The Musculature of <Emphasis Type="Italic">Testudinella patina</Emphasis> (Rotifera: Flosculariacea), Revealed with CLSM

The musculature of Testudinella patina was visualized using phalloidin-linked fluorescent dye by confocal laser scanning microscopy. The conspicuous broad retractors appear to be made up of five separate fibers, of which three anchor in the neck region whereas two extend into the corona. Besides the broad retractors, a total of five paired longitudinal retractors are present and all of them extend into the corona. Incomplete circular muscles are found in groups in the neck region and in the medial and posterior parts of the trunk. The foot musculature comprises eight thin ventral foot muscles and six thicker dorsal foot muscles that all extend from the foot basis to the distal part of the foot. At the basis of the foot, each of the dorsal foot muscles anchors on a smaller, S-shaped subterminal foot muscle. The foot musculature furthermore comprises one pair of paraterminal foot muscles that each anchors basally on a subterminal foot muscle, extends into the most proximal part of the foot and attaches on one of the dorsal foot muscles. The visceral musculature is composed of extremely delicate fibers and is restricted to an area around and posterior to the foot opening. The presence of incomplete circular muscles supports that these muscles are a basal trait for Rotifera, whereas the morphology of the broad retractors and foot muscles is much more specialized and may be autapomorphic for Testudinella or alternatively for this genus and its closest relatives. The present results stress that revealing muscles by staining may produce new information from even well-investigated species, and that this information may contribute to a better understanding of functional as well as phylogenetic aspects of rotifer biology.     

Ultrastructure of tentacles in the sipunculid worm <Emphasis Type="Italic">Thysanocardia nigra</Emphasis> Ikeda, 1904 (Sipuncula)

The ultrastructure of the tentacles was studied in the sipunculid worm Thysanocardia nigra. Flexible digitate tentacles are arranged into the dorsal and ventral tentacular crowns at the anterior end of the introvert of Th. nigra. The tentacle bears oral, lateral, and aboral rows of cilia; on the oral side, there is a longitudinal groove. Each tentacle contains two oral tentacular canals and an aboral tentacular canal. The oral side of the tentacle is covered by a simple columnar epithelium, which contains large glandular cells that secrete their products onto the apical surface of the epithelium. The lateral and aboral epithelia are composed of cuboidal and flattened cells. The tentacular canals are lined with a flattened coelomic epithelium that consists of podocytes with their processes and multiciliated cells. The tentacular canals are continuous with the radial coelomic canals of the head and constitute the terminal parts of the tentacular coelom, which shows a highly complex morphology. Five tentacular nerves and circular and longitudinal muscle bands lie in the connective tissue of the tentacle wall. Similarities and differences in the tentacle morphology between Th. nigra and other sipunculan species are discussed.Original Russian Text Copyright © 2005 by Biologiya Morya, Maiorova, Adrianov.     

Musculature and nervous system of<Emphasis Type="Italic"> Gnathostomula peregrina</Emphasis> (Gnathostomulida) shown by phalloidin labeling,immunohistochemistry, and cLSM,and their phylogenetic significance

Musculature and nervous system of Gnathostomula peregrina (Gnathostomulida, Scleroperalia) were reconstructed from whole animals by immunohistochemistry and confocal laser scanning microscopy. The F-actin muscular subset, stained with FITC-labeled phalloidin, consists of: (1) eleven pairs (four ventral, one ventrolateral, one dorsolateral, five dorsal) of longitudinal muscles; (2) two types of diagonal muscles (thin fibers throughout the body, and slightly thicker fibers of which seven pairs occur ventrally and two pairs dorsally); (3) evenly spaced thin circular fibers that gird the posterior half of the body, continuing less prominently into the anterior half; and (4) a complex pharyngeal and genital musculature. Dorsoventral muscles are absent. The organization of the FMRFamidergic nervous system shows: (1) a central nervous system with a frontal ganglion and one pair of longitudinal nerves ending in a terminal commissure, and one median ventral nerve; (2) eight to ten unipolar perikarya above, and up to ten bipolar perikarya in front of the brain; (3) a total of five (one unpaired, two paired) longitudinal nerves of the peripheral nervous system with two to four accompanying perikarya; and (4) a buccal ganglion of the stomatogastric nervous system with six to eight perikarya above the pharyngeal bulbus. Our results reveal the musculature and nervous system of Gnathostomula to be more complex than hitherto reported.     

<Emphasis Type="Italic">In vitro</Emphasis> propagation of eight species or subspecies of <Emphasis Type="Italic">Turbinicarpus (Cactaceae)</Emphasis>

Summary In vitro propagation systems by means of areole activation were developed for Turbinicarpus laui, T. lophophoroides, T. pseudopectinatus, T. schmiedickeanus subsp. flaviflorus, T. schmiedickeanus subsp. klinkerianus, T. schmiedickeanus subsp. schmiedickeanus, T. subterraneus, and T. valdezianus. In vitro-germinated seedlings were used as a primary source of explants. Multiple shoot formation from areoles was achieved for three explant types (apical, lateral, and transverse), cultured on Murashige and Skoog (MS) basal medium supplemented with 3% sucrose, 10 gl⁻¹ agar and several treatments with cytokinins. Efficiencies were in the range from 7.8 shoots per explant in T. valdezianus up to 19.7 shoots per explant in T. pseudopectinatus, using the best treatment for each species and in a single proliferation cycle. Four of the studied species responded best when 6-benzylaminopurine (3.3–8.8μM) was used, while 6-(γ,γ-dimethylallylamino)purine (19.7–24.6μM) showed better results in two species. The two remaining species showed no significant differences in their response to both cytokinins. Regarding explant type, the best results were obtained with transverse cuts for five species, with apical explants for one species, and the two remaining species showed no significant differences among the explants tested. Rooting of the in vitro-generated shoots was achieved most efficiently on half- or full-strength MS basal medium. Rooting frequencies were in the range from 54.2 to 94.2%, and the frequency of survival of the plants once transferred to soil was 91.6% on average.     

Surface structure and frictional properties of the skin of the Amazon tree boa <Emphasis Type="Italic">Corallus hortulanus</Emphasis> (Squamata,Boidae)

The legless locomotion of snakes requires specific adaptations of their ventral scales to maintain friction force in different directions. The skin microornamentation of the snake Corallus hortulanus was studied by means of scanning electron microscopy and the friction properties of the skin were tested on substrates of different roughness. Skin samples from various parts of the body (dorsal, lateral, ventral) were compared. Dorsal and lateral scales showed similar, net-like microornamentation and similar friction coefficients. Average friction coefficients for dorsal and lateral scales on the epoxy resin surfaces were 0.331 and 0.323, respectively. In contrast, ventral scales possess ridges running parallel to the longitudinal body axis. They demonstrated a significantly lower friction coefficient compared to both dorsal and lateral scales (0.191 on average). In addition, ventral scales showed frictional anisotropy comparing longitudinal and perpendicular direction of the ridges. This study clearly demonstrates that different skin microstructure is responsible for different frictional properties in different body regions.     

Structure and direction of jaw adductor muscles as herbivorous adaptations in <Emphasis Type="Italic">Neotoma mexicana</Emphasis> (Muridae,Rodentia)

The herbivorous adaptations of the jaw adductor muscles in Neotoma mexicana were clarified by a comparative study with an unspecialized relative, Peromyscus maniculatus. In P. maniculatus, the anterior part of the deep masseter arises entirely from the lateral side of an aponeurosis, i.e., superior zygomatic plate aponeurosis, whereas N. mexicana has an additional aponeurosis for this part of the muscle, and the fibers attach on both sides of the superior zygomatic plate aponeurosis. Although the structure of the temporalis muscle is nearly identical in the two genera, a clear aponeurosis of origin occurs only in N. mexicana. These characteristics allow fibrous tissues to be processed with a large occlusal force. The deep masseter, internal pterygoid, and external pterygoid muscles of N. mexicana incline more anterodorsally than those of P. maniculatus. The transverse force component of these muscles relative to whole muscle force is smaller in N. mexicana than in P. maniculatus, with the exception of the internal pterygoid. The anterior part of the temporalis muscle of N. mexicana is specialized to produce occlusal pressure. These findings suggest that in N. mexicana a large anterior force is required to move the heavy mandible, due to the hypsodont molars, against frictional force from food, and that the posterior pull of the temporalis, which adjusts the forward force by the other jaw adductor muscles to a suitable level, need not be large for the mandibular movement.     

The musculature of horsehair worm larvae (<Emphasis Type="Italic">Gordius aquaticus</Emphasis>,<Emphasis Type="Italic"> Paragordius varius</Emphasis>, Nematomorpha): F-actin staining and reconstruction by cLSM and TEM

The musculature of larvae of Gordius aquaticus was investigated by laser-scanning microscopy and compared to transmission electron microscopic data for the larva of Paragordius varius. In the anterior portion of the body, the preseptum, four different muscle groups can be distinguished: (1) 12 anterior parietal muscles in the body wall, (2) six oblique muscles that function as retractors of the introvert, (3) six proboscideal muscles, which function as retractors for the proboscis, and (4) six muscles associated with spines of the outermost of the three rings of spines. The posterior portion of the body, the postseptum, possesses four pairs of longitudinal muscle strands in G. aquaticus, the postseptal parietal muscles, that are located dorsolaterally and ventrolaterally. These are not clearly visible in P. varius, where instead three pairs of parietal muscles are present. Additional small muscles are associated with the terminal spines and with the duct running from the pseudointestine to the body wall. All fibers show a cross-striated pattern although this striation is less obvious at the ends of the fibers.     

Observations on the skulls of sturgeons (Acipenseridae): shared similarities of <Emphasis Type="Italic">Pseudoscaphirhynchus kaufmanni</Emphasis> and juvenile specimens of <Emphasis Type="Italic">Acipenser stellatus</Emphasis>

A novel hypothesis uniting Acipenser stellatus and Pseudoscaphirhynchus as sister groups has recently been proposed based on analysis of DNA sequences. In this paper I compare specimens of A. stellatus and P. kaufmanni, and show that they share several putatively derived similarities in the structure of their skulls, including: the presence of large spines on the dermal bones of skull; lateral extrascapular bones that enclose the confluence of the posttemporal, supratemporal, and otic sensory canals; elongate dorsal rostral bones; border rostral bones distinct in shape from dorsal rostral bones; greatly enlarged jugal that lacks a median flange; rostral canal bones that loop posteriorly at the anterior commissure of the rostral sensory canals; and the presence of an elongate, flat, and broad posterior ventral rostral bone. These similarities support a close relationship between A. stellatus and Pseudoscaphirhynchus, but still remain to be critically tested.     

Two New Species of Goby of the Genus <Emphasis Type="Italic">Astrabe</Emphasis> from Japan

Two species of goby belonging to the genus Astrabe are described from Japan as new species, A. flavimaculata and A. fasciata. A. flavimaculata is distinguishable from A. lactisella, the type species and hitherto the only known species of the genus, in that it has no protrusion on the upper posterior part of the dermal fold along the upper margin of the eye, fewer scales in a longitudinal row, predorsal scales, scales on the belly, a narrower white transverse band across the base of the pectoral fins, and in life yellow markings on a dark brown ground colour except for the white transverse band across the base of the pectoral fins. A. fasciata is distinguishable from A. lactisella in that it has fewer scales in a transverse row, a narrower scaled area on the lateral side of the body, a narrower white transverse band across the base of the pectoral fins, and a white transverse band across the anterior part of the 1st dorsal fin extending to the ventral side of the body.     

<Emphasis Type="Italic">In vitro</Emphasis> organogenesis of <Emphasis Type="Italic">Pelecyphora aselliformis</Emphasis> erhenberg (cactaceae)

Summary In vitro propagation of Pelecyphora aselliformis, a Mexican cactus which is considered rare and is highly valued in the commercial market, was initiated using seeds as explants. The longitudinal explants from seedlings germinated in vitro were cultivated on Murashige and Skoog medium containing 8.8 μM benzyladenine (BA) or 4.6 μM kinetin at pH 7.0. After 120 d, each explant gave rise to five shoots and this number of shoots increased 20–25% after subculture. The hyperhydricity was similar in both media, but callus formation was lower on the medium with BA. The shoot development, in terms of epicotyl length, and fresh and dry weight after 6 wk, was also recorded. The epicotyl length was similar on shoot-forming media but the quality of shoots was better on media containing BA. In about 1 yr, 500–600 well-defined shoots were obtained. The rooting of shoots was very slow and a vigorous radical system was observed after 1 yr of culture.     

The tyrosine <Emphasis Type="Italic">O</Emphasis>-prenyltransferase SirD catalyzes <Emphasis Type="Italic">O</Emphasis>-, <Emphasis Type="Italic">N</Emphasis>-, and <Emphasis Type="Italic">C</Emphasis>-prenylations

Recently, the prenyltransferase SirD was found to be responsible for the O-prenylation of tyrosine in the biosynthesis of sirodesmin PL in Leptosphaeria maculans. In this study, the behavior of SirD towards phenylalanine/tyrosine and tryptophan derivatives was investigated. Product formation has been observed with 12 of 19 phenylalanine/tyrosine derivatives. It was shown that the alanine structure attached to the benzene ring and an electron donor, e.g., OH or NH₂, at its para-position are essential for the enzyme activity. Modifications were possible both at the side chain and the benzene ring. Enzyme products from seven phenylalanine/tyrosine derivatives were isolated and characterized by MS and NMR analyses including HSQC and HMBC and proven to be O- or N-prenylated derivatives at position C4 of the benzene rings. K _M values of six selected derivatives were found in the range of 0.10–0.68 mM. Catalytic efficiencies (K _cat/K _M ) were determined in the range of 430–1,110 s⁻¹·M⁻¹ with l-tyrosine as the best substrate. In addition, 7 of 14 tested tryptophan analogs were also accepted by SirD and converted to C7-prenylated derivatives, which was confirmed by comparison with products obtained from enzyme assays using a 7-dimethylallyltryptophan synthase 7-DMATS from Aspergillus fumigatus.     

Expression of a<Emphasis Type="Italic"> SoxB</Emphasis> and a<Emphasis Type="Italic"> Wnt2/13</Emphasis> gene during the development of the mollusc<Emphasis Type="Italic"> Patella vulgata</Emphasis>

We cloned and analysed the expression of a SoxB gene (PvuSoxB) in the marine mollusc, Patella vulgata. Like its orthologues in deuterostomes, after an early broad ectodermal distribution, PvuSoxB expression only persists in cells competent to form neural structures. In the post-gastrulation larva, PvuSoxB is expressed in the prospective neuroectoderm in the head and in the trunk. No expression can be seen dorsally, around the mouth and the anus, or along the ventral midline. We also report the expression of a Wnt2/13 orthologue (PvuWnt2) in Patella. After gastrulation, PvuWnt2 is expressed in the posterior part of the mouth, along the ventral midline and around the anus. This expression seems to be complementary to that of PvuSoxB in the trunk. We suggest the existence of a fundamental subdivision of the Patella trunk ectoderm into midline (mouth, midline, anus) and more lateral structures.Edited by D. Tautz     

Poorly known stonefly species, <Emphasis Type="Italic">Isoperla pseudornata</Emphasis> and <Emphasis Type="Italic">Kaszabia nigricauda</Emphasis> (Plecoptera,Perlodidae), from the Eastern Palaearctic

The previously unknown male of Isoperla pseudornata Zhiltzova and nymph of Kaszabia nigricauda (Navás) of the family Perlodidae are described. Sternite VIII of the male of I. pseudornata has no vesicle, the posterior margin is covered medially with strong short brownish setae. Sternite IX forms a subgenital plate. The everted aedeagus is upcurved, turned towards the dorsum, with two rounded lobes and one finger-shaped apical lobe terminating with a membranous funnel depressed medially. The aedeagus membrane is covered with small fine pointed sclerotized spinules; spinules absent on the tip of the funnel. The funnel basally and the finger-shaped apical lobe ventrally are covered with fine pointed sclerotized spinules and rounded sclerotized formations. I. pseudornata differs from the other Eastern Palaearctic congeners in the shape of spots on the head, pronotum, and abdominal tergites. The head of K. nigricauda nymph bears a large brown M-shaped spot forming along the anterior margin two fine stripes widened on the frontoclypeus; the interocular area bears a small pale spot merged with the large transverse W-shaped spot on the occiput. The lacinia is narrowed to the apex, bidentate; the tip of the galea reaches the base of the subapical tooth. The pronotum bears a pale cruciform medial spot, the lateral fields are dark, the lateral margins are pale. The meso-and metanotum exhibit a pale complicated pattern widened to the base of the wing pads. The legs are pale. The abdominal tergites are brown, with a pale transverse stripe widened laterally on the last tergites. Tergite X bears a large pale heart-shaped spot medially. The cerci bear a silky hair fringe dorsally; in the apical part, the apical whorl of cerci has short setae; one ventral and one dorsal seta are longer than the others; the dorsal setae are longer than ventral ones.     

Nervous and muscle system development in <Emphasis Type="Italic">Phascolion strombus</Emphasis> (Sipuncula)

Recent interpretations of developmental gene expression patterns propose that the last common metazoan ancestor was segmented, although most animal phyla show no obvious signs of segmentation. Developmental studies of non-model system trochozoan taxa may shed light on this hypothesis by assessing possible cryptic segmentation patterns. In this paper, we present the first immunocytochemical data on the ontogeny of the nervous system and the musculature in the sipunculan Phascolion strombus. Myogenesis of the first anlagen of the body wall ring muscles occurs synchronously and not subsequently from anterior to posterior as in segmented spiralian taxa (i.e. annelids). The number of ring muscles remains constant during the initial stages of body axis elongation. In the anterior-posteriorly elongated larva, newly formed ring muscles originate along the entire body axis between existing myocytes, indicating that repeated muscle bands do not form from a posterior growth zone. During neurogenesis, the Phascolion larva expresses a non-metameric, paired, ventral nerve cord that fuses in the mid-body region in the late-stage elongated larva. Contrary to other trochozoans, Phascolion lacks any larval serotonergic structures. However, two to three FMRFamide-positive cells are found in the apical organ. In addition, late larvae show commissure-like neurones interconnecting the two ventral nerve cords, while early juveniles exhibit a third, medially placed FMRFamidergic ventral nerve. Although we did not find any indications for cryptic segmentation, certain neuro-developmental traits in Phascolion resemble the conditions found in polychaetes (including echiurans) and myzostomids and support a close relationship of Sipuncula and Annelida.     

<Emphasis Type="Italic">In vitro</Emphasis> propagation of endangered <Emphasis Type="Italic">Mammillaria</Emphasis> genus (Cactaceae) species and genetic stability assessment using SSR markers

In vitro propagation protocols were established for endangered species of cacti Mammillaria hernandezii, M. dixanthocentron, and M. lanata. In vitro-germinated seedlings were used as the explant source. Three explant types were evaluated as apical, basal, and lateral stem sections. Shoot multiplication was achieved using Murashige and Skoog (MS) medium supplemented with benzyladenine, kinetin, meta-topolin, and thidiazuron in equimolar concentrations (0.0, 0.4, 1.1, 2.2, 4.4, and 8.9 μM). Shoot regeneration was obtained primarily in the lateral stem section explants. In M. hernandezii, an average of 7.4 shoots was regenerated in MS medium with 2.2 μM meta-topolin. M. dixanthocentron and M. lanata averaged 16.7 and 17.9 shoots/explant, respectively, in MS medium supplemented with 1.1 μM meta-topolin. Rooting occurred in MS medium without growth regulators. Three in vitro culture cycles were performed to validate the propagation protocols and to verify genetic stability. Shoots were collected in each cycle and genomic DNA was extracted. Amplified microsatellites were used to compare each genotype with its respective donor plant. Polymorphic information content analysis showed low levels of intra-clonal polymorphisms—M. hernandezii 0.04 and M. dixanthocentron and M. lanata both 0.12. More than 95% of the plants were successfully acclimatized in the greenhouse. After 12 months, plants of M. hernandezii reached the flowering stage; M. dixanthocentron and M. lanata flowered at 24 mo.     

Micropropagation of pitaya (<Emphasis Type="Italic">Hylocereus undatus</Emphasis> Britton et rose)

Summary A procedure for micropropagation of pitaya using thidiazuron (TDZ) and naphthaleneacetic acid (NAA) is described. Explants were excised from young joints of mature plants and cultured on Murashige and Skoog medium (MS) containing 0.5 μM NAA and 0.5 μM TDZ. Shoots produced from these first explants were cut up to produce secondary explants, either by decapitation or by longitudinal division into three parts. The decapitated and longitudinal explants were cultured on MS supplemented with 0.5 μM NAA and either 0.01, 0.09, 0.5, or 0.9 μM TDZ. Decapitated explants produced more shoots at higher frequency that the longitudinal explants. For both types of secondary explants, most shoots were developed from the distal parts. Shoots produced from secondary explants were rooted in MS and then transferred to soil where they produced normal plants.     

The Inflorescence Stem Fibers of <Emphasis Type="Italic">Arabidopsis thaliana Revoluta</Emphasis> (<Emphasis Type="Italic">ifl1</Emphasis>) Mutant

Arabidopsis thaliana is gradually gaining significance as a model for wood and fiber formation.revolute/ifl1 is an important mutant in this respect. To better characterize the fiber system of therevolute/ifl1 mutant, we grew plants of two alleles (rev-9 in Israel andrev-1 in the USA) and examined the fiber system of the inflorescence stems using both brightfield and polarized light. Microscopic examination of sections of plants belonging to the two different alleles clearly revealed that, contrary to previous views, in 18 (13 in Israel and 5 in Ohio) out of 30 stems (20 in Israel and 10 in Ohio) the mutant produced the primary wavy fiber system of the inflorescence stems. Our findings are further supported by the fact that fibers are seen in the figures published in other studies of the mutant even when it was stated that there were no fibers. The impression of a total lack of the wavy band of fibers is in many cases just a result of poorly lignified secondary walls. This specific gene that reduces lignification in fibers is of great significance for biotechnological developments for the paper industry and thus for the global economy and ecology. We propose thatrevoluta, the first name given to this mutant (Talbert and others 1995), is more appropriate thanifl1. Online publication: 7 April 2005     

Increase of CGRP Expression in Motor Endplates Within Fore and Hind Limb Muscles of the Degenerating Muscle Mouse (<Emphasis Type="Italic">Scn8a</Emphasis><Superscript><Emphasis Type="Italic">dmu</Emphasis></Superscript>)

The distribution of calcitonin gene-related peptide (CGRP) was examined in skeletal muscles of fore and hind limb as well as in oral and cranio-facial regions of the degenerating muscle (dmu) mouse, which harbours a null mutation in the voltage-gated sodium channel gene Scn8a. In limb, oral and cranio-facial muscles of wild type mice, only a few motor endplates contained CGRP-immunoreactivity. However, many CGRP-immunoreactive motor endplates appeared in the triceps brachii muscle, the biceps brachii muscle, the brachialis muscle, and the gastrocnemius muscle of dmu mice. CGRP-immunoreactive density of motor endplates in the skeletal muscles was also elevated by the mutation. In these muscles, the atrophy of muscle fibers could be detected and the density of cell nuclei in the musculature increased. In the flexor digitorum profundus muscle, the flexor digitorum superficialis muscle, and the soleus muscle as well as in oral and cranio-facial muscles, however, the distribution of CGRP-immunoreactivity was barely affected by the mutation. The morphology of muscle fibers and the distribution of cell nuclei within them were also similar in wild type and dmu mice. In the lumbar spinal cord of dmu mice, CGRP-immunoreactive density of spinal motoneurons increased. These findings suggest that the atrophic degeneration in some fore and hind limb muscles of dmu mice may increase CGRP expression in their motoneurons.