Phylogeographical, ecological and biological patterns shown by nuclear (ssrRNA and gGAPDH) and mitochondrial (Cyt b) genes of trypanosomes of the subgenus Schizotrypanum parasitic in Brazilian bats

The genetic diversity and phylogeographical patterns of Trypanosoma species that infect Brazilian bats were evaluated by examining 1043 bats from 63 species of seven families captured in Amazonia, the Pantanal, Cerrado and the Atlantic Forest biomes of Brazil. The prevalence of trypanosome-infected bats, as estimated by haemoculture, was 12.9%, resulting in 77 cultures of isolates, most morphologically identified as Trypanosoma cf. cruzi, classified by barcoding using partial sequences from ssrRNA gene into the subgenus Schizotrypanum and identified as T. cruzi (15), T. cruzi marinkellei (37) or T. cf. dionisii (25). Phylogenetic analyses using nuclear ssrRNA, glycosomal glyceraldehyde 3-phosphate dehydrogenase (gGAPDH) and mitochondrial cytochrome b (Cyt b) gene sequences generated three clades, which clustered together forming the subgenus Schizotrypanum. In addition to vector association, bat trypanosomes were related by the evolutionary history, ecology and phylogeography of the bats. Trypanosoma cf. dionisii trypanosomes (32.4%) infected 12 species from four bat families captured in all biomes, from North to South Brazil, and clustered with T. dionisii from Europe despite being separated by some genetic distance. Trypanosoma cruzi marinkellei (49.3%) was restricted to phyllostomid bats from Amazonia to the Pantanal (North to Central). Trypanosoma cruzi (18.2%) was found mainly in vespertilionid and phyllostomid bats from the Pantanal/Cerrado and the Atlantic Forest (Central to Southeast), with a few isolates from Amazonia.     

Genome size in Hieracium subgenus Hieracium (Asteraceae) is strongly correlated with major phylogenetic groups

Background and Aims

Hieracium subgenus Hieracium is one of the taxonomically most intricate groups of vascular plants, due to polyploidy and a diversity of breeeding systems (sexuality vs. apomixis). The aim of the present study was to analyse nuclear genome size in a phylogenetic framework and to assess relationships between genome size and ploidy, breeding system and selected ecogeographic features.

Methods

Holoploid and monoploid genome sizes (C- and Cx-values) of 215 cultivated plants from 89 field populations of 42 so-called ‘basic’ Hieracium species were determined using propidium iodide flow cytometry. Chromosome counts were available for all analysed plants, and all plants were tested experimentally for their mode of reproduction (sexuality vs. apomixis). For constructing molecular phylogenetic trees, the external transcribed spacer region of nuclear ribosomal DNA was used.

Key Results

The mean 2C values differed up to 2·37-fold among different species (from 7·03 pg in diploid to 16·67 in tetraploid accessions). The 1Cx values varied 1·22-fold (between 3·51 and 4·34 pg). Variation in 1Cx values between conspecific (species in a broad sense) accessions ranged from 0·24% to 7·2%. Little variation (not exceeding the approximate measurement inaccurracy threshold of 3·5%) was found in 33 species, whereas variation higher than 3·5% was detected in seven species. Most of the latter may have a polytopic origin. Mean 1Cx values of the three cytotypes (2n, 3n and 4n) differed significantly (average of 3·93 pg in diploids, 3·82 pg in triploids and 3·78 pg in tetraploids) indicating downsizing of genomes in polyploids. The pattern of genome size variation correlated well with two major phylogenetic clades which were composed of species with western or eastern European origin. The monoploid genome size in the ‘western’ species was significantly lower than in the ‘eastern’ ones. Correlation of genome size with latitude, altitude and selected ecological characters (light and temperature) was not significant. A longitudinal component was only apparent for the whole data set, but absent within the major lineages.

Conclusions

Phylogeny was the most important factor explaining the pattern of genome size variation in Hieracium sensu stricto, species of western European origin having significantly lower genome size in comparison with those of eastern European origin. Any correlation with ecogeographic variables, including longitude, was outweighed by the divergence of the genus into two major phylogenetic lineages.Key words: Apomixis, chromosome numbers, Compositae, genome size, hawkweeds, Hieracium subgenus Hieracium, mode of reproduction, nuclear DNA content, phylogeny, polyploidy     

Anthocyanin content of Tulipa species and cultivars and its impact on tepal colours

Flowers of tulips (17 species and 25 cultivars) were subjected to qualitative and relative quantitative examination for anthocyanins. Altogether five anthocyanins were identified as the 3-O-(6″-O-α-rhamnopyranosyl-β-glucopyranoside) of delphinidin (1), cyanidin (2) and pelargonidin (3), and the 3-O-[6″-O-(2‴-O-acetyl-α-rhamnopyranosyl)-β-glucopyranoside] of cyanidin (4) and pelargonidin (5). The pigments 1–5 represented 7%, 43%, 12%, 2% and 31%, respectively, of the total anthocyanin amount in the tepals of the Tulipa species, and 20%, 37%, 30%, 6% and 4%, respectively, in the cultivar tepals. Nearly 50% of the samples contained acetylated anthocyanins. The colours of the freeze-dried tepals described by the CIELab coordinates, hue angle (h_ab), saturation (C*), and lightness (L*) together with the anthocyanin content were subjected to multivariate analysis. All tepals classified with hue angles described as “blue nuances” were from cultivars. They contained 1 as the major anthocyanin, and no or just traces of pelargonidin derivatives. The species and cultivars having “magenta nuances” showed similar anthocyanin content with increased relative proportions of 2 at the expense of 1. Orange coloured tepals were to a large extent correlated with high relative proportions of the pelargonidin derivatives, 3 and 5. Acetylation of anthocyanins furnished a weak colour effect opposite to the bluing effect previously reported for anthocyanins with aromatic acyl groups. All six species belonging to the section Eichleres (subgenus Tulipa) were after principal component analysis grouped closely together. They were characterized by high concentrations of the pelargonidin derivatives 3 and 5, and orange petal nuances. However, within section Tulipa (subgenus Tulipa), considerable anthocyanin variation was observed. Species in the subgenus Eriostemones were generally characterized by the two anthocyanins 1 and 2, and no pelargonidin derivatives.     

Leaf flavonoid glycosides as chemosystematic characters in Ocimum

Thirty-one accessions of nine species belonging to three subgenera of Ocimum (basil, family Lamiaceae) were surveyed for flavonoid glycosides. Substantial infraspecific differences in flavonoid profiles of the leaves were found only in O. americanum, where var. pilosum accumulated the flavone C-glycoside, vicenin-2, which only occurred in trace amounts in var. americanum and was not detected in cv. Sacred. The major flavonoids in var. americanum and cv. Sacred, and also in all other species investigated for subgenus Ocimum, were flavonol 3-O-glucosides and 3-O-rutinosides. Many species in subgenus Ocimum also produced the more unusual compound, quercetin 3-O-(6″-O-malonyl)glucoside, and small amounts of flavone O-glycosides. The level of flavonol glycosides produced was reduced significantly in glasshouse-grown plants, but levels of flavone glycosides were unaffected. A single species investigated from subgenus Nautochilus, O. lamiifolium, had a different flavonoid glycoside profile, although the major compound was also a flavonol O-glycoside. This was identified as quercetin 3-O-xylosyl(1‴→2″)galactoside, using NMR spectroscopy. The species investigated from subgenus Gymnocimum, O. tenuiflorum (=O. sanctum), was characterised by the accumulation of flavone O-glycosides. These were isolated, and identified as the 7-O-glucuronides of luteolin and apigenin. Luteolin 5-O-glucoside was found in all nine species of Ocimum studied, and is considered to be a key character for the genus.     

Evolution of mixed-linkage (1 -> 3, 1 -> 4)-β-D-glucan (MLG) and xyloglucan in Equisetum (horsetails) and other monilophytes

Background and Aims

Horsetails (Equisetopsida) diverged from other extant eusporangiate monilophytes in the Upper Palaeozoic. They are the only monilophytes known to contain the hemicellulose mixed-linkage (1 → 3, 1 → 4)-β-d-glucan (MLG), whereas all land plants possess xyloglucan. It has been reported that changes in cell-wall chemistry often accompanied major evolutionary steps. We explored changes in hemicelluloses occurring during Equisetum evolution.

Methods

Hemicellulose from numerous monilophytes was treated with lichenase and xyloglucan endoglucanase. Lichenase digests MLG to di-, tri- and tetrasaccharide repeat-units, resolvable by thin-layer chromatography.

Key Results

Among monilophytes, MLG was confined to horsetails. Our analyses support a basal trichotomy of extant horsetails: MLG was more abundant in subgenus Equisetum than in subgenus Hippochaete, and uniquely the sister group E. bogotense yielded almost solely the tetrasaccharide repeat-unit (G4G4G3G). Other species also gave the disaccharide, whereas the trisaccharide was consistently very scarce. Tetrasaccharide : disaccharide ratios varied interspecifically, but with no consistent difference between subgenera. Xyloglucan was scarce in Psilotum and subgenus Equisetum, but abundant in subgenus Hippochaete and in the eusporangiate ferns Marattia and Angiopteris; leptosporangiate ferns varied widely. All monilophytes shared a core pattern of xyloglucan repeat-units, major XEG products co-chromatographing on thin-layer chromatography with non-fucosylated hepta-, octa- and nonasaccharides and fucose-containing nona- and decasaccharides.

Conclusions

G4G4G3G is the ancestral repeat-unit of horsetail MLG. Horsetail evolution was accompanied by quantitative and qualitative modification of MLG; variation within subgenus Hippochaete suggests that the structure and biosynthesis of MLG is evolutionarily plastic. Xyloglucan quantity correlates negatively with abundance of other hemicelluloses; but qualitatively, all monilophyte xyloglucans conform to a core pattern of repeat-unit sizes.     

Unexpectedly high diversity of trypanosomes in small sub-Saharan mammals

The extremely species-rich genus Trypanosoma has recently been divided into 16 subgenera, most of which show fairly high host specificity, including the subgenus Herpetosoma parasitizing mainly rodents. Although most Herpetosoma spp. are highly host-specific, the best-known representative, Trypanosoma lewisi, has a cosmopolitan distribution and low host specificity. The present study investigates the general diversity of small mammal trypanosomes in East and Central Africa and the penetration of invasive T. lewisi into communities of native rodents. An extensive study of blood and tissue samples from Afrotropical micromammals (1528 rodents, 135 shrews, and five sengis belonging to 37 genera and 133 species) captured in the Central African Republic, Ethiopia, Kenya, Malawi, Mozambique, Tanzania, and Zambia revealed 187 (11.2%) trypanosome-positive individuals. The prevalence of trypanosomes in host genera ranged from 2.1% in Aethomys to 37.1% in Lemniscomys. The only previously known trypanosome detected in our dataset was T. lewisi, newly found in Ethiopia, Kenya, and Tanzania in a wide range of native rodent hosts. Besides T. lewisi, 18S rRNA sequencing revealed 48 additional unique Herpetosoma genotypes representing at least 15 putative new species, which doubles the known sequence-based diversity of this subgenus, and approaches the true species richness in the study area. The other two genotypes represent two new species belonging to the subgenera Ornithotrypanum and Squamatrypanum. The trypanosomes of white-toothed shrews (Crocidura spp.) form a new phylogroup of Herpetosoma, unrelated to flagellates previously detected in insectivores. With 13 documented species, Ethiopia was the richest region for trypanosome diversity, which corresponds to the very diverse environments and generally high biodiversity of this country. We conclude that besides T. lewisi, the subgenus Herpetosoma is highly host-specific (e.g., species parasitizing the rodent genera Acomys and Gerbilliscus). Furthermore, several newly detected trypanosome species are specific to their endemic hosts, such as brush-furred mice (Lophuromys), dormice (Graphiurus), and white-toothed shrews (Crocidura).     

Chromosomal and molecular studies of cryptic species of the subgenus <Emphasis Type="Italic">Terricola</Emphasis> (Rodentia,Arvicolinae, <Emphasis Type="Italic">Microtus</Emphasis>) in the Caucasian region: Analysis of new records

Using chromosomal and molecular (cytb) markers, the specific identity of 78 individuals of cryptic species of voles of the subgenus Terricola was ascertained. The animal samples were obtained at 13 localities of the Caucasian region, from Krasnodar krai to North Ossetia in the Greater Caucasus and in the Stavropol Upland (forest island in the steppe) in Ciscaucasia, which had not been covered earlier in genetic studies. In the total sample, two chromosomal forms (cryptic species), namely, Microtus (Terricola) majori (2n = 54, NF = 60) and M. (T.) daghestanicus (2n = 54, NF = 58), were identified. The specific identity of most animals tested was confirmed by karyological means, and for individuals from two localities on the south slopes of the Greater Caucasus (Adlerskii district of Krasnodar krai), it was confirmed exclusively by means of molecular (cytb) markers. The last two records were used for evaluation of the differential role of sibling-species of the subgenus Terricola in circulation of the pathogen in mountain loci of hemorrhagic fever with renal syndrome (HFRS) in the Western Caucasus. For the first time, using the example of M. (T.) majori from the Ciscaucasia, which was compared with those of the mountain part of the species’ natural habitat, the role of isolation factors in morphogenesis of the subgenus Terricola was studied. As a whole, the results obtained specify the character of the geographical distribution and biotope attachment of cryptic species of the subgenus Terricola in the Caucasian region.     

From open areas to forests? The evolutionary history of Philodendron subgenus Meconostigma (Araceae) using morphological data

Due to its wide distribution covering three of the largest Neotropical biomes, Philodendron subgenus Meconostigma is an interesting model to discuss the diversification of Neotropical plants. The aim of this study was to test a previous hypothesis that the Eastern and Southern species of P. subg. Meconostigma have plesiomorphic gynoecial structure while in Amazonian species they are apomorphic. To this end, we conducted an analysis of maximum parsimony with generalized frequency coding method using a matrix with 59 morphological characters and 90% of the species of P. subg. Meconostigma. The phylogenetic reconstruction suggests that the subgenus is monophyletic and originates from open areas of Cerrado. Four morphological synapomorphies support the monophyly of the subgenus and seven synapomorphies support four minor clades within it. Our results also include characterization of three new gynoecial subtypes (A1, A2 and A3) within the subgenus. Subtype A2 (undeveloped stylar body with long stylar canals, absent central stylar dome, shallow compitum) is basal and represents the ancestral gynoecium in the group. These findings suggest that the balance between pollen uptake and accessibility of the locules were decisive to drive gynoecium evolution in the subgenus Meconostigma.     

An unusual new species of the genus Brachypalpus Macquart (Diptera: Syrphidae) from Eastern Asia

Brachypalpus (Brachypalpus) longifacies sp. nov. is described and illustrated from Eastern Asia (Russian Far East and Japan). It differs from other species of the nominative subgenus by several unusual characters, i.e. face in profile weakly convex dorsally and concave ventrally, antennae very elongated, and abdomen with reddish transversal maculae on 2nd tergum (male) or 2nd and 3rd terga (female). A key is given for known species of the subgenus Brachypalpus.LSID: urn:lsid:zoobank.org:pub:634CB6E2-B2C1-40EC-8811-BF3DD4BDE69A     

Divergence of genomic DNA in species of the subgenus Camptochironomus (Diptera, Chironomidae) differing in their cytogenetic similarity levels

The variation and divergence of genomic DNA in four species of the subgenus Camptochironomus (C. tentans, C. dilutus, C. pallidivittatus, and C. setivalva) differing in the level of their cytological similarity were analyzed using the RAPD (Randomly Amplified Polymorphic DNA) method. A high level of variation in the RAPD markers was found in the species studied. Genetic distances (GD) were assessed between natural C. tentans populations, between different species of the camptochironomus sibling species group (C. tentans, C. dilutus, and C. pallidivittatus), and between these species and C. setivalva which is outside this sibling species group. The GD values obtained characterize the levels of genomic differentiation among natural populations (GD = 0.248), among sibling species (GD = 0.635), and between incipient species (GD = 0.784) of the subgenus Camptochironomus. The degree of genomic DNA divergence between sibling and incipient species in the subgenus Camptochironomus was found to be lower than that in the genus Chironomus. The rate of genomic DNA divergence appears to be lower than the rate of chromosomal divergence in species of the subgenus Camptochironomus.     

Molecular phylogeny of the subgenus Ceratotropis (genus Vigna, Leguminosae) reveals three eco-geographical groups and Late Pliocene-Pleistocene diversification: evidence from four plastid DNA region sequences

Background and Aims

The subgenus Ceratotropis in the genus Vigna is widely distributed from the Himalayan highlands to South, Southeast and East Asia. However, the interspecific and geographical relationships of its members are poorly understood. This study investigates the phylogeny and biogeography of the subgenus Ceratotropis using chloroplast DNA sequence data.

Methods

Sequence data from four intergenic spacer regions (petA-psbJ, psbD-trnT, trnT-trnE and trnT-trnL) of chloroplast DNA, alone and in combination, were analysed using Bayesian and parsimony methods. Divergence times for major clades were estimated with penalized likelihood. Character evolution was examined by means of parsimony optimization and MacClade.

Key Results

Parsimony and Bayesian phylogenetic analyses on the combined data demonstrated well-resolved species relationships in which 18 Vigna species were divided into two major geographical clades: the East Asia–Southeast Asian clade and the Indian subcontinent clade. Within these two clades, three well-supported eco-geographical groups, temperate and subtropical (the East Asia–Southeast Asian clade) and tropical (the Indian subcontinent clade), are recognized. The temperate group consists of V. minima, V. nepalensis and V. angularis. The subtropical group comprises the V. nakashimae–V. riukiuensis–V. minima subgroup and the V. hirtella–V. exilis–V. umbellata subgroup. The tropical group contains two subgroups: the V. trinervia–V. reflexo-pilosa–V. trilobata subgroup and the V. mungo–V. grandiflora subgroup. An evolutionary rate analysis estimated the divergence time between the East Asia–Southeast Asia clade and the Indian subcontinent clade as 3·62 ± 0·3 million years, and that between the temperate and subtropical groups as 2·0 ± 0·2 million years.

Conclusions

The findings provide an improved understanding of the interspecific relationships, and ecological and geographical phylogenetic structure of the subgenus Ceratotropis. The quaternary diversification of the subgenus Ceratotropis implicates its geographical dispersal in the south-eastern part of Asia involving adaptation to climatic condition after the collision of the Indian subcontinent with the Asian plate. The phylogenetic results indicate that the epigeal germination is plesiomorphic, and the germination type evolved independently multiple times in this subgenus, implying its limited taxonomic utility.     

Veronica: Acylated flavone glycosides as chemosystematic markers

HPLC/DAD and LC–MS of an extract of Veronica spicata (subgenus Pseudolysimachium, Plantaginaceae) revealed the presence of six 6-hydroxyluteolin glycosides acylated with phenolic acids, three of which are new compounds and which we called spicosides. A flavonoid survey of seven more species belonging to V. subgenus Pseudolysimachium and eight species of V. subgenus Pentasepalae showed that all the Pseudolysimachium species and four of the Pentasepalae species produced 6-hydroxyflavone glycosides, whereas the remaining four Pentasepalae species contained acetylated 8-hydroxyflavone glycosides instead. Spicosides appeared to be common in subgenus Pseudolysimachium (detected in five out of eight species), but we did not find them in subgenus Pentasepalae. Previously, acetylated 8-hydroxyflavone glycosides have been isolated from or detected in eight species of V. subgenus Pentasepalae (in 13 species altogether including the new results) and seven species of V. subgenus Pocilla. However, 6-hydroxyflavone glycosides occur in some other species of these subgenera. The results are discussed in an evolutionary context.     

A comparison of mandibular gland volatiles from ants of the bull horn acacia,Acacia collinsii

GC–MS analyses of dichloromethane extracts of the mandibular glands from three obligate symbiotic Psuedomyrmex ant species of Acacia collinsii from Costa Rica: Pseudomyrmex flavicornis (synonym Pseudomyrmex belti), Psuedomyrmex spinicola, and Psuedomyrmex nigrocincta, showed distinct differences in the 16 ketones, 15 alcohols, 2 aldehydes and 2 carboxylic acids that were identified. Different compounds were the major component from each species: P. flavicornis, 3-octanone; P. spinicola, 4-methyl-3-heptanone; and P. nigrocincta, 3-methyl-2-hexanol. The secretion of P. flavicornis contained 10 compounds not found in the other species, including the two terpene alcohols, citronellol and geraniol. The secretions of P. spinicola and P. nigrocincta had 12 compounds in common, that were not found in P. flavicornis' secretion. The similarity of the mandibular gland secretion of P. spinicola and P. nigrocincta may indicate that they are more closely related to each other than either is to P. flavicornis. The components from the mandibular gland of Crematogaster rochai, another ant associated with this acacia, are 2- and 3-methylbutanoic acid, 3-octanone, 3-octanol, 6-methyl-3-octanol and 3-nonanone.     

Molecular phylogeny of black flies (Diptera: Simuliidae) from Thailand, using ITS2 rDNA

The sequences of the second internal transcribed spacer (ITS2) of ribosomal DNA (rDNA) were determined for 40 black fly species from Thailand, belonging to 4 subgenera of the genus Simulium, namely Gomphostilbia (12 species), Nevermannia (5 species), Montisimulium (1 species), Simulium sensu stricto (21 species), and an unknown subgenus with one species (Simulium baimaii). The length of the ITS2 ranged from 247 to 308 bp. All black fly species had high AT content, ranging from 71 to 83.8%. Intraindividual variation (clonal variation) occurred in 13 species, ranging from 0.3 to 1.1%. Large intrapopulation and interpopulation heterogeneities exist in S. feuerboni from the same and different locations in Doi Inthanon National Park, northern Thailand. Phylogenetic relationships among 40 black fly species were examined using PAUP (version 4.0b10) and MrBAYS (version 3.0B4). The topology of the trees revealed two major monophyletic clades. The subgenus Simulium and Simulium baimaii were placed in the first monophyletic clade, whereas the subgenera Nevermannia + Montisimulium were placed as the sister group to the subgenus Gomphostilbia in the second monophyletic clade. Our results suggest that S. baimaii belongs to the malyschevi-group or variegatum-group in the subgenus Simulium. The molecular phylogeny generally agrees with existing morphology-based phylogenies.     

Evolutionary History and Novel Biotic Interactions Determine Plant Responses to Elevated CO2 and Nitrogen Fertilization

A major frontier in global change research is predicting how multiple agents of global change will alter plant productivity, a critical component of the carbon cycle. Recent research has shown that plant responses to climate change are phylogenetically conserved such that species within some lineages are more productive than those within other lineages in changing environments. However, it remains unclear how phylogenetic patterns in plant responses to changing abiotic conditions may be altered by another agent of global change, the introduction of non-native species. Using a system of 28 native Tasmanian Eucalyptus species belonging to two subgenera, Symphyomyrtus and Eucalyptus, we hypothesized that productivity responses to abiotic agents of global change (elevated CO₂ and increased soil N) are unique to lineages, but that novel interactions with a non-native species mediate these responses. We tested this hypothesis by examining productivity of 1) native species monocultures and 2) mixtures of native species with an introduced hardwood plantation species, Eucalyptus nitens, to experimentally manipulated soil N and atmospheric CO₂. Consistent with past research, we found that N limits productivity overall, especially in elevated CO₂ conditions. However, monocultures of species within the Symphyomyrtus subgenus showed the strongest response to N (gained 127% more total biomass) in elevated CO₂ conditions, whereas those within the Eucalyptus subgenus did not respond to N. Root:shoot ratio (an indicator of resource use) was on average greater in species pairs containing Symphyomyrtus species, suggesting that functional traits important for resource uptake are phylogenetically conserved and explaining the phylogenetic pattern in plant response to changing environmental conditions. Yet, native species mixtures with E. nitens exhibited responses to CO₂ and N that differed from those of monocultures, supporting our hypothesis and highlighting that both plant evolutionary history and introduced species will shape community productivity in a changing world.     

Evolution of Iris subgenus Xiphium based on chromosome numbers, FISH of nrDNA (5S, 45S) and trnL–trnF sequence analysis

The subgenus Xiphium is one of the six infrageneric divisions of the genus Iris. Chromosome numbers of six of the seven Xiphium species are known. Here the aim was to infer genetic and phylogenetic relationships based on chromosome numbers, chromosome markers and plastid sequences. Chromosomal locations of 5S and 45S rDNA loci were determined in 19 populations of the 7 species by fluorescence in situ hybridization (FISH). Additionally, the trnL–trnF plastid spacer was sequenced and a phylogenetic analysis performed. Based on chromosome markers, subgenus Xiphium species were classified into four groups that differed in the number and locations of both types of nrDNA: (1) I. tingitana (2n = 28), I. filifolia (2n = 30, 34) and I. xiphium (2n = 34), (2) I. juncea (2n = 32) and I. boissieri (2n = 36), (3) I. serotina (2n = 34) and (4) I. latifolia (2n = 42). Although the trnL–trnF phylogeny was not fully resolved, the sequence analysis showed a well-supported subgroup of I. filifolia, I. tingitana and I. xiphium, as well as I. juncea. FISH physical maps of the Iris subgenus Xiphium taxa are species dependent. I. filifolia, I. tingitana and I. xiphium are very closely related species and share cytogenetic characteristics. Disploidy appears to have been central in the evolution of this subgenus, given a series of chromosome numbers (2n = 28, 30, 32, 34, 36, 42) and our phylogenetic results. Clear differences were found among European and African populations of I. filifolia. A different taxonomic treatment of I. filifolia is supported for populations on both sides of the Strait of Gibraltar.     

Two new species and a new subgenus of toothed Brachyhypopomus electric knifefishes (Gymnotiformes,Hypopomidae) from the central Amazon and considerations pertaining to the evolution of a monophasic electric organ discharge

We describe two new, closely related species of toothed Brachyhypopomus (Hypopomidae: Gymnotiformes: Teleostei) from the central Amazon basin and create a new subgenus for them. Odontohypopomus, new subgenus of Brachyhypopomus, is diagnosed by (1) small teeth present on premaxillae; (2) medialmost two branchiostegal rays thin with blades oriented more vertically than remaining three rays; (3) background color in life (and to lesser extent in preservation) distinctly yellowish with head and sides peppered with small, widely spaced, very dark brown stellate chromatophores that greatly contrast with light background coloration; (4) a dark blotch or bar of subcutaneous pigment below the eye; (5) electric organ discharge waveform of very long duration (head-positive phase approx. 2 milliseconds or longer, head-negative phase shorter or absent) and slow pulse repetition rate (3–16 Hz). The type species of the new subgenus, Brachyhypopomus (Odontohypopomus) walteri sp. n., is diagnosed by the following additional character states: (1) subcutaneous dark pigment at base of orbit particularly prominent, (2) body semi-translucent and nearly bright yellow background coloration in life, (3) a biphasic electric organ discharge (EOD) waveform of very long duration (between 3.5 and 4 milliseconds at 25° C) with head-positive first phase significantly longer than second head-negative phase in both sexes. Brachyhypopomus (Odontohypopomus) bennetti sp. n. is diagnosed by two character states in addition to those used to diagnose the subgenus Odontohypopomus: (1) a deep electric organ, visible as large semi-transparent area, occupying approximately 14–17% body depth directly posterior to the abdominal cavity in combination with a short, but deep, caudal filament, and (2) a monophasic, head-positive EOD waveform, approximately 2.1 milliseconds in duration in both sexes. These are the only described rhamphichthyoid gymnotiforms with oral teeth, and Brachyhypopomus bennetti is the first Brachyhypopomus reported to have a monophasic (head-positive) EOD waveform. Unlike biphasic species, the waveform of its EOD is largely unaffected by tail damage from predators. Such injuries are common among specimens in our collections. This species’ preference for floating meadow habitat along the major channels of the Amazon River basin may put it at particularly high risk of predation and “tail grazing.”     

基于mtDNA和rDNA基因序列的中国按蚊属塞蚊亚属种类的系统发育研究（双翅目：蚊科）

【目的】应用mtDNA和rDNA基因特征重建中国按蚊属塞蚊亚属已知种类的系统发育关系, 以阐明亚属内各蚊种的亲缘关系。【方法】对采自中国的按蚊属塞蚊亚属Anopheles (Cellia) 20种蚊的mtDNA-COⅡ和 rDNA-28S-D3序列进行测定和分析, 以按蚊属按蚊亚属Anopheles (Anopheles)的中华按蚊An. (An.) sinensis和赫坎按蚊An. (An.) hyrcanus为外群, 采用COⅡ和D3单基因, 以及“COⅡ+D3”联合数据组以邻接法（NJ）、 最大简约法（MP）、 最大似然法（ML）和贝叶斯法（BI）等重建这些种类的系统发育树。【结果】 mtDNA-COⅡ和rDNA-28S-D3序列的长度范围分别为685 bp和375～410 bp, 在塞蚊亚属蚊种间的遗传距离分别为0.015～0.117和0.003～0.111。各系统树显示外群被合理分开,除在COⅡ树中新塞蚊系为并系外,各系均聚为单系群,新迈蚊系和迈蚊系亲缘关系最近。联合数据组构建的系统合意树显示中国塞蚊亚属各蚊种形成4支,除伪威氏按蚊与多斑按蚊种团未聚为单系群外,其他各种团和复合体成员种均分别聚在一起,各分支的置信值均大于50%。【结论】本研究获得的分子系统发育树清楚地显示了中国按蚊属塞蚊亚属各种类及系之间的系统发育关系, 对其分类和防治研究具有参考价值。     

The systematic value of nuclear genome size for “all” species of Tulipa L. (Liliaceae)

Nuclear genome size, as measured by flow cytometry with propidium iodide, was used to investigate the relationships within the genus Tulipa L. (Liliaceae). More than 400 accessions representing 123 taxa from mainly wild-collected plants were investigated. Most species of Tulipa have the same basic chromosome number, 2n = 2x = 24. However, the somatic DNA 2C value (2C) is shown to range from 32 to 69 pg for the diploids. The largest genome contains roughly 3.4 × 10¹⁰ more base pairs than the smallest and has chromosomes that are more than twice as large. These large differences in the amount of nuclear DNA predict that the hybrids, if any arise, are usually sterile. Depending on the size of the total genome, 1 pg amounts to several thousand genes. Moreover, genome sizes are evaluated here in combination with available morphological, geographical, and molecular data. Therefore, the taxonomy proposed here is not a single-character taxonomy based on genome size alone. The genus Tulipa, as here determined, has 87 species, 29 more than accepted by van Raamsdonk et al. [Acta Hort (ISHS) 430:821–828, 1997], but including 25 species that were not available to them. Of these 87 species, 28 were not seen by Hall (The genus Tulipa, The Royal Horticultural Society, London, 1940) in a living state and placed by him in an addendum. Species of the subgenus Clusianae (Baker) Zonn. differ strongly in nuclear DNA content (DNA 2C value), 32 versus 40–68 pg for all other tulips, and are placed here in a separate subgenus. Also Orithyia, the only group with a style and with only 38–39 pg is placed in a separate subgenus. Therefore, all tulips are attributed to four subgenera, Clusianae (Baker) Zonn., Tulipa, Eriostemones Raamsd., and Orithyia (D. Don) Baker and divided further into 12 sections. Seven of the eight series of section Eichleres (A.D. Hall) Raamsd. are now placed in four sections: (1) section Lanatae (Raamsd.) Zonn., mainly confined to species from the Pamir-Alay and including series Lanatae Raamsd., (2) section Multiflorae (Raamsd.) Zonn. (including series Glabrae Raamsd.), (3) section Vinistriatae (Raamsd.) Zonn. (including series Undulatae Raamsd.), and (4) section Spiranthera Vved. ex Zonn. and Veldk. Triploids, tetraploids, and pentaploids were found in several species. DNA content confirmed the close relationships of the species within the different sections. The rather similar looking and therefore often confused T. armena Boiss. (51.8 pg), T. systola Stapf (56.3 pg), and T. julia K., Koch (61.6 pg) could be clearly distinguished. The same is true for T. biebersteiniana Schult. f. (56.9 pg), T. sylvestris ssp. australis (Link) Pamp. (62.0 pg), and T. primulina Baker (64.6 pg). T. doerfleri Gand. and T. whittalli (Dykes) Hall could be placed as polyploid forms of T. orphanidea Boiss. ex Heldr. On the basis of DNA content, a systematic association between T. julia K. Koch and the triploid T. aleppensis Boiss. and between T. systola Stapf and the triploid T. praecox Tenore was suggested. The new species T. lemmersii Zonn., Peterse, and de Groot is described, and four possible new species are indicated. Genome size as measured by using flow cytometry may conveniently be used to produce systematic data. It is applicable even in the case of dormant bulbs or sterile plants for monitoring the trade in bulbous species.