Dynamics of heat-induced thermal stress resistance and Hsp70 expression in the springtail, Orchesella cincta.

The relationship between thermal resistance and expression of inducible heat shock proteins, especially Hsp70, depends on the species and temperature treatments. The induction of Hsp70 has been shown to be essential for heat stress survival in a number of species, yet the maximum protein expression levels do not coincide with peak survival after heat hardening in Drosophila. 2. Here we study the functional relationship between heat-induced expression of the heat shock protein Hsp70, and thermal resistance in adult Orchesella cincta by comparing thermal resistance (survival of 37·4 °C for 60 min) with Hsp70 gene and protein expression levels, all three measured at time points 2, 4, 6, 23, 27, 49 h after a heat hardening treatment (35·4 °C for 60 min). 3. Thermotolerance increased over time after heat hardening until 49 h after exposure when the experiment ended. On the other hand the expression of hsp70 messenger RNA reached a peak within the first 2 h and then sharply decreased after 6 h. Within 23 h hsp70 expression was back to control levels. 4. Surprisingly, protein levels of Hsp70 followed thermotolerance and reached the highest levels 49 h after heat hardening. A significant positive association was found between thermotolerance and Hsp70 protein levels, but not with hsp70 mRNA levels. 5. Our results support a strong correlation between Hsp70 expression levels and thermal resistance following a heat hardening treatment. They also show that gene and protein expression follow different dynamics, a difference that may be important for our understanding of the role of candidate genes in functional studies. © 2009 British Ecological Society

Genetic variation in heat resistance and HSP70 expression in inbred isofemale lines of the springtail Orchesella cincta.

The functional relationship between thermotolerance and heat shock protein (HSP) expression is well described. However, the intraspecific variation in HSP70 expression and heat resistance is not fully understood. Some studies have shown a correlation between thermotolerance and HSP70 expression levels at the intraspecific level, whereas others have not, and results depend on species, developmental stage and environment. The present study extends such correlative studies to species from soil ecosystems, where we have characterized the variation in thermotolerance and HSP70 expression in 18 inbred isofemale lines of the springtail Orchesella cincta. The results show ample genetic variation among lines in heat shock resistance as well as in HSP70 protein levels and hsp70 mRNA expression. However, we did not detect any significant positive correlations between HSP70 expression and thermotolerance. These results indicate that the variation in HSP70 expression and heat resistance at the intraspecific level is dependent on species and/or ecosystem and further studies are needed to clarify this relationship. © Inter-Research 2010

Evidence for multiple origins of Wolbachia infection in springtails.

Bacteria of the genus Wolbachia (Rickettsiae) are endosymbionts found in many species of invertebrates. The traditional division of Wolbachia lines into four supergroups has recently been challenged with the finding of a fifth supergroup. Until now, this E supergroup exists of a single host species of the order Collembola (springtails) and is hypothesized to form a 'missing link' between Wolbachia lines in higher arthropods and nematodes. We have sequenced the 16S rDNA, the ftsZ and the wsp genes of a Wolbachia lineage present in the sexually reproducing collembolan Orchesella cincta. We tested the hypothesis on the origin of Wolbachia in Collembola by adding this new Wolbachia lineage and a lineage found in the collembolan Mesophorura macrochaeta to the Wolbachia phylogenetic tree. The addition of these two species clearly shows that Wolbachia is not monophyletic in Collembola. We conclude that the origin of Wolbachia in Collembola is a combination of vertical and horizontal transfer between coexisting species. © 2004 Elsevier GmbH. All rights reserved

Genetic structure in Orchesella cincta (Collembola): strong subdivision of European populations inferred from mtDNA and AFLP markers.

Population genetic structure is determined both by current processes and historical events. Current processes include gene flow, which is largely influenced by the migration capacity of a species. Historical events are, for example, glaciation periods, which have had a major impact on the distribution of many species. Species with a low capacity or tendency to move about or disperse often exhibit clear spatial genetic structures, whereas mobile species mostly show less spatial genetic differentiation. In this paper we report on the genetic structure of a small, wingless arthropod species (Orchesella cincta: Collembola) in Europe. For this purpose we used mtDNA COII sequences and AFLP markers. We show that large genetic differences exist between populations of O. cincta, as expected from O. cincta's winglessness and sedentary lifestyle. Despite the fact that most variability was observed within populations (59%), a highly significant amount of AFLP variation (25%) was observed between populations from northwestern Europe, central Europe and Italy. This suggests that gene flow among regions is extremely low, which is additionally supported by the lack of shared mtDNA alleles between regions. Based on the genetic variation and sequence differences observed we conclude that the subdivision occurred long before the last glaciation periods. Although the populations still interbreed in the lab, we assume that in the long term the genetic isolation of these regions may lead to speciation processes

Narcotic mechanisms of acute toxicity of chlorinated anilines in Folsomia candida (Collembola) revealed by gene expression analysis.

In order to clarify the mechanisms of reproductive toxicity in a QSAR approach, the transcriptional signatures upon the 2 day exposure to the 28 days EC₅₀ of a series of increasingly chlorinated aniline compounds and 1,2,3,4-tetrachlorobenzene were measured in Folsomia candida. In general, the transcriptional patterns associated with all compounds revealed toxicity at the cellular membranes and hence components of narcosis type I, but a principal component analysis revealed a deviating response by the pentachloroaniline and 2,3,5,6-tetrachloroaniline exposure. Moreover the expression of a subset of mainly biotransformation related genes showed a significant relationship with the logK(ow,) which stresses the presence of narcosis type I. This was confirmed by GO term enrichment at the level of cellular component. Genes involved in the three phases of xenobiotic biotransformation exhibited strict compound specific transcription patterns, which may reflect biotransformation processes in F. candida. Additional toxic mechanisms were especially observed for the 2,3,5,6-tetrachloroaniline, which possible works as an uncoupler or inhibitor of electron transport systems, which is revealed by the up-regulation of genes that encode different members of the electron transport chain. The aniline and 2,3,4-trichloroaniline exposure caused the induction of genes in the ROS defense system. Additional toxicity mechanisms were less clear, but they include the attack of microbial pathogens for the six other compounds and for 2,3,5,6-tetrachloroaniline an effect on mitochondrial protein folding

Traits underpinning desiccation resistance explain distribution patterns of terrestrial isopods

Predicted changes in soil water availability regimes with climate and land-use change will impact the community of functionally important soil organisms, such as macro-detritivores. Identifying and quantifying the functional traits that underlie interspecific differences in desiccation resistance will enhance our ability to predict both macro-detritivore community responses to changing water regimes and the consequences of the associated species shifts for organic matter turnover. Using path analysis, we tested (1) how interspecific differences in desiccation resistance among 22 northwestern European terrestrial isopod species could be explained by three underlying traits measured under standard laboratory conditions, namely, body ventral surface area, water loss rate and fatal water loss; (2) whether these relationships were robust to contrasting experimental conditions and to the phylogenetic relatedness effects being excluded; (3) whether desiccation resistance and hypothesized underlying traits could explain species distribution patterns in relation to site water availability. Water loss rate and (secondarily) fatal water loss together explained 90 % of the interspecific variation in desiccation resistance. Our path model indicated that body surface area affects desiccation resistance only indirectly via changes in water loss rate. Our results also show that soil moisture determines isopod species distributions by filtering them according to traits underpinning desiccation resistance. These findings reveal that it is possible to use functional traits measured under standard conditions to predict soil biota responses to water availability in the field over broad spatial scales. Taken together, our results demonstrate an increasing need to generate mechanistic models to predict the effect of global changes on functionally important organisms. © 2012 Springer-Verlag Berlin Heidelberg

Comparative population analysis of metallothionein promoter alleles suggests stress-induced microevolution in the field.

We investigate a model system for microevolution of transcriptional regulation: metallothionein expression in springtails. A previous survey of the metallothionein promoter in Orchesella cincta (Collembola) revealed nine alleles with differential basal activities and responses to cadmium and oxidative stress. In this study, 23 woodlands, with a divergent degree of pollution, were sampled, and heavy metals were measured. When grouped to their contamination degree, they were discriminated best on the pmtD2 metallothionein promoter allele frequency, which was higher in populations from heavily polluted sites. Taken together with previous work showing high inducibility of the pmtD2 promoter allele by Cd in a reporter assay, this suggests a fitness advantage of the pmtD2 allele in polluted sites. Redundancy analysis revealed associations between allele frequencies and specific metals in the environment, resulting in a subdivision between pollution associated alleles and others. A positive relationship between the pmtD2 allele frequency and the Cd content of the soil as well as between pmtE and Ni in the litter emerged. An increase of genetic diversity was observed with increasing Pb in the soil, reached through substitution of the pmtA1 allele, suggesting balancing selection. Our results illustrate that environmental factors can exert selection on promoter polymorphisms and cause adaptation through altered transcriptional regulation. © 2008 American Chemical Society