Spermatozoon Ultrastructure of Two Holothurian Species of the Genus Cucumaria (Dendrochirotida,Holothuroidea) from the Sea of Japan

The ultrastructure of spermatozoa of Cucumaria japonica and a congeneric morphologically similar deep-sea species was studied. The spermatozoa of both C. japonica and C. conicospermium are similar to those of other holothurians: the acrosome is composed of an acrosomal granule and periacrosomal material; the centrioles lie at an acute angle to one another; and the proximal centriole is connected to the nuclear envelope by a flagellar rootlet. The spermatozoa of C. japonica differ from those of C. conicospermium in the shape of the head and the dimensions and position of the acrosome. In C. japonica, the acrosome is completely embedded in the nuclear fossa and measures 0.7 m. In C. conicospermium, only one-third of the acrosome is embedded in the nuclear fossa; this acrosome measures 1.3 m. A correlation between the structure of the sperm acrosome and that of the egg envelope is discussed.     

Ultrastructure of spermatozoa of Peregrinus maidis (Homoptera,Delphacidae)

Summary The spermatozoa of Peregrinus maidis Ashm. are thread-like, approximately 650 long and 1 wide including the head (approximately 28 ).The main part of the spermatozoa consists of two mitochondria derivatives, a central body between them, the axial filament complex, and a newly found element consisting of two wing-shaped bodies. Each mitochondrion derivative shows a peripheral and an inner part. The peripheral part is formed by cristae arranged perpendicularly to the long axis of the spermatozoon. The cristae are approximately 70 Å wide. The dense layers between them measure approximately 280 Å. The inner part of the mitochondrion derivative shows a crystalline array, formed by sub-units of approximately 100 Å diameter. The wing-shaped bodies consist of tubular elements.The head has an elongated nucleus with an electron transparent space inside. At the anterior end of the nucleus lies a tapered acrosome. This appears fibrous and parts of the acrosome fibers seem to run along the nucleus. Acknowledgements. The authors wish to thank Dr. G. H. Bergold for suggestions and support, Drs. J. André, D. W. Fawcett, P. Maillet and G. F. Meyer for very helpful discussion. They are also grateful to Mr. O. Suárez for assistance in the preparation of the organs of P. maidis and to Mrs. M. de Pingarrón for technical assistance.     

Lemdana latifi n. sp. (Filarioidea: Onchocercidae) from the red jungle fowl (Gallus gallus spadiceus)

Lemdana latifi n. sp. was found in connective tissues around the trachea and crop and in the body-cavity of seven of 14 Malayan red jungle fowl Gallus gallus spadiceus. The new species is described and illustrated. Morphologically it is most closely related to Lemdana pavonica and Lemdana francolini. Lemdana latifi is distinguished from the eight valid species of Lemdana by the mean spicular ratio of 1.7:1; the right spicule with a right margin 18–29% (15–31 m; mean 24 m) longer than the left margin; the distal half of the left spicule twisted and S-shaped; and the absence of unpaired papillae at tip of male tail. The new species has smaller adults, a shorter left spicule and a shorter glandular oesophagus than those of L. pavonica and a wider male, shorter spicules and a longer muscular oesophagus than those of L. francolini. The male of L. latifi is 7–9 (8.1)mm long, the left spicule 164–215 (184)m long and the right spicule 98–117 (108)m long. The female is 17–23 (21)mm in length. Sheathed microfilariae from blood smears are 78–100 m long and those from the uterus are 89–103 m long. This is the sixth valid species of Lemdana in the Phasianidae.     

Spermatozoa and spermatogenesis in the northern quahaug <Emphasis Type="Italic">Mercenaria mercenaria</Emphasis> (Mollusca,Bivalvia)

We studied the ultrastructure of spermatogenesis and spermatozoa in the northern quahaug, the clam Mercenaria mercenaria. Spermatogenetic cells gradually elongate. Mitochondria gradually fuse and increase in size and electron density. During spermatid differentiation, proacrosomal vesicles migrate towards the presumptive anterior pole of the nucleus and eventually form the acrosome. The spermatozoon of M. mercenaria is of a primitive type. It is composed of head, mid-piece, and tail. The acrosome shows a subacrosomal space with a short conical contour. The slightly curved nucleus of the spermatozoon contains fine-grained dense chromatin. The middle piece consists of a centriolar complex which is surrounded by four mitochondria. The flagellum has a standard “9 + 2” microtubular structure. The ultrastructure of spermatozoa and spermatogenesis of M. mercenaria shares a number of features with other species of the family Veneridae. M. mercenaria may be a suitable model species for further investigations into the mechanisms of spermatogenesis in the Bivalvia.     

Ultrastructure of Sperm in <Emphasis Type="Italic">Mercenaria stimpsoni</Emphasis> and <Emphasis Type="Italic">Mactra chinensis</Emphasis> (Mollusca: Bivalvia) from the Sea of Japan

The ultrastructure of the sperm of the common bivalve species Mercenaria stimpsoni and Mactra chinensis from Peter the Great Bay is described. The sperm structure is typical for animals with external insemination. The sperm consists of a head, middle part, and flagellum. The sperm head of M. stimpsoni has a curved crescent form and includes the nucleus and acrosome; the head length is 9.8 μm. The acrosome is subdivided to the acrosome granule and the periacrosomal material. There are 4 mitochondria of about 0.8 μm in size in the middle part of the spermatozoon. The mitochondria surround the centriolar apparatus, which consists of proximal and distal centrioles located at a right angle. The axoneme originates from the distal centriole. The sperm of M. chinensis is barrel-shaped, with a head length of 3.2 μm. The acrosome is relatively larger, and its height is 1–1.2 μm. There are also 4 mitochondria 0.6–0.8 μm in the middle part of the spermatozoon. The sperm structure of the described species is typical of the families to which the mollusks belong, with insignificant variations.     

The ultrastructure of the spermatozoon of Lernaeocera branchialis (Copepoda : Pennellidae)

Spermatozoa are produced by males of the fish-parasitic copepod, Lernaeocera branchialis, on the flounder (Platichthys flesus) host and packaged into spermatophores. Mature spermatozoa from spermatophores have been investigated with light microscopy and t.e.m. and s.e.m. techniques. Each is a filiform cell up to 30 µm long and 1 µm in diameter, being symmetrically tapered at each end to produce a javelin-shaped configuration. Surface s.e.m. appearances suggest that the cell is helically twisted. The spermatozoon possesses no flagellum, no nuclear envelope, no mitochondria and no orthodox acrosome. For most of its length the cell has a four-lobed appearance in transverse section and contains little more than a cylindrical sleeve of pseudomembranous material with which is associated finely filamentous or granular material which is assumed to be nuclear chromatin. The spermatozoon is immobile in seawater.The ultrastructure of L. branchialis spermatozoa has been compared with that of eight other copepod species.     

Gamete Structure and Fertilization in the Barents Sea Sponge Leucosolenia complicata

The ultrastructure of male and female gametes of asconoid sponge Leucosolenia complicata(Calcispongiae, Calcaronea), a hermaphrodite species that reproduces in autumn, is described. The mature sponge's oocytes were up to 70 m in diameter, had no coatings, and contained a nucleus about 31 m in diameter with large nucleoli (up to 6.6 m). There were vacuoles with fibrillar contents typical of calcareous sponges in ooplasm. During vitellogenesis, a cluster of a great number of nurse cells developed above each oocyte from transformed choanocytes. Mature spermia of L. complicatalooked like orbicular cells about 2.5 m in diameter, with no acrosome or tail. The spermium nucleus (diameter about 2.2 m) was formed by incompletely condensed chromatin and was surrounded with a thin layer of cytoplasm of nonuniform thickness. In the thick layer of cytoplasm beyond the ribosomes, there were two or three mitochondria, dictyosomes, and electron-dense protein bodies lying freely under the nucleus. Fertilization occurred with the aid of a carrier cell. During spawning (mass release of spermia), any nurse cell complex can seize a spermium and transform into a carrier cell in situ. The transformation of a seized spermium into a spermiocyst was connected with the rapid isolation of the spermium nucleus from the protein body. Fertilization began with the penetration of the protein body into the oocyte cytoplasm. Only after this did the spermium's nucleus penetrate into the oocyte.     

Sperm ultrastructure in two species of the polychaete genusHarmothoe (Polynoidae)

The structure of spermatozoa is described for two species of polynoid polychaete,Harmothoe imbricata andHarmothoe impar, from material fixed and examined by both scanning and transmission electron microscopy. The two species undergo spermiogenesis within discrete testes. The testis ofH. imbricata is shown to have a layer of epithelial cells which possess an outer cuticular layer and a microvillous inner surface. Spermatocytes of both species are spherical but there are marked differences in the shape and size of the spermatozoa of the two species.H. impar has a classical primitive spermatozoon with a rounded head (2 μm long) and a button-shaped acrosome. Fully differentiated spermatozoa ofH. imbricata are modified from the primitive form by having a long head (10 μm length) with a pointed acrosome about 6 μm in length. Spermatozoa ofH. imbricata have a ring of up to fourteen mitochondria around a centrally inserted flagellum at the posterior whereasH. impar has a ring of four or five spherical mitochondria. Spermiogenesis is well synchronised inH. imbricata but all developmental stages can be found simultaneously in the testis ofH. impar. The differences in sperm structure of the two species may be related to differences in breeding biology which are hitherto unknown.     

Establishment and characterization of Rubus tissue culture systems for in vitro bioassays against phytotoxins from Rubus fungal pathogens

The Rubus species R. parviflorus, R. spectabilis and R. strigosus interfere with conifer seedling establishment on forest regeneration sites in Canada and the United States. As a first step towards microbial metabolite-based control, callus and cell suspension cultures of the Rubus species were developed as a bioassay system to detect phytotoxic compounds that may have relevance in a vegetation control context. Rapidly growing friable callus and suspension cultures were obtained from leaf disks of the three weedy Rubus species using similar culture media conditions (modified Murashige and Skoog) but required different plant growth regulators (R. parviflorus, 4.5 M 2,4-D; R. spectabilis, 26.9 M NAA/0.5 M zeatin; and R. strigosus, 12.4 M picloram). Cell growth and health attributes including callus circumference, degree of browning and suspension culture cell viability as measured by the TTC vital stain assay were developed and were rapid and convenient to use. We have established Rubus tissue culture systems that will make it possible for large scale screening of phytotoxic metabolites.     

两种石龙子精子超微结构的比较

利用透射电镜研究多线南蜥和印度蜓蜥附睾精子的超微结构。两种卵胎生石龙子的精子具有一些有鳞类精子的共同特征,即具有顶体囊、顶体下锥、单个核前穿孔器和核喙,无核内管,纤维鞘伸入中段,与双联微管3和8相邻的外周致密纤维具双份纤维结构。多线南蜥和印度蜓蜥精子超微结构的种间差异主要表现在:多线南蜥精子核前方的顶体下锥电子密度较小,顶体囊具单侧嵴,横切面上可见非连续的致密体环或11个线粒体;印度蜓蜥无单侧嵴,横切面上可见连续的致密体环或12个线粒体。迄今未发现石龙子科精子的独征,但该科不同类群的顶体囊、顶体下腔、核前方的顶体下锥电子致密程度、核肩、纵切面线粒体与致密体的排列方式、横切面致密体环形状和线粒体等精子超微结构特征有一定程度的差异。这些差异可为研究石龙子科系统发生提供辅助信息。     

Plant regeneration via somatic embryogenesis in Styrian pumpkin: cytological and biochemical investigations

Somatic embryo formation was induced from cotyledon explants of Styrian pumpkin (Cucurbita pepo L. subsp. pepo var. styriaca Greb.) by using a solid MS medium supplemented with 16.11M NAA and 4.44M BA or 26.85M NAA and 13.32M BA. The callus proliferation was more efficient on medium supplemented with 26.85M NAA and 13.32M BA. In contrast, the embryogenic response was higher on medium with lower concentrations of growth regulators (16.11M NAA and 4.44M BA). The time needed for embryo induction did not depend on medium composition. Embryos in globular stage were transferred to three different maturation media, containing 2.89M GA₃ in combination with 0.54M NAA, 11.42M IAA and growth regulator-free medium. The germination rate was the highest when embryos were cultured on medium with 11.42M IAA. Plantlets grown on this medium achieved maturity suitable for transplantation into soil within 9 to 10weeks. The regenerated plants were successfully transferred into field and developed fertile flowers and set fruits. Biochemical analysis showed significant lower total glutathione levels among in vitro grown plantlets compared to seedlings grown in soil. When the plantlets were transferred into soil, they reached a normal size within a month and the glutathione concentration was comparable to seed-derived plants at the same developmental stage. Transmission electron microscopy was used to investigate possible differences in the ultrastructure of cells from callus cultures, and leaf cells of regenerated and seed-derived plants. Differences in the ultrastructure were found within chloroplasts which contained only single thylakoids, large starch grains and small plastoglobuli in callus cells in comparison to leaf cells, which possessed a well developed thylakoid system, small starch grains and large plastoglobuli.     

The Spermatozoa of the Thysanuran Insects Petrobius brevistylis Carp. and Lepisma saccharina L.

The spermatozoa of Petrobius and Lepisma share a few general insect features (filamentous shape, two mitochondria, compact acrosome vesicle, bilateral symmetry) but differ fundamentally with regard to specializations. In Petrobius, a long coiled acrosome, a coiled nucleus, and a “body” with axonema, two mitochondria, and a pair of lateral bodies follow each other in normal sequence. In Lepisma the acrosome is a small vestige in the spoon-shaped anterior end, the centriole is dislocated anteriorly, and nucleus, two mitochondria and axonema run like parallel filaments through most of the spermatozoon. The centriole adjunct develops into a postnuclear body in Lepisma but forms a pair of complicated “lateral bodies” in Petrobius. It is concluded that ancestral forms must have had fairly primitive spermatozoa and that specialization has proceeded independently within each evolutionary line.     

Comparative sperm ultrastructure of<Emphasis Type="Italic"> Neodasys ciritus</Emphasis> and<Emphasis Type="Italic"> Musellifer delamarei</Emphasis>, two species considered to be basal among Chaetonotida (Gastrotricha)

The spermatozoa of two species supposed to be basal to Gastrotricha Chaetonotida, Neodasys ciritus and Musellifer delamarei, were studied in order to supply further elements to the understanding of sperm evolution in Chaetonotida, a group in which a fully parthenogenetic reproduction is dominant. Two considerably different sperm patterns were found: the spermatozoon of N. ciritus has a simple, conical acrosome, a short, condensed nucleus, few conventional mitochondria randomly arranged along the sperm head, and a 9×2+2 flagellum perpendicular to the sperm major axis. The spermatozoon of M. delamarei is a filiform cell with a simple acrosome, a partially condensed nucleus, four mitochondria at the nuclear base, and a flagellum with a 9×2+2 axoneme and large accessory fibers. Some sperm features of M. delamarei are comparable to those of Xenotrichulidae, the only other Chaetonotida producing conventional spermatozoa, whereas the sperm of N. ciritus appears different from all the other patterns known among Gastrotricha, thus knowledge of it does not help in solving the problem of the discussed phylogenetic position of the genus.     

The evolution of sperm ultrastructure among Boidae (Serpentes)

We investigate the evolution of sperm ultrastructure of three species of Boidae (Epicrates cenchria, Boa constrictor amarali, and Corallus hortulanus). Spermatozoa of these species are filiform consisting of a head region, containing the nucleus and acrosome complex, a midpiece, and a tail region subdivided into principal piece and endpiece. Multilaminar membranes and extracellular microtubules were observed next to the plasma membrane of the spermatozoa. The following differences were observed among the species: ridge on acrosome surface in Boa constrictor amarali (absent in Epicrates cenchria and Corallus hortulanus), stopper-like perforatorium base plate in Boa constrictor amarali and Epicrates cenchria (absent in Corallus hortulanus), rounded mitochondria in transverse section in Epicrates cenchria and Corallus hortulanus (irregular in Boa constrictor amarali). We mapped sperm characters onto two phylogenies based on morphological (Kluge in Misc Publ Mus Zool Univ Michigan 178:1–58, 1991) and molecular (Austin in Copeia 2:341–352, 2000) data, using a number of squamate species as outgroups. We identified 31 unambiguous character transformations in the morphological phylogeny and 30 in the molecular phylogeny, but only 13 and 12 transformations, respectively, are possible synapomorphies. We identified novel sperm synapomorphies, which were common between the morphological and molecular phylogenies: absence of perforatorium base plate and mitochondria arranged as sinuous tubes in oblique section (Serpentes), acrosome vesicle not subdivided and fibers 3 and 8 at the anteriormost region of principal piece (Boidae), and absence of an electron dense structure inside the proximal centriole (Elapidae + Colubridae). Our results suggest greater agreement between sperm ultrastructure and gross anatomical characters. In addition, we found no tendency for more homoplasies in the sperm head than in the flagellum, as recorded in previous studies.     

A comparative analysis of testicular sperm morphology in fossorial and surface-living skinks in South Africa

We described for the first time the spermatozoan ultrastructure of the fully pentadactyl surface-living skink Trachylepis punctatissima, and limbless fossorial skink Acontias meleagris. The spermatozoa of both species follow the general patterns observed within the Squamata. However, several important differences were detected between the two species in the head region (shape of the anterior acrosome, size of acrosome and nucleus) and especially in the midpiece (size of the midpiece, the presence of regular rows of dense bodies, size and number of mitochondria and beginning of the fibrous sheath). Both species shared more characters with the Sphenomorphus + Egernia group than with the Eugongylus group proposed by Jamieson, Oliver, and Scheltinga (Acta Zoologica, 77, 85). Differences in the spermatozoan ultrastructure between T. punctatissima and A. meleagris could be due to distinct ecological and physiological requirements for fertilization, but additional research on these genera and within the Scincidae is required to disentangle this hypothesis, and to disentangle the phylogenetic relationships among skinks.     

Thermus kawarayensis sp. nov., a new member of the genus Thermus, isolated from Japanese hot springs

A long-rod-shaped thermophilic microorganism, strain KW11, was isolated from a hot springs located in the Kawarayu, Gunma, Japan. Cloning and preliminary sequence analysis of 16S rDNA showed that this isolate belongs to the genus Thermus. The cells were 10–20 m long, about 0.8 m in diameter, and produced no pigment in contrast with most of the Thermus species previously reported. KW11 was an aerobic heterotroph and grew at temperatures ranging from 40–73°C, with optimal growth occurring at 68°C. The pH range for growth was from 5.8–8.9, with optimal growth around pH 7. KW11 was sensitive to ampicillin, penicillin G, kanamycin, and streptomycin. The G+C content of DNA was 69 mol%. The main fatty acids were 16:0 (52.9%), iso-15:0 (22.1%), and iso-17:0 (15.6%). The 16S rDNA sequence of KW11 showed 96.0, 95.8, and 95.4% similarity with the sequences of T. aquaticus, T. igniterrae, and T. thermophilus, respectively, and less than 95% with other Thermus species. The physiological differences and phylogenetic evidence indicated that strain KW11 represents T. kawarayensis, a novel species of the genus Thermus. The type strain is isolate KW11^T (JCM12314, DSM16200).     

Spermiogenesis and spermatozoa ultrastructure of Aonides oxycephala (Annelida: Spionidae) from the Sea of Japan

Adults of Aonides oxycephala, common inhabitants of shallow boreal waters in the Atlantic and Pacific Oceans, release gametes into the water where fertilization and lecithotrophic larval development occur. During spermiogenesis, the acrosomal vesicle migrates from the posterior to the anterior end of the spermatid and the number of mitochondria reduces from six in early spermatids to four in mature spermatozoa. Each spermatozoon has an ovoid head with the acrosome 1.4?±?0.1?µm long and 1.6?±?0.1?µm wide and the nucleus 1.7?±?0.1?µm long and 2.3?±?0.1?µm in diameter, four spherical mitochondria, two centrioles oriented perpendicular to each other, putative glycogen in the shape of dense granules in the midpiece, and a flagellum with 9?×?2?+?2 organization of microtubules. The acrosome is a complex heterogeneous structure with five ordered layers of different electron densities, lying in a shallow depression on the anterior end of the nucleus. The nucleus is barrel-shaped (truncated ovoid) with the centriolar fossa housing the distal and proximal centrioles. Spermiogenesis and ultrastructure of spermatozoa of A. oxycephala are similar to those of another free spawning spionid, Marenzelleria viridis. Aonides and Marenzelleria have not, however, been considered as closely related taxa; thus, similarity in the morphology of their sperm might result from convergence or parallelism.     

Ultrastructure of spermatids and spermatozoa in the cyclostomatous bryozoan Tubulipora (Bryozoa,Cyclostomata)

Summary Differentiation of spermatids to mature spermatozoa in the bryozoan Tubulipora liliacea was studied by transmission electron microscopy. The spermatozoon of Tubulipora is of a filiform, modified type, and has evolved from the primitive type as an adaptation to a specialized biology of fertilization. The head of the spermatozoon consists of a small, conical acrosome capping an elongated, cylindrical, anteriorly tapering nucleus. A basal invagination in the nucleus contains the proximal portion of the axoneme and a dense attachment matrix. The flagellar axoneme has the typical 9+2 structure. Four elongated rodshaped mitochondria with typical cristae surround the axoneme in the cylindrical middle piece. Granular electron-dense material is accumulated in the form of four columns alternating with four long cylindrical mitochondria. The mitochondrial middle piece is separated externally from the tail region by an involution of the plasma membrane. The tail region contains a cytoplasmic sheath with accessory fibers surrounding the axoneme. Nine outer, coarse fibers extend posteriorly paralleling the nine doublets of the axoneme. The coarse fibers develop from electron-dense plate-like structures associated with the doublets of the axoneme. A characteristic feature in spermiogenesis is that spermatozoa develop in tetrads. There seem to be significant differences in spermatozoan ultrastructure between the three bryozoan classes Stenolaemata, Gymnolaemata, and Phylactolaemata. The differences indicate different lines of evolution of fertilization biology in these groups.Abbreviations used in the figures a acrosome - av acrosomal vesicles - ax axoneme - c coarse fiber - d electron dense rod - m mitochondrion - mp middle piece - Scale bars=0.5 m - mt microtubule - n nucleus - ne nuclear envelope - p nuclear protrusion - pm plasma membrane - t tail     

Size-fractionated biomass and productivity of phytoplankton and particulate organic carbon in the southern ocean

During the austral summer of 1989/1990, surface samples were obtained of size-fractionated biomass, and the productivity of phytoplankton, its cell abundance, the composition of the dominant species, the concentration of particulate organic carbon (POC) and the related environmental surface parameters were measured in a large-scale survey primarily of the Atlantic and Indian Sectors. The results showed that the southern atlantic sector is the most fertile; chlorophylla (Chla) concentration averaged over 2 μg l⁻¹, average cell abundance was about 41.0 × 10³ cell l⁻¹, and average POC concentration was also the highest (>100 μg l⁻¹), but was lower in the Drake Passage and the southern Indian sector. The results for size-fractionated Chla showed that netplankton (>20 μm) in the South Atlantic Ocean, having abundant nutrients, accounted for the highest proportion (average 65%) of biomass. In the infertile South Indian Ocean, picoplankton (<2 μm) accounted for the highest proportion, averaging 47%. The results for size-fractionated productivity showed that the contribution of picoplankton to total productivity was the largest in the South Atlantic Ocean and Drake Passage, those of nanoplankton (2–20 μm) and netplankton being about equal. The relatively high photosynthesis assimilation number of picoplankton demonstrates their importance in the marine ecosystems of Antarctic water. In comparison with the Antarctic water, the subantarctic and subtropical waters are infertile.