Contrasting covariation of above- and belowground invertebrate species across plant genotypes

1. Invertebrate species generally do not respond independently to genotypic variation in plants, giving rise to clusters of species that naturally associate with or avoid certain genotypes. This covariation causes coevolution to be diffuse rather than pairwise. Studies on this topic, however, have never considered the belowground invertebrate community, leaving a critical gap in our understanding. 2. We investigated the covariation among naturally colonising above- and belowground invertebrate species across six genetically distinct populations of the dune grass Ammophila arenaria. After having grown from seed in a common garden, plants were randomised in a single field site to exclude all but broad-sense genetic variation. 3. Strong positive covariation across genotypes among both above- and belowground invertebrates was detected, while correlations between these two groups were negative. This clustering of above- and belowground species matched well with order level taxonomy. Host range, trophic level and food type on the other hand did not correspond well with the clusters. Within the cluster of aboveground fauna, subsequent groupings were not related to any phylogenetic or ecological characteristic, although correlations within these subgroups were very high. We furthermore demonstrated significant differences in multiple invertebrate species occurrence between plant genotypes, in general as well as at the above- and belowground level. 4. The observed strong covariation suggests diffuse coevolution between A. arenaria and its associated invertebrate species. The trade-off between root and shoot invertebrates could however hamper directional selection on resistance to either group. 5. Our results clearly demonstrate the need for studies of plant-animal interactions to include the belowground fauna, as this might drastically alter our general conception of how plants and their associated animal communities interact and how these interactions shape the process of evolution.     

Nematodes surfing the waves: long‐distance dispersal of soil‐borne microfauna via sea swept rhizomes

Dispersal mechanisms of soil‐borne microfauna have hitherto received little attention. Understanding dispersal mechanisms of these species is important to unravel their basic life history traits, biogeography, exchange of individuals between populations, and local adaptation. Soil‐borne nematodes and root‐feeding nematodes in particular occupy a key position in soil‐food webs and can be determinants for plant growth and vegetation structure and succession. However, their dispersal abilities have been scarcely addressed, predominantly focusing on species of agricultural importance. Still, root‐feeding nematodes are usually considered as being extremely limited and bound to the rhizosphere of plants. We investigated a mechanism for long distance dispersal of root‐feeding nematodes associated to two widespread coastal dune grasses. The nematodes are known to be crucial for the functioning of these grasses. We experimentally tested the hypothesis that root‐feeding nematodes are able to move across long distances inside rhizome fragments that are dispersed by seawater. We also tested the survival capacities of the host plants in seawater. Our study demonstrates that root‐feeding nematodes and plants are able to survive immersion in seawater, providing a mechanism for long distance dispersal of root feeding nematodes together with their host plant. Drifting rhizome fragments enable the exchange of plant material and animals between dune systems. These results provide new insights to understand the ecology of dune vegetation, the interaction with soil‐borne organisms and more importantly, re‐set the scale of spatial dynamics of a group of organisms considered extremely constrained in its dispersal capacities.     

Gamma-secretase-regulated proteolysis of the Notch receptor by mitochondrial intermediate peptidase

Notch is a transmembrane receptor that controls a diverse array of cellular processes including cell proliferation, differentiation, survival, and migration. The cellular outcome of Notch signaling is dependent on extracellular and intracellular signals, but the complexities of its regulation are not well understood. Canonical Notch signaling involves ligand association that triggers sequential and regulated proteolysis of Notch at several sites. Ligand-dependent proteolysis at the S2 site removes the bulk of the extracellular domain of Notch. Subsequent γ-secretase-mediated intramembrane proteolysis of the remaining membrane-tethered Notch fragment at the S3 site produces a nuclear-destined Notch intracellular domain (NICD). Here we show that following γ-secretase cleavage, Notch is proteolyzed at a novel S5 site. We have identified this S5 site to be eight amino acids downstream of the S3 site. Biochemical fractionation and purification resulted in the identification of the S5 site protease as the mitochondrial intermediate peptidase (MIPEP). Expression of the MIPEP-cleaved NICD (ΔNICD) results in a decrease in cell viability and mitochondria membrane potential. The sequential and regulated proteolysis by γ-secretase and MIPEP suggests a new means by which Notch function can be modulated.     

Flowering phenology and reproduction of a forest understorey plant species in response to the local environment

Temperate forest understorey plants are subjected to a strong seasonality in their optimal growing conditions. In winter and early spring, low temperatures are suboptimal for plant growth while light becomes limited later in spring season. We can thus expect that differences in plant phenology in relation to spatiotemporal environmental variation will lead to differences in reproductive output, and hence selection. We specifically studied whether early flowering, a paradoxical pattern that is observed in many plant species, is an adaptive strategy, and whether selection for early flowering was confounded with selection for flower duration or was attributable to environmental variables. We used Geum urbanum as a study species to investigate the effect of relevant environmental factors on the species’ flowering phenology and the consequences for plant reproductive output. We monitored the phenology of four to six plants in each of ten locations in a temperate deciduous forest (Belgium). We first quantified variation in flowering time within individuals and related this temporal variation to individual flower reproductive output. Then, we studied inter-individual variation here-in and linked this to reproduction at the plant level, hence studying the selection differential. We found that flowering within individual plants of Geum urbanum was spread over a long period from June to October. Reproductive output of individual flowers, measured as total seed mass per flower, declined during the season. We found no indication for selection for early flowering but rather for longer flower duration. Larger plants had an earlier flowering onset and a higher seed mass, which suggests that these factors covary and are condition dependent. None of the studied environmental variables could explain plant size, although soil pH and to a lesser extent light availability had a positive direct effect on seed mass per plant. Finally, we suggest that the high intra-individual variation in flowering time, which might be a risk spreading strategy of the plant in the presence of seed predation, limits the potential for selection on flowering phenology.

     

Interdemic variation of cannibalism in a wolf spider (Pardosa monticola) inhabiting different habitat types

Abstract.  1. Cannibalism was investigated in the wolf spider Pardosa monticola (Clerck) using spiders collected from four populations with varying densities, inhabiting two different coastal dune habitat types. Sampled individuals were paired randomly and tested immediately for their cannibalism propensity.
2. The occurrence of cannibalism was found to be influenced by the size (cephalothorax width) of both the smaller and the larger spider of a pair. Larger size differences enhanced cannibalism.
3. Cannibalism rates were not significantly different in spiders from high-density compared with low-density populations. Cannibalism rates showed, however, large variability between habitat types, with higher rates in spiders from dune grasslands than from dune slacks. This is suggested to result from differences in prey availability throughout the growing season between both habitat types.
4. Different size classes of spiders did not use different microhabitats, indicating that microhabitat segregation as a cannibalism-avoidance behaviour is absent in this species.     

Bridging the gap between the Natura 2000 regional conservation status and local conservation objectives

Natura 2000 targets the sustainable conservation of Europe's biodiversity. An important cornerstone of Natura 2000 is the Habitats Directive, which is currently implemented across European member states. However, straightforward implementation is not obvious since the favourable conservation status of habitats and species needs to be achieved at the member state level, while conservation objectives need to be formulated at the protected site level. To bridge this gap, we propose to start from regional conservation objectives before site level objectives are formulated. These regional conservation objectives have the advantage of providing a framework according to which conservation objectives can be allocated both within and outside the protected sites of the Natura 2000 network. Especially since they all contribute to the national or regional conservation status. Recently, Flanders (northern Belgium) has adopted this approach and has quantified conservation objectives at the regional scale. As the current regional conservation status of habitats and species is mostly unfavourable, regional conservation objectives entail a drastic increase in area (42%) for habitats, and active conservation measures for 78% of the species. We are convinced that the method outlined here, may substantially contribute to a helpful discussion about implementing and streamlining Natura 2000 across European member states.     

Transference of function shapes organ identity in the dove tree inflorescence

? An important evolutionary mechanism shaping the biodiversity of flowering plants is the transfer of function from one plant organ to another. To investigate whether and how transference of function is associated with the remodeling of the floral organ identity program we studied Davidia involucrata, a species with conspicuous, petaloid bracts subtending a contracted inflorescence with reduced flowers. ? A detailed ontogeny enabled the interpretation of expression patterns of B-, C- and E-class homeotic MADS-box genes using qRT-PCR and in situ hybridization techniques. We investigated protein-protein interactions using yeast two-hybrid assays. ? Although loss of organs does not appear to have affected organ identity in the retained organs of the reduced flowers of D. involucrata, the bracts express the B-class TM6 (Tomato MADS box gene 6) and GLOBOSA homologs, but not DEFICIENS, and the C-class AGAMOUS homolog, representing a subset of genes also involved in stamen identity. ? Our results may illustrate how petal identity can be partially transferred outside the flower by expressing a subset of stamen identity genes. This adds to the molecular mechanisms explaining the diversity of plant reproductive morphology.     

Subtle population structure and male-biased dispersal in two <Emphasis Type="Italic">Copadichromis</Emphasis> species (Teleostei,Cichlidae) from Lake Malawi,East Africa

Marine protected areas are generally designed and managed on the basis of the presence and extent of specific habitat types or the habitats of important species. However, it has become clear that in addition to including these ‘structural’ elements of marine systems, management strategies should incorporate a consideration of the functional aspects of the ecosystems. Biological traits analysis (BTA) has been successfully used to describe ecological functioning in marine benthic systems. BTA uses a number of biological characteristics expressed by the taxa present as indicators of key ecosystem functions. Two expert workshops were used to examine the potential for the application of BTA in the designation and management of MPAs. They concluded that BTA represented the best tool currently available for quantifying ecological functioning and agreed on 10-key ecological functions delivered by marine benthic communities. Twenty-four biological traits were also identified by the workshops as indices of these ten functions. In order to demonstrate the practical utility of the approach, BTA using these traits, was applied to a dataset covering benthos from within and around the proposed Eddystone Special Area of Conservation (SW England). The case study demonstrated that with the type of data normally available from conservation assessment type surveys, and a knowledge of the relevant biological traits, it is possible to use a consideration of ecological functioning to set boundaries for the MPA and to inform the site management objectives. The use of structure and function information to inform the designation process and subsequent management of marine protected areas is discussed. Guest editors: J. Davenport, G. Burnell, T. Cross, M. Emmerson, R. McAllen, R. Ramsay & E. Rogan Challenges to Marine Ecosystems     

A new FRAP/FRAPa method for three-dimensional diffusion measurements based on multiphoton excitation microscopy

We present a new convenient method for quantitative three-dimensionally resolved diffusion measurements based on the photobleaching (FRAP) or photoactivation (FRAPa) of a disk-shaped area by the scanning laser beam of a multiphoton microscope. Contrary to previously reported spot-photobleaching protocols, this method has the advantage of full scalability of the size of the photobleached area and thus the range of diffusion coefficients, which can be measured conveniently. The method is compatible with low as well as high numerical aperture objective lenses, allowing us to perform quantitative diffusion measurements in three-dimensional extended samples as well as in very small volumes, such as cell nuclei. Furthermore, by photobleaching/photoactivating a large area, diffusion along the optical axis can be measured separately, which is convenient when studying anisotropic diffusion. First, we show the rigorous mathematical derivation of the model, leading to a closed-form formula describing the fluorescence recovery/redistribution phase. Next, the ability of the multiphoton FRAP method to correctly measure absolute diffusion coefficients is tested thoroughly on many test solutions of FITC-dextrans covering a wide range of diffusion coefficients. The same is done for the FRAPa method on a series of photoactivatable green fluorescent protein solutions with different viscosities. Finally, we apply the method to photoactivatable green fluorescent protein diffusing freely in the nucleus of living NIH-3T3 mouse embryo fibroblasts.