Species delimitation in the Acomys cahirinus–dimidiatus complex (Rodentia, Muridae) inferred from chromosomal and morphological analyses

Our earlier chromosome banding studies of Acomys cahirinus and Acomys dimidiatus (the latter long considered to be a subspecies of the former) revealed that, despite very close diploid numbers (36 vs. 38), these taxa possess sharply different karyotypes and undoubtedly belong to different species. In this context, the taxonomic status and the relationship between the two chromosomal forms in Sinai (2 n  = 36) and Israel (2 n  = 38), chromosomally homozygous across a vast range except for a very narrow hybrid zone, remain poorly documented. Neither of these forms have previously been studied by chromosome banding; thus, the exact nature of chromosomal differences as well as the species to which these forms should be assigned remain unknown. Here, we present the data on comparative G-banding analysis and morphometrics of Acomys from Israel, Sinai, and Saudi Arabia, and a hybrid obtained in laboratory crosses between latter two. The analysis revealed that karyotype of Acomys from Israel is identical to that of Acomys from Saudi Arabia and both are different from that of Acomys from Sinai by one Robertsonian fusion. Therefore, karyotypically, all three are very different from A. cahirinus . It follows from the study that Sinai and probably Arabian peninsula and Minor Asia must be excluded from geographical distribution of A. cahirinus , which is limited from West Sahara to Egypt along Nile river (except Sinai). Furthermore, the synthesis of chromosomal and recent molecular data suggests a phylogeographical scenario explaining the modern distribution of Acomys in the Sinai and Arabian peninsulas and permits the update of the taxonomic status of these populations.  © 2007 The Linnean Society of London, Biological Journal of the Linnean Society , 2007, 91 , 203–214.     

Systematics of the tribe Podalyrieae (Fabaceae) based on DNA, morphological and chemical data

Phylogenetic relationships among nine genera and 28 species of the southern African tribe Podalyrieae were estimated from sequences of the internal transcribed spacer (ITS) of nuclear ribosomal DNA as well as morphological and chemical data. Morphological and ITS sequence data produced cladograms with similar topologies, both supporting the monophyly of Podalyrieae (excluding Hypocalyptus ). The combined data sets indicate that subtribe Xiphothecinae are monophyletic, but embedded within Podalyriinae. The high degree of congruence between previous taxonomic hypotheses and those based on DNA data provides further evidence for the utility of ITS sequences in studying phylogeny.  © 2002 The Linnean Society of London , Botanical Journal of the Linnean Society , 2002, 139 , 159–170.     

Contributions to the karyological study of the genus Ranunculus L. subgenus Batrachium (DC.) A Gray from the Iberian Peninsula

DIOSDADO. J. C, PASTOR, J. E. & VALDÉS, B. 1993. Contributions to the karyological study of the genus Ranunculus L. subgenus Batrachium (DC.) A. Gray from the Iberian Peninsula. The somatic and gametic chromosome numbers and detailed chromosome morphology are presented for eight taxa of aquatic ranunculi from the Iberian Peninsula. Within the subgenus Batrachium diploid and tetraploid levels have been observed (2 n = 16, 32) the commoner of which is the diploid. From the karyological data relationships have been established between the subgenus Batrachium and subgenus Ranunculus.     

A morphological study of the Petunia integrifolia complex (Solanaceae)

BACKGROUND AND AIMS: Petunia inflata has been treated taxonomically in various ways: it has been described as an independent species, treated as a synonym of P. integrifolia, and also regarded as a subspecies of P. integrifolia. The present study was designed to resolve the ambiguity involving the P. integrifolia complex (P. integrifolia plus P. inflata). METHODS: Tentative identification (either integrifolia group or inflata group) was carried out in the field based on the observation of live specimens at the restricted type localities. The accuracy of the tentative identification was later tested with principal component and cluster analyses of data obtained by measuring 21 morphological characters on cultivated live specimens sourced from 113 natural populations of the P. integrifolia complex in Argentina, Brazil, Paraguay and Uruguay. KEY RESULTS: There was a clear, statistically significant gap between the morphological measurements of the two groups, ensuring the accuracy of identification carried out in the field except for a probable hybrid swarm. Previously, the condition of the pedicel in the fruiting state was considered an important character distinguishing between these two groups; however, the condition of the pedicel was rather variable in the integrifolia group. The two groups were found to have geographically distinct distributions: the integrifolia group occurred in southern regions, whereas the inflata group occurred in northern regions. CONCLUSIONS: Based on the available evidence, it is suggested that the two groups are allopatric species, P. integrifolia and P. inflata, in agreement with the opinion of Fries (1911).     

The blackburni/murchisona species complex in Australian Pseudotetracha (Coleoptera: Carabidae: Cicindelinae: Megacephalini): evaluating molecular and karyological evidence

Our phylogenetic analysis of three endemic species of the Australian tiger beetle genus Pseudotetracha (Fleutiaux, 1864) from South Australia used sequences of two fragments of the mitochondrial genes 16S rRNA and cytochrome oxidase III. A matrix for each gene and two combined matrices were constructed. We compared these three riparian species, together with data from nine taxa of this genus available in GenBank, using parsimony and Bayesian methods. These molecular results are in agreement with the phylogenetic hypothesis for the blackburni/murchisona species complex previously proposed based on morphology, whereas other recent molecular analyses have questioned the existence of this species complex. In all of our analyses, samples of P. blackburni divided into two statistically supported clades, one of which is more closely related to P. mendacia and P. pulchra than to the other P. blackburni clade. This suggests the existence of a cryptic new species. Additionally, we analysed chromosomes of the second metaphase cells of members of the two clades. The observations showed different karyotypes as blackburni‐1 has two types of second meiotic metaphase cells with 11 and 12 chromosomes, whereas in blackburni‐2, all cells have 12 chromosomes, adding evidence for the putative existence of two species.     

The first record of a natural hybrid of the roach Rutilus rutilus and nase Chondrostoma nasus in the Danube River Basin, Czech Republic: morphological, karyological and molecular characteristics

Morphological (meristic and morphometric traits), karyological and molecular (microsatellites, cytochrome b ) analyses were performed to characterize a hybrid of the roach Rutilus rutilus and nase Chondrostoma nasus . Meristic and morphometric traits were different between hybrid and both parental species. The number of chromosomes found in hybrid specimen indicated that this individual represents the post-F₁ generation of hybrids and the microsatellite analysis of the hybrid showed the presence of variants typical for R. rutilus and C. nasus .     

A reappraisal of the genus Megacaryon (Boraginaceae,Lithospermeae) based on molecular,morphological, and karyological evidence

The systematic position and phylogenetic relationships of Echium orientale, a rare endemic of the mountains of northern Turkey, were elucidated based on morphological, molecular, and karyological evidence.

Using nuclear and plastid DNA sequences, we found that this species is not included in Echium, but is rather at least as divergent from it as other related lineages, namely the South African Lobostemon and Echiostachys. Pollen characters revealed a strong affinity with Onosma, especially in the non-reticulate ornamentation of the tectum. Fruit morphology is unique in the genera of the Echium ‘alliance' (e.g. Echium, Pontechium, Lobostemon, and Echiostachys), because of the large size and the smooth, glossy nutlet surface as in most species of Onosma. Karyological observations show that this species is diploid with 2n = 12, and differing from both Lobostemon or Echium, where nearly all species have higher numbers (usually 2n = 14 and 2n = 16, respectively). Combined with a ‘megaherb' habit, the weak zygomorphism of the large flowers and a restricted range in the humid mountain forests of the Black Sea region, fruit, pollen and chromosome characters suggest an isolated position for E. orientale in the Echium alliance. We therefore propose its placement in the monotypic Megacaryon, a genus originally described by Boissier, but largely disregarded by later botanists. An original specimen is designated as generic lectotype.     

Karyological and taxonomic notes on Cytisus Desf. Sect. Spartopsis Dumort. and Sect. Alburnoides DC. (Genisteae, Leguminosae) from the Iberian Peninsula and Morocco

Karyological information on Cytisus species indicates at least two chromosome numbers for most of the taxa. This instability is, a striking karyological feature of Cytisus . Chromosome numbers of taxa in Sect. Spartopsis and Sect. Alburnoides, both well represented in Morocco and the Iberian Peninsula, are presented here. We provide the first data on chromosome numbers for the Moroccan taxa: C. grandiflorus subsp. barbarus , and subsp. haplophyllus (n = 23, 2 n = 46) , C. maurus (2 n = 48), C. megalanthus ( n = 23), C. arboreus subsp. arboreus , subsp. baeticus , and subsp. catalaunicus (2 n = 50), C. valdesii ( n = 23 ). New populations from the Iberian Peninsula have been counted: C. grandiflorus subsp. grandiflorus (2 n = 46), C. scoparius subsp. scoparius ( n = 23) , C. striatus subsp. eriocarpus ( n = 23, 2 n = 46), C. multiflorus (n = 23), C. oromediterraneus ( n = 23, 24). Our data confirm the instability of the chromosome number in Cytisus . The presence of B chromosomes in C. valdesii and C. megalanthus , as well in other species, is discussed in relation to this instability and previous data. We suggest that instability of the chromosome number within a taxon, and even in the same population, may be related to the breakage of A chromosomes and the appearance of B chromosomes.     

Karyotype,sex determination and male meiosis in three benthic water bugs (Hemiptera: Nepomorpha: Aphelocheiridae)

We report here the first chromosome numbers for benthic water bugs (Nepomorpha: Aphelocheiridae). All three studied species, Aphelocheirus aestivalis (Fabricius, 1794), A. murcius Nieser and Millán, 1989 and Aphelocheirus sp. from Ebro River (northern Spain), have karyotype 2n = 22 + XX/X(0) and inverted sequence of the X chromosome divisions in male meiosis. The similarity and difference in cytogenetic traits between Aphelocheiridae and other families of Nepomorpha are shortly discussed.     

A karyological study of three species of Scincidae (Reptilia)

In this paper the Authors decribe the karyogram of three species of Scincidae (Chalcides chalcides chalcides, Chalcides ocellatus tiligugu, and Mabuya striata). The diploid number of these species is 2n=28. It is not possible to subdivide the chromosome set in micro- and macrochromosomes or to recognize the heterochromosomes morphologically. Ch. ocellatus and M. striata have very similar karyograms; that of Ch. ch. chalcides is different in that chromosomes 6, 7, 8, 9, and 10 are acrocentric. Pericentric inversion is probably involved in the karyotypic evolution of these species.     

Rocky Mountain species of Paronychia (Caryophyllaceae): A morphological,cytological, and chemical study

Data derived from a morphological, cytological, flavonoid, and greenhouse study indicate thatParonychia pulvinata (2n = 32) andP. sessiliflora (2n = 64) should be distinguished in specific rank rather than in subspecific rank as recently proposed. Additionally, new evidence is brought to bear onP. depressa var.brevicuspis, which is shown to be synonymous withP. sessiliflora.     

Allopolyploid origin of Cardamine silana (Brassicaceae) from Calabria (southern Italy): karyological, morphological and molecular evidence

Endemic Cardamine silana from Calabria (southern Italy) previously reported to be related to C. raphanifolia was found to be hexaploid. Morphological characters and AFLP data were analysed to evaluate the degree of differentiation of C. silana from closely related taxa and to find parental taxa of this polyploid. Cardamine apennina from the C. pratensis group was examined as one putative parent, as indicated in previous studies of nuclear ITS sequences, along with other related taxa based on both cpDNA and ITS sequences. Both multivariate morphometric analyses of quantitative characters and evaluation of qualitative morphological characters showed: (1) closest position of C. silana to two diploids: C. acris from the Balkan Peninsula and C. apennina from Central Italy; (2) good extent of morphological separation of C. silana from related taxa; and (3) within C. acris subspecies, C. acris ssp. vardousiae from Central Greece as closest to C. silana . Neighbour-joining tree and PCoA ordinations of AFLP data, as well as patterns of AFLP bands sharing, corroborated results of multivariate morphometrics. This evidence supports an allopolyploid origin of C. silana , with C. apennina and C. acris as parental taxa. Its origin may be dated to Pleistocene glacial events, because of the presumably wider geographical distributions of its parental taxa during more humid periods at that time.  © 2005 The Linnean Society of London, Botanical Journal of the Linnean Society , 2005, 148 , 101–116.     

A new species of Haplophyllum A. Juss. (Rutaceae) from the Iberian Peninsula: evidence from morphological, karyological and molecular analyses

BACKGROUND AND AIMS: The discovery of a new species, Haplophyllum bastetanum F.B. Navarro, V.N. Suárez-Santiago & Blanca sp. nov., in the south-east of Spain has prompted the comparative study of species of the Iberian Peninsula, and others related, through morphological, cytogenetic, molecular, distributional and ecological characterization. METHODS: The morphological study involved a quantitative analysis of the species present in the Iberian Peninsula and a comparative analysis of the morphological characteristics between H. bastetanum and other related species. Mitotic analyses were made with root meristems taken from germinating seeds. Phylogenetic analyses of the internal transcribed spacer sequences of nuclear ribosomal DNA were performed using neighbour-joining (NJ) and maximum-parsimony methods. KEY RESULTS: Haplophyllum bastetanum is a diploid species (2n = 18) distinguished primarily for its non-trifoliate glabrous leaves, lanceolate sepals, dark-green petals with a dorsal band of hairs, and a highly hairy ovary with round-apex locules. The other two Iberian species (H. linifolium and H. rosmarinifolium) are tetraploid (2n = 36) and have yellow petals. Both phylogenetic methods generated a well-supported clade grouping H. linifolium with H. rosmarinifolium. In the NJ tree, the H. linifolium-H. rosmarinifolium clade is a sister group to H. bastetanum, while in the parsimony analysis this occurred only when the gaps were coded as a fifth base and the characters were reweighted according to the rescaled consistency index. This latter group is supported by the sequence divergence among taxa. CONCLUSIONS: The phylogenies established from DNA sequences together with morphological and cytogenetic analyses support the separation of H. bastetanum as a new species. The results suggest that the change in the number of chromosomes may be the key mechanism of speciation of the genus Haplophyllum in the Iberian Peninsula. An evolutionary scheme for them is propounded.     

A comparative karyological study of the blue-breasted quail (Coturnix chinensis, Phasianidae) and California quail (Callipepla californica, Odontophoridae)

We conducted comparative chromosome painting and chromosome mapping with chicken DNA probes against the blue-breasted quail (Coturnix chinensis, CCH) and California quail (Callipepla californica, CCA), which are classified into the Old World quail and the New World quail, respectively. Each chicken probe of chromosomes 1-9 and Z painted a pair of chromosomes in the blue-breasted quail. In California quail, chicken chromosome 2 probe painted chromosomes 3 and 6, and chicken chromosome 4 probe painted chromosomes 4 and a pair of microchromosomes. Comparison of the cytogenetic maps of the two quail species with those of chicken and Japanese quail revealed that there are several intrachromosomal rearrangements, pericentric and/or paracentric inversions, in chromosomes 1, 2 and 4 between chicken and the Old World quail. In addition, a pericentric inversion was found in chromosome 8 between chicken and the three quail species. Ordering of the Z-linked DNA clones revealed the presence of multiple rearrangements in the Z chromosomes of the three quail species. Comparing these results with the molecular phylogeny of Galliformes species, it was also cytogenetically supported that the New World quail is classified into a different clade from the lineage containing chicken and the Old World quail.     

A comparison of morphological and chemical fruit traits between two sites with different frugivore assemblages

Large-scale comparisons might reveal matching between fruit traits and frugivore assemblages that might be cryptic on a local scale. Therefore, we compared morphological (colour, size, husk thickness) and chemical fruit traits (protein, nitrogen, sugar, lipid, tannin and fibre content) between Malagasy and South African tree communities with different frugivore communities. In Madagascar, where lemurs are important seed dispersers, we found more tree species with fruit colour classified as primate fruits. In contrast, in South Africa we found more tree species with fruits classified as bird coloured. This correlated with the greater importance of frugivorous birds in South Africa vs. Madagascar. Additionally, we found higher sugar concentrations in fruits from the South African tree community and higher fibre content in fruits from the Malagasy tree community. However, fibre content could be related to differences in abiotic conditions between the two study sites. This suggests that fruit colour more than other morphological and chemical fruit traits, reflects food selection by the different frugivore assemblages of those two sites.     

A karyological study of Asphodelus L. (Asphodelaceae) from the Western Mediterranean

The following aspects of Asphodelus karyology are analysed: base number, polyploidy, chromosome size, chromosome morphology, satellited chromosomes, structural heteromorphism, karyotype asymmetry and karyotype evolution. The base number 0 ×= 14 is common to all species except for A. refractus , which has the derived ×= 13. Three ploidy levels occur, often in the same species; diploid, tetraploid and hexaploid, with 2n = 28, 56 and 84. Chromosomes are generally small to medium-small, with the occasional presence of medium-large chromosomes. The most frequent chromosome types are metacentric of type m and submetacentric. Metacentric chromosomes of type M occur only in sections. Verineopsis, Verinea and Plagiasphodelus ; subtelocentric chromosomes occur only in sections Asphodelus and Plagiasphodelus. There is a wide variability in relation to the number of satellited chromosomes, relative to ploidy level. There are usually two to four in diploids, four to eight in tetraploids and usually six, exceptionally up to 12, in the hexaploid. Satellites are present on the shortest arm, exceptionally on the longest arm. There is a high degree of structural heteromorphism in practically all the species which affects satellited and non satellited chromosomes. Karyotype asymmetry is generally of type 2B. Inter-and intra-chromosomal differences are estimated by the A1 and A2 indexes. Both indices vary in the karyotype evolution of the genus, with a decrease of A1 and an increase of A2. The role of polyploidy, hybridization, asymmetry and decrease of chromosome size in the evolution of Asphodelus is discussed.     

A karyological study of the spirurid nematode Mastophorus muris (Nematoda: Spirocercidae)

The karyotype of Mastophorus muris (Gmelin, 1790) comprises four pairs of small autosomal chromosomes and two larger sex X chromosomes in females or one X chromosome in males (2n = 8 + XX/XO). All pairs of chromosomes possess rather uniform morphology without distinct primary or secondary constrictions. No heterochromatin bands were found by C-banding analysis. The absolute chromosome length ranges from 4.02 to 2.24 microns. The mean total length of the haploid complement is 14.34 microns. The course of gametogenesis represents a typical pattern common in the order Spirurida. The recently available karyotypes of spirurid nematodes have been reviewed.     

Cytogenetic evidence for a complex of species within the taxon Anopheles maculatus (Diptera: Culicidae)

Two largely independent studies of chromosomes from natural populations of Anopheles maculatus provide evidence for several genetic species within the taxon. (1) Polytene chromosome variation shows four different rearrangements of arm 2 and three rearrangements of the X chromosome. There is strong evidence for three species. Two allopatric populations represent either dramatic geographic variation for two independent inversion systems within one of the genetic species, or represent two additional species. Their species status remains unresolved by this work. (2) Heterochromatic variation occurs in both X and Y chromosomes as revealed by Giemsa-banding of mitotic chromosomes from larval brains. The distribution and association of these various sex chromosomes give further evidence of a species complex. A preliminary correlation of these two kinds of chromosomal variation is given.     

Multiple simultaneous speciation in killifishes of the Cynolebias adloffi species complex (Cyprinodontiformes, Rivulidae) from phylogeography and chromosome data

The annual killifishes of the genus Cynolebias Steindachner, 1876, include one of the well-supported clades composed of the Cynolebias adloffi species complex of the Bañados del Este Biosphere Reserve Site, from the Atlantic coastal wetlands in Uruguay. One member of this clade, Cynolebias charrua , has been considered as an intergradation complex of populations between C. adloffi and C. viarius by showing high levels of morphological variability. Systematic affinities between both taxa have been historically discussed. In the present study, a phylogeographic approach based on mitochondrial cytochrome b haplotype analysis and chromosome data are presented. According to the present data a scenario of a multiple simultaneous speciation process and perhaps reticulation events is possible in the C. adloffi species complex, followed by divergence associated with Quaternary marine transgressions in this region. This hypothetic scenario could explain the remarkable genetic diversity and the lack of phylogenetic resolution among clades in C. charrua , as well as the disconcordance in the phylogenetic relationships among Cynolebias from different data sets. Several parameters of DNA polymorphism and chromosome data have also suggested that populations of C. charrua from southern Laguna Merin, in eastern Uruguay and southern Brazil, could represent a relictual group from a widespread ancestral population. Chromosome rearrangements like pericentric inversions in isolated populations could have triggered cladogenetic events from this highly polymorphic population. Present analysis suggests that the metapopulation could represent the management units in these endangered killifishes and that the populations of C. charrua from the Laguna Merin basin could be included in a high-priority conservation programme.     

A morphological and chemical study of Populus acuminata Rydberg

Evidence from morphology, flavonoid chemistry, and field observations suggests thatPopulus acuminata is of hybrid origin. The putative parents areP. angustifolia, the narrow leaf cottonwood, and deltoid leaved plants that are assigned toP. sargentii (P. deltoides var.occidentalis), P. fremontii, orP. wislizenii (P. fremontii var.wislizenii). Populus angustifolia exhibits a series of flavonol glycosides (kaempferol, quercetin, and myricetin) in its leaves. By contrast, the major leaf flavonoids of the broad leaved plants are flavone glycosides (apigenin and luteolin).Populus acuminata is intermediate between the suspected parents in morphological features. Additionally, the leaves of mostP. acuminata plants contain the exact summation of the flavonoid compounds characteristic of the putative parents. A diploid chromosome number of 2n = 38 was obtained for six plants, which confirms the one previous report for the species. Meiosis was regular in all cases. Correlated data indicate that the majority of plants ofP. acuminata represent F₁ hybrids and that complex hybridization is not common. Evidence from morphological and chemical studies is presented to show that in certain instances backcrossing to both parents has occurred. Results gathered in this study show thatP. ×andrewsii is undoubtedly “typical”P. acuminata, but the type specimen is from a sucker shoot, and thus has been interpreted as a backcross toP. sargentii. Populus acuminata var.rehderi is not considered worthy of taxonomic recognition.