Proteomic and phenotypic profiling of the amphibian pathogen Batrachochytrium dendrobatidis shows that genotype is linked to virulence

Population genetics of the amphibian pathogen Batrachochytrium dendrobatidis ( Bd ) show that isolates are highly related and globally homogenous, data that are consistent with the recent epidemic spread of a previously endemic organism. Highly related isolates are predicted to be functionally similar due to low levels of heritable genetic diversity. To test this hypothesis, we took a global panel of Bd isolates and measured (i) the genetic relatedness among isolates, (ii) proteomic profiles of isolates, (iii) the susceptibility of isolates to the antifungal drug caspofungin, (iv) the variation among isolates in growth and phenotypic characteristics, and (v) the virulence of isolates against the European common toad Bufo bufo . Our results show (i) genotypic differentiation among isolates, (ii) proteomic differentiation among isolates, (iii) no significant differences in susceptibility to caspofungin, (iv) differentiation in growth and phenotypic/morphological characters, and (v) differential virulence in B. bufo . Specifically, our data show that Bd isolates can be profiled by their genotypic and proteomic characteristics, as well as by the size of their sporangia. Bd genotypic and phenotypic distance matrices are significantly correlated, showing that less-related isolates are more biologically unique. Mass spectrometry has identified a set of candidate genes associated with inter-isolate variation. Our data show that, despite its rapid global emergence, Bd isolates are not identical and differ in several important characters that are linked to virulence. We argue that future studies need to clarify the mechanism(s) and rate at which Bd is evolving, and the impact that such variation has on the host–pathogen dynamic.     

Conservation and ecological implications of the use of space by chilla foxes and free‐ranging dogs in a human‐dominated landscape in southern Chile

We investigated whether domestic dogs (Canis lupus familiaris) influenced the use of space by chilla foxes (Lycalopex griseus) in southern Chile and tested the hypothesis that dogs interfere with chillas. We used scent stations and occurrence of scats within systematically placed plots to assess habitat use by both species and to test whether dogs were associated to the use of space by chillas. Activity data were obtained from captures for both species, and telemetry for foxes. Diet of both species was analysed in order to tease out the potential existence of exploitation competition. We found that, when active, chillas used prairies more and native forest less than expected according to availability, whereas inactive chillas (during the day) preferred native forest, the only habitat type that dogs did not use. The odds of dog occurrence increased at shorter distance to human houses, whereas the inverse pattern was observed for chillas. Poisson models showed that the number of chilla visits to scent stations was negatively correlated with the number of dog visits. We observed dogs persecuting and/or killing chillas which was also corroborated by local people that used dogs to prevent chilla‐related poultry losses, supporting the idea that dogs harass foxes actively. The analysis of scats showed that dogs fed mainly on house food and domestic ruminant carcasses whereas foxes fed on hare, hens, mice and wild birds, thus suggesting that exploitation competition is not a strong alternative hypothesis as a proximate cause for the patterns observed. We conclude that dogs seem to constrain the use of space by wild carnivores via interference.     

Two closely linked tomato HKT coding genes are positional candidates for the major tomato QTL involved in Na+/K+ homeostasis

The location of major quantitative trait loci (QTL) contributing to stem and leaf [Na⁺] and [K⁺] was previously reported in chromosome 7 using two connected populations of recombinant inbred lines (RILs) of tomato. HKT1;1 and HKT1;2, two tomato Na⁺‐selective class I‐HKT transporters, were found to be closely linked, where the maximum logarithm of odds (LOD) score for these QTLs located. When a chromosome 7 linkage map based on 278 single‐nucleotide polymorphisms (SNPs) was used, the maximum LOD score position was only 35 kb from HKT1;1 and HKT1;2. Their expression patterns and phenotypic effects were further investigated in two near‐isogenic lines (NILs): 157‐14 (double homozygote for the cheesmaniae alleles) and 157‐17 (double homozygote for the lycopersicum alleles). The expression pattern for the HKT1;1 and HKT1;2 alleles was complex, possibly because of differences in their promoter sequences. High salinity had very little effect on root dry and fresh weight and consequently on the plant dry weight of NIL 157‐14 in comparison with 157‐17. A significant difference between NILs was also found for [K⁺] and the [Na⁺]/[K⁺] ratio in leaf and stem but not for [Na⁺] arising a disagreement with the corresponding RIL population. Their association with leaf [Na⁺] and salt tolerance in tomato is also discussed.     

DIGITAL IMAGE ANALYSIS OF CHROMATIN FIBRE PHENOTYPE AFTER ‘IN SITU’ DIGESTION WITH RESTRICTION ENDONUCLEASES

Restriction Endonucleases (REs) may recognize, cleave and remove DNA from fixed chromatin producing specific chromosome banding patterns. However, the modifications produced in the chromatin fibre are not easy to evaluate and compare. The aim of the present investigation was to visualize differences resulting in the texture of the chromatin fibre from metaphase chromosomes after each digestion using digital image analysis (DIA) facilities. To this purpose, metaphase chromosomes derived from a L-929 mouse cell line were digested with different REs (AluI,HpaII andHaeIII). Since light microscopy does not permit the observation of the chromatin fibre, DIA was performed on digitalized images of metaphase chromosomes under electron microscopy. The application of a LUT (Look Up Table) within the DIA software assigns a colour to each grey level of a digital image. The results obtained using a particular LUT, which permits the discrimination of specific chromatin fibre phenotypes resulting from each digestion, are reported and compared with those obtained under the light microscope.     

Positive effects of native shrubs on three specially protected cacti species in Durango,México

The spatial association between the specially protected cacti Coryphanta durangensis, Echinocereus longisetus and Peniocereus greggii and potential nurse plants was evaluated, as was their relative position to the sun under the crown of the latter in the southern Chihuahuan Desert. The soil temperature under potential nurse plants was lower than under direct sunlight. There was more nitrogen and organic matter in the soil under Prosopis laevigata trees than in soil under direct sunlight. There were 68 plants of C. durangensis, 59 plants of E. longisetus and 157 of P. greggii. Only one individual of C. durangensis and one of E. longisetus grew outside the shade of the crown of other plants. Echinocereus longisetus was not associated with any particular species, but grew more often than expected by chance in the northern segment of the crown (i.e. the area most shaded in the afternoon). Coryphanta durangensis and P. greggii grew more often under P. laevigata than expected by chance. The effect of other plants, and P. laevigata in particular, on the facilitation of growth of protected cacti species should be considered in management plans of the Chihuahuan Desert, where mesquite (P. laevigata) is often harvested for charcoal production.     

Chromosomes tell half of the story: the correlation between karyotype rearrangements and genetic diversity in sedges,a group with holocentric chromosomes

Chromosome rearrangements may affect the rate and patterns of gene flow within species, through reduced fitness of structural heterozygotes or by reducing recombination rates in rearranged areas of the genome. While the effects of chromosome rearrangements on gene flow have been studied in a wide range of organisms with monocentric chromosomes, the effects of rearrangements in holocentric chromosomes—chromosomes in which centromeric activity is distributed along the length of the chromosome—have not. We collected chromosome number and molecular genetic data in Carex scoparia, an eastern North American plant species with holocentric chromosomes and highly variable karyotype (2n = 56–70). There are no deep genetic breaks within C. scoparia that would suggest cryptic species differentiation. However, genetic distance between individuals is positively correlated with chromosome number difference and geographic distance. A positive correlation is also found between chromosome number and genetic distance in the western North American C. pachystachya (2n = 74–81). These findings suggest that geographic distance and the number of karyotype rearrangements separating populations affect the rate of gene flow between those populations. This is the first study to quantify the effects of holocentric chromosome rearrangements on the partitioning of intraspecific genetic variance.     

Long‐distance abscisic acid signalling under different vertical soil moisture gradients depends on bulk root water potential and average soil water content in the root zone

To determine how root‐to‐shoot abscisic acid (ABA) signalling is regulated by vertical soil moisture gradients, root ABA concentration ([ABA]_root), the fraction of root water uptake from, and root water potential of different parts of the root zone, along with bulk root water potential, were measured to test various predictive models of root xylem ABA concentration [RX‐ABA]_sap. Beans (Phaseolus vulgaris L. cv. Nassau) were grown in soil columns and received different irrigation treatments (top and basal watering, and withholding water for varying lengths of time) to induce different vertical soil moisture gradients. Root water uptake was measured at four positions within the column by continuously recording volumetric soil water content (θ_v). Average θ_v was inversely related to bulk root water potential (Ψ_root). In turn, Ψ_root was correlated with both average [ABA]_root and [RX‐ABA]_sap. Despite large gradients in θ_v, [ABA]_root and root water potential was homogenous within the root zone. Consequently, unlike some split‐root studies, root water uptake fraction from layers with different soil moisture did not influence xylem sap (ABA). This suggests two different patterns of ABA signalling, depending on how soil moisture heterogeneity is distributed within the root zone, which might have implications for implementing water‐saving irrigation techniques.     

Effect of overwintering adults on juvenile survival of Calomys venustus (Muridae: Sigmodontinae)

Abstract The aim of this paper was to test the hypothesis that the presence of overwintering Calomys venustus males or females at the beginning of the breeding period reduced the survival of juveniles belonging to the first cohort of each year (C1). The study was carried out in four 0.25‐ha enclosures (two control and two experimental), each situated on natural pasture. This study was conducted between August 1997 and February 1999 and had two parts: removal of overwintering males (ROM) and removal of overwintering females (ROF). For the ROM treatment, overwintering males were removed immediately after C1 juveniles were born, in the ROF treatment overwintering females were removed after the C1 were weaned. Weekly censuses were taken for 3 successive days over 6 weeks. Juvenile survival was examined through a survivorship schedule. The survivorship values were compared by sex and treatment (removal of overwintering males or females) using repeated‐measures anova . Removal of overwintering males and females did not improve the survival of C1 juveniles. This result could be related to the delay in maturation of C1 juvenile males.     

Climate change and fitness components of a migratory bird breeding in the Mediterranean region

The increase in spring temperatures in temperate regions over the last two decades has led to an advancing spring phenology, and most resident birds have responded to it by advancing their onset of breeding. The pied flycatcher (Ficedula hypoleuca) is a long‐distance migrant bird with a relatively late onset of breeding with respect to both resident birds and spring phenology in Europe. In the present correlational study, we show that some fitness components of pied flycatchers are suffering from climate change in two of the southernmost European breeding populations. In both montane study areas, temperature during May increased between 1980 and 2000 and an advancement of oak leafing was detected by using the normalized difference vegetation index (NDVI) to assess tree phenology. This might result in an advancement of the peak in availability of caterpillars, the main prey during the nestling stage. Over the past 18 yr, the time of egg laying and clutch size of pied flycatchers were not affected by the increase in spring temperatures in these Mediterranean populations. However, this increase seems to have an adverse effect on the reproductive output of pied flycatchers over the same period. Our data suggest that the mismatch between the timing of peak food supply and nestling demand caused by recent climate change might result in a reduction of parental energy expenditure that is reflected in a reduction of nestling growth and survival of fledged young in our study populations. The data seem to indicate that the breeding season has not shifted and it is the environment that has shifted away from the timing of the pied flycatcher breeding season. Mediterranean pied flycatchers were not able to advance their onset of breeding, probably, because they are constrained by their late arrival date and their restricted high altitude breeding habitat selection near the southern border of their range.