Mitochondrial DNA and karyotypic data confirm the presence of Mus indutus and Mus minutoides (Mammalia,Rodentia, Muridae,Nannomys) in Botswana

We use a combination of cytochrome b sequence data and karyological evidence to confirm the presence of Mus indutus and Mus minutoides in Botswana. Our data include sampling from five localities from across the country, including one site in northwestern Botswana where both species were captured in syntopy. Additionally, we find evidence for two mitochondrial lineages of M. minutoides in northwestern Botswana that differ by 5% in sequence variation. Also, we report that M. minutoides in Botswana have the 2n=34 karyotype with the presence of a (X.1) sex-autosome translocation.     

Phylogeography of Mus (Nannomys) minutoides (Rodentia,Muridae) in West Central African savannahs: singular vicariance in neighbouring populations

We studied the phylogeography of the strict savannah pygmy mice Mus (Nannomys) minutoides in West Central Africa. A total of 846 base pairs of the cytochrome b sequence were obtained for 66 individuals collected in Gabon, Cameroon, Republic of Congo and Central African Republic. These sequences were compared to those of M. minutoides from other African countries and to eight other species of the genus Mus. We performed maximum likelihood, Bayesian and nested clade analyses, as well as neutrality tests and time estimates. We show that M. minutoides is a well‐differentiated monophyletic species that separated from other pygmy mice 1.17 Myr ago. A distinct West Central African M. minutoides clade diverged early from the other African populations of the species, with a more recent common ancestor dating 0.14 Myr. West Central African populations are globally homogeneous, despite the present fragmentation of savannahs by the rain forest. However, our analyses show an unexpected vicariance between geographically close savannahs, embedded in the rain forest in Central Gabon. One of these populations is genetically more similar to very distant peripheral populations than to three closely neighbouring populations situated on both sides of the Ogooué River. A non‐river geographical barrier probably persisted in this area, durably isolating these local populations. This hypothesis about the history of the savannah landscape should be testable through the biogeographical analysis of other strict savannah small mammal species.     

Evolution of the major histocompatibility complex: a hundred-fold amplification of MHC class I genes in the African pigmy mouse Nannomys setulosus

The genome of the African murine rodent Nannomys setulosus was found to harbor several thousand major histocompatibility complex (MHC) class I genes instead of the 30–40 genes found in conventional laboratory mice, which are mostly of Mus musculus domesticus origin. Other genes of N. setulosus, either functionally or physically linked to class I genes, are not amplified. Amplified genes derive from as few as three ancestors and amplification has likely occured after the divergence of the two Nannomys species, N. setulosus and N. minutoides, which took place about three million years ago. Amplified genes are mostly pseudogenes. Statistical analysis of dinucleotide frequencies leads us to propose that inactivation of the genes has occured through the repeat induced mutation process, a possible newcomer in the evolution of the MHC.The nucleotide sequence data reported in this paper have been submitted to the EMBL nucleotide data library and have been assigned the accession numbers X61685 and X61686. Correspondence to : G. Gachelin.     

Molecular Evidence for Multiple Origins of the European Spined Loaches (Teleostei,Cobitidae)

We present a phylogenetic investigation of the Northern Clade, the major monophyletic clade within the freshwater fish family Cobitidae, one of the most prominent families of freshwater fishes found in Asian and European waters. Phylogenetic reconstructions based on the cytochrome b and RAG-1 genes show the genera Microcobitis, Sabanejewia, Koreocobitis and Kichulchoia as monophyletic groups. These reconstructions also show a Cobitis sensu lato and a Misgurnus sensu lato group. The Cobitis sensu lato group includes all species of Cobitis, Iksookimia, Niwaella and Kichulchoia, while the Misgurnus sensu lato group includes Misgurnus, Paramisgurnus and Koreocobitis. Although the monophyly of both the Cobitis sensu lato and Misgurnus sensu lato groups is supported, relationships within the groups are incongruent with current generic definitions. The absence of monophyly of most genera included in the Cobitis sensu lato group (Cobitis, Iksookimia and Niwaella) or their low genetic differentiation (Kichuchoia) supports their consideration as synonyms of Cobitis. Molecular phylogenies indicate that the Asian species of Misgurnus experienced a mitochondrial introgression from a lineage of Cobitis. We also find two nuclear haplotypes in the same Cobitis species from the Adriatic area that, in the absence of morphological differentiation, may indicate molecular introgression. Most lineages within the Northern Clade consist of species found in East Asia. However, some lineages also contain species from Europe and Asia Minor. The phylogenetic relationships presented here are consistent with previous studies suggesting an East Asian origin of the Northern Clade. According to the current distributions and phylogenetic relationships of the Misgurnus sensu lato and Cobitis clade lineages, particularly of M. fossilis and C. melanoleuca, the range expansion of East Asian species into Europe was most likely via Siberia into Northern and Central Europe. Phylogenetic analyses also show that the Cobitis sensu lato group consists of two clear subgroups (I and II), each presenting geographical differences. Subgroup I is distributed exclusively in East Asian drainages with an Eastern European offshoot (C. melanoleuca), whereas Subgroup II includes species widespread throughout Europe (including the Mediterranean), Asia Minor, the Black Sea and the Caucasus, with some lineages related to species restricted to East Asia.     

Syphacia (Syphacia) ohtaorum n. sp. (Nematoda: Oxyuridae) from Mus caroli on Okinawa Island,Japan

Syphacia (Syphacia) ohtaorum n. sp. is described from Mus caroli on Okinawa Island, Japan. It is closest to Syphacia (Syphacia) megaloon Quentin, 1966 from Mus minutoides and M. setulosus of Africa, but is distinguished in having a slimmer body, distinct lateral alae in the male, many eggs in the female and a smaller egg-size.     

Speciation slowing down in widespread and long-living tree taxa: insights from the tropical timber tree genus Milicia (Moraceae)

The long generation time and large effective size of widespread forest tree species can result in slow evolutionary rate and incomplete lineage sorting, complicating species delimitation. We addressed this issue with the African timber tree genus Milicia that comprises two morphologically similar and often confounded species: M. excelsa, widespread from West to East Africa, and M. regia, endemic to West Africa. We combined information from nuclear microsatellites (nSSRs), nuclear and plastid DNA sequences, and morphological systematics to identify significant evolutionary units and infer their evolutionary and biogeographical history. We detected five geographically coherent genetic clusters using nSSRs and three levels of genetic differentiation. First, one West African cluster matched perfectly with the morphospecies M. regia that formed a monophyletic clade at both DNA sequences. Second, a West African M. excelsa cluster formed a monophyletic group at plastid DNA and was more related to M. regia than to Central African M. excelsa, but shared many haplotypes with the latter at nuclear DNA. Third, three Central African clusters appeared little differentiated and shared most of their haplotypes. Although gene tree paraphyly could suggest a single species in Milicia following the phylogenetic species concept, the existence of mutual haplotypic exclusivity and nonadmixed genetic clusters in the contact area of the two taxa indicate strong reproductive isolation and, thus, two species following the biological species concept. Molecular dating of the first divergence events showed that speciation in Milicia is ancient (Tertiary), indicating that long-living tree taxa exhibiting genetic speciation may remain similar morphologically.     

The tropical African legume Scorodophloeus clade includes two undescribed Hymenostegia segregate genera and Micklethwaitia,a rare,monospecific genus from Mozambique

Legume subfamily Caesalpinioideae accommodates approximately 2250 species in 171 genera which traditionally are placed in four tribes: Caesalpinieae, Cassieae, Cercideae and Detarieae. The monophyletic tribe Detarieae includes the Amherstieae subclade which contains about 55 genera. Our knowledge of the relationships among those genera is good in some cases but for many other genera phylogenetic relationships have been unclear. The non-monophyletic nature of at least two amherstioid genera, Cynometra and Hymenostegia has also complicated the picture. During the course of a multi-disciplinary study of Hymenostegia sensu lato, which includes phylogenetic analyses based on matK and trnL data, we have recovered the “Scorodophloeus clade”, an exclusively tropical African clade of four genera which includes the eponymous genus Scorodophloeus, two undescribed generic segregates of Hymenostegia sensu lato, and the previously unsampled rare monospecific genus Micklethwaitia from Mozambique. Zenkerella is suggested as a possible sister genus to the Scorodophloeus clade. A distribution map is presented of the seven species that belong to the Scorodophloeus clade.     

Molecular phylogenetic status of the Bulgarian marbled polecat (Vormela peregusna,Mustelidae, Carnivora), revealed by Y chromosomal genes and mitochondrial DNA sequences

In this study, we investigated molecular phylogenetic status of the marbled polecat (Vormela peregusna) from Bulgaria, using sequences of two Y-chromosomal genes (SRY and ZFY). The phylogenetic tree inferred using combined sequences of both genes indicated that the marbled polecat was split from genera Lutra, Neovison and Mustela after genus Martes was diverged in family Mustelidae. In addition, we analyzed molecular phylogeography of the Bulgarian population of the marbled polecat, using cytochrome b and control region sequences of mitochondrial DNA (mtDNA). The phylogenetic tree of cytochrome b indicated that the haplotypes of the Bulgarian population comprised two haplogroups, which were the most ancestral clades. Additionally, the control region phylogeny showed that the haplotypes of Bulgaria formed two haplogroups: one was the most ancestral clade, and the other was the derivative clade. One individual with the most ancestral cytochrome b clade had a control region haplotype of the derivative clade. Thus, this study revealed that the most ancestral lineages of the marbled polecat are included in the population of Bulgaria. The Bulgarian population could be a remnant lineage from a basal for the species, which in Pleistocene occupied a relatively large area related to the Balkan-Caucasian.     

Novel Arenavirus Sequences in Hylomyscus sp. and Mus (Nannomys) setulosus from C?te d'Ivoire: Implications for Evolution of Arenaviruses in Africa

This study aimed to identify new arenaviruses and gather insights in the evolution of arenaviruses in Africa. During 2003 through 2005, 1,228 small mammals representing 14 different genera were trapped in 9 villages in south, east, and middle west of Côte d''Ivoire. Specimens were screened by pan-Old World arenavirus RT-PCRs targeting S and L RNA segments as well as immunofluorescence assay. Sequences of two novel tentative species of the family Arenaviridae, Menekre and Gbagroube virus, were detected in Hylomyscus sp. and Mus (Nannomys) setulosus, respectively. Arenavirus infection of Mus (Nannomys) setulosus was also demonstrated by serological testing. Lassa virus was not found, although 60% of the captured animals were Mastomys natalensis. Complete S RNA and partial L RNA sequences of the novel viruses were recovered from the rodent specimens and subjected to phylogenetic analysis. Gbagroube virus is a closely related sister taxon of Lassa virus, while Menekre virus clusters with the Ippy/Mobala/Mopeia virus complex. Reconstruction of possible virus–host co-phylogeny scenarios suggests that, within the African continent, signatures of co-evolution might have been obliterated by multiple host-switching events.     

Molecular phylogenetics of Reig's short-tailed opossum (Monodelphis reigi) and its distributional range extension into Guyana

Reig's short-tailed opossum (Monodelphis reigi) was recently described morphologically, based on a single specimen from southeastern Venezuela. It was considered most similar to M. adusta, which is allopatrically distributed in the Andes and surrounding areas, but there has not been an explicit study of systematic relationships with other species of Monodelphis. We report the first occurrence from Guyana of this rare species that was previously known by only the holotype. In a molecular phylogenetic analysis of mitochondrial cytochrome b sequence variation, it groups within a well-supported monophyletic clade that includes M. adusta, M. handleyi, M. osgoodi, and M. peruviana. M. adusta was found to be the sister taxon to the rest of the adusta-complex. M. reigi is the basal taxon (sister species) of remaining adusta-complex forms. This corroborates earlier morphological studies suggesting close affinity of these taxa. As presently known, M. reigi is endemic to the highland regions (>1,000 m asl) of the Guiana Shield of northern South America, and is the only taxon within the M. adusta species complex that does not occur in the Andes or adjacent lowland regions. Based on previous molecular dating of Didelphidae, this suggests that there was a dispersal event from the Andes to the Guiana Highlands in the Miocene that gave rise to M. reigi.     

Morphological cranial diversity contributes to phylogeny in soft-furred sengis (Afrotheria, Macroscelidea)

Despite the well-supported Macroscelidea phylogeny proposed at the end of the 1960s, several systematic arrangements have been suggested in the last 20 years, raising doubts about the phylogeny of the Macroscelidinae; sengi inter-specific relationships are still debated to this day. The main issue of concern involves the supposed Elephantulus diphyly. To solve this persisting debate about sengi phylogeny, we examined the cranium ventral surface of 13 species using geometric morphometric techniques and neighbour-joining algorithms. This study supported the idea that the ventral side of the sengi cranium has the potential to provide important signals for reconstructing the Macroscelidea phylogeny. The phylogenetic signals seemed to differentiate between two major clades in the sengi radiation. In the first clade, the two monospecific genera (Petrodromus and Macroscelides), the two African Horn species (Elephantulus revoilii and E. rufescens), and the only North African species (E. rozeti) were clustered together. The second clade includes the remnant south-central African Elephantulus species. Our results were in agreement with both mitochondrial and nuclear data, confirmed that there is no Elephantulus monophyly and highlighted the close relationship between Petrodromus and E. rozeti. It appears that all the soft-furred sengi species are organised in two evolutionary lines: an old monophyletic clade, comprising only Elephantulus species, and a new polyphyletic clade, including P. tetradactylus, M. proboscideus, and E. rozeti. This requires a taxonomic and nomenclatural rearrangement within Macroscelidinae, where the phylogenetic position of the remnant 4 (of 12) Elephantulus species has yet to be fully defined.     

How to get off the mismatch at the generic rank in African Podostemaceae?

The Podostemaceae are highly enigmatic plants which are restricted to submerged river-rock habitats. The availability of new material of nine taxa from continental Africa prompted this new study. Five species belonging to the genera Dicraeanthus, Leiothylax, Letestuella, Macropodiella, and Stonesia and another four species of the large genera Inversodicraea sensu stricto and Ledermanniella sensu stricto have been analysed for the first time. New anatomical and developmental data are described and illustrated by use of microtome sections and scanning electron microscopy. In parallel, phylogenetic analyses of all available sequence data of African Podostemaceae have been conducted using three plastid markers (matK, trnD-trnT, rpoB-trnC). Inversodicraea cf. bosii appears basal within the continental African clade. The remaining taxa are distributed in three, rather poorly supported, major clades which are consistent with their morphology: (1) the Inversodicraea clade is characterised by stem scales and contains members of the former Ledermanniella subg. Phyllosoma with either pollen-monads or dyads; (2) the Ledermanniella-Monad group consisting of Leiothylax, Letestuella, Macropodiella, Stonesia, and Ledermanniella species—all taxa without stem scales but showing pollen as single grains, with Monandriella linearifolia being basal to this clade; (3) the Ledermanniella-Dyad clade including Djinga, Dicraeanthus, and Ledermanniella species without stem scales but with pollen dyads. To reduce the polyphyly of Ledermanniella sensu lato (i.e. sensu C. Cusset) we propose restricting Ledermanniella to the species of the former subgenus Ledermanniella, resurrecting Monandriella as monotypic genus, and accepting the genus name Inversodicraea for members of Ledermanniella subg. Phyllosoma.     

Morphological variability,chromosome polymorphism,and problems of identifying individual species of gobies of the group <Emphasis Type="Italic">Ponticola</Emphasis> (Gobiidae) and assessing taxonomic relations of local populations

In connection with the problems of identification of certain species of the genus Ponticola, and especially their juveniles, morphological and genetic studies of gobies collected in the northwestern part of the Black Sea basin were carried out using literature data and museum collections. Karyological data indicate a distinct species status of western (P. odessicus) and eastern (P. eurycephalus) mushroom gobies; the range of the latter includes not only the Sea of Azov but also waters of the Black Sea coast of the Crimean Peninsula; these conclusions are supported by preliminary molecular genetic studies. Based on the obtained results of intraspecific morphological variability of species, we propose morphological characteristics for identification of adult P. syrman and P. odessicus, as well as a specific set of characteristics to identify juvenile (body length less than 50 mm) P. syrman, P. kessleri, and P. odessicus. We reveal differences in the karyotype structure of P. syrman from populations of the Sasyk Lake and the Don River, which do not differ in terms of external morphological characteristics and variability of cytochrome b haplotypes. The need for further research of phylogeography and morphological and karyological variability of P. syrman sensu lato is assumed.     

Molecular diversity and phylogeography of Andricus curtisii (Hymenoptera,Cynipidae) in Turkey

A 433-bp fragment of the mitochondrial cytochrome b gene and the entire nuclear ITS2 region were sequenced in an oak gall wasp species, Andricus curtisii, which represents 21 populations across its range in Turkey. Forty cytochrome b haplotypes and 63 ITS2 alleles were discovered among the 161 individuals studied. Remarkable genetic variation was detected in the species, with an average haplotype diversity (h) of 0.72 and 0.76 and an average nucleotide diversity (π) of 0.06 and 0.01 for the cyt b gene and the ITS region, respectively. Most populations were genetically distinct, possessing unique cytochrome b haplotypes. Maximum likelihood and Bayesian phylogenetic analyses and application of the haplotype network to both datasets revealed that the most basally located haplotypes/alleles were from the eastern part of Turkey. The detection of older sequences in the phylogenetic trees of the easterly located populations implied an eastern origin of the species. The application of a conventional insect mitochondrial DNA clock to the dataset suggested a splitting of ingroup haplotypes from the outgroup lineages predating the Pleistocene epoch and the formation of two major haplogroups that coincided with the oscillation of the early Pleistocene glaciations.     

Systematics of Serinus canaries and the status of Cape and Yellow-crowned Canaries inferred from mtDNA and morphology

The taxonomy of, and phylogenetic relationships among, African canaries typically assigned to the genus Serinus sensu lato (including the putative genera Alario, Pseudochloroptila, Serinops, Ochrospiza, Dendrospiza and Crithagra) were investigated, based on 823bp of the mitochondrial cytochrome b gene. Two clades emerged: (1) Palaearctic and Afrotropical taxa, including Serinus serinus, S. canaria, A. alario and the S. canicollis complex (S. c. canicollis, thompsonae and flavivertex), and (2) taxa endemic to the Afrotropics comprising Pseudochloroptila, Serinops, Ochrospiza, Poliospiza, Dendrospiza, Poliospiza and Crithagra spp. However, only clade one has jackknife and bootstrap support values above 50. The two clades were separated by Carduelis taxa, suggesting that Serinus sensu lato is not monophyletic. As it stands, the phylogeny does not support the recognition of Alario and Serinops and possibly Ochrospiza, Dendrospiza and Pseudochloroptila (Cape siskins) as distinct genera. Alario is sister to Serinus canicollis whereas the others are intermingled in a clade comprised primarily by Crithagra spp. The use of Serinus is confined to the Palaearctic canaries and allied species, and Crithagra for the strictly Afrotropical clade. Since the cytochrome b sequence divergence between the allopatric Cape (S. [c.] canicollis) and Yellow-crowned (S. [c.] flavivertex) canaries (2.8%) was similar to differences between other closely-related species and there are marked plumage differences between these taxa, we suggest that they be elevated to full species rank.     

Evolutionary and dispersal history of Eurasian house mice Mus musculus clarified by more extensive geographic sampling of mitochondrial DNA

We examined the sequence variation of mitochondrial DNA control region and cytochrome b gene of the house mouse (Mus musculus sensu lato) drawn from ca. 200 localities, with 286 new samples drawn primarily from previously unsampled portions of their Eurasian distribution and with the objective of further clarifying evolutionary episodes of this species before and after the onset of human-mediated long-distance dispersals. Phylogenetic analysis of the expanded data detected five equally distinct clades, with geographic ranges of northern Eurasia (musculus, MUS), India and Southeast Asia (castaneus, CAS), Nepal (unspecified, NEP), western Europe (domesticus, DOM) and Yemen (gentilulus). Our results confirm previous suggestions of Southwestern Asia as the likely place of origin of M. musculus and the region of Iran, Afghanistan, Pakistan, and northern India, specifically as the ancestral homeland of CAS. The divergence of the subspecies lineages and of internal sublineage differentiation within CAS were estimated to be 0.37–0.47 and 0.14–0.23 million years ago (mya), respectively, assuming a split of M. musculus and Mus spretus at 1.7 mya. Of the four CAS sublineages detected, only one extends to eastern parts of India, Southeast Asia, Indonesia, Philippines, South China, Northeast China, Primorye, Sakhalin and Japan, implying a dramatic range expansion of CAS out of its homeland during an evolutionary short time, perhaps associated with the spread of agricultural practices. Multiple and non-coincident eastward dispersal events of MUS sublineages to distant geographic areas, such as northern China, Russia and Korea, are inferred, with the possibility of several different routes.     

Phylogeography of the Golden Jackal (Canis aureus) in India

The golden jackal (Canis aureus) is one of the most common and widely distributed carnivores in India but phylogeographic studies on the species have been limited across its range. Recent studies have observed absence of mitochondrial (mt) DNA diversity in European populations while some North African populations of golden jackal were found to carry gray wolf (Canis lupus lupaster) mtDNA lineages. In the present study, we sequenced 440 basepairs (bp) of control region (CR) and 412 bp of cytochrome b (cyt b) gene of mtDNA from 62 golden jackals sampled from India (n = 55), Israel (n = 2) and Bulgaria (n = 5), to obtain a total of eighteen haplotypes, comprising sixteen from India and one each from Israel and Bulgaria. Except for three previously described haplotypes represented by one cyt b and one CR haplotype both from India, and one CR haplotype from Bulgaria, all haplotypes identified in this study are new. Genetic diversity was high in golden jackals compared to that reported for other canids in India. Unlike the paraphyletic status of African conspecifics with the gray wolf, the Indian (and other Eurasian) golden jackal clustered in a distinct but shallow monophyletic clade, displaying no evidence of admixture with sympatric and related gray wolf and domestic dog clades in the region. Phylogeographic analyses indicated no clear pattern of genetic structuring of the golden jackal haplotypes and the median joining network revealed a star-shaped polytomy indicative of recent expansion of the species from India. Indian haplotypes were observed to be interior and thus ancestral compared to haplotypes from Europe and Israel, which were peripheral and hence more derived. Molecular tests for demographic expansion confirmed a recent event of expansion of golden jackals in the Indian subcontinent, which can be traced back ~ 37,000 years ago during the late Pleistocene. Our results suggest that golden jackals have had a potentially longer evolutionary history in India than in other parts of the world, although further sampling from Africa, the Middle East and south-east Asia is needed to test this hypothesis.     

Le caryotype et la spermatogénèse de Mus setulosus (bandes Q,C, G et coloration argentique)

The karyotype and the spermatogenesis of Mus setulosus (G, Q, C bands and Ag-staining). — The G, Q and C banding patterns of Mus setulosus are described. In the male five Ag-NORs were detectable, located at the distal extremities of the chromosomes. The meiotic divisions of Mus setulosus were studied by silver-staining and C-banding. The X and Y chromosomes remain close to each other during leptotene, zygotene and pachytene; they do not establish and end-to-end or side-by-side association. During diplotene, diakinesis and metaphase I, they most often are separated. At pachytene the bivalents 5 and 7 lie near each other and form a heterochromatic vesicle. The r-RNA synthesis declines at the end of pachytene and stops during diplotene, diakinesis, metaphase I and II. During these four stages the centromeres are Ag-positive. A comparative analysis of G-bands revealed a great number of analogies between Mus musculus and M. setulosus, which confirms the hypothesis that M. setulosus originated from a Mus of the musculus group. Nevertheless the existence of non-homologous pairs and the differences in the distribution and the size of heterochromatic blocks show that the derivation of the karyotype of setulosus from that of musculus imply several complex rearrangements, the nature of which could not be determined despite the refinement of the actual techniques.     

New insights into the taxonomy and phylogeny of social voles inferred from mitochondrial cytochrome b sequences

We sequenced the entire cytochrome b gene in Microtus paradoxus from Turkmenistan and Microtus socialis from Crimea and Kalmykia. Phylogenetic relationships among social voles were reconstructed by the inclusion into analyses of a further 23 published haplotypes belonging to six species. The two probabilistic methods which were used in phylogenetic analyses, the Bayesian inference and Maximum Likelihood, yielded very similar results. Both trees showed two highly divergent lineages which were further subdivided into seven species. The socialis lineage encompassed four species (M. socialis, M. irani, M. anatolicus, and M. paradoxus), and the remaining three species clustered into the guentheri lineage (M. guentheri, M. hartingi, M. dogramacii). The ranges for nucleotide divergences between seven species of social voles (4.95–9.28% and 4.18–8.81% for mean and net divergences, respectively) mainly exceeded 4.3%, which is frequently regarded as the conservative cut-off between sibling species in the specious genus Microtus.