Fat frogs,mobile genes: unexpected phylogeographic patterns for the ornate chorus frog (Pseudacris ornata)

The southeastern coastal plain of the United States is a region marked by extraordinary phylogeographic congruence that is frequently attributed to the changing sea levels that occurred during the glacial‐interglacial cycles of the Pleistocene epoch. A phylogeographic break corresponding to the Apalachicola River has been suggested for many species studied to date that are endemic to this region. Here, we used this pattern of phylogeographic congruence to develop and test explicit hypotheses about the genetic structure in the ornate chorus frog (Pseudacris ornata). Using 1299 bp of mtDNA sequence and seven nuclear microsatellite markers in 13 natural populations of P. ornata, we found three clades corresponding to geographically distinct regions; one spans the Apalachicola River (Southern Clade), one encompasses Georgia and South Carolina (Central Clade) and a third comprises more northerly individuals (Northern Clade). However, it does not appear that typical phylogeographic barriers demarcate these clades. Instead, isolation by distance across the range of the entire species explained the pattern of genetic variation that we observed. We propose that P. ornata was historically widespread in the southeastern United States, and that a balance between genetic drift and migration was the root of the genetic divergence among populations. Additionally, we investigated fine‐scale patterns of genetic structure and found the spatial scale at which there was significant genetic structure varied among the regions studied. Furthermore, we discuss our results in light of other phylogeographic studies of southeastern coastal plain organisms and in relation to amphibian conservation and management.     

New microsatellite loci for the European bushcricket,Ephippiger ephippiger (Orthoptera: Tettigoniidae)

Ephippiger ephippiger is a model organism for studies of sexual selection and phylogeography but little is known about fine‐scale population structure. Available microsatellite loci have null allele problems so we used an enrichment technique to isolate 21 new microsatellite loci for E. ephippiger. We present primer pairs for 10 polymorphic loci (3–11 alleles per locus). Observed heterozygosities at polymorphic loci ranged from 0.118 to 0.787, but several were significantly lower than expected.     

The scheduling of spawning with the moult cycle in Northern krill (Crustacea: Euphausiacea): a strategy for allocating lipids to reproduction

Summary

Euphausiids moult and grow throughout their life, which implies sharing of resources between growth and reproduction for adult krill. In the Northern krill, Meganyctiphanes norvegica (M. Sars), female krill produce eggs cyclically. Spawning moult cycles alternate with vitellogenic moult cycles for lipid yolk accumulation. Histology shows that lipids are associated with the R cells of the digestive gland in both sexes, with the yolk platelets of mature oocytes and with the fat body cell membranes and blood lacunae in reproducing females. Mature female krill can have a total lipid content twice as high as males, mostly due to accumulation in the ovary, the fat body and the haemolymph. In contrast, in males, as well as in non-reproducing females, the highest percentage of lipids is found in the digestive gland and the haemolymph. In Meganyctiphanes norvegica, the most abundant lipid fractions are polar lipids and triglycerides, the latter being relatively low in reproducing female gonad and fat body. Triglycerides are believed to be a pure energy source and polar lipids are essential for membrane development in embryos. The fatty acid content and composition of the triglyceride and polar lipid fractions in females are different from males, related to both reproductive and dietary processes. Higher levels of polyunsaturated fatty acids (PUFA) in the polar lipid fraction were found in reproductive females. During the non-reproductive season, the converse was found, indicating the specific role PUFA and other fatty acids play in growth and egg production. Adaptive processes linked to reproduction were studied comparatively in three populations of the Northern krill—Clyde Sea (W, Scotland), Kattegat (E, Denmark), Ligurian Sea (Mediterranean)—all differing considerably in climatic and trophic conditions. Such adjustments in lipid synthesis and storage are viewed as reproductive strategies developed by the Northern krill in response to different environmental conditions.     

Co-sedimentation of Actin, Tubulin andMembranes in the Cytoskeleton Fractions from Peas and Mouse 3T3 Cells

Pea stem tissue (Pisum sativum L. var. Alaska) was homogenizedin a recently-developed cytoskeleton-stabilizing buffer, CSB,(Abeand Da vies, 1991) and homogenates electrophoresed and blottedon to membranes. Blots probed individually withantibodies toactin, alpha-tubulin, and beta-tubulin, revealed bands withapparent molecular weights of 42, 46, and 48–50 kDa,respectively.Blots probed with all three antibodies simultaneously revealedall three bands which could be distinguished in thesame lane.Homogenates of mouse 3T3 cells yielded an actin band at about42 kDa, but both alpha- and beta-tubulin appeared atabout 50kDa and thus could not be distinguished on blots probed simultaneously.This ‘triple-blotting technique’ was, therefore,suitablefor pea tissue, but not for mouse tissue. In pea tissue, sedimentabletubulin and actin were found maximally in the 4000 xg pelletand less in successive 15000 and l00000xg pellets. Both EGTAand Mg²⁺ which had been found earlier to beessential for stabilityof the actin cytoskeleton as revealed by fluorescence microscopy,were essential for co-sedimentation of actinand tubulin. Incontrast to the results with pea stems, only the actin componentof the cytoskeleton could be isolated from mouse 3T3 cells usingCSB. Pea tissue was homogenized in CSB without PTE and the resultingcytoskeletal pellets resuspended in actin- or tubulin-solubilizingbuffers with and without PTE. In the absence of PTE, the bufferssolubilized their appropriate cytoskeletal protein, but littleof the other protein, while in the presence of PTE both proteinswere quite effectively solubilized by both buffers. Incontrast,in CSB with or without PTE, both proteins remained in the sedimentablefraction. These results, taken together withother evidence,indicate that microtubules, as well as microfilaments are importantcomponents of the sedimentable cytoskeletonfraction of peasand that the membrane system is intimately involved in organizationof the cytoskeleton in peas. Key words: Actin, tubulin, membranes, detergent, Ca²⁺, Mg²⁺, cytoskeleton     

Acclimation of leaf hydraulic conductance and stomatal conductance of Pinus taeda (loblolly pine) to long-term growth in elevated CO2 (free-air CO2 enrichment) and N-fertilization

We investigated how leaf hydraulic conductance (K_leaf) of loblolly pine trees is influenced by soil nitrogen amendment (N) in stands subjected to ambient or elevated CO₂ concentrations (CO₂^a and CO₂^e, respectively). We also examined how K_leaf varies with changes in reference leaf water potential (Ψ_leaf‐ref) and stomatal conductance (g_s‐ref) calculated at vapour pressure deficit, D of 1 kPa. We detected significant reductions in K_leaf caused by N and CO₂^e, but neither treatment affected pre‐dawn or midday Ψ_leaf. We also detected a significant CO₂^e‐induced reduction in g_s‐ref and Ψ_leaf‐ref. Among treatments, the sensitivity of K_leaf to Ψ_leaf was directly related to a reference K_leaf (K_leaf‐ref computed at Ψ_leaf‐ref). This liquid‐phase response was reflected in a similar gas‐phase response, with g_s sensitivity to D proportional to g_s‐ref. Because leaves represented a substantial component of the whole‐tree conductance, reduction in K_leaf under CO₂^e affected whole‐tree water use by inducing a decline in g_s‐ref. The consequences of the acclimation of leaves to the treatments were: (1) trees growing under CO₂^e controlled morning leaf water status less than CO₂^a trees resulting in a higher diurnal loss of K_leaf; (2) the effect of CO₂^e on g_s‐ref was manifested only during times of high soil moisture.     

The relationship between limb morphology, kinematics, and force during running: the evolution of locomotor dynamics in lizards

Terrestrial locomotion occurs via the hierarchical links between morphology, kinematics, force, and center-of-mass mechanics. In a phylogenetically broad sample of seven lizard species, we show that morphological variation drives kinematic variation, which, in turn, drives force variation. Species with short limbs use a short stride–high frequency strategy when running at steady-speed and to change speeds. This link between morphology and kinematics results in relatively small vertical forces during the support phase of the stride cycle. Conversely, species with long limbs use a long stride–low frequency strategy, resulting in large vertical forces during the support phase. In view of these findings, we suggest that limb length may predict locomotor energetics in lizards because energetics are largely determined by vertical forces and stride frequency. Additionally, we propose an energetic trade-off with both long- and short-limbed species paying the most energy to move, whereas intermediate-limbed species move using less energy. Finally, when these traits are mapped onto a lizard phylogeny, we show that locomotor functional morphology exhibits both deep phylogenetic effects and contemporary patterns of evolutionary convergence. Overall, the present study provides a foundation for testing hypotheses regarding the integration and evolution of functional traits in lizards and animals in general.  © 2009 The Linnean Society of London, Biological Journal of the Linnean Society , 2009, 97 , 634–651.     

Identification and functional characterization of effectors in expressed sequence tags from various life cycle stages of the potato cyst nematode Globodera pallida

In this article, we describe the analysis of over 9000 expressed sequence tags (ESTs) from cDNA libraries obtained from various life cycle stages of Globodera pallida . We have identified over 50 G. pallida effectors from this dataset using bioinformatics analysis, by screening clones in order to identify secreted proteins up-regulated after the onset of parasitism and using in situ hybridization to confirm the expression in pharyngeal gland cells. A substantial gene family encoding G. pallida SPRYSEC proteins has been identified. The expression of these genes is restricted to the dorsal pharyngeal gland cell. Different members of the SPRYSEC family of proteins from G. pallida show different subcellular localization patterns in plants, with some localized to the cytoplasm and others to the nucleus and nucleolus. Differences in subcellular localization may reflect diverse functional roles for each individual protein or, more likely, variety in the compartmentalization of plant proteins targeted by the nematode. Our data are therefore consistent with the suggestion that the SPRYSEC proteins suppress host defences, as suggested previously, and that they achieve this through interaction with a range of host targets.     

Environmental variation and behaviour: resource availability during ontogeny and feeding performance in salamander larvae (Ambystoma texanum)

1. The effects of resource availability during ontogeny on subsequent feeding performance were investigated in larvae of the small-mouthed salamander ( Ambystoma texanum ).
2. Salamander larvae were reared individually in either high or low prey density treatments for 7 weeks prior to intermediate prey density foraging trials. Larvae from the low prey density treatment were on average 35% smaller in body size than individuals from the high prey density treatment.
3. Resource availability during development influenced larval feeding rates and altered the relationship between body size and three feeding performance measures (attack rates, capture success and feeding rates). Feeding rates in predation trials were also positively correlated with growth rate early in the larval period (until the end of week 5).
4. These results suggest that the environment to which developing organisms are exposed can have significant effects on subsequent behaviour, and that small-mouthed salamander larvae may show state-dependent changes in feeding behaviour in response to differences in long-term feeding history. Additionally, differences in feeding performance may influence the probability of survival to the adult stage for organisms that utilize ephemeral habitats.     

Host-plant susceptibility to the carrot fly, Psila rosae. 3. The role of oviposition preferences and larval performance

The acceptability of various plant species to ovipositing carrot flies was weakly, but significantly correlated with the host's suitability for larval development. Both adult host-plant preferences and larval performance as determined in laboratory experiments explained a part of the variation in susceptibility among the various test plants observed in the field. Across the whole set of plant species examined, antixenosis contributed more substantially to resistance than antibiosis, while the reverse seemed to be true for carrot cultivars.     

Morphological convergence during pregnancy among predator and nonpredator populations of the livebearing fish Brachyrhaphis rhabdophora (Teleostei: Poeciliidae)

Predation can drive morphological divergence in prey populations, although examples of divergent selection are typically limited to nonreproductive individuals. In livebearing females, shape often changes drastically during pregnancy, reducing speed and mobility and enhancing susceptibility to predation. In the present study, we document morphological divergence among populations of nonreproductive female livebearing fish (Brachyrhaphis rhabdophora) in predator and nonpredator environments. We then test the hypothesis that shape differences among nonreproductive females are maintained among reproductive females between predator and nonpredator environments. Nonreproductive females in predator environments had larger caudal regions and more fusiform bodies than females in nonpredator environments; traits that are associated with burst speed in fish. Shape differences were maintained in reproductive females, although the magnitude of this difference declined relative to nonreproductive females, suggesting morphological convergence during pregnancy. Phenotypic change vector analysis revealed that females in predator environments became more similar to females in nonpredator environments in the transition from nonreproductive to reproductive. Furthermore, the level of reproductive allocation affected shape similarly between predator environments. These results suggest a life‐history constraint on morphology, in which predator‐driven morphological divergence among nonreproductive B. rhabdophora is not maintained at the same level during pregnancy. © 2011 The Linnean Society of London, Biological Journal of the Linnean Society, 2011, 104 , 386–392.