菌进行了形态学描述。     

2,4-Diacetylphloroglucinol suppresses zoosporogenesis and impairs motility of Peronosporomycete zoospores

2,4-Diacetylphloroglucinol (DAPG) produced by Pseudomonas fluorescens, shows toxicity to many microorganisms including fungi, bacteria, and peronosporomycetes. Zoosporogenesis and motility of zoospores are critical for a complete disease cycle and pathogenicity of the peronosporomycete phytopathogens. The aim of this study was to test the effects of DAPG and its derivatives on zoosporogenesis and motility of zoospores of a downy mildew pathogen, Plasmopara viticola, and a damping-off pathogen, Aphanomyces cochlioides. In both cases, DAPG inhibited zoosporogenesis (5 μg/ml) and the motility of zoospores (10 μg/ml) in a dose-dependent manner. Generally, zoospores became immotile shortly after exposure to DAPG followed by lysis. However, a fraction of DAPG treated A. cochlioides zoospores formed round cystospores instead of lysis and then germinated with excessively-branched germ tubes. All derivatives of DAPG had similar inhibitory activities but at varying doses. Among them, 2,4-dipropylphloroglucinol exerted the highest inhibitory activity against both zoosporogenesis and motility of zoospores. This revealed that the degree of hydrogen atoms substitution in the benzene ring by acyl groups and the length of substituted acyl groups were related to the level of bioactivity. This is the first report of inhibitory activities of DAPG and its derivatives against zoosporogenesis and motility of zoospores of two important peronosporomycete phytopathogens.     

Observations of in vitro gametangial copulation and oosporogenesis in Lagenidium giganteum

An in vitro liquid culture oospore production method yielding 5 × 10³ oospores/ml was used to follow the sequential events of gametangial copulation and oospore formation in Lagenidium giganteum. Observations were made with Nomarski differential interference microscopy and scanning electron microscopy. After septation and division of fungal thalli into a chain-like series of links, certain individual subthalli differentiated into gametaniga, oogonia, and antheridia. Antheridia issued a fertilization tube which made contact with, and fused to a single oogonium. Copulative behavior was relatively synchronous and necessitated physical contact between thalli. Sexual reproduction was manifested by the migration and condensation of gametes. Plasmogamy was achieved following the introduction of the male gamete into the oogonium. The fused gametes gave rise to a zygote. Small amounts of periplasm remained in the oogonium. Zygote maturation into a fully developed oospore was characterized by the deposition of a multilaminated oospore wall, the coalescence of lipids into a highly refractive central reserve globule surrounded by a layer of fine-grained cytoplasm.     

布氏轮藻(Chara braunii)生殖器官环境扫描电镜的观察

本文采用环境扫描电镜和光学显微镜,对布氏轮藻(Chara braunii)的藏卵器和藏精器进行了仔细观察。结果表明：藏卵器外的管细胞(包围细胞或螺旋细胞)在卵细胞受精以后,不但没有钙的沉淀、加厚,而且还呈现出逐渐萎缩,最终消失。因此,在古轮藻的研究上,常提到的包围细胞或螺旋细胞,实际上并非是细胞结构,而是受精卵的外壁。     

Fungal sporocarps from the Carboniferous: An unusual specimen of Traquairia

Small spherical structures commonly termed sporocarps are widespread elements in Carboniferous permineralized peat. While sporocarps today are generally interpreted as fungal, their precise systematic affinities largely remain unknown. A specimen of the sporocarp-type Traquairia from the Lower Coal Measures (Lower Pennsylvanian) of Great Britain demonstrates a heretofore unknown feature in the form of a preformed aperture from which emerges a fascicle of radially oriented structures that are constricted sub-distally; transverse septa are present in the constricted areas of some of the structures. The outgrowths are morphologically similar to conidiophores bearing terminal conidia of certain extant fungi in the order Entomophthorales, and thus might suggest affinities of Traquairia with the Zygomycota. Alternatively, the outgrowths might represent sporulating hyphae of a parasitic or saprotrophic blastocladalean fungus or a peronosporomycete that used Traquairia as a nutrient source.     

美洲绵霉超微结构的初步研究

对美洲绵霉(Achlya americana Humphrey)有性器官的壁结构,细胞核相及一些细胞器的结构进行了初步的超微结构研究,我们发现藏卵器壁由两层组成,卵孢子壁至少由三层组成,而藏卵器横隔膜则由两层壁和中间一层原生质体组成。在细胞核分裂过程中发现有明显的染色体的形成和存在,这是第一次在电子显微镜下看到卵菌的染色体,并对染色体的存在进行了讨论。     

Dynamic intracellular localization of Dazl protein during <Emphasis Type="Italic">Xenopus</Emphasis> germline development

Xenopus dazl encoding an RNA-binding protein has been identified as a component of the germ plasm and is involved in the migration and differentiation of the primordial germ cells (PGCs). Here, we investigated the intracellular localization of Dazl in germline cells throughout the lifetime of Xenopus. In early embryogenesis, Dazl was detected initially in the germ plasm and then translocated to a perinuclear region. Then, it was detected within the nucleus in PGCs. Dazl was observed only in the cytoplasm in PGCs when sex differentiation began in the gonads. Dazl was distributed in both the nucleus and cytoplasm of the primary oogonium and spermatogonium, but only in the cytoplasm of the secondary oogonium and spermatogonium. In spermatocytes, Dazl was distributed throughout cytoplasm and localized at the spindles and cytoplasm during meiosis. Then, it was detected as speckles in the nucleus in the round spermatid. The dynamic intracellular localization suggests that Dazl is a multifunctional protein regulating RNA metabolism required for Xenopus germline development.     

Ultrastructural observation of oogenesis in the crustacea amphipoda Orchestia gammarellus (Pallas)

The oogenesis of the Crustacea Amphipoda Orchestia gammarellus can be divided in five stages taking into consideration both the oocyte ultrastructure and the physiology of the ovary. The primary oogonium (12 μm in diameter) is lodged within the germinative zone: after division, the daughter cell (or secondary oogonium) leaves this area and enters meiotic prophase. Stage I is represented by the oocyte with visible chromosomes (12–18 μm in diameter) the cytoplasmic ultrastructure of which is comparable to that of the oogonium. Stage II or previtellogenesis is characterized by a considerable growth of the oocyte (18–80 μm in diameter) which becomes enriched in ribosomes and vesicles of the rough endoplasmic reticulum; the oocyte does not yet contain any vitelline reserve (proteinaceous and lipid). Stage III or primary vitello-genesis (80–160 μm in diameter) is typified by the synthetic activity of the rough endoplasmic reticulum, corresponding to an endogenous accumulation of proteinaceous yolk. Stage IV or secondary vitellogenesis (160–800 μm in diameter) only appears during the period of reproduction; by means of endocytosis the oocyte accumulates yolk spheres in addition to lipid droplets, the origin of which is uncertain; towards the end of vitellogenesis, cortical granules become a feature that is noted for the first time in Crustacea. The last stage or maturation (800 μm in diameter) starts right before or immediately after the exuviation of the female and ends with fertilization.     

Fossil calibrations and molecular divergence time estimates in centrarchid fishes (Teleostei: Centrarchidae)

Molecular clock methods allow biologists to estimate divergence times, which in turn play an important role in comparative studies of many evolutionary processes. It is well known that molecular age estimates can be biased by heterogeneity in rates of molecular evolution, but less attention has been paid to the issue of potentially erroneous fossil calibrations. In this study we estimate the timing of diversification in Centrarchidae, an endemic major lineage of the diverse North American freshwater fish fauna, through a new approach to fossil calibration and molecular evolutionary model selection. Given a completely resolved multi-gene molecular phylogeny and a set of multiple fossil-inferred age estimates, we tested for potentially erroneous fossil calibrations using a recently developed fossil cross-validation. We also used fossil information to guide the selection of the optimal molecular evolutionary model with a new fossil jackknife method in a fossil-based model cross-validation. The centrarchid phylogeny resulted from a mixed-model Bayesian strategy that included 14 separate data partitions sampled from three mtDNA and four nuclear genes. Ten of the 31 interspecific nodes in the centrarchid phylogeny were assigned a minimal age estimate from the centrarchid fossil record. Our analyses identified four fossil dates that were inconsistent with the other fossils, and we removed them from the molecular dating analysis. Using fossil-based model cross-validation to determine the optimal smoothing value in penalized likelihood analysis, and six mutually consistent fossil calibrations, the age of the most recent common ancestor of Centrarchidae was 33.59 million years ago (mya). Penalized likelihood analyses of individual data partitions all converged on a very similar age estimate for this node, indicating that rate heterogeneity among data partitions is not confounding our analyses. These results place the origin of the centrarchid radiation at a time of major faunal turnover as the fossil record indicates that the most diverse lineages of the North American freshwater fish fauna originated at the Eocene-Oligocene boundary, approximately 34 mya. This time coincided with major global climate change from warm to cool temperatures and a signature of elevated lineage extinction and origination in the fossil record across the tree of life. Our analyses demonstrate the utility of fossil cross-validation to critically assess individual fossil calibration points, providing the ability to discriminate between consistent and inconsistent fossil age estimates that are used for calibrating molecular phylogenies.     

Morphological review ofPelvetia andSilvetia (Fucaceae, Phaeophyta) with an emphasis on phylogenetic relationships

We compared the morphology of all four members ofPelvetia andSilvetia (Fucaceae, Phaeophyta), with an emphasis on phylogenetic relationships.Silvetia is segregated fromPelvetia because it has two, longitudinally divided eggs in the oogonium. In contrast, the eggs of the genusPelvetia are transversally divided. A cladistic analysis, based on 17 morphological features, shows thatPelvetia is closely related toHesperophycus andPelvetiopsis, as are three species ofSilvetia. We can infer from the cladistic tree and biogeographic information that some silvetian ancestor populations from the northern Pacific region likely evolved toS. babingtonii in northern Japan and then moved to Korea and California (USA), whereS. siliquosa andS. compressa, respectively, diverged. Our morphological study corroborates the DNA-based phytogeny and the ensuing taxonomy for the two genera. These results demonstrate the necessity for systematically revising the family Fucaceae to emphasize egg development, rather than egg number, in the oogonium, as a diagnostic character.