Formation of the Micropyle in Tilapia mossambica Peters and Stigmatogobius javanicus (Blkr.)

The formation of the micropyle in Tilapia mossambica and Stigmatogobius javanicus, two fresh water teleosts is described. In T. mossambica the micropyle is formed by a single modified follicular cell, while in S. javanicus it is formed by a cellular mass derived from the follicle.     

POLLINATION DROP IN RELATION TO CONE MORPHOLOGY IN PODOCARPACEAE: A NOVEL REPRODUCTIVE MECHANISM

Observation of ovulate cones at the time of pollination in the southern coniferous family Podocarpaceae demonstrates a distinctive method of pollen capture, involving an extended pollination drop. Ovules in all genera of the family are orthotropous and single within the axil of each fertile bract. In Microstrobus and Phyllocladus ovules are erect (i.e., the micropyle directed away from the cone axis) and are not associated with an ovule-supporting structure (epimatium). Pollen in these two genera must land directly on the pollination drop in the way usual for gymnosperms, as observed in Phyllocladus. In all other genera, the ovule is inverted (i.e., the micropyle is directed toward the cone axis) and supported by a specialized ovule-supporting structure (epimatium). In Saxegothaea there is no pollination drop and gametes are delivered to the ovule by pollen tube growth. Pollination drops were observed in seven of the remaining genera. In these genera the drop extends over the adjacent bract surface or cone axis and can retain pollen that has arrived prior to drop secretion (“pollen scavenging”). The pollen floats upward into the micropylar cavity. The configuration of the cone in other genera in which a pollination drop has not yet been observed directly suggests that pollen scavenging is general within the family and may increase pollination efficiency by extending pollination in space and time. Increased pollination efficiency may relate to the reduction of ovule number in each cone, often to one in many genera, a derived condition. A biological perspective suggests that animal dispersal of large seeds may be the ultimate adaptive driving force that has generated the need for greater pollination efficiency.     

Ovulate cone,pollination drop,and pollen capture in Sequoiadendron (Taxodiaceae)

In Sequoiadendron ovules are borne inside the ovulate cone, and pollination drops secreted from these ovules collect pollen. We examined: (1) the relation between ovular position and pollen capture; (2) pollen behavior when in contact with a pollination drop; and (3) ultrastructure of ovules during pollination drop secretion. During wet periods a water sheet forms on the surface of the cone due to bract shape and wettability. Pollination drops persist inside the wetted cone, and pollen capture resumes immediately after drying. Pollen landing on a pollination drop is taken inside the drop and carried into the micropyle when the drop contracts. Several notable ultrastructural features appear in the nucellus, integument, chalaza, and bract lamina during pollination-drop secretion. The abaxial surface of the lamina is covered by a membrane that may contribute to the wettable nature of the surface.     

澳洲杨(大戟科)胚珠及其种子附属结构的个体发育研究

以澳洲杨的胚珠及其种子为材料,运用光学显微镜和电镜扫描的方法对其从胚珠发生直到种子成熟的个体发育过程和结构进行观察,同时与鸭脚西番莲的种子附属结构的发育过程进行对比研究。结果表明:(1)澳洲杨的珠孔类型为外珠孔类型,种子附属结构起源于珠孔而非珠柄,其为种阜,而非假种皮。(2)鸭脚西番莲的珠孔类型为内珠孔类型,种子附属结构起源于珠柄,并且最终将珠孔包被,其为真正的假种皮结构。通过种阜与假种皮的不同个体发育过程,建立了大戟科种阜与假种皮的不同发育模式,并对种子附属结构的生物学功能及其暗示的不同植物进化路径进行了讨论。     

CONE AND OVULE ONTOGENY IN TAXODIUM AND GLYPTOSTROBUS (TAXODIACEAE – CONIFERALES)

Seed cones in Taxodium distichum and Glyptostrobus pensilis occupy the position of permanent shoots and are initiated in the summer preceding spring pollination. Morphological features are similar in the two genera, reflecting their close taxonomic relationship. Ovule complexes originate as two (rarely more) ovule primordia in the axil of each fertile bract but without any indication of a preceding discrete ovuliferous scale. When the nucellus, integument, and micropyle are well developed, a series of up to ll abaxial lobes forms at the base of each ovule pair. They become fused by basal growth. After pollination the common basal meristem of lobes and bract extends by intercalary growth to form the conspicuous “ovuliferous scale” of the mature cone; the lobes enlarge and exceed the ovules. Despite the topographic similarity in the cones of both genera, there are differences in vasculature such that the vascular traces to the axillary complex originate directly from the axial cylinder in Glyptostrobus but from the bract trace in Taxodium. The complex vasculature of the mature cone develops late and primarily as an expression of intercalary growth.     

Sperm penetration and transformation of sperm entry site in eggs of the freshwater teleost Rhodeus ocellatus ocellatus

Eggs of bony fishes are enveloped by an egg envelope (chorion) in which a micropyle is present near the animal pole. Therefore, sperm penetration into the eggs is limited to the sperm entry site (SES), a region of plasma membrane just beneath the micropyle. In rose bitterling eggs, the SES transforms from a tuft of microvilli into a swollen mass (SM) that continues to plug the micropyle after sperm penetration. The present observations using the rose bitterling Rhodeus ocellatus ocellatus were conducted to examine: 1) whether or not sperm penetration is necessary for formation of the SM and 2) whether or not actin microfilaments are involved in the formation of the SM. Water activation without sperm transformed the SES from a tuft of microvilli into the SM, although it took a longer time for the transformation and the SMs were smaller than in the case of inseminated eggs. The SES presumably has the ability to transform into the SM upon activation of eggs in the present species. Cytochalasin B, which acts on actin microfilaments, did not prevent formation of the SM, irrespective of insemination or activation. The present observations suggest that sperm penetration is not necessary for SM formation and actin microfilaments do not participate in SM formation. © 1996 Wiley-Liss, Inc.     

Concentration-dependent stimulation and inhibition of growth cone behavior and neurite elongation by protein kinase inhibitors KT5926 and K-252a

We examined the concentration- and time-dependent effects of two related protein kinase inhibitors, KT5926 and K-252a, on neurite formation and nerve growth cone migration of chick embryo sensory neurons. The effects of these drugs on neurite formation over an 18-h period were dissimilar. KT5926 stimulated neurite formation at concentrations between 100 and 500 nM and inhibited neurite formation at 5 μM. K-252a had no stimulatory effects on neurite formation, and it inhibited neurite formation at concentrations above 50 nM. This difference may occur because K-252a inhibits activation of the nerve growth factor receptor trk A, while KT5926 does not inhibit trk A. Both drugs, however, had similar immediate effects on growth cone migration. Growth cone migration and lamellipodial spreading were rapidly stimulated by 500 nM concentrations of KT5926 and K-252a. At 2 μM levels of either drug, growth cone spreading was still stimulated, but growth cone migration was inhibited by both drugs. These results show that changes in protein phosphorylation/dephosphorylation can rapidly regulate the cellular machinery that is responsible for driving growth cone migration and neurite elongation. The different effects of 2 μM concentrations of either KT5926 or K-252a on growth cone spreading versus migration suggests that the actin-dependent protrusive motility of the growth cone leading margin is regulated differently by changes in protein phosphorylation and dephosphorylation than the cytoskeletal mechanism that drives neurite elongation. © 1997 John Wiley & Sons, Inc. J Neurobiol 33: 161–171, 1997     

Micropyle of some salmonins and coregonins

Synopsis The micropyle of seven salmonins and four coregonins were examined by a scanning electron microscope. In addition, the micropyle of Coregonus pidschian is described for the first time. The micropyles and the surface pattern may be used for identification of eggs. The studied micropyle belongs to two different types: 1. micropyle with flat pit and long canal (type II) and 2. micropyle with canal but without pit (type III). The sides of the micropyle canal are often reinforced by annular or helicoid thickening.     

New Species of Eimeria (Protozoa: Eimeriidae) from Malaysian Rats*

SYNOPSIS. Four new species of Eimeria were found in a survey of 255 rats of 14 species in Malaysia. E. tikusi n. sp. and E. edwardsi n. sp. are described from Edwards' rat Rattus edwardsi. The ellipsoidal, single-layered oocysts of E. tikusi average 30.3 by 24.4 μ. A micropyle is absent; a polar granule is present. Ovoid sporocysts average 14.2 by 9.8 μ. A sporocyst residuum and Stieda body are present. The ovoid, 2-layered oocysts of E. edwardsi average 29.1 by 21.8 μ. A micropyle is present; a polar granule is absent. Ellipsoidal to ovoid sporocysts average 14.5 by 6.5 μ. A sporocyst residuum is present; Stieda body is small or absent. E. surifer n. sp. is described from the red spiny rat Rattus surifer. Its ellipsoidal 3-layered oocysts average 34.7 by 24.8 μ. A micropyle is absent; a polar granule is present. The ellipsoidal sporocysts average 15.4 by 9.5 μ. A sporocyst residuum, Stieda body and sub-Stieda body are present. E. sabani n. sp. is described from the long-tailed giant rat R. sabanus. Its ellipsoidal 2-layered oocysts average 28.5 by 21.7 μ. A micropyle is absent; a polar granule is present. The ellipsoidal-to-ovoid sporocysts average 11.9 by 8.0 μ. A sporocyst residuum and Stieda body are present.     

The surface structure of Antarctic fish eggs and its use in identifying fish eggs from the Southern Ocean

Summary The surface structure of eggs of Notothenia neglecta, Nototheniops larseni, Nototheniops nudifrons (Nototheniidae), Champsocephalus gunnari and Chaenocephalus aceratus (Channichthyidae), collected near Elephant Island (Atlantic sector of the Southern Ocean) and of Notothenia rossii rossii (Nototheniidae) from Kerguelen waters (Indian sector of the Southern Ocean) was studied by scanning electron microscopy (SEM). Eggs of N. larseni differed from the other species by containing a micropyle with two micropyle pits. External morphological characteristics of the egg envelope such as the ultrastructure of the zona radiata, its interpore distances and the diameter of the micropyle were species-specific and could aid in the identification of fish eggs collected from the Southern Ocean.     

Fine structures of the micropyles of pelagic eggs of some marine fishes

The micropyles and their surrounding structures of pelagic fish eggs of nine species were examined and compared using a scanning electron microscope (SEM). The surface of the egg membrane possesses uniformly distributed pores. However,Lateolabrax japonicus possesses a number of small knobs. The region of the micropyle of fish eggs examined is characterized by a circular elevation in which the pores are larger than those on the other parts of the egg membrane. In contrast,Kareius bicoloratus egg has no elevation and possesses uniform pores to the edge of the micropylar canal. The funnel-like structure is seen in the opening of the micropyle inSardinops melanostictus, Saurida elongata, Sillago japonica, Parapristipoma trilineatum, Hypodytes rubripinnis andK. bicoloratus. These specific features of the micropyle may be used for the identification of pelagic eggs.     

A new stomach worm,Obeliscoides pentalagi n. sp. (Nematoda; Trichostrongyloidea) of Ryukyu rabbits,Pentalagus furnessi (Stone, 1900)

Summary A new trichostrongylid nematode, Obeliscoides pentalagi n. sp., from the stomach of the Ryukyu Rabbit, Pentalagus furnessi, collected on Amami-ohshima Island, south Japan is described. O. pentalagi n. sp. resembles O. leporis Schulz, 1931 and O. travassosi Liu & Wu, 1941, but differs in the relative size and morphology of the dorsal ray and genital cone of the males. The number and distribution of longitudinal cuticular ridges of O. pentalagi n. sp. is similar to those of O. cuniculi multistriatus Measures & Anderson, 1983, but it differs in the morphology of the spicules, dorsal ray and genital cone in males, and in the morphology of the prevulval lobe of the females. The discovery of O. pentalagi further supports the hypothesis of a Eurasian origin of the genus Obeliscoides (Graybill, 1923) Graybill, 1924 (Durette-Desset, 1983; Durette-Desset & Chabaud, 1977; Measures & Anderson, 1983a, b, c, 1984). The number and distribution of longitudinal cuticular ridges and variation of the prevulval lobe in females of O. leporis Schulz, 1931 are also examined and compared with those of other species.     

Ultrastructural features of the egg envelope of silver carp, <Emphasis Type="Italic">Hypophthalmichthys molitrix</Emphasis> (Osteichthyes,Cyprinidae)

Synopsis We investigated the ultrastructure of the egg membrane surface (unfertilized egg) of Hypophthalmichthys molitrix using scanning electron microscopy. The eggs of silver carp like most other teleosts are surrounded by a relatively thick egg membrane, and have a type III micropyle at the animal pole. The micropyle is almost circular in shape and the micropyle canal is located in its center. The micropyle region is not flat. Round or oval accessory pores are also observed in the canal. The surface of zona radiata is wavy and uneven with a uniform distribution of almost round pores with lips. Microvilli like structure are found in the pore opening region.     

The evolution of fertilization modes independent of the micropyle in Fagales and `pseudoporogamy'

In contrast to a majority of angiosperms showing porogamous fertilization, several fagalean families such as Betulaceae and Casuarinaceae are known to show chalazogamy, where fertilization is effected by a pollen tube passing through the chalaza instead of the micropyle. Our developmental study of pollen-tube growth in pistils of Myrica rubra (Myricaceae, Fagales) further shows that pollen tubes reached the nucellus before the micropyle is formed by the integument. Since fertilized ovules appeared as if the pollen tube had passed through the micropyle for fertilization, we propose the new term `pseudoporogamy' to this mode. By mapping diverse modes of fertilization, dependent or independent of the micropyle, onto a phylogenetic tree of Fagales, it appears that fertilization mode evolved from porogamy to chalazogamy and then further from chalazogamy to pseudoporogamy. Possible reasons for the evolution of fertilization modes independent of the micropyle in Fagales are discussed.     

Two new species ofOdilia (Nematoda: Heligmonellidae) from Australian rodents,with comments onO. bainae Beveridge & Durette-Desset, 1992

Odilia tasmaniensis n. sp.,O. praeputialis n. sp. andO. bainae Beveridge & Durette-Desset, 1992 from the small intestine of Australian murids are described and illustrated. The two new species are distinguished from the other eight species in the genus, namelyO. mackerrasae (Mawson, 1961),O. brachybursa (Mawson, 1961),O. emanuelae (Mawson, 1961),O. melomyos (Mawson, 1961),O. polyrhabdote (Mawson, 1961),O. uromyos (Mawson, 1961),O. mawsonae (Durette-Desset, 1969) andO. bainae Beveridge & Durette-Desset, 1992.O. tasmaniensis n. sp. fromRattus lutreolus is characterised by the longitudinal cuticular ridges being continuous, the presence of 18 ridges in cross-section in the middle region of the body, the joined distal ends of the spicules forming a curved bluntly rounded tip, a short genital cone with a single ventral papilla and a pair of laterally curving dorsal raylets and the posterior end of the female tapering sharply.O. praeputialis n. sp. fromZyzomys woodwardi is characterised by continuous longitudinal cuticular ridges, the presence of 22–35 ridges in cross-section in the middle of the body, the sharply pointed joined distal tips of the spicules, a complex genital cone with a flat membraneous proconus, a ventral papilla projecting from an extension of the body wall, a pair of short straight dorsal raylets and the presence of a praepuce on the posterior end of the female.O. bainae is characterised by the longitudinal cuticular ridges being continuous, the presence of 17–22 ridges in cross-section in the middle region of the body, the joined distal ends of the spicules surrounded in an oval transparent cap, a long genital cone with a single ventral papilla and a pair of laterally curving dorsal raylets, and the absence of a praepuce on the posterior end of the female.Rattus lutreolus and the pseudomyine rodentsPseudomys higginsi andMastacomys fuscus are new host records for this species.     

Structural aspects of in vitro pollen tube growth and micropylar penetration inGasteria verrucosa (Mill.) H. Duval andLilium longiflorum Thunb.

Summary In vitro penetration of the micropyle of freshly isolatedGasteria verrucosa ovules by pollen tube was monitored on agar medium. 40–60% of the micropyles were penetrated, comparable with in vivo penetration percentages. When germinated on agar,Gasteria pollen tube elongation lasts for up to 8 h while plasma streaming continues for about 20–24 h. The generative cell divides between 7 and 20 h after germination, and after 20 h the pollen tube arrives at one of the synergids. The sperm cells arrive after 22 h. The whole process takes more time in vitro than in vivo. In fast growing pollen tubes, a pulsed telescope-like growth pattern of tube elongation is observed. The formation of pollen tube wall material precedes tube elongation and probably prevents regular enlargement of the pollen tube tip-zone. Rapid stretching of the new pollen tube wall material follows, probably due to gradually increased osmotic pressure and the use of lateral wall material below the tip. The stretching ceases when the supplies of plasma membrane and excretable wall material are exhausted. Multiple pollen tube penetration of the micropyle occurs in vitro as it does in vivo. Most pollen tube growth ceases within the micropyle but, if it continues, the pollen tubes curl. Inside the micropyle the pollen tube shows haustorial growth. At the ultrastructural level, the wall thickening of in vitro pollen tubes is quite similar to that in vivo. Before transfer of pollen tube cytoplasm a small tube penetrates one of the synergids. Sperm nuclei with condensed chromatin are observed in the pollen tube and the synergid. In vivo prometaphase nuclei are found in the most chalazal part of a synergid, against the egg cell nucleus and nucleus of the central cell at a later stage. Using media forLilium ovule culture,Gasteria ovules were kept alive for at least 6 weeks. Swelling of the ovule depends on pollen tube penetration. The conditions for fertilization to occur after in vitro ovular pollination seem to be present.     

Envelope surface ultrastructure and specific gravity of artificially fertilized Pacific cod Gadus macrocephalus eggs

The envelope surface ultrastructure and specific gravity of artificially fertilized eggs of the Pacific cod Gadus macrocephalus were examined. The unfertilized, demersal and slightly adhesive eggs of G. macrocephalus were almost spherical and had no oil globules. Wrinkled envelope surface with elaborated hexagonal reticulated patterns and type I micropyle were observed under a scanning electron microscope. The adhesiveness of the eggs was lost at the blastodermal‐cap stage after fertilization. The micropylar canal was sealed by secretion of the perivitelline fluid, and the entire surface became rough. Numerous bacilli were deposited at the micropyle and the outer envelope surface at the late germ‐ring stage and at the embryo five‐eighths around the yolk stage. The micropyle was completely deformed at the embryo seven‐eighths around the yolk stage. The specific gravity of the fertilized G. macrocephalus eggs ranged from c. 1·0316 to 1·0454. These values, however, sharply decreased towards the end stages of egg development to produce pelagic larvae. The ultrastructural changes in the micropyle and envelope surface of the G. macrocephalus eggs protected the embryo from microorganism infections and mechanical stress during the long incubation period. The adhesiveness and specific gravity of the eggs influenced their dispersion potential.     

New species of Eimeria Schneider, 1875 and Isospora Schneider, 1881 (Apicomplexa: Eimeriidae) from the short-crested flycatcher Myiarchus ferox (Gmelin) (Passeriformes: Tyrannidae) in South America

In the current study, two new coccidian species (Protozoa: Apicomplexa: Eimeriidae) obtained from short-crested flycatcher Myiarchus ferox (Gmelin) are reported from Brazil. Isospora feroxis n. sp. has o?cysts which are spheroidal to sub-spheroidal, 18.7 × 18.0 μm, with a smooth and bi-layered wall, c. 1.2 μm. The micropyle and o?cyst residuum are absent, but two polar granules are present. Its sporocysts are broadly ellipsoidal and 11.7 × 8.5 μm. Stieda and substieda bodies are present. A sporocyst residuum is present and the sporozoites have a refractile body and nucleus. O?cysts of Eimeria sicki n. sp. are spheroidal to sub-spheroidal, 30.3 × 28.5 μm, with a smooth and bi-layered wall, c. 1.3 μm. The micropyle, o?cyst residuum and polar granule are absent. Its sporocysts are ellipsoidal, 18.4 × 10.0 μm. Stieda and substieda bodies are present. A sporocyst residuum is present and sporozoites have a refractile body and nucleus.     

Sperm Incorporation Independent of Fertilization Cone Formation in the Danio Egg

The responses of the egg to insemination in a modified Fish Ringer's solution (FRS) were examined in eggs of the zebrafish ( Brachydanio rerio ) primarily by scanning electron microscopy. FRS is a physiological saline which temporarily inhibits parthenogenetic activation of the egg for 5–8 min. Spermatozoa were collected in a small volume of water and pipetted over eggs in FRS. Eggs inseminated in FRS typically incorporated the fertilizing sperm within 3–4 min. Inseminated cells showed an absence of a fertilization cone and no cortical granule exocytosis. The deep conical depression in the egg surface beneath the micropyle remained unaltered. Control eggs inseminated in tank water developed a large fertilization cone during sperm incorporation. Occasionally, eggs inseminated in water were observed to incorporate the entire sperm head prior to egg activation. Our results corroborate earlier findings showing that in the zebrafish, cortical granule exocytosis, fertilization cone formation and elevation of the sperm entry site are not triggered by the fertilizing sperm in experimental conditions (18, 19). Furthermore, sperm incorporation requires neither egg activation nor formation of a fertilization cone in this fish.