Molecular phylogeny of North American Branchiobdellida (Annelida: Clitellata)

Branchiobdellidans, or crayfish worms, are ectosymbiotic clitellate annelids associated primarily with freshwater crayfishes. The main objectives of our study were to infer a molecular phylogeny for the North American Branchiobdellida, examine its congruence with morphology-based hypotheses of relationships at the subfamily and genus level, and use our dataset to assess consistency of GenBank-archived branchiobdellidan sequences. We used nucleotide sequence data from two mtDNA genes (COI and 16S rDNA) and three nuclear genes (28S rDNA, 18S rDNA, and ITS1) to estimate phylogenetic relationships among 47 described and one undescribed species of Branchiobdellida. We recovered a monophyletic branchiobdellidan clade with generally short branch lengths, suggesting that a large portion of the taxon has likely undergone a recent and rapid radiation in North America. Results from our phylogenetic analyses indicate that current taxonomic groupings are largely unsupported by the molecular data. All four subfamilies are either paraphyletic or polyphyletic, and only three of seven sampled non-monotypic genera were monophyletic. We found a high rate (49%) of inconsistency in GenBank-archived sequences, over 70% of which can be attributed to field- or laboratory-based error.     

Molecular phylogeny of lugworms (Annelida, Arenicolidae) inferred from three genes

Arenicolids comprise a group of four genera in which about 30 nominal species are described. Whereas the biology of many arenicolids is well known, the phylogenetic relationships of these worms are inadequately studied. A close relationship of Arenicolidae and Maldanidae is generally accepted. The phylogenetic relationships of arenicolid taxa were reconstructed based on sequence data of the mitochondrial 16S rRNA gene, the nuclear 18S rRNA gene, and a small fraction of the nuclear 28S rRNA gene. Members of all described arenicolid genera are included in the data set. Phylogenetic analyses were conducted using Maximum Likelihood, Bayesian inference, and Maximum Parsimony. The monophyly of the Maldanidae, as well as of the Arenicolidae is supported by all conducted analyses. Two well supported major clades are highest ranked sister taxa in the Arenicolidae: one containing all Abarenicola species and one containing Arenicola, Arenicolides, and Branchiomaldane. Evidence is given for a closer relationship between the two investigated Branchiomaldane species and Arenicolides ecaudata in the combined analysis. In the light of the molecular data the best explanation for structural and morphological observations is that Branchiomaldane evolved by progenesis.     

Molecular phylogeny of naidid worms (Annelida: Clitellata) based on cytochrome oxidase I

Naidids are tiny, primarily freshwater oligochaete annelids which reproduce asexually by fission. We investigated the phylogenetic relationships within this group by sequencing 1224 bp of the mitochondrial gene cytochrome oxidase I (COI) from 26 species of naidids (representing 13 of the 23 genera currently recognized), as well as from four tubificids, their closest allies. Although not completely concordant, maximum parsimony and Bayesian inference analyses agreed in several important respects, with no well-supported conflicts. Our study, the first detailed molecular investigation of naidid relationships, suggests that naidids fall into two groups, one comprised of the genus Pristina, and another comprised of all other genera sampled. The clear division of naidids into these two groups best matches an early, simple classification of the group by Lastockin (1924); the more recent classifications proposed by Sperber (1948) and Nemec and Brinkhurst (1987) are not as consistent with our results. We note that our study suggests the genus Stylaria is comprised of two distinct species, Stylaria lacustris and Stylaria fossularis, rather than merely two morphotypes of a single species. Based on our phylogenetic results, we suggest that pigmented eyes evolved only once among naidids but must have been lost multiple times, and that the elongation of the prostomium into a proboscis evolved at least twice independently. The simplest form of fission, architomy (fragmentation), occurs in two of the most basally branching naidid genera, and may represent the plesiomorphic condition for naidids.     

Molecular systematics of polychaetes (Annelida)

Hydrobiologia - Some progress has been made in the field of molecular systematics of polychaetes over the past couple of years. In particular, phylogenetic analyses of sequence data from the 18S...     

The Nearctic species of Orthoplema (Hymenoptera: Ichneumonidae)

Abstract. The species of the genus Orthopelma Taschenburg in North America are reviewed. Members are endoparasites of the cynipids Diastrophus on Rubus and Diplolepis on Rosa. The Nearctic species ar occidentale, califomicum and erythropa , described by Ashmead, ovale Provancher, and the Holarctic species mediator Thunberg. The names coloradense Ashmead and diastrophi Ashmead are newly placed in synonymy with occidentale Ashmead. Relationships of the species are discussed.     

Systematic revision of Nearctic species of Achalcus Loew (Diptera: Dolichopodidae) with comments on their phylogeny, ecology and zoogeography

The species of the genus Achalcus Loew from the Nearctic Region are revised and a key to males and females is provided. Achalcus utahensis (Harmston & Miller) and A. oregonensis (Harmston & Miller) are redescribed, and A. californicus sp.n., A. dytei sp.n. and A. similis sp.n. are described. Together with eight of the nine known Palaearctic species of the genus, they represent a monophyletic species group, which is characterized by the presence of five pairs of dorsocentral bristles and the incorporation of the epandrial setae onto the shaft of the epandrial lobe. Like their Palaearctic congeners, most Nearctic species occur in damp habitats, such as coastal forests and riparian vegetation. Most specimens were collected in the Pacific coastal states from British Columbia to California, but one species occurs in Utah, and one of six species which were not described because males are lacking, was discovered in Ontario.     

Molecular phylogeny of Caloboletus (Boletaceae) and a new species in East Asia

Caloboletus has recently been erected to accommodate Boletus calopus and its allies. Our phylogenetic analyses based on four gene markers (ITS, nrLSU, tef1-α and rpb1) recognized six distinct species within the genus. Morphologically, the genus is characterized by the combination of the yellow hymenophore, smooth spores that are olive-brown in deposit, an interwoven pileipellis, a bluish color-change and a distinctly bitter taste of the context. A new species was described and a key to the six species of Caloboletus is provided. Members of Caloboletus are mainly distributed in subtropical and temperate zones in the Holarctic and are often associated with plants of Pinaceae and Fagaceae.     

Morphological and molecular evidence of the phylogeny of Nereidiform polychaetes (Annelida)

The phylogeny of Nereidiformia is assessed in a parsimony analysis of combined morphological and DNA data, with special focus on previously questioned relationships between Chrysopetalidae and Hesionidae, between Pilargidae and Hesionidae, and the affinities of Hesionides and Microphthalmus. A 660 bp segment of the mtDNA cytochrome c oxidase subunit I gene was sequenced for two chrysopetalids, one nereidid, one pilargid, one pisionid, two hesionids, plus the two questionable hesionids Hesionides arenaria and Microphthalmus sp. Phylogenetic resolution was poor for the cytochrome c oxidase subunit I gene data alone, but the combined analysis yielded partially robust topologies, suggesting that nereids are the sister group to chrysopetalids, and that pilargids, Hesionides and Microphthalmus do not belong within the hesionids     

Molecular phylogeny and adaptive radiation of the endemic Hawaiian Plantago species (Plantaginaceae)

Insular oceanic islands provide excellent opportunities for the study of evolutionary processes and adaptive radiation. The Hawaiian Plantago radiation comprises six endemic taxa showing considerable inter- and intraspecific morphological and ecological diversity. The rDNA internal (ITS) and external (ETS) transcribed spacers and two recently described chloroplast spacers, ndhF-rpl32 and rpl32-trnL, were sequenced to study phylogenetic relationships within this morphologically complex group. Phylogenetic analysis provided strong evidence for the monophyly of Hawaiian Plantago, suggesting that the lineage arose from a single long-distance dispersal event. Inconsistencies between nuclear and chloroplast phylogenies suggest a history of hybridization. The basal, unresolved dichotomy of the combined phylogeny is consistent with rapid phenotypic diversification of the major lineages early in the history of this group. Speciation has largely occurred allopatrically, with divergence a result of intraisland ecological shifts between bog and woodland habitats and interisland dispersal events. Most interisland colonizations were from older to younger islands with initial colonization of Kaua'i. In our analysis, P. pachyphylla is paraphyletic and taxonomic separation of the distinct morphotypes of this species appears justified. Furthermore, the apparent hybrid ancestry and unique morphology and habitat of the endangered P. princeps var. longibracteata support its recognition at the specific rank.     

Sperm ultrastructure of three Syllinae (Annelida, Phyllodocida) species with considerations on syllid phylogeny and Syllis vittata reproductive biology

Phylogeny of Syllidae is under debate due to new studies based on molecular and morphological data. The noticeable taxonomic diversity of syllids (about 700 listed species) is also mirrored in the array of reproductive strategies as well as in sperm morphology, counting a display of forms already supposed to reflect phylogenetic relationships between the species. The sperm ultrastructure of Syllis gerlachi, S. prolifera and S. vittata is herein presented and compared to the Syllinae species studied previously. Moreover, the egg structure and the gamete allocation within stolons of S. vittata are particularly investigated. Both male germinal cells at different level of maturation and oocytes were found in the same individual of S. vittata, suggesting simultaneous hermaphroditism. The ultrastructural analysis revealed that the observed spermatozoa belong to the ect-aquasperm type resembling those of the similar studied species (Syllis sp., S. pigmentata and S. krohni). Differences in the acrosome structure and nucleus shape are in accordance with a recent phylogenetic reconstruction and suggest a trend in the evolution of spermatozoa in Syllinae toward the development of the apical part. However, further molecular and ultrastructural analyses are needed to support this hypothesis. This is the first record of simultaneous hermaphroditism within Syllinae.     

Molecular phylogeny and distribution of three Apodemus species (Muridae, Rodentia) in Serbia

Phylogenetic relationships among three species of the genus Apodemus ( A. flavicollis, A. sylvaticus and A. agrarius ) from seven different localities in Serbia were estimated using arbitrarily primed PCR (AP-PCR) profiling, which allowed clear distinction between the sibling species A. flavicollis and A. sylvaticus . Eight oligonucleotide primers out of the 22 screened, gained a total of 397 amplified bands. The average pairwise genetic distances within Apodemus species ranged from 0.51 between A. flavicollis and A. sylvaticus , to 0.74 between A. sylvaticus and A. agrarius. The large genetic distances obtained among populations of A. flavicollis (0.13) indicate that this part of the Balkan Peninsula was probably a refuge during the last glaciation. Furthermore, differences obtained between populations south and north from the river Danube suggest that this species survived the Last Glacial Maximum in at least two separate refuges. Although our data for molecular markers are from a limited number of localities, extended analyses of C-band distribution on chromosomes from previously studied localities south of the rivers Danube and Sava, indicate that distribution of A. sylvaticus is limited to northern Serbia. Therefore, it is possible that this part of the Balkan Peninsula was not a refuge for this species during the last glaciation.     

Molecular phylogeny and morphology reveal a new species of Amyloporia (Basidiomycota) from China

Amyloporia pinea sp. nov. is described and illustrated on the basis of collections from southern China. Morphology and phylogenetic analysis of rDNA ITS sequences support this new species. Morphologically, it is characterized by resupinate, annual basidiocarps, cream to yellowish buff pore surface when fresh, which becomes yellowish brown to clay-buff upon drying, a dimitic hyphal system with clamped generative hyphae and inamyloid skeletal hyphae, fusoid cystidioles, and cylindrical basidiospores; moreover, it causes a brown rot. Molecular phylogeny inferred from ITS sequence data suggested a close relationship between A. pinea and Amyloporia crassa sensu lato. Antrodia subxantha has amyloid skeletal hyphae, and grouped within the Amyloporia clade, hence, it is transferred to Amyloporia, and a new combination Amyloporia subxantha is proposed.     

Molecular phylogeny of Siberian glossiphoniidae (Hirudinea)

A molecular phylogenetic analysis based on gene fragments of the first subunit of the mitochondrial cytochrome-c-oxidase (COI) from Siberian glossiphoniids and phylogenetically relative Palearctic species demonstrated that Siberian representatives do not belong to the species Glossiphonia complanata, as was believed previously, and have an autonomous taxonomic status. Furthermore, the detected genetic distances make it possible to accept the mtCOI fragment as a suitable marker for the molecular identification of the species within the Glossiphoniidae (Hirudinea).     

Molecular phylogeny of chloroplast DNA of Nicotiana species

Amplified fragment length polymorphism (AFLP) analysis of chloroplast DNA was used to study the relationships within the genus Nicotiana. Resulting phylogenetic tree, reconstructed using the UPGMA method, generally agreed with the existing taxonomic classification based on morphological and cytogenetic data, as well as sequence comparison of the internal transcribed spacer of the nuclear ribosomal DNA.     

Molecular phylogeny and ultrastructure of Selenidium serpulae (Apicomplexa, Archigregarinia) from the calcareous tubeworm Serpula vermicularis (Annelida, Polychaeta, Sabellida)

Archigregarines are dynamic single‐celled parasites that inhabit the intestinal systems of marine invertebrates, especially suspension feeding and deposit feeding polychaetes. Certain archigregarines in the genus Selenidium have retained several plesiomorphic characters, and improved knowledge of these species is expected to shed considerable light onto the earliest stages in apicomplexan evolution. Although archigregarines are related to some of the most notorious parasites known (e.g., Cryptosporidium and Plasmodium), current knowledge of the group is meagre. In an attempt to improve our understanding of archigregarine diversity and evolution, I have characterised the general ultrastructure and molecular phylogenetic position of Selenidium serpulae (Lankester) Caullery and Mesnil. The parasites were isolated from the intestines of the calcareous tubeworm Serpula vermicularis (Polychaeta) collected in the eastern Pacific Ocean. The trophozoites (extracellular feeding stages) were spindle‐shaped and capable of slow and continuous bending, and coiling, especially when dislodged from the host epithelium. The trophozoite surface was composed of 19–23 longitudinal folds, prominent transverse folds and a robust, trilayered pellicle subtended by a single row of microtubules, each surrounded by an electron transparent sheath. Putative mitochondria were observed, but they were inconspicuous and apparently highly reduced, a condition that is indicative of anaerobic metabolism. The small subunit (SSU) rDNA sequence from S. serpulae was very closely related to two (short) sequences derived from an environmental PCR survey of an oxygen‐depleted hydrothermal vent system in the Gulf of California, namely C1‐E017 (AY046619) and C2‐E016 (AY046806). This result suggested that archigregarines with a morphology and lifecycle much like that in S. serpulae are thriving in this oxygen poor ecosystem. In addition, phylogenetic analyses of a larger SSU rDNA dataset (excluding the shorter environmental sequences) indicated that the nearest sister lineage to S. serpulae was S. vivax, a bizarre tape‐like gregarine found in the intestines of sipunculids. This relationship was bolstered by comparative ultrastructural data and helped to illustrate that the diversification and biogeographical distribution of archigregarine parasites is probably more extensive than usually assumed.     

Molecular phylogeny of Gossypium species by DNA fingerprinting

Total genomic DNA from 31 available Gossypium species, three subspecies and one interspecific hybrid, were analysed to evaluate genetic diversity by RAPD, using 45 random decamer primers. A total of 579 amplified bands were observed, with 12.9 bands per primer, of which 99.8% were polymorphic. OPJ-17 produced the maximum number of fragments while the minimum number of fragments was produced with primer OPA-08. Cluster analysis by the unweighted paired group method of arithmetic means (UPGMA) showed six main clusters. Cluster ’A’ consisted of two species and one subspecies of the A-genome, with a 0.78–0.92 Nei’s similarity range. Cluster B, composed of all available tetraploid species and one interspecific hybrid, showed the same sister cluster. Nei’s similarity ranged from 0.69 to 0.84. The B-genome formed the UPGMA sister cluster to the E-genome species. Cluster ’C’ consisted of five Gossypium species of which three belong to the B-genome, with Nei’s similarity values of 0.81 to 0.86. Although there was considerable disagreement at lower infra-generic ranks, particularly among the D- genome (diploid New World species) and C-genome (diploid Australian species) species. The sole F-genome species Gossypium longicalyx was resolved as a sister group to the D-genome species. Gossypium herbaceum and G. herbaceum Africanum showed the maximum Nei’s similarity (0.93). Minimum similarity (0.29) was observed between Gossypium trilobum and Gossypium nelsonii. The average similarity among all studied species was 50%. The analysis revealed that the interspecific genetic relationship of several species is related to their centre of origin. As expected, most of the species have a wide genetic base range. The results also revealed the genetic relationships of the species Gossypium hirsutum to standard cultivated Gossypium barbadense, G. herbaceum and Gossypium arboreum. These results correspond well with previous reported results. The level of variation detected in closely related genotypes by RAPD analysis indicates that it may be a more efficient marker than morphological marker, isozyme and RFLP technology for the construction of genetic linkage maps. Received: 2 January 2000 / Accepted: 12 February 2000     

New insights into polychaete phylogeny (Annelida) inferred from 18S rDNA sequences

Annelid systematics and the ingroup relationships of polychaete annelids are matter of ongoing debates in recent analyses. For the investigation of sedentary polychaete relationships a molecular phylogenetic analysis was conducted based on 94 sequences of 18S rDNA, including unpublished sequences of 13 polychaete species. The data set was analyzed with maximum parsimony and maximum likelihood methods, as wells as Bayesian inference. As in previous molecular analyses the monophyly of many traditional polychaete families is confirmed. No evidence has been found for a possible monophyly of Canalipalpata or Scolecida. In all analyses a placement of the Echiura as a derived polychaete ingroup with a close relationship to the Capitellidae is confirmed. The orbiniids appear paraphyletic with regard to Questa. Travisia is transferred from Opheliidae to Scalibregmatidae. The remaining opheliids include a yet undescribed ctenodrilid species from Elba, whereas the other investigated ctenodrilid Ctenodrilus serratus groups with the Cirratulidae and shows a close affinity to the cirratulid genus Dodecaceria. A common ancestry of Branchiomaldane and Arenicola, which has been predicted on morphological data, is confirmed by the analysis and a sistergroup relationship between Arenicolidae and Maldanidae is also recovered. These results support our assumption that on the basis of a broader taxon sampling the phylogenetic position of controversially discussed taxa can be inferred by using 18S rDNA sequence data.     

Molecular phylogeny,taxonomy, and distribution of French Unio species (Bivalvia,Unionidae)

A plethora of unionid names was established in the nineteenth century by the “Nouvelle Ecole”. Although naiad morphological plasticity is well documented, the currently recognized fauna, with 17 species and subspecies included in the French checklist for the Unio genus, is still based upon morphological characters only. Insights have been provided from molecular data elsewhere in Europe and North Africa, but the French fauna remains unstudied. We present a molecular phylogeny of the Unio genus in France based on COI, 16S and 28S genes; taking up all available data in Europe plus 273 specimens collected in all main French drainages. The results show that there are either three valid species in France, with U. pictorum and U. mancus synonymized, or five, with the subspecies U. crassus courtillieri elevated to species level. Subspecies were generally not recovered, which questions the evolutionary units tacitly implied by subspecific names. Although sampling topotypes is the most reliable way to evaluate the status of a nominal subspecies, major human-induced changes in aquatic hydrosystems challenge the method. Nevertheless, operational taxonomy has to rely on ground-truthed data and we propose to reduce the actual number of valid taxa in France to the seven observed operational taxonomic units.