Functional anatomy of male copulatory organs of Pomacea canaliculata (Caenogastropoda, Ampullariidae)

This study was aimed to investigate the functional morphology of copulation and sperm transfer in the invasive snail Pomacea canaliculata. Three-dimensional renderings of the male copulatory apparatus were made and showed elaborate systems for innervation and for hemolymph supply and drainage. A key component of the male copulatory apparatus is the penial sheath, which shows three specialized glands; the medial and distal glands may participate in adherence to the mantle cavity wall of the female during copulation. The outer gland has an epithelium composed of columnar cells with branched microvilli, mucous goblet cells and large granular secretory cells containing intragranular crystalloids, which produce an exocrine secretion during copulation. The interaction of male/female copulatory organs was studied in dissections of snap-frozen pairs. Sperm are left in the sperm pit, at the end of the pallial spermiduct. Afterwards, the muscular action of the penial bulb takes the sperm up to the vermiform penis, which slides from the penial pouch into the central groove of the penial sheath, and it later emerges through a T-shaped sulcus of this structure and enters the female vagina. Then, it climbs through the capsule duct, and its tip reaches the proximity of the seminal receptacle. A model of copulation and sperm transfer is presented on the basis of the new findings and on published literature.     

External structures used during attachment and sperm transfer in tubificids (Annelida,Oligochaeta)

The genital region of seven species of Tubificidae has been studied by SEM (Scanning Electron Microscopy). The form and the position of penial and spermathecal chaetae, male and spermathecal pores and other special structures have been examined. Peristodrilus montanus shows a special system to hold the partner: the penial chaetae anchor in an elaborated structure of the body wall formed between the spermathecal pores, the `anchorage bridge'. Protuberodrilus tourenqui has a long glandular porophore with the male pores at the tip, allowing contact with the spermathecal pores which are located in deep, close to the mid-ventral line of the body. The grooved and strongly curved penial chaetae of Rhyacodrilus falciformis seem to be used both for attachment and for sperm transfer, entering into the lateral spermathecal pores. The embrace of the partners, as suggested by observations on Psammoryctides barbatus, Potamothrix bavaricus, Potamothrix hammoniensis and Potamothrix heuscheri, seems to be another important mechanism to fix contact between male and spermathecal pores and allow sperm transfer. The spermathecal chaetae could be interpreted as piercing chaetae with a chemical or mechanical stimulating role. Sensitive cilia near the penial chaetae seem to be used by the three rhyacodrilines studied to find the correct anchorage place. There is a great variety of structures which appear to be used for attachment and sperm transfer in tubificids, and consequently their role in the evolution of the whole family may be profound.     

The male copulatory apparatus in an opisthobranch mollusc, Runcina

The copulatory apparatus of a primitive opisthobranch, Runcina, is described. The apparatus is comprised of the following organs: the spermatic bulb, the prostrate, the penis and the penial sac. The spermatic bulb wall consists of cuboidal epithelium with forked microvilli and densely arranged cilia. Prior to copulation the interior is tightly packed with sperm. The prostate is lined with alternating glandular and supporting cells, the latter being compressed but with a mushroom-shaped apex bearing a few forked microvilli and many cilia. The glandular cells produce differing secretions, each cell producing a single type. Large paracrystalline structures enclosed in cells close to the penial area are particularly striking. Considerable amounts of the secretory products are accumulated in the protruding cell apices. One type of inclusion is found at a later stage, packed around the sperm mass within the spermatophore; its function, and the fate of the other secretions is not yet clear. The epithelium of the penis is of more columnar structure, covered with forked microvilli and extremely long cilia which are anchored by long rootlets in the cells. Some of the cells contain large electron-dense secretory granules, others hold accumulations of small secretory vesicles in their apices. It seems likely that these contribute towards the outer layer of spermatophore. The wall of the penial sac is lined by one to two rows of flat-cuboidal cells bearing sparse forked microvilli and cilia.     

Cutaneous application of an accessory-gland secretion after sperm exchange in a terrestrial slug (Mollusca: Pulmonata)

Competition for fertilisation in hermaphroditic animals seems to have led to many odd behaviours and complex morphologies involved in the transfer of accessory-gland products to the partner. Terrestrial slugs of the genus Deroceras show remarkably elaborate and interspecifically diverse penis morphologies and mating behaviours. Most species have an appending penial gland, which in Deroceras panormitanum consists of a few long fingers that are everted after sperm exchange and laid onto the partner’s back. To investigate whether this gland transfers a secretion onto the partner’s skin, we killed slugs at different mating stages and studied their penial glands and skin histologically. Two types of secretion granules appeared at a very early stage of courtship, and the penial gland was already filled 15 min into the courtship. At copulation, the gland everted this secretion onto the partner’s body, where it remained for at least 50 min. No lysis of skin tissue or other effects on the skin were observed. The slugs tried to lick the received secretion off their own body, and some droplets were observed to be shed with the body mucus. Our results indicate the external application of a glandular substance that could function as either a pheromone or allohormone. The behaviours of the recipients suggest sexual conflict, although mutual interest cannot be ruled out.     

Isolation of Sperms from the Pollen Tubes of Flowering Plants during Fertilization

Sperm cells have been isolated from pollen tubes growing in style segments of the dicotlyledon Rhododendron macgregoriae and the monocotyledon Gladiolus gandavensis by the in vivo/in vitro method at various stages of fertilization. Pollen tubes emerged from the cut end of the style into agar medium, and more than 95% contained sperm cells. Sperm cells were released from the pollen tubes by osmotic shock or by placing styles in wall-degrading enzymes: 0.5% macerozyme and 1% cellulase. The isolated sperms were ellipsoidal protoplasts of diameter about 2 × 3 micrometers in Gladiolus and about 3 × 4 micrometers in Rhododendron. After isolation, a proportion of the sperm cells occurred in pairs linked at one end by finger-like connections. The pairs of isolated sperms were dimorphic in terms of surface area and volume. By cutting the styles at various positions and times after pollination, the potential exists to detect changes in sperm gene expression associated with fertilization.     

Evaluation of pollen tube growth ability in Petunia species having different style lengths

Pollen tube growth is essential for the fertilization process in angiosperms. When pollen grains arrive on the stigma, they germinate, and the pollen tubes elongate through the styles of the pistils to deliver sperm cells into the ovules to produce the seeds. The relationship between the growth rate and style length remains unclear. In previous studies, we developed a liquid pollen germination medium for observing pollen tube growth. In this study, using this medium, we examined the pollen tube growth ability in Petunia axillaris subsp. axillaris, P. axillaris subsp. parodii, P. integrifolia, and P. occidentalis, which have different style lengths. Petunia occidentalis had the longest pollen tubes after 6 h of culture but had a relatively shorter style. Conversely, the pollination experiments revealed that P. axillaris subsp. parodii, which had the longest style, produced the longest pollen tubes in vivo. The results revealed no clear relationship between the style lengths and the growth rate of pollen tubes in vitro. Interspecific pollinations indicated that the styles affected pollen tube growth. We concluded that, in vitro, the pollen tubes grow without being affected by the styles, whereas, in vivo, the styles significantly affected pollen tube growth. Furthermore, interspecific pollination experiments implied that the pollen tube growth tended to be suppressed in the styles of self-incompatibility species. Finally, we discussed the pollen tube growth ability in relation to style lengths.     

Semi in vivo pollen tube growth of Aechmea fasciata

With semi in vivo pollen tube growth assays, stigmas are pollinated in vivo and, after a fixed time interval, the styles are isolated from the ovary and placed on culture medium in vitro. Semi in vitro pollination includes isolation of the stigma and style complex, followed by pollination and placing the stylar end on nutrient medium. After semi in vivo pollination more and longer pollen tubes protruded from the cut end of the styles into medium, in comparison to semi in vitro pollination. Medium with 3 g l^–1 agar was better than that with 6 g l^–1 agar for pollen tube growth after the tubes emerged from the cut style. Semi in vitro pollination of the reversed style indicated that pollen tube growth was not influenced by the direction of the style. Fructose and glucose inhibited pollen tube growth compared to sucrose. Swollen tips characterized tube growth inhibition. After semi in vivo pollination all generative nuclei had divided to give two sperm nuclei. The average distance between the last sperm nucleus and the pollen tube tip as well as the distance between the two sperm nuclei diminished in growing pollen tubes between 24 and 48 h after pollination. The arrangements between the vegetative and the generative nuclei did not differ in semi in vivo and in vitro cultured pollen tubes of Aechmea fasciata. This information is important to explain why fertilization rate is low after placental pollination in comparison to placental grafted style pollination of Aechmea fasciata. The data may also contribute to the improvement of in vitro fertilization methods in Bromeliaceae and other higher plants.     

Functional morphology of the mammiliform penial glands ofLittorina saxatilis (Gastropoda)

Summary The functional morphology of the mammiliform penial glands ofLittorina saxatilis has been investigated with both light and electron microscopy. These penial glands line the ventral edge of the penis and orient with the female mantle during copulation. Secretions are released from the penial glands to this interface where they probably function in adhesion. The penial gland secretions comprise heterogeneous granules as well as apocrine and mucous secretions. The heterogeneous granules are produced in separate multicellular glands arranged in a series of lobes that lie outside a thick smooth muscle layer enclosing the lumen. Each glandular lobe is surrounded by a thin layer of smooth muscle. Secretions are transported in individual cellular processes that pass through the thick smooth muscle layer and empty into the lumen. Surrounding the lumen is an epithelium containing apocrine secretory cells as well as occasional goblet-type, mucous cells. The combined action of the muscles forces secretions out of the lumen through the penial papilla, onto the external surface of the mammiliform penial gland. Longitudinal muscles extend into the penial papilla enabling its protrusion or retraction. Retraction of the penial papilla following secretion release is thought to create negative pressure beneath the penial gland producing suction adhesion. The visco-elastic properties of the penial gland secretion are qualitatively different from foot mucus and may represent specialization to an adhesive function.     

Introduction to Physidae (Gastropoda: Hygrophila); biogeography, classification, morphology

Physidae, a world-wide family of freshwater snails with about 80 species, are reclassified by progressive characters of the penial complex (the terminal male reproductive system): form and composition of penial sheath and preputium, proportions and structure of penis, presence or absence of penial stylet, site of pore of penial canal, and number and insertions of penial retractor muscles. Observation of these characters, many not recognized previously, has been possible only by the technique used in anesthetizing, fixing, and preserving. These progressive characters are the principal basis of 23 genera, four grades and four clades within the family. The two established subfamilies are divided into seven new tribes including 11 new genera, with diagnoses and lists of species referred to each. Proposed as new are: in Aplexinae, Austrinautini, with Austrinauta g.n. and Caribnautu harryi g.n., nom.nov.; Aplexini; Amecanautini with Amecanauta jaliscoensis g.n., sp.n., Mexinauta g.n., and Mayabina g.n., with M. petenensis, polita, sanctijohannis, tempisquensis spp.nn., Tropinauta sinusdulcensis g.n., sp.n.; and Stenophysini, with Stenophysa spathidophallus sp.n.; in Physinae, Haitiini, with Haitia moreleti sp.n.; Physini, with Laurentiphysa chippevarum g.n., sp.n., Physa mirollii nom.nou.; and Physellini, with Chiapaphysa g.n., and C. grijalvae, C. pacifica spp.nn., Utahphysa g.n., Archiphysa g.n., with A. ashmuni, A. sonomae spp.nn., Physella hemphilli sp.n., and Ultraphysella sinaloae g.n., sp.n. The simplest reproductive system is found in Austrinauta of the Aplexinae; its penial complex approaches that in the related family Lymnaeidae. Within Physinae a close approximation is found in Haitia. By these two genera the two subfamilies are drawn close together. Four grades of progressive complexity are recognized: (I) penial sheath entirely muscular; (II) penial sheath with both glandular and muscular tissue; (III) penis with penial stylet or other specialization of the tip of the penis; and (IV) pore of penial canal lateral rather than terminal as in the lower grades. In both subfamilies there are clades with glandular tissue in the penial sheath, a penial sheath subdivided into two parts, and tip of penis specialized in various ways. These clades are formalized as new tribes. Of 23 genera of Physidae, 17 occur in Pacific drainages of North and Central America, eight of these restricted to the region. Concentration of primitive genera along the Pacific coast from Mexico to Costa Rica conforms to previous observations that primitive pulmonate families are concentrated within, or along the continental margins of, the Pacific Ocean. An ancestral origin of Physidae along an ancient eastern Pacific coast is probable. From this region the several lineages have spread north, south and east in the Americas, and through Siberia to Europe. Although Physinae have fewer genera than Aplexinae (11 v. 12), they have more species (47 v. 34). Greater land area in the temperate zone has provided more opportunity for speciation of Physinae, in contrast to the generally tropical and warm-temperate range of Aplexinae. Furthermore, 10 species of Physinae are localized in individual lakes, whereas Aplexinae are not lake-dwellers. Both well-developed egg strings and capsular strings are found in the spawn of Sibirenauta elongatus. These structures have been known in Lymnaeidae, but not hitherto in Physidae; they are a link with some marine groups, such as Siphonariidae. Spiral color bands and white streaks in the shell of Mexinauta recall those in Lancidae (Lymnaeacea), whereas the radula of Physidae is like that of Chilinidae. Physidae thus show affinities to various basal stocks of aquatic pulmonates; no clear-cut sister-group can be recognized. Most species have a restricted range; out of 55 with sufficiently detailed information for analysis, 25 are limited to a single 1 degrees x 1 degrees quadrangle. Only a few species are widespread, on one or even two continents. Accordingly, more species of Physidae are threatened by habitat destruction than in other families of Hygrophila with generally wider distributions. Other features are a key to genera; catalog of more than 430 names applied to living Physidae, with original reference, type locality, and location of type specimens; summary of museums with types; and glossary.     

The morphology of styles and stigmas inVicia (Leguminosae), and its systematic implications

The morphology and development of styles and stigmas ofVicia, one of the five genera of tribe Vicieae, were studied by scanning electron microscopy to contribute to a revision of infrageneric classification. Among the 22 species examined, variations were found as to: (1) whether a boundary between stigma and style is V-shaped when viewed from the adaxial side or not; (2) whether stylar hairs sprout on all sides of the style or only on the abaxial side; (3) whether a hairy area of the style connects with the stigma area first at lateral sides or at an adaxial side; (4) whether styles are tapered or bottle-neck-shaped; (5) whether styles are straight or bend abaxially. Based on these variations, five morphological types of styles and stigmas were provided with a diagnostic key. Comparisons with related tribes Cicereae, Coronilleae, Galegeae, Hedysareae, Loteae and Trifolieae as possible outgroups suggested that the V-shaped boundary between stigma and style is a synapomorphy of some species withinVicia, and that a dorsally compressed and abaxially tufted style, which was distributed in separate subgenera and sections sensu Kupicha but is now known to occur together with the V-shaped boundary, has evolved only one time in the genus, rather than more than once as supposed earlier.     

New insights into the functional morphology of the male copulatory apparatus of bullid gastropods

In this study, semi-thin sections stained with histochemical techniques and transmission electron microscopy were used to obtain new data about the morphology and function of the male copulatory apparatus of the cephalaspidean gastropod Bulla striata. The apparatus comprises a vestibule, a penial papilla and a prostate consisting of a coiled unbranched tube ending in a blind caecum. The penial papilla and the coiled tubular prostate are enclosed by a muscular sheath, which is continuous with the muscular tissue of the vestibule. The epithelium lining the lumen of the vestibule is formed by ciliated and mucus-secreting cells. Two new types of subepithelial secretory cells were discovered in this region. The penial papilla is a muscular structure without secretory cells in the epithelium lining the narrow lumen. The tubule that constitutes the prostate possesses a muscular wall and can be divided in three distinct regions: a non-secretory duct connected to the penial papilla, a glandular region rich in large secretory cells and the terminal caecum containing just a few small secretory cells. In the terminal blind caecum, the muscular sheath is fused with the muscular wall of the tubular prostate. Large numbers of spermatozoa were found in the glandular region and in the terminal caecum of the prostate. A new functional mechanism is proposed to explain penial eversion during copulation. This differs from a previous hypothesis in two main aspects: (1) existence of a permanent penial papilla in mature animals acting as a functional penis and (2) functional role of vestibule during copulation, which everts and surrounds the penial papilla, while the latter protrudes outwards.     

Observations on graphing paleozoological data: Suggestions for better graphs

Paleozoologists have long used graphs of diverse styles to describe, analyze, and summarize their data. Some of these graphs provide excellent visual representations of complex data and are readily deciphered. Other graph styles require close study to be interpreted. Ease of visual decoding of information contained in a graph – graph perception – varies from graph style to graph style. Historical instances of graphing paleozoological data indicate some difficult to decipher graph styles have been used for at least a century. Graphs with three-dimensions, moiré effects, superimposed lines, or segmented bars, or which demand simultaneous decipherment of position and magnitude, are ill-advised. Temporal trends in data are best graphed following the principle of superposition such that data from old material is graphed at the bottom and data from younger material is graphed at the top of the diagram.     

从水稻花粉管分离精细胞(简报)

精细胞的分离是植物生殖工程的一个重要组成部分,是目前被子植物有性生殖研究的一个活跃领域[1,2]。随着精细胞分离技术的完善和分离出精细胞的植物类型的增加,目前对精细胞的分子生物学研究已有一些进展,主要是精细胞特异蛋白的分离[3,4]和cDNA文库的构建以及一些精细胞特异基因的分离[5,6]。     

Observations on the anatomy of Hermaea variopicta (Opisthobranchia: Ascoglossa)

The alimentary system, reproductive system and central nervous system of Hermaea variopicta (Costa) are described for the first time. Its gut resembles that of Hermaea bifida (Montagu), type species of the genus; the radula is considered in detail. Its diaulic reproductive system is described in morphological and functional terms. The central nervous system has three ganglia on the visceral loop. The anatomical evidence supports the opinion of Lemche & Thompson (1974) that Hermaeopsis Costa, former genus of H. variopicta, should be discarded. However, the division of the genus Hermaea into two subgenera, Hermaea and Placida, is not recommended. H. variopicta appears to be more primitive than H. bifida. In the discussion, the origin of hermaeid and stiligerid lines from a common ancestral stock is suggested, to which is added a new theory of the evolution of stiligerid penial styles.     

Morph-specific patterns of variation in stigma height in natural populations of distylous Jasminum fruticans

The purpose of this study was to describe morph-specific patterns of stigma–anther separation and style curling in the distylous Jasminum fruticans . We also examined whether variation in floral traits is correlated with variation in seed production. Stigma–anther separation is more variable in short-styled plants than in long-styled plants. In all populations studied, some plants bear flowers that lack any stigma–anther separation. These plants have the pollen characteristics (size and number) and compatibility relations of short-styled plants. Comparison with other distylous species illustrates that the variability of stigma–anther separation in short-styled plants is a novel finding for a distylous species. Long-styled plants have greater stigma–anther separation than short-styled plants, styles are often curled and protrude from the corolla, and anthers are placed well within the corolla tube. The frequency of long-styled plants with curled styles and the mean degree of style curling were significantly correlated with style length. Short-styled plants have larger corollas than long-styled plants and never have curled styles. Morph ratios are always 50:50 in natural populations. Mean values of each floral trait in the two morphs were significantly correlated among populations. There were no consistent differences in fecundity of the two morphs nor any correlation between floral traits and seed set for each morph in natural populations. We discuss the potential causes and significance of the two morph-specific patterns we describe; reduced stigma–anther separation in short-styled plants and the presence of curled styles in long-styled plants.     

The internal anatomy of Stiliger fuscatus (A. Gould, 1870) (Opisthobranchia: Sacoglossa)

The central nervous system of Stiliger fuscatus has two ganglia on the visceral loop. The superficial buccal musculature includes an ascus muscle; its contraction drives the lower limb of the radula in the action of piercing an algal wall. The reproductive system is described in functional terms and related to those of other species of the family Stiligeridae. The vagina does not open to the exterior and the penial style is of the penetrant type; fertilization is by hypodermic impregnation. The system could be called pseudo-diaulic and this term is explained in the additional notes.     

Redescriptions of Phallodrilus parthenopaeus Pierantoni and P. obscurus Cook (Oligochaeta, Tubificidae)

The type species of Phallodrilus Pierantoni, 1902, P. parthenopaeus Pierantoni, 1902; is redescribed on the basis of new material from W Scotland. The species is characterized by its specific spermathecal and penial setae, its very slender atria, and its long spermathecal ducts. Sexually fully mature specimens of P. obscurus Cook, 1969 are described for the first time, on the basis of material from New Jersey (E USA). The new material shows that this species possesses 5 to 9 penial setae in each ventral bundle of segment XI, and that its atria are coated with a relatively thick musculature.     

How are speech styles elaborated for presenting individual personality while bearing in mind social norms? Certain cases in Beppu City, Oita Prefecture, Japan

The purpose of this paper is to examine how speech styles are elaborated for presenting individual personality. Through the narratives of ten selected interviewees, some social norms internalised by them emerged. Age, status and career are associated to the joo/ge (upper/lower) and the senpai/koohai (senior/junior) relationships. Whether it is the first meeting and the length of relationship are associated to shin/so (intimacy/distance) and uchi/soto (in-group/out-group). In particular the notion of shin/so takes a role for adjusting speech styles. According to the narratives of individual episodes, there are several factors of determination of speech style, which are not necessarily following social norms. The way of constructing relationships, keeping power relations, protection and so on are influenced by regional characteristics, which include factors of determination of speech style such as generation, gender, solidarity, and jibunrashisa (like myself, personality) and so forth.     

Evolution of the Sacoglossa (Mollusca, Opisthobranchia) and the ecological associations with their food plants

Evolution in the opisthobranch order Sacoglossa has been closely linked to their specialized suctorial herbivorous habits. All shelled Sacoglossa (about 20% of the species) feed on one algal genus, Caulerpa. The non-shelled Sacoglossa have 'radiated' to other diets, mainly siphonalean or septate green algae (Class Ulvophyceae). Comparing the phylogeny of sacoglossan genera with the phylogeny of the Ulvophyceae indicates that co-speciation may have taken place at the basal node of the Sacoglossa, and that host switching has taken place several times in the two non-shelled clades. It is suggested that the most important evolutionary process has been speciation by 'resource-tracking'; the resource tracked is most probably cell wall composition of the algal prey. The fossil record of extant sacoglossan genera dates back to the Eocene and, based on the fossil record of siphonalean green algae, the Sacoglossa most likely appeared in the Cretaceous. It is hypothesized that the ancestral sacoglossan was epifaunal, suctorial and herbivorous, and the 'ancestral' food plant was not Caulerpa, but filamentous, calcified, now extinct, Udoteaceae.     

Stylish lengths: Mate choice in flowers

The styles of flowers may represent an arena for pollen competition in the race to fertilize ovules. Accordingly, selection should favour a longer ‘race’ to better discriminate among variable pollen by increasing style length. Sampling across a taxonomically diverse range of wild and outcrossed species, we found that the distribution of style lengths within plants were skewed towards longer styles, as predicted. In self-pollinated domesticated species, where discrimination among pollen is less important, we found no such pattern. We conclude that style length is under directional selection towards longer styles as a mechanism for mate choice among pollen of variable quality.