Genetic variation of an endangered Malagasy frog, <Emphasis Type="Italic">Mantella cowani</Emphasis>, and its phylogeographic relationship to the widespread <Emphasis Type="Italic">M. baroni</Emphasis>

We investigated the degree and distribution of the genetic variation, and phylogeography, of two species of Malagasy poison frogs, Mantella cowani and M. baroni. The former is critically endangered due to its restricted distribution, habitat destruction and overcollection for the pet trade. Analysis of 526 bp of mtDNA (cytochrome b) resulted in separate haplotype networks for the two species, and discovered hybridization at a single locality. The two networks confirm the status of M. baroni and M. cowani as separate evolutionary species and units for conservation. Within both mitochondrial haplotype networks, specimens from different localities shared numerous identical haplotypes, even those from the most distant sample sites of M. baroni. Most populations were characterized by high haplotype diversity and no haplotype clades exclusive to geographical regions were observed. Protection of a few large populations of these species is therefore likely to conserve much of the mtDNA genetic diversity found in the entire species. While M. baroni is widespread and occurs in many nature reserves, we recommend efficient legal protection of some M. cowani habitats to protect this species against extinction.     

Molecular phylogeny of Malagasy poison frogs,genus Mantella (Anura: Mantellidae): homoplastic evolution of colour pattern in aposematic amphibians

We studied the evolution of colour pattern in Malagasy poison frogs, genus Mantella, a group of diurnal and toxic frogs endemic to Madagascar. Based on a phylogeny reconstructed using 1130 bp of the mitochondrial 16S rRNA gene, the genus can be divided into five species groups. Within some of these groups, interspecific genetic divergences were very low (1.2–2.8% sequence divergence) while colour patterns were markedly different. In contrast, Mantella madagascariensis and M. baroni, two species which show extremely similar dorsal coloration patterns, were not included in the same clade. This conclusion was supported by high bootstrap values and by significant rejection of alternative topologies using KH-tests. Analysis of colour patterns and tentative reconstruction of ancestral states yielded five character states shared by these two species but not by their respective sister species, M. aurantiaca and M. nigricans. Considering these detailed similarities as symplesiomorphic therefore requires the assumption of multiple reversals in other species, whereas a homoplastic colour evolution in the sympatric M. madagascariensis andM. baroni appears as most parsimonious. This parallelism may have been triggered by MÜllerian mimicry. However, additional data is necessary to support this hypothesis.     

Decoupling litter barrier and soil moisture influences on the establishment of an invasive grass

Background and aims

Through recruitment, plants establish in novel environments. Recruitment also is the stage where plants undergo the highest mortality. We investigate the recruitment niche for Microstegium vimineum, an annual grass from East Asia spreading throughout eastern North American forests.

Methods

Current observational and greenhouse research indicates that M. vimineum recruitment may be inhibited by leaf litter and promoted by soil moisture; we use field studies to experimentally test how these factors influence M. vimineum germination, seedling survival and reproduction. Specifically, we introduce M. vimineum seeds into forest microhabitats with experimentally varied levels of soil moisture and leaf litter.

Results

Soil moisture increases M. vimineum germination regardless of leaf litter thickness and ameliorates seedling mortality in deep leaf litter. Seed production per m² increases with watering, reflecting higher germination and survival, whereas per capita seed production increases with leaf litter thickness, reflecting density-dependent limits on seed production.

Conclusions

The interactive effects of varied levels of soil moisture and leaf litter thickness on key M. vimineum life history stages highlight the need to consider multiple drivers, such as rainfall and local forest disturbance, when assessing how soil properties influence the establishment of invasive plants.     

Progeny-testing of full-sibs IBD in a SSC2 QTL region highlights epistatic interactions for fatness traits in pigs

Background

Tetraena mongolica (Zygophyllaceae), an endangered endemic species in western Inner Mongolia, China. For endemic species with a limited geographical range and declining populations, historical patterns of demography and hierarchical genetic structure are important for determining population structure, and also provide information for developing effective and sustainable management plans. In this study, we assess genetic variation, population structure, and phylogeography of T. mongolica from eight populations. Furthermore, we evaluate the conservation and management units to provide the information for conservation.

Results

Sequence variation and spatial apportionment of the atp B- rbc L noncoding spacer region of the chloroplast DNA were used to reconstruct the phylogeography of T. mongolica. A total of 880 bp was sequenced from eight extant populations throughout the whole range of its distribution. At the cpDNA locus, high levels of genetic differentiation among populations and low levels of genetic variation within populations were detected, indicating that most seed dispersal was restricted within populations.

Conclusions

Demographic fluctuations, which led to random losses of genetic polymorphisms from populations, due to frequent flooding of the Yellow River and human disturbance were indicated by the analysis of BEAST skyline plot. Nested clade analysis revealed that restricted gene flow with isolation by distance plus occasional long distance dispersal is the main evolutionary factor affecting the phylogeography and population structure of T. mongolica. For setting a conservation management plan, each population of T. mongolica should be recognized as a conservation unit.     

Asymmetric introgression between Magnolia stellata and M. salicifolia at a site where the two species grow sympatrically

In order to understand the ongoing evolutionary relationships between species, it is important to elucidate patterns of natural hybridization. In the zone where two species are sympatrically distributed, we examined 274 individuals of Magnolia stellata, Magnolia salicifolia, and their putative hybrids by means of 16 nuclear and three chloroplast microsatellite markers. Hybrid classes of individuals were estimated by admixture analyses. Morphological traits were also investigated for 64 of the 274 individuals. Admixture analyses revealed that 66 of the 274 individuals were classified as hybrids, comprising 17 F₁ and 19 F₂ individuals, 27 backcrosses to M. salicifolia, and 3 individuals of unknown origin. Morphological data from the 64 individuals agreed well with their genetic admixture rates. Spatial locations of F₁ and F₂ hybrids at the study site were intermediate between the two purebred species, indicating that the site preferences of hybrids are intermediate. The occurrences of F₂ and backcross hybrids indicate that F₁ hybrids are fertile. The chloroplast DNA haplotypes of all F₁ hybrids corresponded to those detected in M. salicifolia, so that maternal parents of the F₁ hybrids were all M. salicifolia. Furthermore, no hybrid individuals derived from a backcross to M. stellata were detected. These results suggest that the direction of hybridization and the subsequent introgression have been quite asymmetric and that the introgression occurred from M. stellata into M. salicifolia.     

Hirudinella ventricosa (Pallas, 1774) Baird, 1853 represents a species complex based on ribosomal DNA

Digeneans in the genus Hirudinella de Blainville, 1828 (Hirudinellidae) from three species of pelagic fishes, Acanthocybium solandri (Cuvier), Makaira nigricans Lacépède and Thunnus albacares (Bonnaterre), and one benthic fish, Mulloidichthys martinicus (Cuvier), from the Gulf of Mexico are investigated using comparison of ribosomal DNA. Four species are identified based on molecular differences: Hirudinella ventricosa (Pallas, 1774) Baird, 1853 from A. solandri, Hirudinella ahi Yamaguti, 1970 from T. albacares, and two unidentified but distinct species of Hirudinella, herein referred to as Hirudinella sp. A (from both M. nigricans and M. martinicus) and Hirudinella sp. B from M. nigricans. Additionally, H. ahi, based tentatively on morphological identification, is reported from Thunnus thynnus (Linnaeus). This represents the first record of a hirudinellid from M. martinicus and the first record of H. ahi from T. thynnus. A phylogeny of some Hemiurata Skrjabin & Guschanskaja, 1954 using partial fragments of the 28S rDNA sequences is consistent with earlier phylogenies and the position of the Hirudinellidae Dollfus, 1932 is well-supported as a derived group most closely related to the Syncoeliidae Looss, 1899.     

N and P colimitation of N2-fixing and N-supplied fynbos legumes from the Cape Floristic Region

Background and aims

Legume species in the fynbos vegetation of the Cape Floristic Region, that fix N₂ in soils with low P, may have evolved for enhanced acquisition and efficient use of P. It was hypothesized that N₂-fixing and combined-N supplied (N-supplied) A. linearis, P. calyptrata and C. genistoides are adapted to low P and would be relatively unresponsive to increased P of 100 μM.

Methods

18 legume species were evaluated for their nodulation response to low P availability. The N X P interaction was then examined in A. linearis, P. calyptrata and C. genistoides reliant on either N₂-fixation or 300 μM N (NH₄NO₃), and receiving 0.1, 1.0, 10 and 100 μM P (NaH₂PO₄).

Results

In the species selection experiment, A. linearis, P. calyptrata and C. genistoides, with the greatest nodule fresh weight (FW) and nodule FW to root FW ratio, were the most prolific nodulating species. In the N X P experiment, with low P supply, the biomass of N₂-fixing P. calyptrata and C. genistoides was consistently greater than that of N-supplied plants. In contrast, with high P supply of 100 μM P, all N-supplied plants accumulated more biomass than the corresponding N₂-fixing plants. High P-use efficiency, poor down-regulation of P uptake and P storage was evident in A. linearis and P. calyptrata.

Conclusion

The growth response to P and the significant N X P interactions indicate that N₂-fixing and N-supplied plants were not adapted to low P, but rather colimited by both N and P.     

Introgression of bacterial wilt resistance from eggplant to potato via protoplast fusion and genome components of the hybrids

Key message

Bacterial wilt resistant somatic hybrids were obtained via protoplast fusion between potato and eggplant and three types of nuclear genomes were identified in the hybrids through GISH and SSR analysis.

Abstract

Cultivated potato (Solanum tuberosum L.) lacks resistance to bacterial wilt caused by Ralstonia solanacearum. Interspecific symmetric protoplast fusion was conducted to transfer bacterial wilt resistance from eggplant (S. melongena, 2n = 2x = 24) into dihaploid potato (2n = 2x = 24). In total, 34 somatic hybrids were obtained, and of these, 11 rooted and were tested for genome components and resistance to race 1 of R. solanacearum. The hybrids exhibited multiple ploidy levels and contained the dominant nuclear genome from the potato parent. Three types of nuclear genomes were identified in the hybrids through genomic in situ hybridization (GISH) and simple sequence repeat (SSR) analysis, including (1) the potato type of the tetraploids in which eggplant chromosomes could not be detected by GISH but their nuclear DNA was confirmed by SSR, (2) the biased type of the hexaploids in which the chromosome dosage was 2 potato:1 eggplant, and (3) the chromosome translocation type of the mixoploids and aneuploids that was characterized by various rates of translocations of nonhomologous chromosomes. Cytoplasmic genome analysis revealed that mitochondrial DNA of both parents coexisted and/or recombined in most of the hybrids. However, only potato chloroplast DNA was retained in the hybrids speculating a compatibility between cpDNA and nuclear genome of the cell. The pathogen inoculation assay suggested a successful transfer of bacterial wilt resistance from eggplant to the hybrids that provides potential resistance for potato breeding against bacterial wilt. The genome components characterized in present research may explain partially the inheritance behavior of the hybrids which is informative for potato improvement.     

Comparative studies on the photosynthetic responses of three freshwater phytoplankton species to temperature and light regimes

The photosynthetic responses of Microcystis aeruginosa, Scenedesmus obliquus, and Cyclotella meneghiniana to temperature and light regimes were investigated. M. aeruginosa had a higher specific growth rate at 30°C than at 14 and 20°C. Its specific growth rate was the maximum among the three species at 30°C. This suggests that M. aeruginosa could predominate in a water body having high temperature. When exposed to high light, M. aeruginosa showed lower maximal photosystem II (PSII) quantum yield (Φ_M), operational PSII quantum yield ( $ \Phi_{\text{M}}^{\prime } $ ), and active reaction centers per excited cross section (RC/CS_m) than S. obliquus and C. meneghiniana. Moreover, after 2?h low light recovery at 14°C and 20°C, the recovery of Φ_M, $ \Phi_{\text{M}}^{\prime } $ and RC/CS_m in M. aeruginosa were less than the other two species. This indicates that the capacity of high light adaptation of M. aeruginosa is the lowest among the studied species at 14–20°C. When exposed to high light, C. meneghiniana had higher Φ_M and $ \Phi_{\text{M}}^{\prime } $ lost and induced higher nonphotochemical quenching at 14–20°C. This suggests that C. meneghiniana developed a higher resistance to high light under low growth temperatures. M. aeruginosa showed the lowest light compensation point among these three species, which indicates that it could utilize low light more efficiently than the other two species. Cyclic electron flow around PSII may play a role in the photoprotective mechanism of all these three species.     

The interrelations of radiologic findings and mechanical ventilation in community acquired pneumonia patients admitted to the intensive care unit: a multicentre retrospective study

Background

Burkholderia cepacia complex (BCC) bacteria are highly virulent, typically multidrug-resistant, opportunistic pathogens in cystic fibrosis (CF) patients and other immunocompromised individuals. B. vietnamiensis is more often susceptible to aminoglycosides than other BCC species, and strains acquire aminoglycoside resistance during chronic CF infection and under tobramycin and azithromycin exposure in vitro, apparently from gain of antimicrobial efflux as determined through pump inhibition. The aims of the present study were to determine if oxidative stress could also induce aminoglycoside resistance and provide further observations in support of a role for antimicrobial efflux in aminoglycoside resistance in B. vietnamiensis.

Findings

Here we identified hydrogen peroxide as an additional aminoglycoside resistance inducing agent in B. vietnamiensis. After antibiotic and hydrogen peroxide exposure, isolates accumulated significantly less [³H] gentamicin than the susceptible isolate from which they were derived. Strains that acquired aminoglycoside resistance during infection and after exposure to tobramycin or azithromycin overexpressed a putative resistance-nodulation-division (RND) transporter gene, amrB. Missense mutations in the repressor of amrB, amrR, were identified in isolates that acquired resistance during infection, and not in those generated in vitro.

Conclusions

These data identify oxidative stress as an inducer of aminoglycoside resistance in B. vietnamiensis and further suggest that active efflux via a RND efflux system impairs aminoglycoside accumulation in clinical B. vietnamiensis strains that have acquired aminoglycoside resistance, and in those exposed to tobramycin and azithromycin, but not hydrogen peroxide, in vitro. Furthermore, the repressor AmrR is likely just one regulator of the putative AmrAB-OprM efflux system in B. vietnamiensis.     

Methylene blue for malaria in Africa: results from a dose-finding study in combination with chloroquine

Background

Atovaquone is part of the antimalarial drug combination atovaquone-proguanil (Malarone^®) and inhibits the cytochrome bc₁ complex of the electron transport chain in Plasmodium spp. Molecular modelling showed that amino acid mutations are clustered around a putative atovaquone-binding site resulting in a reduced binding affinity of atovaquone for plasmodial cytochrome b, thus resulting in drug resistance.

Methods

The prevalence of cytochrome b point mutations possibly conferring atovaquone resistance in Plasmodium falciparum isolates in atovaquone treatment-naïve patient cohorts from Lambaréné, Gabon and from South Western Ethiopia was assessed.

Results

Four/40 (10%) mutant types (four different single polymorphisms, one leading to an amino acid change from M to I in a single case) in Gabonese isolates, but all 141/141 isolates were wild type in Ethiopia were found.

Conclusion

In the absence of drug pressure, spontaneous and possibly resistance-conferring mutations are rare.     

Growth, carboxylate exudates and nutrient dynamics in three herbaceous perennial plant species under low, moderate and high phosphorus supply

Background and aims

Australian herbaceous native species have evolved in phosphorus (P) impoverished soils. Our objective was to explore shoot and root adaptations of two of these species with potential to be developed as pasture plants, at low, moderate and high P supply after 4 and 7?weeks of growth.

Methods

A glasshouse experiment examined the effect of 5, 20 and 80?mg?P?kg^?1 air-dry soil on growth, rhizosphere carboxylate content, and mineral nutrition of two Australian native perennials, Kennedia nigricans (Fabaceae) and Ptilotus polystachyus (Amaranthaceae), and the exotic Medicago sativa (Fabaceae).

Key results

Leaf P concentrations at P80 were 6, 14 and 52?mg?P?g^?1 leaf dry weight for M. sativa, K. nigricans and P. polystachyus, respectively. As soil P concentration increased, rhizosphere carboxylate content decreased for M. sativa, increased and then decreased for K. nigricans and was unchanged for P. polystachyus. For all species, the contribution of malic acid declined at the second harvest. For all species and P treatments, the amount of rhizosphere carboxylates per unit root length decreased as root length of a plant increased. Plant P content was determined more by P uptake rate per unit root length and time than by root length. Uptake of Mo for all species, and uptake of K, Mg and Mn for P. polystachyus, increased with soil P concentration. Uptake of Fe and S was higher when the content of carboxylates in the rhizosphere was higher.

Conclusion

Root physiological adaptations (i.e. rhizosphere carboxylate content and P-uptake rate) are more important than morphological adaptations (i.e. root length and diameter) to enhance the uptake of P and cations.     

Taxonomic and biostratigraphic re-evaluation ofPerisphinctes internispinosus Krantz, 1926 (Upper Jurassic, Ammonoidea)

The lectotype ofPerisphinctes internispinosus Krantz, 1926, type species ofWindhauseniceras Leanza, 1945, from the upper Middle Tithonian of Cerro Lotena (Neuquén-Mendoza Basin, Argentina) is designated. The type series, housed in the Paläontologisches Institut der Universität Bonn, was originally illustrated by hand-drawings of two of the eleven specimens. The lectotype is illustrated by a photograph for the first time. The specimen is probably a large microconch; the most likely corresponding macroconch appears to beH. aff.steinmanni (Steuer, 1897) from Cerro Lotena.Windhauseniceras internispinosum is the index and guide species of theInternispinosum Biozone, upper Middle Tithonian, most probablyPonti Zone in age. The occurrence in Arroyo del Yeso (Mendoza) below a succession ofMicracanthoceras species includingM. mirum (Leanza, 1945) which is very close toM. microcanthum, strongly supports this correlation. The species occurs throughout the Neuquén-Mendoza Basin and probably also in the Tarapacá Basin, Northern Chile.     

Canalization effect in the coagulation cascade and the interindividual variability of oral anticoagulant response. a simulation Study

Background

The aim of the study was to find a simple intravenous glucose tolerance test (IVGTT) that can be used to estimate insulin sensitivity.

Methods

In 20 healthy volunteers aged between 18 and 51 years (mean, 28) comparisons were made between kinetic parameters derived from a 12-sample, 75-min IVGTT and the M_bw (glucose uptake) obtained during a hyperinsulinemic euglycemic glucose clamp. Plasma glucose was used to calculate the volume of distribution (V _d) and the clearance (CL) of the injected glucose bolus. The plasma insulin response was quantified by the area under the curve (AUC_ins). Uptake of glucose during the clamp was corrected for body weight (M_bw).

Results

There was a 7-fold variation in M_bw. Algorithms based on the slope of the glucose-elimination curve (CL/V _d) in combination with AUC_ins obtained during the IVGTT showed statistically significant correlations with M_bw, the linearity being r² = 0.63-0.83. The best algorithms were associated with a 25-75^th prediction error ranging from -10% to +10%. Sampling could be shortened to 30-40 min without loss of linearity or precision.

Conclusion

Simple measures of glucose and insulin kinetics during an IVGTT can predict between 2/3 and 4/5 of the insulin sensitivity.     

The performance of DGT versus conventional soil phosphorus tests in tropical soils - An isotope dilution study

Aims

The aim of the present study was to compare lead accumulation and tolerance among heavy metal hyperaccumulating and non-hyperaccumulating metallophytes.

Methods

To this purpose, we compared Pb tolerance and accumulation in hydroponics among calamine and non-calamine populations of Silene vulgaris, Noccaea caerulescens, and Matthiola flavida. We established the effects of Ca on Pb tolerance and accumulation in M. flavida, and measured exchangeable soil Pb and Ca at two calamine sites.

Results

Results revealed that calamine populations of S. vulgaris and N. caerulescens were Pb hypertolerant, but the calamine M. flavida population was not. Pb hyperaccumulation capacity was exclusively found in one of the calamine N. caerulescens populations.

Conclusions

1) Pb hypertolerance is sometimes lacking in metallophyte populations from strongly Pb-enriched soil, probably due to a relatively high level of exchangeable soil Ca, 2) Ca effectively counteracts Pb uptake and Pb toxicity, 3) The tendency to hyperaccumulate Pb is a population-specific phenomenon in N. caerulescens, 4) Pb hypertolerance in N. caerulescens is not necessarily associated with a tendency to hyperaccumulate Pb, 5) apparent natural Pb hyperaccumulation in M. flavida is not reproducible in hydroponics, probably due to the absence of air-born contamination in laboratory experiments.     

Fertile introgression products generated via somatic hybridization between wheat and Thinopyrum intermedium

Key message

Fertile hybrids were produced with genetic material transferred from Th. intermedium into a wheat background and supply a source of genetic variation to wheat improvement.

Abstract

Both symmetric and asymmetric somatic hybrids have been obtained from the combination of wheatgrass (Thinopyrum intermedium) and bread wheat (Triticum aestivum). Two wheat protoplast populations, one derived from embryogenic calli and the other from a non-regenerable, rapidly dividing cell line, were fused with Th. intermedium protoplasts which had been (or not been) pre-irradiated with UV. Among the 124 regenerated calli, 64 could be categorized as being of hybrid origin on the basis of plant morphology, peroxidase isozyme, RAPD DNA profiling and karyological analysis. Numerous green plantlets were regenerated from 13 calli recovered from either the symmetric hybrid (no UV pre-treatment) or the asymmetric one (30 s UV irradiation). One of these hybrid plants proved to be vigorous and self-fertile. The regenerants were all closer in phenotype to wheat than to Th. intermedium. Genomic in situ hybridization analysis showed that the chromosomes in the hybrids were largely intact wheat ones, although a few Th. intermedium chromosome fragments had been incorporated within them.     

Mentzelia canyonensis (Loasaceae), a new species endemic to the Grand Canyon, Arizona, U.S.A.

We describe Mentzelia canyonensis, a new species endemic to the Grand Canyon. Based on floral and fruit forms, shoot system architecture, and phylogenetic analyses of nuclear ribosomal DNA, we infer that M. canyonensis belongs to the ‘subshrubby’ clade of Mentzelia section Bartonia, which includes the morphologically similar M. hualapaiensis, M. oreophila, M. puberula, and M. tiehmii. The new species is most similar to M. hualapaiensis, which is also known only from the Grand Canyon. These two species share leaf shape, subshrubby habits, corolla color, and fruit shape, although they differ in staminode presence and especially in leaf and corolla size. Principal component analyses of variation among floral and vegetative traits demonstrates that M. canyonensis and M. hualapaiensis share highly overlapping floral character space, but M. canyonensis occupies unique vegetative character space compared to similar species. A dichotomous key is provided to assist with identification of, and differentiation among, species of Mentzelia section Bartonia that occur in the Grand Canyon region.     

Xylem sap calcium concentrations do not explain liming-induced inhibition of legume gas exchange

Background and aims

Liming is considered normal agricultural practise for remediating soil acidity and improving crop productivity; however recommended lime applications can reduce yield. We tested the hypothesis that elevated xylem sap Ca²⁺ limited gas exchange of Phaseolus vulgaris L. and Pisum sativum L. plants that exhibited reduced shoot biomass and leaf area when limed.

Methods

We used Scholander and whole-plant pressure chamber techniques to collect root and leaf xylem sap, a calcium-specific ion-selective electrode to measure xylem sap Ca²⁺, infra-red gas analysis to measure gas exchange of limed and unlimed (control) plants, and a detached leaf transpiration bioassay to determine stomatal sensitivity to Ca²⁺.

Results

Liming reduced shoot biomass, leaf area and leaf gas exchange in both species. Root xylem sap Ca²⁺ concentration was only increased in P. vulgaris and not in P. sativum. Detached leaves of both species required 5 mM Ca²⁺ supplied to via the transpiration stream to induce stomatal closure, however, maximum in vivo xylem sap Ca²⁺ concentrations of limed plants was only 1.7 mM and thus not high enough to influence stomata.

Conclusion

We conclude that an alternative xylem-borne antitranspirant other than Ca²⁺ decreases gas exchange of limed plants.