MP Trees Research Articles

Phylogenetic analysis of the genus Cylicocyclus (Nematoda: Strongylidae) based on nuclear ribosomal sequence data

Seven species of Cylicocyclus Ihle, 1922 (Nematoda: Strongylidae) were collected from donkeys from Henan Province, China. Five samples of each species were selected for sequencing. Sixteen different internal transcribed spacer (ITS) sequences representing the seven species of Cylicocyclus were obtained. Sequence differences in the first internal transcribed spacer (ITS-1) among species was lower than that of the second internal transcribed spacer (ITS-2). Phylogenetic analyses were conducted using the combined ITS-1 and ITS-2 data sets from the present study and using reference sequences from the GenBank database. The MP and ML trees were similar in topology. The phylogenetic trees were divided into two clades. Clade I included 8 species of Cylicocyclus; within this group, Cylicocyclus leptostomus (Kotlan, 1920) is nested between different samples of Cylicocyclus ashworthi (LeRoux, 1924), suggesting C. ashworthi may represent a species complex. Clade II included Cylicocyclus elongatus (Looss, 1900) and Cylicocyclus ultrajectinus (Ihle, 1920); however, these two species always clustered with the comparative species (Petrovinema poculatum (Looss, 1900) and Poteriostomum imparidentatum Quiel, 1919), suggesting that C. elongatus and C. ultrajectinus represent members of other genera.

Cacopsylla fraudatrix sp.n. (Hemiptera: Psylloidea) recognised from testis structure and mitochondrial gene COI

Cacopsylla fraudatrix Labina & Kuznetsova sp.n., collected from Vaccinium myrtillus in Bieszczady Mountains (S. Poland) is described and compared with the most closely related species, C. myrtilli (developing on V. myrtillus) and C. ledi (on Ledum palustre) as well as with a distantly related species C. vaccinii (on V. bracteatum). The identity of the new species is supported by the structure of testes consisting of two follicles each, in contrast to four in C. myrtilli and C. ledi, and by a molecular phylogenetic analysis based on a 714 bp fragment of the mitochondrial COI gene. C. fraudatrix sp. n. forms a distinct clade in MP, ML and BI trees and differs from C. myrtilli by 28 fixed nucleotide substitutions (uncorrected p-distance = 3.92%) and from C. ledi by 26 fixed nucleotide substitutions (uncorrected p-distance = 3.64%).

Resolving and dating the phylogeny of Cornales – Effects of taxon sampling, data partitions, and fossil calibrations

The order Cornales descends from the earliest split in the Asterid clade of flowering plants. Despite a few phylogenetic studies, relationships among families within Cornales remain unclear. In the present study, we increased taxon and character sampling to further resolve the relationships and to date the early diversification events of the order. We conducted phylogenetic analyses of sequence data from 26S rDNA and six chloroplast DNA (cpDNA) regions using parsimony (MP), maximum likelihood (ML), and Bayesian inference (BI) methods with different partition models and different data sets. We employed relaxed, uncorrelated molecular clocks on BEAST to date the phylogeny and examined the effects of different taxon sampling, fossil calibration, and data partitions. Our results from ML and BI analyses of the combined cpDNA sequences and combined cpDNA and 26S rDNA data suggested the monophyly of each family and the following familial relationships ((Cornaceae–Alangiaceae)–(Curtisiaceae–Grubbiaceae))–(((Nyssaceae–Davidiaceae)–Mastixiaceae)–((Hydrostachyaceae–(Hydrangeaceae–Loasaceae))). These relationships were strongly supported by posterior probability and bootstrap values, except for the sister relationship between the N–D–M and H–H–L clades. The 26S rDNA data and some MP trees from cpDNA and total evidence suggested some alternative alignments for Hydrostachyaceae within Cornales, but results of SH tests indicated that these trees were significantly worse explanations of the total data. Phylogenetic dating with simultaneous calibration of multiple nodes suggested that the crown group of Cornales originated around the middle Cretaceous and rapidly radiated into several major clades. The origins of most families dated back to the late Cretaceous except for Curtisiaceae and Grubbiaceae which may have diverged in the very early Tertiary. We found that reducing sampling density within families and analyzing partitioned data sets from coding and noncoding cpDNA, 26S rDNA, and combined data sets produced congruent estimation of divergence times, but reducing the number and changing positions of calibration points resulted in very different estimations.

Systematics of Grammitid Ferns (Polypodiaceae): Using Morphology and Plastid Sequence Data to Resolve the Circumscriptions of <I>Melpomene</I> and the Polyphyletic Genera <I>Lellingeria</I> and <I>Terpsichore</I>

Recent phylogenetic analyses of grammitid ferns (Polypodiaceae) have demonstrated that some genera recognized within this clade are not monophyletic. We focus on resolving the circumscriptions of some large Neotropical genera by focusing on a clade, identified in previous analyses, that includes the monophyletic genus Melpomene, plus portions of two polyphyletic genera, Lellingeria and Terpsichore. Phylogenetic analyses using chloroplast markers atpB, rbcL, and trnL-F, along with 109 morphological characters, resolve this ingroup as monophyletic and sister to a clade that includes Ceradenia, Enterosora, and Zygophlebia. We use morphological characters included in the analysis to circumscribe clades. Melpomene is clearly monophyletic, but forms a polytomy with Lellingeria s. l. in a strict consensus of MP trees. Ingroup species of Terpsichore form three well supported monophyletic groups that together are paraphyletic with respect to Melpomene plus Lellingeria. Two clades corresponding to species groups of Terpsichore will be recognized as new genera.

Polypteridae (Actinopterygii: Cladistia) and DANA-SINEs insertions

SINE sequences are interspersed throughout virtually all eukaryotic genomes and greatly outnumber the other repetitive elements. These sequences are of increasing interest for phylogenetic studies because of their diagnostic power for establishing common ancestry among taxa, once properly characterized. We identified and characterized a peculiar family of composite tRNA-derived short interspersed SINEs, DANA-SINEs, associated with mutational activities in Danio rerio, in a group of species belonging to one of the most basal bony fish families, the Polypteridae, in order to investigate their own inner specific phylogenetic relationships. DANA sequences were identified, sequenced and then localized, by means of fluorescent in situ hybridization (FISH), in six Polypteridae species ( Polypterus delhezi, P. ornatipinnis, P. palmas, P. buettikoferi P. senegalus and Erpetoicht h ys calabaricus) After cloning, the sequences obtained were aligned for phylogenetic analysis, comparing them with three Dipnoan lungfish species ( Protopterus annectens, P. aethiopicus, Lepidosiren paradoxa), and Lethenteron reissneri (Petromyzontidae)was used as outgroup. The obtained overlapping MP, ML and NJ tree clustered together the species belonging to the two taxonomically different Osteichthyans groups: the Polypteridae, by one side, and the Protopteridae by the other, with the monotypic genus Erpetoichthys more distantly related to the Polypterus genus comprising three distinct groups: P. palmas and P. buettikoferi, P. delhezi and P. ornatipinnis and P. senegalus. In situ hybridization with DANA probes marked along the whole chromosome arms in the metaphases of all the Polypteridae species examined.

Phylogenetic Position of Tarsius bancanus Based on Partial Cytochrome b DNA Sequences

This study was carried out to ascertain the molecular phylogenetic position of Tarsius bancanus among Malaysian primates based on the partial of Cytochrome b (cyt b) gene sequences. A total of five samples of Tarsius bancanus from Sabah, Malaysia, were used in this study. Several other Malaysian primates were also included in the analysis (Leaf monkeys (Presbytis and Trachypithecus), Macaques (Macaca), Siamang (Symphalangus) and Slow loris (Nycticebus). We also included DNA sequences of several prosimians (Galago, Cheirogaleus, Daubentonia, Indri, Avahi, Lemur and Lepilemur) from GenBank. In addition, one individual of orang utan (Pongo pygmaeus) and human (Homo sapiens) were used as outgroups to root the tree. All taxa were analysed using character method (Maximum Parsimony, MP) and distance method (Neighbor-Joining, NJ). From the 375 examined characters, 43.2% were constant characters while 4.8% characters were parsimony uninformative whereas 52.0% characters were parsimony informative. Tree topologies discriminated three major clades in which primitive primates, Old World Monkeys and Anthropoids belongs to their own monophyletic clades. Both MP and NJ trees showed that T. bancanus was placed in primitive primates group.

Molecular phylogeny of Nothoholosticha (Protozoa, Ciliophora, Urostylida) and systematic relationships of the Holosticha-complex

The marine ciliate Nothoholosticha is characterized by having a Holosticha-like ciliature pattern but without migratory frontoterminal cirri. However, its systematic position and its relationship to other members of the Holosticha-complex have not yet been resolved. In order to gain deeper insights into these relationships, the small subunit rRNA (SSrRNA) gene and the rRNA internal transcribed spacer and 5.8SrRNA (ITS1–5.8S-ITS2) region of two marine Anteholosticha species, as well as the ITS1–5.8S-ITS2 region of N. fasciola, were sequenced, and molecular trees (BI, ML, NJ and MP trees) were constructed. Although our analyses failed to conclusively resolve the phylogeny of this assemblage, certain conclusions could be drawn. Firstly, Nothoholosticha is a valid genus that is more closely related to Pseudokeronopsis than to other Holosticha-complex genera. Secondly, sequence analyses and phylogenetic trees of several Anteholosticha species revealed a high molecular diversity, which does not support the monophyly of this genus. Thirdly, the current assignment of certain well-known genera, e.g. Holosticha, Anteholosticha, Apokeronopsis, Parabirojimia, Psammomitra, Diaxonella, Metaurostylopsis and Thigmokeronopsis, to the families Bakuellidae (sensu Berger 2006), Urostylidae (sensu Berger 2006) or Holostichidae (sensu Berger 2006) is challenged by the molecular data presented here. And fourthly, the families Holostichidae and Pseudokeronopsidae (sensu Lynn 2008) are probably paraphyletic, and their systematic assignments await further evaluation.

Contracaecum gibsoni n. sp. and C. overstreeti n. sp. (Nematoda: Anisakidae) from the Dalmatian pelican Pelecanus crispus (L.) in Greek waters: genetic and morphological evidence

Two new species of Contracaecum Railliet & Henry, 1912, previously referred to as C. multipapillatum sp. A and C. multipapillatum sp. B by Nascetti et al. (1990) from the Dalmatian pelican Pelecanus crispus (L.) in the Ambracian Gulf off Greece, are described as C. gibsoni n. sp. and C. overstreeti n. sp., respectively. Morphological analysis and the differential diagnosis of genetically recognised male specimens of C. gibsoni and C. overstreeti with respect to C. multipapillatum (von Drasche, 1882) (sensu lato) from Egretta alba (L.) in northern Colombia and other morphologically related Contracaecum spp. enabled the detection of differences between the two species in a number of characters, including spicule length and the shape of its tip, and the arrangement of the proximal and distal papillae on the male tail. Accordingly, formal descriptions are presented for C. gibsoni n. sp. and C. overstreeti n. sp. from P. crispus. The genetic characterisation of the two taxa is based on 20 allozyme loci and sequence analyses (519 bp) of the mtDNA cox2 gene. Reproductive isolation was demonstrated between these two taxa, which sympatrically infect the same definitive host, and fixed allele differences between the two species were found at some (Aat-2, PepC-1, PepC-2 and Pgm-1) of the 20 allozyme loci analysed. Their genetic divergence, estimated at the allozyme level, was D ( Nei ) = 0.31. The genetic relationships of C. gibsoni and C. overstreeti with respect to C. multipapillatum (s. l.) collected from E. alba in Colombia, as well as with other congeners from fish-eating birds which had previously been genetically characterised using the same genetic markers, i.e. C. rudolphii A and C. rudolphii B of Bullini et al. (1986), C. septentrionale Kreis, 1955, C. microcephalum (Rudolphi, 1809), C. bioccai Mattiucci et al., 2008, C. pelagicum Johnston & Mawson, 1942 and C. micropapillatum (Stossich, 1890), were inferred from mtDNA cox2 sequence analysis. The MP, NJ and BI trees obtained were congruent in depicting C. gibsoni and C. overstreeti as closely related species but quite distinct from each other and forming a subclade with specimens of C. multipapilllatum (s. l.) from E. alba (L.) in Colombia. This subclade was also found to be distinct from the remainder of the taxa considered.

Systematics of Scorpaeniformes Species in the Mediterranean Sea Inferred from Mitochondrial 16S rDNA Sequence and Morphological Data

Genetic and morphological divergence and phylogenetic relationships of Scorpaeniformes fish including two genera and six species, Helicolenus dactylopterus, Scorpaena maderensis, Scorpaena porcus, Scorpaena elongata, Scorpaena scrofa, Scorpaena notata, living in the Mediterranean Sea, were investigated with morphological and mitochondrial 16S rDNA sequence data. The mean nucleotide diversity was found to be 0.0792. Average sequence divergence between species of Sebastidae and Scorpaenidae was 8.4%, and 6.4%. between species of the genus Scorpaena. For congeneric comparisions, the lowest genetic divergence (0.7%) was observed between S. porcus and S. notata, and the highest divergence (10.8%) was detected between S. maderensis and S. notata. High levels of nucleotide divergence were detected between species of two families, and the maximum value was found to be 14.5% between H. dactylopterus and S. elongata. The two phylogenetic methods (NJ and MP) identified two major lineages. In the NJ tree S. elongata was the sister group to S. scrofa. S. maderensis was more divergent from these groups. Another lineage contained S. porcus and S. notata. The topology of the MP tree is similar to that of the NJ tree. The pattern and degree of morphological differentiation was not congruent with the genetic differentiation. The Euclidiean distances of morphological data revealed very high morphological divergence between the two families. The highest morphological divergence was observed between H. dactylopterus and S. porcus, and the lowest was detected between S. elongata and S. notata. The present genetic and morphological data support the present monophyletic status of the Scorpaena genus.

Identification of three major Bemisia tabaci biotypes in China based on morphological and DNA polymorphisms

The sweet potato whitefly, Bemisia tabaci, is a complex species, and many of its biotypes are important agricultural pests. The B and Q biotypes are invasive and coexist with the native Cv biotype on vegetable and ornamental crops in China. In this study, these three biotypes were identified based on their morphological characteristics, RAPD–PCR analysis, and DNA sequences of the COI gene. The anterior and posterior wax fringes of the B, Q, and Cv biotypes significantly differed from each other. Based on this morphological characteristic, the three biotypes can easily be distinguished in greenhouses and fields. Genomic DNA RAPD–PCR band patterns also revealed differences between these biotypes using the H 16 primer. The B biotype has three bright DNA bands between 250 and 600 bp, the Q biotype only has one bright band at ∼300 bp, while the Cv biotype has no band between 250 and 500 bp. Both the Cv and Q biotypes have two bright bands at ∼750 and 1000 bp while the B biotype has only one band at ∼1000 bp. Based on the COI gene, the genetic identity between B and Cv, B and Q, and Q and CV was 85.8%, 94.7%, and 86.0%, respectively. The MP tree indicated that the phylogenetic relationship between the B and Q biotypes is much closer than that between the B and Cv or the Q and Cv biotypes. The uses of the morphological, RAPD–PCR, and DNA sequencing methods in biotype identification of B. tabaci are discussed.

Morphological variability and genetic diversity in five species of Skeletonema (Bacillariophyta)

Twelve strains of Skeletonema from Chinese coastal waters and another strain from the West Sea of Korea were examined using light microscopy (LM), scanning and transmission electron microscopy (SEM, TEM), and small subunit (SSU) rDNA sequence analysis. We identified four species of Skeletonema: S. marinoi Sarno and Zingone, S. subsalsum (A.Cleve) Bethge, S. dohrnii Sarno and Kooistra, S. tropicum Cleve, as well as a Skeletonema-like species. These are the first records of S. marinoi, S. dohrnii and S. subsalsum in China. The Skeletonema-like species, MMDL5067, was isolated from the estuary of the Changjiang River. The morphologic and phylogenetic characteristics of this species do not correspond to those of any previously published Skeletonema species. The intercalary fultoportula processes (IFPPs) have long open tubes, and the intercalary rimoportula process (IRP) is short and located inside the rim of the IFPPs. Phylogenetic analysis revealed that MMDL5630 clustered with S. tropicum in the NJ and MP trees, but its IRPs differed from those of previously described specimens of S. tropicum. The IRP is partially open, with an elongated opening at the base of the tube, and is longer than the IFPPs. Some specimens of S. marinoi and S. dohrnii also have longer tubular IRPs than the IFPPs. Compared to previously described specimens of S. marinoi and S. dohrnii, these two specimens we identified have shorter IRPs. Our results reflect the ambiguity and diversity of Skeletonema taxonomy. They are also significant for the future identification and management of red tides caused by Skeletonema in the seas around China.

On the systematics and phylogeography of eight-barbel loaches of the genus Lefua (Cobitoidea: Nemacheilidae): mtDNA typing of L. pleskei

Comparative analysis of personal sequence data for the mtDNA control region (926 to 928 bp) from eight-barbel loaches inhabiting eight localities in the Amur River basin (4) and the Sea of Japan (4) and the GeneBank/NCBI data for the Lefua individuals from the other regions of the world showed that eight-barbel loaches from Primorskii krai water basins were marked by a specific group of mtDNA haplotypes. This finding is considered as supporting the species status of L. pleskei. Genetic distances within L. pleskei are small (on average 0.355) and close to those within L. nikkonis (on average 0.48%). The distances between this species pair are the least (on average 2.15%) among all other pairs compared. In MP, ML, and Bayesian trees, L. pleskei and L. nikkonis haplotypes formed a common clade with high statistically significant support. In all tree variants, L. costata mtDNA haplotypes were located out of the group of interest. A clade consisting of highly diverged lineages of Lefua sp. and L. echigonia haplotypes occupied even more independent position. The mtDNA haplotypes of L. pleskei and L. costata from the Amur River basin were evolutionary young and derived from the haplotypes found in these species from the Sea of Japan (L. pleskei) or the Yellow Sea (L. costata) basins. It is thereby suggested that both species rather recently migrated into the Amur River system. According to the molecular clock data, basal diversification of the eight-barbel loach lineages took place at the end of middle Miocene (about 11 to 12 Myr ago), while divergence of L. pleskei and L. costata ancestral forms probably occurred approximately, 5 Myr ago. Since all main lineages of eight-barbel loaches were found in the Sea of Japan basin (continental coastline and the islands), the divergence order and dispersal patterns of the Lafua species might have been largely determined by the geological development pattern of this water body and the adjacent territories.

Taxonomic identification of dolphin love charms commercialized in the Amazonian region through the analysis of cytochrome b DNA

Previous studies identified the Amazon river dolphin (Inia geoffrensis) as the species frequently traded in public markets of Pará and Amazonas State (Brazil). In order to investigate the geographic origin and the taxonomic identification of these specimens we analysed 21 samples provided by residents of the city of Belém and vicinities (Pará State, Brazil), presumably purchased in the ‘Ver-o-Peso’ Market (Belém) or the Manaus Municipal Market. DNA was extracted from dehydrated genitalia or from genitalia conserved in alcohol and the cytochrome b gene was PCR-amplified. Phylogenetic analyses showed identical topologies in both MP and NJ trees, with Sotalia splitting into two groups, one clade comprising the estuarine dolphin (S. guianensis) and all eight haplotypes from market-samples and another one, comprising the tucuxi (S. fluviatilis) haplotypes from the GenBank. Our findings indicated that the marine species is under a stronger commercial pressure than the Amazonian river dolphin (I. geoffrensis) and the tucuxi (S. fluviatilis) in the region. Therefore population dynamic studies as well as population monitoring should be carried out in order to evaluate the effects of this commercial hunting on the species and its local populations. The marine dolphins have been incidentally captured in gill-nets in the region, suggesting that these activities must also be monitored at the same time that social programmes must be implemented in order to inform and clarify local community and people involved in fishery activity to avoid that more animals might be captured and killed.

A phylogenetic study of Boletus section Boletus in Europe.

A phylogenetic study of the species in Boletus sect. Boletus was undertaken using the molecular markers ITS1-5.8S-ITS2 and GAPDH. Four well-supported lineages, one comprising Boletus edulis s.l., the others referring to B. aereus, B. reticulatus and B. pinophilus have been distinguished. The ML and MP trees of ITS showed remarkably low resolution within the B. edulis clade, and confirmed earlier published results, despite the use of samples from a wider geographical area and different hosts. The results of GAPDH demonstrate clearly that this low resolution must be ascribed to a low genetic variability with the B. edulis clade, and make clear that morphological and ecological characters have been overestimated within this species complex. Boletus edulis is therefore defined as a variable species with a wide morphological, ecological and geographic range, and includes several specific and subspecific taxa described in the literature (e.g. B. betulicola, B. persoonii, B. quercicola and B. venturii). Three other European species (B. aereus, B. pinophilus and B. reticulatus) are well delimited species based on morphology and our genetic data.

Molecular characterization ofGonatocerus tuberculifemur(Ogloblin) (Hymenoptera: Mymaridae), a prospectiveHomalodisca vitripennis(Germar) (Hemiptera: Cicadellidae) biological control candidate agent from South America: divergent clades

We genetically characterized the prospective South American egg parasitoid candidate, Gonatocerus tuberculifemur, of the glassy-winged sharpshooter (GWSS), Homalodisca vitripennis, for a neoclassical biological control program in California. Two molecular methods, inter-simple sequence repeat-polymerase chain reaction DNA fingerprinting and a phylogeographic approach inferred from the mitochondrial cytochrome oxidase subunit I gene (COI), were utilized. Five geographic populations from South America were analyzed; in addition, a phylogenetic analysis was performed with several named and one unnamed Gonatocerus species using the COI gene. DNA fingerprinting demonstrated a fixed geographic banding pattern difference in the population from San Rafael, Mendoza Province, Argentina. The COI analysis uncovered haplotype or geographic structure in G. tuberculifemur. A neighbour-joining distance (NJ) and a single most parsimonious tree (MP) clustered the populations into two well-supported distinct clades with strong bootstrap values (97-99% and 92-99%, respectively) with populations from San Rafael clustering into clade 2 and the rest of the populations clustering into clade 1. No haplotype sharing was observed between individuals from the two clades. Phylogenetic analyses performed by NJ and MP methods with 15 Gonatocerus species confirmed species boundaries and again uncovered two distinct clades in G. tuberculifemur with strong bootstrap support (95-100% and 68-100%, respectively). However, the NJ tree supported the morphologically defined relationships better than the MP tree. The molecular evidence in the present study is suggestive of a species level divergence. Because G. tuberculifemur is under consideration as a potential biological control agent for GWSS in California, understanding cryptic variation in this species is critical.

Phylogenetic Relationships Within the Genus Jesogammarus (Crustacea, Amphipoda, Anisogammaridae) Deduced from Mitochondrial COI and 12S Sequences

The genus Jesogammarus contains 16 species in two subgenera, Jesogammarus and Annanogammarus. To examine relationships among species in the genus, a molecular phylogenetic study including eight species of the former subgenus and four of the latter was conducted using partial DNA sequences of the mitochondrial COI and 12S rRNA genes. MP, NJ, and ML trees based on the combined COI and 12S data indicated monophyly of the subgenus Annanogammarus, though the monophyly of Jesogammarus was left unresolved. Consistent with few morphological differences, Jesogammarus (A.) naritai and J. (A.) suwaensis showed low genetic differentiation and did not show reciprocal monophyly, which suggests a close affinity of these taxa.

Phylogenetic relationships of egg parasitoids (Hymenoptera: Eulophidae) and correlated life history characteristics of their Neotropical Cassidinae hosts (Coleoptera, Chrysomelidae)

Egg parasitoids in the family Eulophidae (Hymenoptera) are an important part of the community of insects attacking neotropical leaf beetles in the subfamily Cassidinae. We present a phylogeny of 24 species of oophagous Eulophidae, using the 28S rDNA, the ITS2 rDNA and the cytochrome b genes, applying the NJ, MP, ML and Bayesian tree reconstruction methods on each data set. We ask whether the phylogenetic relationships of the parasitoids are linked with the life history characteristics of their beetle hosts. We show that cladogenesis in the oophagous Eulophidae does correlate with ovipositional behaviour and, to a lesser extent, diet and tribal affinities of their hosts. Additionally using two methods of simultaneous analysis of several gene sets: the Total Evidence method, and the construction of a “supertree” by Matrix Representation Parsimony (MRP), we substantiate the same major phylogenetic relationships within the Eulophidae.

The utility of automated analysis of inter‐simple sequence repeat (ISSR) loci for resolving relationships in the Canary Island species of Tolpis (Asteraceae)

Plants of oceanic islands, often remarkably divergent morphologically from continental relatives, are useful models for studying evolution and speciation because evolution is telescoped in time and space. Prior studies revealed little DNA sequence variation within the clade of ca. 10 Canary Island species of Tolpis, which precluded resolving species relationships. The present study assessed the utility of automated analysis of inter-simple sequence repeat (ISSR) loci for resolving relationships within the clade using 264 individuals from 36 populations of all recognized species and three undescribed morphological variants. Similarity (Dice coefficient) and Fitch parsimony were used to generate neighbor-joining (NJ) and strict consensus trees (MP), respectively. All individuals of the morphologically distinct endemic species formed clusters in both trees. There is also support for clusters of two undescribed variants in the NJ tree. Individuals from a morphologically variable complex consisting primarily of two species are not well resolved at population or species levels. The NJ and MP trees are not congruent at deeper levels, including relationships among species. Results are interpreted in terms of the biology of the species, and the utility of automated analysis of ISSR markers for interpreting patterns of evolution of Tolpis in the Canary Islands is discussed.

Phylogeography of an Australian termite, Amitermes laurensis (Isoptera, Termitidae), with special reference to the variety of mound shapes

In northern Australia, the debris-feeding termite Amitermes laurensis builds tall, wedge-shaped mounds in the northern part of Cape York Peninsula and Arnhem Land, where their habitats are seasonally flooded, and small dome shaped mounds in the southeastern part of Cape York Peninsula, where their habitats are well-drained. Phylogeographic analyses were conducted in 238 individuals from 30 populations using the mitochondrial cytochrome oxidase II (COII) gene. DNA sequences of 50 haplotypes were used to construct NJ, MP and ML trees. Phylogenetic trees for 16 Amitermes species showed monophyly of A. laurensis and the variation of A. laurensis mounds did not strongly correspond to the intraspecific phylogeny. It was observed that mounds with the same shape were constructed by phylogenetically different groups under similar environmental conditions and different mounds shapes were built by phylogenetically closely related groups under the different environmental conditions. Thus, phylogenetically close groups of A. laurensis, in different habitats, may adapt to environmental conditions by constructing different mound shapes. We also investigated the phylogeographic structure of A. laurensis. The significant positive correlation between genetic and geographic distances indicated isolation by distance, reflecting restricted dispersal ability of alates. Although the overall genetic structure of A. laurensis showed isolation by distance, we also identified two exceptions: (i) secondary contacts of genetically divergent lineages in southern Cape York Peninsula, and (ii) low genetic differences between geographically separated populations of Cape York Peninsula and Arnhem Land. Therefore, the phylogeography of A. laurensis may reflect continuous gene flow restricted to short distances and past changes of gene flow associated with the fluctuation of environmental conditions accompanying the changing sea levels in the Quaternary.

Genetic evidence of extensive introgression of short-tailed ground squirrel genes in a hybridization zone of Spermophilus major and S. erythrogenys, inferred from sequencing of the mtDNA cytochrome b gene

We have completely sequenced the mtDNA cytochrome b gene of ground squirrels from the zone of overlapping ranges of Spermophilus major and S. erythrogenys in the Tobol-Ishim interfluve, which is a putative hybridization zone of these species. The results of the sequencing showed extensive introgression of mtDNA genes of the short-tailed ground squirrel S. e. brevicauda, whose haplotype had fully replaced the S. major haplotype. All of the ground squirrels from the Tobol-Ishim interfluve had a variant of the S. e. brevicauda mtDNA haplotype that was specific for this zone. On average, 119 substitutions (10.44%) were found between S. major from Ul'yanovsk oblast and S. e. brevicauda from the northern Kazakhstan, the mean genetic distance (D) between them being 0.115, which conforms to the corresponding parameters for the S. e. brevicauda-S. pygmaeus pair (122 substitutions, D = 118). Insignificant differences (seven substitutions, D = 0.043) were found between the S. major and S. pygmaeus haplotypes, which suggest that these species have similar mitochondrial haplotypes. Five to ten nucleotide substitutions (0.44--0.88%) were detected between the animals from the Tobol--Ishim interfluve and S. e. brevicauda. The mtDNA haplotype divergence D within the genus Spermophilus (ten species) for all codon positions ranged from 0.035 to 0.158. Phylogenetic reconstructions (MP, ML, and NJ trees) showed two well-differentiated clusters with high bootstrap support. However, there was different branching topology within the cluster and their species composition varied. The maximum likelihood tree, ML, differentiating the species into two subgenera, Citellus and Colobotis, most reliably reflected taxonomic relationships of the species from the genus Spermophilus, inferred from morphological and genetic biochemical data. The morphologically pure S. major (subgenus Colobotis) animals, used in the analysis, proved to carry the haplotype of another species, S. pygmaeus (subgenus Citellus). This poses a question on the existence of the specific haplotype of S. major, the reason of its replacement by haplotype of other species, and possible consequences of this phenomenon for survival of the species.