On the Fine Structure of the Nervous System of Tubiluchus philippinensis (Tubiluchidae,Priapulida)

At the mouth tube/introvert border a circumenteric intraepithelial nerve ring occupies a circular ridge protruding into the body cavity. The ring has a centrally located neuropile nearly free of perikarya and two zones of different perikarya above and below the neuropile. Presumably non-neuronal perikarya have an oval nucleus, large heterochromatin clumps and marked filament bundles. Such elements resemble tanycytic glial cells. Two types of presumably neuronal perikarya contain small cytoplasmic granules, similar to those in nerve fibre profiles. One of these neurons has a pale nucleus with a prominent nucleolus, the other a rather inconspicuous nucleus similar to that of the tanycytic cells. The neuronal processes of the fibre ring differ in diameter and contain clear and dense core vesicles, small granules (high or medium electron density) or granules with a dense periphery and a light centre. Sometimes neighbouring processes seem interconnected by electrical synapses. Images suggesting chemical synapses are rare. A large intraepithelial nerve lies in the wall of the introvert and ventral body wall close to the musculature, possibly innervated by this nerve. Frontal of the anus lies an intraepithelial ganglion demonstrating again a central neuropile. two neuronal types and tanycytic elements with filament bundles. Comparative aspects of the characters of the Tubiluchus nervous system are also discussed.     

Cell-cell-recognition during fertilization in Ectocarpus siliculosus (Phaeophyceae)

Two cellular communication systems are involved in sexual recognition of complementary gametes of Ectocarpus siliculosus. The first is mediated via the pheromone ectocarpene and is not species-specific. A second system has been identified which is based on the interaction of carbohydrate and carbohydrate receptor. Cell fusion is inhibited selectively by wheat germ agglutinin (WGA) applied to female gametes or with the complementary N-acetylglucosamine (GlcNAc) applied to male gametes. Both, WGA and GlcNAc showed gametic specificity in their effects. Fertilization was not affected by the lectins GNA, LCA and SBA. The maximum fertilization success was 50.2% and followed Michaelis-Mentenkinetics (I₅₀ = 6 minutes).     

Double fertilization inEphedra trifurca,a non-flowering seed plant: The relationship between fertilization events and the cell cycle

Summary Fertilization inEphedra trifurca was examined with a combination of light and fluorescence microscopy. Developmental analysis clearly indicates that double fertilization events, similar to those described inE. nevadensis, regularly occur during the process of sexual reproduction inE. trifurca. In addition to the typical fusion of a sperm nucleus and egg nucleus, a second fertilization event occurs between the second sperm nucleus from an individual pollen tube and the ventral canal nucleus. Both of the fertilization events take place within the confines of an individual egg cell of the female gametophyte. Microspectrofluorometric data demonstrate that each nucleus involved in a sexual fusion event proceeds through the synthesis phase of the cell cycle and increases its DNA content from 1C to 2C before the process of nuclear fusion is completed. Photometric data also confirm that the product of the second fertilization event is equal in DNA content (4C) to the zygotic nucleus derived from the first fertilization event, and is prepared to enter into mitosis as a fully functional diploid nucleus.Abbreviations DAPI 4;,6-diamidino-2-phenylindole - RFU relative fluorescence units     

Order of fertilization within the ovary in Phaseolus coccineus L. (Leguminosae)

Summary Phaseolus coccineus typically has six linearly arranged ovules per ovary. The three ovules near the stylar end of the fruit (positions one, two, and three) are more likely to produce mature seeds, to produce heavier seeds, and to produce more vigorous progeny than the ovules in positions near the peduncular/basal end of the fruit (ovule positions four, five, and six). We conducted a series of field experiments designed to supplement our understanding of the mechanisms determining these position effects. We found that approximately 98% of the ovules in 752 fruits were fertilized — about 0.6% of the stylar ovules were not fertilized, whereas 3.2% of the basal ovules were unfertilized. Moreover, we found that only about 49% of the ovules in these 752 fruits produced mature seeds. Over 60% of the stylar ovules produced mature seeds, whereas only 37% of the basal ovules produced mature seeds. Consequently, the proportion of fertilized ovules cannot explain the differences in seed maturation among the ovule positions. We found that after 6.5 h most of the fertilized ovules were located in the stylar ovule positions, and that there were no fertilized ovules in ovule positions five and six, indicating that the stylar ovules are fertilized first. When only the fastest growing pollen tubes were permitted to enter the ovary (due to exision of the style), only the ovules at the stylar end were fertilized, indicating that the ovule positions that are fertilized first are indeed fertilized by the fastest growing pollen tubes.On leave from the Escuela de Biologia, Universidad de Costa Rica, Cuidad Universitaria Rodrigo Facio, San Jose, Costa Rica, Central America     

Mechanisms of sulphide tolerance in the peanut worm,Sipunculus nudus (Sipunculidae) and in the lugworm,Arenicola marina (Polychaeta)

Summary The peanut worm Sipunculus nudus and the lugworm Arenicola marina are inhabitants of intertidal flats. Both species may be exposed to H₂S within their habitat. Sulphide concentrations in the vicinity of A. marina burrows are as high as 340 mol · 1^-1, whereas the pore water in sipuncle areas contains much lower sulphide levels of 13 mol · 1^-1 at most. During in vivo sulphide incubations, H₂S increases within the coelomic fluid of both species. In S. nudus the concentration of total sulphide after 8 h is about 40% of that of the incubation medium containing 200 and 1000 mol · 1^-1, respectively, which is partly due to the acidification of the coelomic fluid by 0.2 pH units during anaerobiosis. After 8 h, the sulphide concentration in A. marina was only 15% of that in the incubation medium containing 1000 mol · 1^-1. When oxygen is available, both species oxidize sulphide to thiosulphate, but in A. marina this capability is more pronounced than in S. nudus. If sulphide is not completely oxidized internally both intertidal worms switch to an anaerobic metabolism as indicated by the accumulation of opines and succinate.Abbreviations ATP adenosine triphosphate - HEPES N-2-hydroxyethylpiperazine-N-2-ethanesulphonic acid - HPLC high-performance liquid chromatography - wwt wet weight     

Diel and seasonal vertical distribution of meiobenthic copepods in muddy sediments of a eutrophic lagoon (fish ponds of Arcachon Bay)

The vertical distribution of meiobenthic copepods was investigated within muddy sediments of a eutrophic lagoon (fish ponds of Arcachon Bay, France). The aim of the study was to determine if in muddy sediments, as previously established in sandy sediments, meiobenthic copepods migrate vertically according to the seasons or diel periods. Two experimental approaches were used, viz: a three-season comparison was made of the diel vertical distribution of the harpacticoid Canuella perplexa T. & A. Scott (1893) and secondly the depth distribution of a meiobenthic copepod assemblage was followed for a 24 h period, in shallow water subtidal locations. The harpacticoid C. perplexa vertically migrated through the top three centimeters of the sediment, showing diel and seasonal variations in depth distribution. The differential vertical distributions shown by the dominant meiobenthic populations suggest that emergence into the water column may mainly concern surface dwelling copepods. The physical and biological factors affecting seasonal and diel changes in the copepod assemblage of the fish ponds are discussed.     

Derived morphology in a basal moth: The uniquely specialized sternum V glands of Agathiphaga (Lepidoptera: Agathiphagidae)

The sternum V gland is a notable specialization shared by basal Lepidoptera and most Trichoptera; it is generally present in both sexes or in females only. In both orders the secretory cells usually discharge into a sac-like reservoir from which a duct leads to the opening on sternum V. In contrast, the gland in agathiphagid moths is only present in males and it exhibits several unique specialisations: the glandular portion being the apical (morphologically anterior) part of a long tubular cuticular invagination covered with contiguous ‘type-3’ glandular units; together with the following part of the efferent duct it is coiled like a ball of yarn; the duct expands into a fusiform reservoir, and a separate small sac surrounded by a complex muscular coat is present just inside the gland opening. The principal opener muscle of each gland originates on the contralateral side of sternum VI, hence crossing its counterpart in the midline. Characters of the agathiphagid sternum V gland, the function of which remains unclear (though sex pheromone production may appear most likely), add significantly to the suite of autapomorphies exhibited by this otherwise overall remarkably generalized moth taxon.     

Every sperm is sacred: fertilization in Caenorhabditis elegans

The nematode Caenorhabditis elegans is an attractive model system for the study of fertilization. C. elegans exists as a self-fertilizing hermaphrodite or as a male. This unusual situation provides an excellent opportunity to identify and maintain sterile mutants that affect sperm and no other cells. Analysis of these mutants can identify genes that encode proteins required for gamete recognition, adhesion, signaling, fusion, and/or activation at fertilization. These genes can also provide a starting point for the identification of additional molecules required for fertility. This review describes progress in the genetic and molecular dissection of fertilization in C. elegans and related studies on sperm competition.     

Cultivation of the brown alga Hizikia fusiformis (Harvey) Okamura: controlled fertilization and early development of seedlings in raceway tanks in ambient light and temperature

Commercial cultivation of the dioecious brown macroalga Hizikia fusiformis (Harvey) Okamura in East Asia depends on the supply of young seedlings from regenerated holdfasts or from wild population. Recent development of synchronized release of male and female gametes in tumble culture provides a possibility of mass production of young seedlings via sexual reproduction. In this paper, we demonstrate that controlled fertilization can be efficiently realized in ambient light and temperature in a specially designed raceway tank in which the sperm-containing water has been recirculated. The effective fertilization time of eggs by sperm was found to be within six hours. Fast growth and development of the young seedlings relied on the presence of water currents. Velocity tests demonstrated that young seedlings of 2–3 mm in length could withstand a water current of 190 cm s⁻¹ without detachment. Culture experiments at 24 h postfertilization showed that elongation of both the seedlings and their rhizoids were not hampered by high irradiance up to 600 μmol photons m⁻² s⁻¹. However, growth was slightly retarded if cultured at a temperature of 16 ^∘C compared to other culture temperatures of 22, 25 and 29 ^°C. No seedling detachment was observed after transfer of the young seedlings to raft cultivation in the sea after one and 1.5 months post-fertilization, indicating the feasibility of obtaining large quantity of seedlings in such a system.     

Morphological aspects of fertilization in Phallusia (Ascidia) nigra (Ascidiacea,Tunicata)

The spermatozoa of Phallusia (Ascidia) nigra have an elongated head (approximately 5 m in length) in which a nucleus and a single mitochondrion are located side by side. There is no midpiece. The apex of the head is wedge-shaped. Acrosomal vesicles (approximately 55–65 nm in diameter) and moderately electron-dense material (MEDM) are present between the plasmalemma and the nuclear membranes in the anterior tip of the head. The MEDM occupies a central position and three or four acrosomal vesicles are seen in a line alongside it. The acrosomal vesicles disappear as the sperm makes contact with the surface of the chorion. Gamete fusion most likely occurs between a small process extending from the peripheral margin of the sperm apex and the egg surface, resulting in incorporation of the sperm into the egg from the anterior region of its head.     

Fertilization in landlocked sea lamprey: storage of gametes, optimal sperm: egg ratio, and methods of assessing fertilization success

Ovulated, unfertilized eggs of sea lamprey Petromyzon marinus could be stored for 1 day at 15° C without significant loss of fertilizing ability. After 2 days storage most eggs could still be fertilized. Lamprey semen could be stored up to 1 day. Thereafter, a decrease in sperm fertilizing ability occurred, accompanied with a decrease in sperm motility. Unlike teleost fish, sea lamprey eggs could still be fertilized after 1 h contact with water. This extended time of gamete fertility after release into water may help to account for the reproductive success of this species. Maximal fertilization rates were obtained at a sperm: egg ratio of 50 000, a ratio recommended for studies on fertility of individual males. Assessing fertilization success 3 min after fertilization (at cytoplasmic bleb stage) or 5 h after fertilization (at two–cell embryo) was strongly correlated ( r =0·92 and 0·98) with estimation and fertilization success at hatching. These results offer improvement in artificial fertilization techniques under laboratory conditions and provide new information on the biology of fertilization in sea lamprey.     

Flavonols are not essential for fertilization in Arabidopsis thaliana

Flavonols are plant metabolites suggested to serve a vital role in fertilization of higher plants. Petunia and maize plants mutated in their flavonol biosynthesis are not able to set seed after self-pollination. We have investigated the role of these compounds in Arabidopsis thaliana. Like in all other plant species, high levels of flavonols could be detected in pollen of wild-type A. thaliana. No flavonols were detected in reproductive organs of the A. thaliana tt4 mutant in which the chs gene is mutated. Surprisingly, this mutant did set seed after self-fertilization and no pollen tube growth aberrations were observed in vivo. The role of flavonols during fertilization of Arabidopsis is discussed.Abbreviations CHS chalcone synthase - TLC thin-layer chromatography     

Variations in 15N abundance in a forest fertilization trial: Critical loads of N,N saturation,contamination and effects of revitalization fertilization

¹⁵N abundances of current needles of Norway spruce collected during 23 yrs of a forest fertilization experiment were studied in order to follow ecosystem gains and losses of N. Unlabelled ammonium nitrate at four rates (N0–N3), phosphorus at three rates (P0–P2), and potassium plus other elements including micronutrients at two rates (K0–K1), had been applied to plots in a complete factorial design. Nitrogen had been applied annually at average rates of 0, 34, 68 and 102 kg N ha^-1 yr^-1. Tree growth had responded positively to additions of N, but the response was remarkably more positive to the N2P2K1 treatment. In N1 treatments, δ¹⁵N (‰) declined over time. This was consistent with an earlier study, and should reflect a change in ¹⁵N abundance towards that of fertilizer N (minus discrimination during uptake), which in turn means accretion of most of the N added. As in the earlier study, in which N3 plots lost most of the N added, the present N3 plots showed an increasing δ¹⁵N (‰). This pattern was not significantly affected by additions of P and K plus other elements, although a weak negative effect of P on N accretion was indicated, i.e. there was a tendency δ¹⁵N (‰) to be higher when P was added. This, and another recent result based on an N budget, shows that so-called revitalization fertilization may well increase growth of trees, but also promotes losses of N from the ecosystem. As in the previous study, a decline in δ¹⁵N (‰) on control plots provided evidence of contamination. Given a removal of 100 kg N ha^-1 at stem harvest and a leaching of 2 kg N ha^-1 yr^-1, our data on ¹⁵N suggest that a load of 9 kg N ha^-1 yr^-1 would saturate the ecosystem after 100 years. This load is only about twice the annual deposition at the site.     

Pleosporales in Japan (5): Pleomassaria, Asteromassaria, and Splanchnonema

Eight species in Pleomassariaceae are described and illustrated. They are Pleomassaria maxima, Pl. swidae, Pl. siparia “type A,” Prosthemium canba, Asteromassaria macroconidica, A. pulchra, Splanchnonema mori, and S. argus. Of these, Pl. swidae on twigs of Swida controversa, Pr. canba on Betula ermanii, and A. macroconidica on twigs of Prunus ×yedoensis and an unknown woody plant are new. Three species, Pl. siparia “type A,” A. pulchra, and S. argus, are reported from Japan for the first time. A new combination, S. mori (≡ Massaria mori), is proposed.     

Pione lampa,a bioeroding sponge in a worm reef

A bioeroding sponge was found in a sabellariid worm reef in Florida, USA. The sponge was investigated with standard techniques for species identification (tissue digestion, microscopy techniques, examination of spicules) and was identified as the clionid Pione lampa (De Laubenfels, 1950). This species is extremely similar to Pione vastifica (Hancock, 1849). Morphological and ecological similarities and possible dissimilarities are discussed. Arguments for conspecifity were found to be weaker than evidence of P. lampa and P. vastifica being separate species. The sampled sponge contained asexual reproductive elements: superficial buds and internal gemmules. Buds were very delicate, consisting mainly of a radially-oriented mesh of collageneous threads. They further comprised abundant granular material and acanthose microrhabds. Numerous choanosomal gemmules were found in the sponge body attached to erosion chamber walls. They were subspherical to lenticular and had an aspicular, unstructured, smooth and rigid coat. It enclosed dense cell material and various spicule types. Whereas buds are interpreted to function as dispersal elements, gemmules will primarily ensure survival under adverse conditions such as smothering, exposure to air and high temperatures. Gemmules from this site might occasionally be freed and scattered, since the Florida reef can suffer heavy damage during periods of increased wave activity such as that created during hurricane season. Bioerosion activity of the sponge increases the chance to free gemmules, as the sponge not only etches into calcareous particles cemented into the matrix produced by the worms, but also into the matrix itself. This ability enables the sponge to utilise the reef as substrate. Within the Clionidae, Cliona annulifera and three species of the genus Pione are the only species known to produce typical gemmules. Possible reasons are: (1) reproduction of bioeroding sponges is understudied and asexual bodies may have been overlooked in other species, and (2) clionid gemmules are an adaptation to survive life in risky environments. Sponges of the genus Pioneare comparatively successful in environments in which they are close to their physical limits or in potentially unstable or mobile substrates.     

Borrelia, Coxiella, and Rickettsia in Carios capensis (Acari: Argasidae) from a brown pelican (Pelecanus occidentalis) rookery in South Carolina, USA

Argasid ticks are vectors of viral and bacterial agents of humans and animals. Carios capensis, a tick of seabirds, infests the nests of brown pelicans, Pelecanus occidentalis, and other ground nesting birds along the coast of South Carolina. This tick is associated with pelican nest abandonment and could pose a threat to humans visiting pelican rookeries if visitors are exposed to ticks harboring infectious agents. We collected ticks from a pelican rookery on Deveaux Bank, South Carolina and screened 64 individual ticks, six pools of larvae, and an egg mass for DNA from Bartonella, Borrelia, Coxiella, and Rickettsia by polymerase chain reaction amplification and sequencing. Ticks harbored DNA from “Borrelia lonestari”, a novel Coxiella sp., and three species of Rickettsia, including Rickettsia felis and two undescribed Rickettsia spp. DNA from the Coxiella and two undescribed Rickettsia were detected in unfed larvae that emerged in the laboratory, which implies these agents are transmitted vertically by female ticks. We partially characterize the novel Coxiella by molecular means.