A phylogeny of the Platyhelminthes: towards a total-evidence solution

We advocate a total-evidence approach for the reconstruction of working phylogenies for the Turbellaria and the phylum Platyhelminthes. Few morphology-based character matrices are available in the systematic literature concerning flatworms, and molecular-based phylogenies are rapidly providing the only means by which we can estimate phylogenies cladistically. Character matrices based on gross morphology and ultrastructure are required and should be internally consistent, i.e. character coding should follow a set of a priori guidelines and character duplication and contradiction is avoided. In order to test our molecular phylogenies we need complementary data sets from morphology. To understand morphological homology we need phylogenetic evidence from independent (e.g. molecular) data. Fully complementary morphological and molecular data sets enable us to validate phylogenetic hypotheses and the combination of these sets in phylogenetic reconstruction utilises all statements of homology. Working phylogenies which include all phylogenetic information not only shed light on individual character evolution, but form a strong basis for comparative studies investigating the origin and evolutionary radiation of the taxonomic group under scrutiny. This revised version was published online in July 2006 with corrections to the Cover Date.     

Aspects of the phylogeny of Platyhelminthes based on 18S ribosomal DNA and protonephridial ultrastructure

DNA studies of 23 taxa (20 platyhelminths, 1 nemertean, Homo and Artemia) and electron-microscopic studies of the protonephridia of many platyhelminths (supported by some additional ultrastructural data) have led to the following conclusions: the Neodermata are monophyletic; Temnocephalida and Dalyelliida form one clade and are not the primitive sister group of the Neodermata; Gyrocotylidea, Amphilinidea and Eucestoda form one monophylum; Pterastericolidae and Umagillidae are dalyelliids and not the sister group of the Neodermata; and Proseriata are unlikely to be closely related with the Tricladida. A large taxon consisting of the Proseriata and some other turbellarians may represent the sister group of the Neodermata.     

Phylogeny and a revised classification of the Monogenoidea Bychowsky, 1937 (Platyhelminthes)

A hypothesis (CI=57.3%) on the evolutionary relationships of families comprising the class Monogenoidea is proposed based on 141 character states in 47 homologous series and employing phylogenetic systematics. Based on the analysis, three subclasses, the Polyonchoinea, Polystomatoinea and Oligonchoinea, are recognised. The analysis supports independent origins of the Montchadskyellidae within the Polyonchoinea and of the Neodactylodiscidae and Amphibdellatidae within the order Dactylogyridea (Polyonchoinea); the suborder Montchadskyellinea is raised to ordinal status and new suborders Neodactylodiscinea and Amphibdellatinea are proposed to reflect these origins. The Gyrodactylidea (Polyonchoinea) is supported by three synapomorphies and comprises the Gyrodactylidae, Anoplodiscidae, Tetraonchoididae and Bothitrematidae. The analysis supports recognition of the Polystomatoinea comprising Polystomatidae and Sphyranuridae. Evolutionary relationships within the Oligonchoinea indicate independent origins of three ordinal taxa, the Chimaericolidea (monotypic), Diclybothriidea (including Diclybothriidae and Hexabothriidae) and Mazocraeidea (with five suborders). The suborder Mazocraeinea comprises the Plectanocotylidae, Mazocraeidae and Mazoplectidae, and is characterised by two synapomorphies. The suborder Gastrocotylinea, characterised by presence of accessory sclerites in the haptoral sucker, is divided into two infraorders, the monotypic Anthocotylina infraorder novum and Gastrocotylina. Two superfamilies of the Gastrocotylina are recognised, the Protomicrocotyloidea and Gastrocotyloidea; the Pseudodiclidophoridae is considered incertae sedis within the Gastrocotylina. The suborder Discocotylinea comprises the Discocotylidae, Octomacridae and Diplozoidae and is supported by four synapomorphies. The monotypic Hexostomatinea suborder novum is proposed to reflect an independent origin of the Hexostomatidae within the Mazocraeidea. The terminal suborder Microcotylinea comprises four superfamilies, the Microcotyloidea, Allopyragraphoroidea, Diclidophoroidea and Pyragraphoroidea. The analysis supports incorporation of the Pterinotrematidae in the Pyragraphoroidea and rejection of the monotypic order Pterinotrematidea. The following taxa are also rejected for reasons of paraphyly and/or polyphyly: Articulonchoinea, Bothriocotylea, Eucotylea, Monoaxonematidea, Tetraonchidea, Gotocotyloidea, Anchorophoridae and Macrovalvitrematidae. The Sundanonchidae, Iagotrematidae and Microbothriidae were not included in the analysis because of lack of pertinent information regarding character states.     

Unciliated body surface in three species of the Umagillidae (Dalyellioida,Platyhelminthes)

Anoplodiera voluta Westblad, Seritia elegans (Westblad) and Wahlia macrostylifera Westblad are species of the family Umagillidae living in the intestine of the holothurian Parastichopus tremulus. In all three species, part of the epidermis is unciliated, but unlike unciliated epidermis in the major parasitic flat-worm groups, it is cellular and has intraepithelial nuclei. The surface of the unciliated cells in A. voluta is convex with the cells separated by lateral gaps. The cells have two distinct regions: the basal, organelle-rich part and the apical part which contains few identifiable organelles except vesicles. In W. macrostylifera the unciliated cells have a flat surface and, between them, narrower gaps that in some cases widen to paracellular compartments below the cell surface. Apically the cells contain electron-dense vesicles, often in contact with the surface. S. elegans has unciliated cells separated by gaps. In all these species, apical vesicles indicate secretory activity. Comparison of the epidermis of these three species with the neodermis of the major parasitic flatworm groups within the Neodermata does not support a close relationship of the three to the Neodermata.     

5S rRNA sequences of 12 species of flatworms: implications for the phylogeny of the Platyhelminthes

5S rRNAs from 12 species of free living and parasitic platyhelminthes were sequenced. In the phylogenetic analysis, attention was focused on the statistical estimates of the trees corresponding to existing phylogenetic hypotheses. The available 5S rRNA data agree well with widely accepted views on the relationships between the Acoela, Polycladida, Tricladida, and Neorhabdocoela; our analysis of the published 18S rRNA sequences also demonstrated good correspondence between these views and molecular data. With available 5S rRNA data the hypothesis that the dalyellioid turbellarians is the sister group of the Neodermata is less convincing than the hypotheses proposing the Neodermata as the sister group of the Neorhabdocoela, or of the Seriata, or of the branch uniting them. A relatively low rate of base replacement in parasitic flatworms, probably, accounts for the uncertain position of the Neodermata, while a relatively high rate in planarians may explain a relatively too early divergence of the Tricladida in several published phylogenetic trees constructed from various rRNA data.     

A new species of freshwater triclad (Platyhelminthes: Tricladida) from Australia

A new species of freshwater triclad, Romankenkius libidinosus sp. nov. , is described from New South Wales, Australia. The species is characterized by a large adenodactyl and unique genito-intestinal connections, and represents the first record of the genus Romankenkius outside Tasmania. The taxonomic and phylogenetic implications of several morphological features of the new species are discussed in the light of a previous phylogenetic study.     

Molecular phylogeny of parasitic Platyhelminthes based on sequences of partial 28S rDNA D1 and mitochondrial cytochrome c oxidase subunit I

The phylogenic relationships existing among 14 parasitic Platyhelminthes in the Republic of Korea were investigated via the use of the partial 28S ribosomal DNA (rDNA) D1 region and the partial mitochondrial cytochrome c oxidase subunit 1 (mCOI) DNA sequences. The nucleotide sequences were analyzed by length, G + C %, nucleotide differences and gaps in order to determine the analyzed phylogenic relationships. The phylogenic patterns of the 28S rDNA D1 and mCOI regions were closely related within the same class and order as analyzed by the PAUP 4.0 program, with the exception of a few species. These findings indicate that the 28S rDNA gene sequence is more highly conserved than are the mCOI gene sequences. The 28S rDNA gene may prove useful in studies of the systematics and population genetic structures of parasitic Platyhelminthes.     

Towards a phylogenetic classification of dendrocoelid freshwater planarians (Platyhelminthes): a morphological and eclectic approach

We explore and review the taxonomic distribution of morphological features that may be used as supporting apomorphies for the monophyletic status of various taxa in future, more comprehensive phylogenetic analyses of the dendrocoelid freshwater planarians and their close relatives. Characters examined are: (i) musculature of the pharynx; (ii) anatomy of the eye; (iii) distribution of testis follicles along the longitudinal axis of the body; (iv) epidermis of the body wall; (v) musculature of the ventral body wall; (vi) presence or absence of a penis papilla; (vii) position of the mouth opening; and (viii) anterior adhesive organ. A phylogenetic hypothesis, in the form of a character state tree, is developed. This hypothesis suggests that the Kenkiidae should be resurrected as a separate family, sharing a sistergroup relationship with the Dendrocoelidae. This sistergroup relationship is supported by the shared presence of an apomorphic anterior adhesive organ. Although the shape of the anterior adhesive organ, both within and between kenkiids and dendrocoelids, may differ considerably between species, they are all constructed according to the same principles. The monophyly of the Kenkiidae is supported by the presence of an apomorphic thickening of the marginal zone of the epidermis and presumably also by the presence of prepharyngeal testes. Within the Kenkiidae, the genus Macrocotyla should be considered as a junior synonym of Kenkia, which is characterized by the presence of an apomorphic ‘planariid pharynx‐type 2’. The monophyly of the Dendrocoelidae is suggested by the presence of a dendrocoelid type of pharynx. It is shown that multicellular eye cups with numerous photoreceptor cells are not restricted to the Dugesiidae and the Terricola but are also characteristic for the Dendrocoelidae; the phylogenetic implications of this character state distribution are discussed. It is pointed out that in planarians there may be a relationship between: (i) body size, the size of an eye cup and the number of its constituent pigment cells; and (ii) the size of an eye cup and the number of photoreceptor cells that it contains. Some tentative phylogenetic groups are suggested on the basis of the following presumed apomorphic characters: presence of transverse fibres in the ventral subepidermal musculature, presence of an extra longitudinal muscle layer in the outer pharynx musculature, a mouth opening that has shifted anteriad. It is suggested that the presumed monophyly of the genus Bdellocephala is supported by the apomorphic absence of a penis papilla, combined with a penial lumen provided with many plicae.     

The interrelationships of proseriata (Platyhelminthes: seriata) tested with molecules and morphology

Proseriate flatworms are common members of the interstitial benthic fauna worldwide, predominantly occupying marine environments. As minute animals, having relatively few characters useful for cladistic analysis, they have been difficult to present in a phylogenetic framework using morphology alone. Here we present a new morphological matrix consisting of 16 putatively homologous characters and two molecular data sets to investigate further this major group of free-living members of the Platyhelminthes. Complete 18S rDNA (representing 277 parsimony-informative characters) from 17 ingroup taxa and partial 28S rDNA spanning variable expansion regions D1 to D3 and D1 to D6 (representing 219 and 361 parsimony-informative characters, respectively) from 27 and 14 ingroup taxa, respectively, were determined and aligned as complementary data sets. Morphological and molecular data sets were analyzed separately and together to determine underlying phylogenetic patterns and to resolve conflict between published scenarios based on morphology alone. The monophyly of the Proseriata cannot be confirmed categorically with any of these data sets. However, the constituent taxa are confirmed as basal members of the Neoophora, and a sister group relationship with Tricladida is rejected. Similarly, the monophyly of one of the two subtaxa of the Proseriata, the Lithophora, could not be confirmed with molecules. Concerning intragroup relationships, we could reject one of the two phylogenetic trees formerly proposed, as well as the clade Otoplanidae + Coelogynoporidae. However, a clade Otoplanidae + Archimonocelididae + Monocelididae (to which the Monotoplanidae belong) was supported, and the position of the genus Calviria shifted from the Archimonocelididae to the Coelogynoporidae.     

A molecular framework for the phylogeny of the Pseudocerotidae (Platyhelminthes,Polycladida)

Systematic relationships within the cotylean family Pseudocerotidae were examined using nucleotide sequences of the D3 expansion segment of the 28S rDNA gene. A previously suggested separation of Pseudoceros and Pseudobiceros based on the number of male reproductive systems was confirmed. Regardless of the algorithm employed, Pseudoceros always formed a monophyletic clade. Pseudobiceros appeared to be paraphyletic; however, a constrained maximum parsimony tree was not significantly longer (2 steps, = 0.05). Additionally, the genera Maiazoon, Phrikoceros and Tytthosoceros were validated as taxonomic entities, and their relationships to other genera within the family were determined. Molecular data also supported species separations based on colour patterns. An intraspecific genetic distance of 1.14% was found for Pseudoceros bifurcus, whereas the intrageneric distance was 3.58%. Genetic distances among genera varied, with the closest distance being 2.048% between Pseudobiceros and Maiazoon, and the largest distance (8.345%) between Pseudoceros and Tytthosoceros.     

First report of gastrocotylinean post-oncomiracidia (Platyhelminthes: Monogenoidea: Heteronchoinea) on gills of flyingfish (Exocoetidae), snapper (Lutjanidae), dolphinfish (Coryphaenidae), and amberjack (Carangidae) from the Gulf of Mexico: decoy hosts and the dilution effect

Larvae, identified as post-oncomiracidia of the suborder Gastrocotylinea (Monogenoidea), were collected from formalin-fixed gills excised from six species of marine fishes captured from the Gulf of Mexico off Mississippi and Florida: common dolphinfish, Coryphaena hippurus and pompano dolphinfish, Coryphaena equiselis (both Perciformes, Coryphaenidae); gray snapper, Lutjanus griseus (Perciformes, Lutjanidae); greater amberjack, Seriola dumerili (Perciformes, Carangidae); and Atlantic flyingfish, Cheilopogon melanurus and sailfin flyingfish, Parexocoetus hillianus (both Beloniformes and Exocoetidae). Based on a combination of diagnostic morphological features, the specimens were divided into two basic forms, each of which was further subdivided into two morphotypes. No gastrocotylinean post-oncomiracidium had been reported previously from these hosts. Of the six host species, only C. hippurus serves as a host (unconfirmed) for the adult of a gastrocotylinean species, suggesting that the recorded fishes from the Gulf of Mexico comprise dead-end hosts acting as decoys for the oncomiracidia. These comparatively non-susceptible “decoy hosts” apparently dilute the susceptible fish-host population and by intercepting infective larvae (oncomiracidia) decrease the abundance of parasites on their typical hosts.     

A new species of giant planarian from Lake Baikal,with some remarks on character states in the Dendrocoelidae (Platyhelminthes,Tricladida, Paludicola)

On the basis of newly collected material the subspecies Bdellocephala angarensis bathyalis Timoshkin & Porfirjeva, 1989 is raised to full species status, B. bathyalis Timoshkin & Porfirjeva, 1989. Specimens of this species have been collected in Lake Baikal from depths ranging between 610 and 1060 m. The species is characterized by a light, uniform brown pigmentation, absence of eyes, distinct atrial folds, and large size. It is suggested that two features may be useful in elucidating the phylogenetic relationships between dendrocoelid genera: pharynx musculature, and presence of an extra layer of circular muscle in the ventral subepidermal body musculature. This revised version was published online in July 2006 with corrections to the Cover Date.     

The phylogeny and systematics of European predacious Syrphidae (Diptera) based on larval and puparial stages

The external larval morphology of predacious Syrphidae is examined and compared with that of other groups of syrphids. Predacious larvae form a distinctive, monophyletic group. Relationships among 29 of the 34 European genera and subgenera are analysed using cladistic methods based on larval and puparial characters. Interpretation of the cladogram suggests that, within Syrphinae, a system of five tribes can be recognized and that a major trend has been a shift from ground layer to arboreal larval habitats, with an increasing degree of obligate dependence on soft-bodied Homoptera as prey. Generic diagnoses with biological and taxonomic notes are given and a key to genera based on larvae and puparia is provided.     

Ultrastructure of the female gonad of two temnocephalids (Platyhelminthes,Rhabdocoela)

The female gonad of Temnocephala dendyi and T. minor consists of a single germarium and two rows of vitellaria. It is enveloped by an extracellular lamina and accessory cells. Accessory cells are only peripherally located in the germarium while their cytoplasmic projections also fill the spaces between vitellocytes in the vitellarium. The main feature of oocyte maturation is the appearance of chromatoid bodies and the development of rough endoplasmic reticulum (R.E.R.) and Golgi complexes which appear to be correlated with the production of double-structured egg granules. The egg granules, which are localized in the cortical cytoplasm of mature oocytes, contain glycoproteins, are devoid of polyphenols and are similar in structure and composition to the cortical granules observed in some Digenea and Monogenea. Vitellocytes are typical secretory cells with well-developed R.E.R. and Golgi complexes which are involved in the production of shell globules and yolk. The multigranular pattern and the polyphenolic composition of the shell globules of the temnocephalids investigated are similar to those observed in other rhabdocoels, and in some Prolecithophora and Neodermata. This feature may represent a synapomorphy shared by these taxa. This revised version was published online in July 2006 with corrections to the Cover Date.     

Molecular phylogeny of the Pooideae (Poaceae) based on nuclear rDNA (ITS) sequences

Phylogenetic relationships of the Poaceae subfamily, Pooideae, were estimated from the sequences of the internal transcribed spacer (ITS) region of nuclear ribosomal DNA. The entire ITS region of 25 species belonging to 19 genera representing seven tribes was directly sequenced from polymerase chain reaction (PCR)-amplified DNA fragments. The published sequence of rice, Oryza saliva, was used as the outgroup. Sequences of these taxa were analyzed with maximum parsimony (PAUP) and the neighbor-joining distance method (NJ). Among the tribes, the Stipeae, Meliceae and Brachypodieae, all with small chromosomes and a basic number more than x=7, diverged in succession. The Poeae, Aveneae, Bromeae and Triticeae, with large chromosomes and a basic number of x=7, form a monophyletic clade. The Poeae and Aveneae are the sister group of the Bromeae and Triticeae. On the ITS tree, the Brachypodieae is distantly related to the Triticeae and Bromeae, which differs from the phylogenies based on restriction-site variation of cpDNA and morphological characters. The phylogenetic relationships of the seven pooid tribes inferred from the ITS sequences are highly concordant with the cytogenetic evidence that the reduction in chromosome number and the increase in chromosome size evolved only once in the pooids and pre-dated the divergence of the Poeae, Aveneae, Bromeae and Triticeae.This paper reports factually on available data; however, the USDA neither guarantees nor warrants the standard of the product, and the use of the name by USDA implies no approval of the product to the exclusion of others that may also be suitableThis paper is a cooperative investigation of USDA-ARS and the Utah Agricultural Experiment Station. Logan, Utah 84322. Journal Paper No. 4581     

Comments on a phylogenetic system of the Platyhelminthes

Using the principles and methods of phylogenetic systematics, the phylogenetic relationships among the basic closed descendent communities (monophyla) of the taxon Platyhelminthes are discussed.A phylogenetic system of the Platyhelminthes is presented and several hypothesized autapomorphies of the well-known main subordinated monophyla of free-living and parasitic platyhelminths are named. A monophylum Turbellaria does not exist in reality.     

The aquatic triclads of the Grozet Islands (Platyhelminthes: Tricladida)

A new species of marine planarian, Obrimoposthia aparala sp. nov. , is described from the Crozet Islands and two other species are newly recorded for these islands, viz. 0. wandeli and Synsiphonium ernesti. The only known freshwater triclad from the Crozets, Dugesia seclusa , is redescribed from the type material. The morphology, ecology, and distribution of these four species is discussed.     

Evolution of metatherian and eutherian (mammalian) characters: a review based on cladistic methodology

Cladistic methodology is used to test the hypothesis that three major monophyletic groups exist among living mammals–the oviparous monotremes (Prototheria), and the viviparous marsupials (Metatheria) and placentals (Eutheria). Evaluation is made of the polarity (i.e. the direction of change in a primitive-to-derived sequence) of numerous characters which distinguish some or all of these groups, and of the usefulness of these characters in phylogenetic inference. An attempt is made to establish the state of these characters in the common Late Jurassic-Early Cretaceous therian ancestor of marsupials and placentals.
It is concluded that the most basic division of the Mammalia is the dichotomy into the subclasses Prototheria (including Monotremata, Multituberculata, Triconodonta, Docodonta) and Theria (including Metatheria, Eutheria, Pantotheria and Symmetrodonta). Two major groups exist among living viviparous mammals, the Metatheria and Eutheria; in a cladistic framework these are sister-groups. It is demonstrated that there is no special (sister-group) relationship between monotremes and marsupials, and there is no justification for placing them in a group Marsupionta.     

Mutation scanning methods for the analysis of parasite genes

Molecular variation is widespread in parasite populations, and its analysis has important implications for studying aspects relating to the function and organisation of genes, and the taxonomy, phylogeny and population genetics of parasites. This article reviews some PCR-based mutation scanning techniques that have advantages over currently used DNA methods for the analysis of genetic variation in parasites. The review is technical and describes briefly the principles of relevant techniques, examines some of their advantages and disadvantages and gives several examples for possible applications.     

Ultrastructural study on the spermiogenesis and spermatozoon of the metacercariae of Microphallus primas (Digenea), a parasite of Carcinus maenas

The thread-like spermatozoon of the crab parasite Microphallus primas was studied by electron microscopy. A survey of the head region of the spermatozoon reveals three features hitherto unknown in Platyhelminthes spermatozoa. The first is the aberrant inclusion of the nucleus within one of the two axonemes, limited to the head end region. The second is the coexistence, in the same axoneme, of two patterns, 9 + 0 (doublets without dynein arms) and 9 + 1. The third is the presence of a layer of cortical microtubules running longitudinally from the zone where the nucleus goes from axoneme to the tail region (where the two flagella start). The sequence of events in spermatogenesis is similar to that described for most of the Digenea trematodes, and the spermiogenesis process conforms to a common plan in nearly the whole group.