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Bernabol P.,University of Modena and Reggio Emilia | Bernabol P.,University of Trento | Rebecchi L.,University of Modena and Reggio Emilia | Jousson O.,University of Trento | And 2 more authors.
Cell Stress and Chaperones | Year: 2011

To better understand the physiological capability of cold-stenothermal organisms to survive high-temperature stress, we analyzed the thermotolerance limits and the expression level of hsp70 genes under temperature stress in the alpine midge Pseudodiamesa branickii (Diptera Chironomidae). A lethal temperature (LT100) of 36°C and a lethal temperature 50% (LT50) of 32.2°C were found for the coldstenothermal larvae after short-term shocks (1 h). Additional experiments revealed that the duration of the exposure negatively influenced survival, whereas a prior exposure to a less severe high temperature generated an increase in survival. To investigate the molecular basis of this high thermotolerance, the expression of the hsp70 gene family was surveyed via semi-quantitative reverse transcription-polymerase chain reaction analysis in treated larvae. The constitutive (hsc70) and inducible (hsp70) forms were both analyzed. Larvae of P. branickii showed a significant up-regulation of inducible hsp70 gene with increasing temperatures and an over-expression of both hsp70 and hsc70 by increasing the time of exposure. Different from that was shown in many cold-stenothermal Antarctic organisms, P. branickii was able to activate hsp70 genes transcription (equal to heat shock response) in response to thermal stress. Finally, the unclear relationship between hsp70 expression and survival led us to surmise that genes other than hsp70 and other processes apart from the biochemical processes might generate the high thermaltolerance of P. branickii larvae. These results and future high-throughput studies at both the transcriptome and proteome level will improve our ability to predict the future geographic distribution of this species within the context of global warming. © 2010 Cell Stress Society International.

Bernabo P.,CNR Institute of Neuroscience | Bernabo P.,Center for Integrative Biology | Lunelli L.,Fondazione Bruno Kessler | Quattrone A.,Center for Integrative Biology | And 3 more authors.
Journal of Insect Physiology | Year: 2015

In stressed organisms, strategic proteins are selectively translated even if the global process of protein synthesis is compromised. The determination of protein concentrations in tissues of non-model organisms (thus with limited genomic information) is challenging due to the absence of specific antibodies. Moreover, estimating protein levels quantifying transcriptional responses may be misleading, because translational control mechanisms uncouple protein and mRNAs abundances. Translational control is increasingly recognized as a hub where regulation of gene expression converges to shape proteomes, but it is almost completely overlooked in molecular ecology studies. An interesting approach to study translation and its control mechanisms is the analysis of variations of gene-specific translational efficiencies by quantifying mRNAs associated to ribosomes. In this paper, we propose a robust and streamlined pipeline for purifying ribosome-associated mRNAs and calculating global and gene-specific translation efficiencies from non-model insect's species. This method might found applications in molecular ecology to study responses to environmental stressors in non-model organisms. © 2015 Elsevier Ltd.

Losapio G.,University of Milan | Losapio G.,University of Zurich | Gobbi M.,Section of Invertebrate Zoology and Hydrobiology | Marano G.,University of Milan | And 9 more authors.
Arthropod-Plant Interactions | Year: 2016

Primary successions of glacier forelands are unique model systems to investigate community dynamics and assembly processes. However, successional changes of plant and insect communities have been mainly analysed separately. Therefore, changes in plant–insect interactions along successional gradients on glacier forelands remain unknown, despite their relevance to ecosystem functioning. This study assessed how successional changes of the vegetation influenced the composition of the flower-visiting insect assemblages of two plant species, Leucanthemopsis alpina (L.) Heyw. and Saxifraga bryoides L., selected as the only two insect-pollinated species occurring along the whole succession. In addition, we investigated the links between reproductive output of these plants and pollinator abundance through experimental exclusion of pollinators. Plant community structure changed along the succession, affecting the distribution and the abundance of insects via idiosyncratic responses of different insect functional groups. L. alpina interacted with ubiquitously distributed pollinators, while S. bryoides pollinators were positively associated with insect-pollinated plant species density and S. bryoides abundance. With succession proceeding, insect assemblages became more functionally diverse, with the abundance of parasitoids, predators and opportunists positively related to an increase in plant cover and diversity. The reproductive output of both plant species varied among successional stages. Contrary to our expectation, the obligate insect-pollinated L. alpina showed a reproductive output rather independent from pollinator abundance, while the reproductive output of the self-fertile S. bryoides seemed linked to pollinator abundance. Observing ecological interactions and using functional traits, we provided a mechanistic understanding of community assembly processes along a successional gradient. Plant community diversity and cover likely influenced insect community assembly through bottom-up effects. In turn, pollinators regulate plant reproductive output through top-down control. We emphasise that dynamics of alpine plant and insect communities may be structured by biotic interactions and feedback processes, rather than only be influenced by harsh abiotic conditions and stochastic events. © 2016 Springer Science+Business Media Dordrecht

Grazioli V.,University of Milan | Rossaro B.,University of Milan | Parenti P.,University of Milan Bicocca | Giacchini R.,University of Milan Bicocca | Lencioni V.,Section of Invertebrate Zoology and Hydrobiology
Journal of Limnology | Year: 2016

The metabolic effects of low oxygen content on alcohol-dehydrogenase (ADH) activity and hemoglobin (Hb) concentration were investigated in IV-instar larvae of Chironomus riparius (Diptera: Chironomidae) from an Italian stream. Two series of short-term (48 h) experiments were carried out: exposure to (1) progressive hypoxia (95 to 5% of oxygen saturation) and (2) anoxia (at <5% of oxygen saturation). In (1), Hb amount increased with increasing oxygen depletion up to a critical value of oxygenation (about 70% of oxygen saturation). Below this percentage, the Hb amount declined to values comparable with those present in the control. The respiration rate (R) remained almost constant at oxygen saturation >50% and decreased significantly only after 48 h of treatment (= <5% of oxygen saturation) reaching values <100 μmolO2 gAFDW–1 h–1. ADH activity showed two phases of growth, within the first 14 h and over 18 h of exposure. Overall, we inferred that i) Hb might function as short-term oxygen storage, enabling animals to delay the onset of anaerobiosis; and ii) alcoholic fermentation co-occurs for a short time with aerobic respiration, becoming the prevalent metabolic pathway below 5% of oxygen saturation (<1 mg L–1). These considerations were supported also by results from anoxia exposure (2). In such condition, larvae were visibly stressed, becoming immobile after few minutes of incubation, and ADH reached higher values than in the hypoxia treatment (2.03±0.15 UADH mg prot–1). Overall, this study showed a shift from aerobic to anaerobic activity in C. riparius larvae exposed to poorly oxygenated water with an associated alteration of ADH activity and the Hb amount. Such metabolites might be valid candidate biomarkers for the environmental monitoring of running waters. © 2016, Page Press Publications. All rights reserved.

Pauls S.U.,Biodiversity and Climate Research Center | Alp M.,Leibniz Institute of Freshwater Ecology and Inland Fisheries | Alp M.,CNRS Biological Evolution and Diversity Laboratory | Balint M.,Biodiversity and Climate Research Center | And 14 more authors.
Freshwater Biology | Year: 2014

Summary: Molecular genetic techniques have been used in freshwater biology for more than 30 years. Early work focussed on studies of population structure, systematics and taxonomy. More recently, the range of studies has broadened to include ecology and adaptation. Advances in analytical methods and in technology (e.g. next-generation sequencing) and decreasing costs of data production ensure that the field will continue to develop and broaden in scope. At least three factors make the application of molecular techniques to freshwater biology exciting. First, the highly variable nature of many aquatic habitats makes them excellent models for the study of environmental change on ecological and evolutionary time scales. Second, the mature state of the field of freshwater biology provides an extensive foundation of ecological knowledge of freshwater organisms and their distinct adaptations. Third, the methodological advances allow researchers to focus more on merging molecular and ecological research and less on designing studies around technical limitations. We identified eight research areas in freshwater biology in which the integration of molecular and ecological approaches provides exceptional opportunities. The list is not exhaustive, but considers a broad range of topics and spans the continuum from basic to applied research. The areas identified use a combination of natural, experimental and in silico approaches. With advancing molecular techniques, freshwater biology is in an unusually strong position to link the genetic basis and ecological importance of adaptations across a wide range of taxa, ecosystems and spatiotemporal scales. Our aim was to identify opportunities for the integration of molecular and ecological approaches, to motivate greater collaboration and crossover, and to promote exploitation of the synergies of bridging ecological and evolutionary freshwater research. © 2014 John Wiley & Sons Ltd.

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