Embryogenesis of a springtail,Tomocerus ishibashii (Collembola,tomoceridae): External morphology

The external features of the developing embryos of the springtail, Tomocerus ishibashii, are described. The clypeolabral anlage arises as a single, unpaired swelling. The entognathy is completed by the ventral growth of the tergal anlagen of mandibular, maxillary, and labial segments. These anlagen also form the posterior part of the cranium. The palpi of maxilla and labium are homologous with the telopodites, and proximal parts of these head appendages are homologous with the coxopodites. The sternal element of the labial segment does not participate in the postmentum formation. The anlagen of abdominal appendages appear in the first to the fourth abdominal segments. The first, third, and fourth appendage anlagen form the ventral tube, tenaculum, and furcula, respectively. The fused proximal parts of the first, third, and fourth appendage anlagen are homologous with the coxopodites, and the distal parts which do not fuse are homologous with the telopodites. The anlagen of the second abdominal appendages become flattened and spread over the ventral side of this segment. The ventral structures of the first four abdominal segments are appendicular in origin.     

A new specimen ofWeinbergina opitzi (Chelicerata: Xiphosura) from the Lower Devonian Hunsriick Slate, Germany

A new specimen of the synziphosurine arthropodWeinbergina opitzi is described from the Lower Devonian (Lower Emsian) Hunsrück Slate of Germany (Rhenish Slate Mountains). It is the smallest and only the fifth specimen of this taxon to be described and is preserved in ventral aspect with exceptional preservation of prosomal and opisthosomal appendages. This specimen confirms the presence of a seventh appendage, similar in morphology to the preceding prosomal appendages, associated with opisthosomal segment one. In addition, at least three opisthosomal plates fringed with teeth are confirmed. Correlation of prosomal appendage podomeres betweenWeinbergina and selected chelicerate taxa shows that appendage structure is most similar to eurypterid appendages III–IV and Araneae appendages III–VI. This is in contrast to modern horseshoe crabs which have fewer podomeres in appendages II–V due to an undifferentiated tibiotarsus.      

An unusual short flagellum inMallomonas guttata (Synurophyceae,chrysophyta)

A short flagellum bearing unusual appendages inMallomonas guttata is described. These appendages, ca. 0.2 μm in diameter and 10–20 μm long, 7–10 in number, originate from the distal end of the short flagellum. Their membrane is continuous with the flagellar membrane and no particular structure was observed in appendage lumen.     

The Expression of Beta ({beta}) Keratins in the Epidermal Appendages of Reptiles and Birds

The integuments of extant vertebrates display a variety of epidermalappendages whose patterns, morphology and terminal differentiation(epidermal keratins) depend upon interactions between ectodermal(epidermis) and mesodermal (dermis) tissues. In reptiles andbirds, appendage morphogenesis precedes terminal differentiation.Studies have demonstrated that appendage morphogenesis influencesthe expression of the appendage specific keratin genes. However,little is known about the nature of the structural genes expressedby the epidermal appendages of reptiles. How pattern formationand/or appendage morphogenesis influence terminal differentiationof reptilian appendages is not known. The epidermal appendages of reptiles and birds are characterizedby the presence of both alpha () and beta (ß) typekeratin proteins. Studies have focused on the genes of avianß keratins because they are the major structural proteinsof feathers. The occurrence of ß keratin proteinsin the scales and claws of both birds and reptiles and theirimmunological cross-reactivity suggest that the genes for reptilianß keratins may be homologous with those of birds.In bird appendages, the ß keratins are the productsof a large family of homologous genes. Specific members of thisgene family are expressed during the development of each appendage.Recent sequence analyses of feather ß keratins, fromdifferent orders of birds, demonstrate that there is more diversityat the DNA level than was implied by earlier protein sequencingstudies. Immunological techniques show that the same antibodies thatreact with the epidermal ß keratins of the chicken(Gallus domesticus) react with the epidermal ß keratinsof American alligators (Alligator mississippiensis). Furthermore,a peptide sequence (20 amino acids) from an alligator claw ßkeratin is similar to a highly conserved region of avian claw,scale, feather, and feather-like ß keratins. Theseobservations suggest that the ß keratin genes of avianepidermal appendages have homologues in the American alligator.Understanding the origin and evolution of the ß keratingene families in reptiles and birds will undoubtedly add toour understanding of the evolution of skin appendages such asscales and feathers.     

<Emphasis Type="Italic">Bradyrhizobium japonicum</Emphasis> bacteroid appendages expressed in senescing and argon-treated soybean nodules

Bradyrhizobium japonicum bacteroids in soybean nodules expressed fibrillar appendages during senescence. In both scanning and transmission electron microscopy (SEM and TEM), these structures were observed to connect adjacent bacteroids, and bacteroids to symbiotic membranes. They were 20–25 nm in diameter, 100–2,500 nm in length and were linear, branched, or part of a web-like matrix. Bacteroids expressing appendages were not uniformly distributed, but were abundant within localized regions in the senescing nodule. The root systems of nodulated greenhouse-grown plants flushed with argon induced the appendages at earlier plant ages, and they were more prolific and wide spread than those in untreated nodules. Bradyrhizobium japonicum symbiotic appendages appear to be a response to an environmental niche within senescing nodules.     

Partial co-option of the appendage patterning pathway in the development of abdominal appendages in the sepsid fly Themira biloba

The abdominal appendages on male Themira biloba (Diptera: Sepsidae) are complex novel structures used during mating. These abdominal appendages superficially resemble the serially homologous insect appendages in that they have a joint and a short segment that can be rotated. Non-genital appendages do not occur in adult pterygote insects, so these abdominal appendages are novel structures with no obvious ancestry. We investigated whether the genes that pattern the serially homologous insect appendages have been co-opted to pattern these novel abdominal appendages. Immunohistochemistry was used to determine the expression patterns of the genes extradenticle (exd), Distal-less (Dll), engrailed (en), Notch, and the Bithorax Complex in the appendages of T. biloba during pupation. The expression patterns of Exd, En, and Notch were consistent with the hypothesis that a portion of the patterning pathway that establishes the coxopodite has been co-opted to pattern the developing abdominal appendages. However, Dll was only expressed in the bristles of the developing appendages and not the proximal–distal axis of the appendage itself. The lack of Dll expression indicates the absence of a distal domain of the appendage suggesting that sepsid abdominal appendages only use genes that normally pattern the base of segmental appendages.     

Structure and membrane affinity of new amphiphilic siderophores produced by Ochrobactrum sp. SP18

The coastal α-proteobacterium Ochrobactrum sp. SP18 produces a suite of three citrate-derived, cell-associated amphiphilic siderophores, ochrobactins A–C. The ochrobactins are composed of a citric acid backbone amide-linked to two lysine residues. Each ε-amine of lysine is hydroxylated and acylated forming two hydroxamic acid moieties. One of the acylated appendages of each ochrobactin is (E)-2-decenoic acid. The other acylated appendages for ochrobactins A–C are (E)-2-octenoic acid, octanoic acid and (E)-2-decenoic acid, respectively. The ferric ochrobactin complexes are photoreactive in UV light, producing an oxidized ligand with loss of 46 mass units that can still coordinate Fe(III). The relative partitioning of the apo-ochrobactins, Fe(III) ochrobactins and Fe(III) photoproducts into 1,2-dimyristoyl-sn-glycero-3-phosphocholine vesicles is presented. The ochrobactins are the first example of aerobactin-based siderophores with two fatty acid appendages produced in a suite with varying acyl appendage lengths.Electronic Supplementary Material Supplementary material is available for this article at and is accessible for authorized users.     

A genetic map of bovine Chromosome 7 with an interspecific hybrid backcross panel

A genetic linkage map of bovine Chromosome (Chr) 7 was generated with a Bos taurus×Bos gaurus interspecific hybrid backcross panel. This study included six previously mapped microsatellites and five unmapped expressed genes that were identified by PCR-based restriction fragment length variants (RFLVs). The gene order (from centromere to telomere) and the map distances (in centimorgans) are as follows: cen–BM2607–11.2–LDLR–3.6–AMH,CSF2–11.2–BP41–19–BM6117–19–SPARC–14.4–FGFA–15.5–BM1853–11.2–RASA–18.8–ILSTS006. Previous comparative synteny mapping demonstrated that bovine Chr 7 shares homologous regions with both HSA5q and HSA19p. A break or fusion between AMH and CSF2 in an ancestral chromosome is suggested to account for the current arrangement of these homologous segments in the human and bovine genomes. In this study, we demonstrate that a short proximal portion of BTA7 is homologous with HSA19p, while a larger distal portion of BTA7 is homologous with human Chr 5q. The orientation of these conserved human segments on BTA7 is also demonstrated. Our data show that the linear order of genes has not been conserved within the homologous region of HSA5 and BTA7, and one chromosomal translocation or inversion is proposed to account for this difference. Received: 11 June 1996 / Accepted: 9 November 1996     

High synteny and colinearity among Eucalyptus genomes revealed by high-density comparative genetic mapping

Understanding genome differentiation is important to compare and transfer genomic information between taxa, such as from model to non-model organisms. Comparative genetic mapping can be used to assess genome differentiation by identifying similarities and differences in chromosome organization. Following release of the assembled Eucalyptus grandis genome sequence (January 2011; ), a better understanding of genome differentiation between E. grandis and other commercially important species belonging to the subgenus Symphyomyrtus is required. In this study, comparative genetic mapping analyses were conducted between E. grandis, Eucalyptus urophylla, and Eucalyptus globulus using high-density linkage maps constructed from Diversity Array Technology and microsatellite molecular markers. There were 236–393 common markers between maps, providing the highest resolution yet achieved for comparative mapping in Eucalyptus. In two intra-section comparisons (section Maidenaria–E. globulus and section Latoangulatae–E. grandis vs. E. urophylla), ∼1% of common markers were non-syntenic and within chromosomes 4.7–6.8% of markers were non-colinear. Consistent with increasing taxonomic distance, lower synteny (6.6% non-syntenic markers) was observed in an inter-section comparison between E. globulus and E. grandis × E. urophylla consensus linkage maps. Two small chromosomal translocations or duplications were identified in this comparison representing possible genomic differences between E. globulus and section Latoangulatae species. Despite these differences, the overall high level of synteny and colinearity observed between section Maidenaria–Latoangulatae suggests that the genomes of these species are highly conserved indicating that sequence information from the E. grandis genome will be highly transferable to related Symphyomyrtus species.     

Inclusion appendages associated with the intraerythrocytic rickettsial parasiteAnaplasma marginale are composed of bundled actin filaments

Summary Anaplasma marginale, a tick-borne rickettsia that infects erythrocytes of cattle, occurs within a parasitophorous vacuole or inclusion body. A tail-like inclusion appendage, composed of multiple filaments, occurs in association with the inclusion body membrane. The composition and function of the inclusion appendage have not been determined. In this study, theA. marginale inclusion appendage in bovine erythrocytes was found to be composed of actin filaments as determined by labeling with rhodamine-conjugated phalloidin. Electron microscopy studies revealed that theA. marginale inclusion appendages differed from F-actin tails reported previously in association with other pathogens in eukaryotic cells because these highly ordered structures were organized into regularly occurring striations, and the appendages were adhered directly to the parasitophorous vacuole membrane. In addition, actin appendages have not been described previously in erythrocytes. The potential role of the inclusion appendage associated withA. marginale in bovine erythrocytes and recently fed ticks is discussed.     

The controversial origin of the abdominal appendage‐like processes in immature insects: Are they true segmental appendages or secondary outgrowths? (Arthropoda hexapoda)

In this article, I review the major characteristics of different types of appendage‐like processes that develop at the abdominal segments of many immature insects, and I discuss their controversial morphological value. The main question is whether the abdominal processes are derived from segmental appendages serially homologous to thoracic legs, or whether they are “secondary” outgrowths not homologous with true appendages. Morphological and embryological data, in particular, a comparison with the structure and development of the abdominal appendages in primitive apterygote hexapods, and data from developmental genetics, support the hypothesis of appendicular origin of many of the abdominal processes present in the juvenile stages of various pterygote orders. For example, the lateral processes, such as the tracheal gills in aquatic nymphs of exopterygote insects, are regarded as derived from lateral portions of appendage primordia, homologous with the abdominal styli of apterygotan insects; these processes correspond either to rudimentary telopodites or to coxal exites. The ventrolateral processes, such as the prolegs of different endopterygote insect larvae, appear to be derived from medial portions of the appendicular primordia; they correspond to coxal endites. These views lead to the rejection of Hinton's hypothesis (Hinton [1955] Trans R Entomol Soc Lond 106:455–545) according to which all the abdominal processes of insect larvae are secondary outgrowths not derived from true appendage anlagen. J. Morphol. 2012. © 2012 Wiley Periodicals, Inc.     

Characterization of a Novel <Emphasis Type="Italic">Bacillus thuringiensis</Emphasis> Phenotype Possessing Multiple Appendages Attached to a Parasporal Body

Bacillus thuringiensis is a bacterium best known for its production of crystal-like bodies comprised of one or more Cry-proteins, which can be toxic to insects, nematodes or cancer cells. Although strains of B. thuringiensis have occasionally been observed with filamentous appendages attached to their spores, appendages in association with their parasporal bodies are extremely rare. Herein we report the characterization of Bt1-88, a bacterial strain isolated from the Caribbean that produces a spore–crystal complex containing six long appendages, each comprised of numerous thinner filaments approximately 10 nm in diameter and 2.5 μm in length. Each of the multi-filament appendages was attached to a single, small parasporal body located at one end of the bacterial spore. Biochemical tests, 16S rDNA gene sequencing, and the identification of two Cry proteins by partial protein sequencing (putatively Cry1A and Cry2A), unambiguously identified Bt1-88 as a strain of B. thuringiensis. Bt1-88 represents the second reported strain of B. thuringiensis possessing a parasporal body/appendage phenotype characterized by one or more long appendages, comprised of numerous filaments in association with a parasporal body. This finding suggests that Bt1-88 is a member of a new phenotypic class of B. thuringiensis, in which the parasporal body may perform a novel structural role through its association with multi-filament appendages.     

Assimilation of inorganic nitrogen by a mycobiont isolated from Pisonia grandis R. Br. (Nyctaginaceae) mycorrhiza

 Single isolates of a mycobiont isolated from Pisonia grandis R. Br., Pisolithus tinctorius (Pers.) Coker & Couch and Tylospora fibrillosa (Burt.) Donk were compared with regard to their relative abilities to produce key enzymes of inorganic nitrogen assimilation. Nitrate reductase (NR) activities in the P. grandis mycobiont and T. fibrillosa were significantly lower than in P. tinctorius. While specific activities for glutamate dehydrogenase (GDH) were higher in P. tinctorius than the other two fungi following NH₄ ⁺ pre-treatment, glutamine synthetase (GS) activity did not differ significantly between the three fungi. In all three fungi, specific activities for GS were significantly higher than for GDH. NR activity was expressed in all three fungi regardless of the nitrogen source in the medium, but in P. tinctorius diminished following continued exposure to either NO₃ ^–, NH₄ ⁺, glutamine or NO₃ ^– + glutamine. The data are discussed in relation to nitrogen utilisation by the P. grandis mycobiont. Accepted: 16 October 1997     

Muscular architecture of <Emphasis Type="Italic">Milnesium tardigradum </Emphasis>and <Emphasis Type="Italic">Hypsibius </Emphasis>sp. (Eutardigrada,Tardigrada) with some data on <Emphasis Type="Italic">Ramazottius oberhaeuseri</Emphasis>

The architecture of the musculature of the eutardigrade species Milnesium tardigradum Doyère, 1840, Hypsibius sp. and Ramazzottius oberhaeuseri (Doyère in Ann Sci Nat Zool Sér 2(14):269–369, 1840) is investigated by phalloidin staining and confocal laser scanning microscopy. There are methodological problems in staining eutardigrades due to physiological alterations under stress (anhydrobiosis) and due to penetration problems of the cuticle. It is helpful to fix specimens in the state of asphyxy, where animals are stretched following an oxygen shortage in their environment. The musculatures of all three species correspond in their general architecture, but differ in detail, such as in the number of muscles. All muscles are isolated muscle strands. There are on each body side two dorsal and one ventral muscle strands, in addition to a system of dorsoventral, lateral and lateroventral muscles. Seven median ventral attachment points give rise to dorsoventral, ventrolateral and appendage muscles. The appendages receive several muscles originating dorsally and ventrally. The number of muscles and the arrangement differ in each appendage. The fourth appendage shows the greatest differences with a far smaller number of muscles compared to other species. The musculature shows comparably few strict segmental patterns, for example, the musculature of each appendage differs from the other ones. By comparison with literature data on the same species and data of Macrobiotus hufelandi it can be shown that eutardigrades have a roughly comparable muscular architecture, but that there are several differences in detail. Dedicated to Professor Westheide on the occasion of his 70th birthday.     

Parasegmental appendage allocation in annelids and arthropods and the homology of parapodia and arthropodia

The new animal phylogeny disrupts the traditional taxon Articulata (uniting arthropods and annelids) and thus calls into question the homology of the body segments and appendages in the two groups. Recent work in the annelid Platynereis dumerilii has shown that although the set of genes involved in body segmentation is similar in the two groups, the body units of annelids correspond to arthropod parasegments not segments. This challenges traditional ideas about the homology of "segmental" organs in annelids and arthropods, including their appendages. Here I use the expression of engrailed, wingless and Distal-less in the arthropod Artemia franciscana to identify the parasegment boundary and the appendage primordia. I show that the early body organization including the appendage primordia is parasegmental and thus identical to the annelid organization and by deriving the different adult appendages from a common ground plan I suggest that annelid and arthropod appendages are homologous structures despite their different positions in the adult animals. This also has implications for the new animal phylogeny, because it suggests that Urprotostomia was not only parasegmented but also had parasegmental appendages similar to extant annelids, and that limb-less forms in the Protostomia are derived from limb-bearing forms.     

Homeotic evidence for the appendicular origin of the labrum in Tribolium castaneum

The ontogeny of the insect labrum, or upper lip, has been debated for nearly a century. Recent molecular data suggest a segmental appendage origin of this structure. Here we report the first arthropod mutation associated with a homeotic transformation of the labrum. Antennagalea-5 (Ag(5)) transforms both antennal and labral structures to resemble those of gnathal appendages in Tribolium castaneum. This labral transformation suggests that the labrum is a fused structure composed of two pairs of appendage endites, and is serially homologous to the gnathal appendages.     

Appendage-restricted gene induction using a heated agarose gel for studying regeneration in metamorphosed Xenopus laevis and Pleurodeles waltl

Amphibians and fish often regenerate lost parts of their appendages (tail, limb, and fin) after amputation. Limb regeneration in adult amphibians provides an excellent model for appendage (limb) regeneration through 3D morphogenesis along the proximodistal, dorsoventral, and anteroposterior axes in mammals, because the limb is a homologous organ among amphibians and mammals. However, manipulating gene expression in specific appendages of adult amphibians remains difficult; this in turn hinders elucidation of the molecular mechanisms underlying appendage regeneration. To address this problem, we devised a system for appendage-specific gene induction using a simplified protocol named the “agarose-embedded heat shock (AeHS) method” involving the combination of a heat-shock-inducible system and insertion of an appendage in a temperature-controlled agarose gel. Gene expression was then induced specifically and ubiquitously in the regenerating limbs of metamorphosed amphibians, including a frog (Xenopus laevis) and newt (Pleurodeles waltl). We also induced gene expression in the regenerating tail of a metamorphosed P. waltl newt using the same method. This method can be applied to adult amphibians with large body sizes. Furthermore, this method enables simultaneous induction of gene expression in multiple individuals; further, the data are obtained in a reproducible manner, enabling the analysis of gene functions in limb and tail regeneration. Therefore, this method will facilitate elucidation of the molecular mechanisms underlying appendage regeneration in amphibians, which can support the development of regenerative therapies for organs, such as the limbs and spinal cord.     

Comparative analysis of leg and antenna development in wild-type and homeotic<Emphasis Type="Italic"> Drosophila melanogaster</Emphasis>

The insect leg and antenna are thought to be homologous structures, evolved from a common ancestral appendage. The homeotic transformations of antenna to leg in Drosophila produced by mutation of the Hox gene Antennapedia are position-specific, such that every particular antenna structure is transformed into a specific leg counterpart. This has been taken to suggest that the developmental programmes of these two appendages are still similar. In particular, the mechanisms for the specification of a cell's position within the appendage would be identical, only their interpretation would be different and subject to homeotic gene control. Here we explore the degree of conservation between the developmental programmes of leg and antenna in Drosophila and other dipterans, in wild-type and homeotic conditions. Most of the appendage pattern-forming genes are active in both appendages, and their expression domains are partially conserved. However, the regulatory relationships and interactions between these genes are different, and in fact cells change their expression while undergoing homeotic transformation. Thus, the positional information, and the mechanisms which generate it, are not strictly conserved between leg and antenna; and homeotic genes alter the establishment of positional clues, not only their interpretation. The partial conservation of pattern-forming genes in both appendages ensures a predictable re-specification of positional clues, producing the observed positional specificity of homeotic transformations.