生长在超干旱环境下的3种相思树种表现出异常低的叶片、树枝、树干、根中δ13C含量

生长在超干旱环境下的3种相思树种表现出异常低的叶片、树枝、树干、根中δ¹³C含量 在植物生理生态学中,叶片中碳13(¹³C)含量负值较少(富集),表明叶片处于通过气孔的气体交换减少,比如在干旱胁迫下。此外,与叶片相比,13C在非光合组织中的负值也较少。然而,对从叶片(光合器官)到树枝、树干和根(非光合器官)中的δ ¹³C数值的关系知之甚少,特别是缺少在关联密切的多个树种间或者不同器官间,以及对生长在极端高温和干旱胁迫下的树木中进行测定。本研究测定了3种近缘沙漠相思树种(Acacia tortilis、A. raddiana和A. pachyceras)从叶片到根的¹³C含量。我们在以色列南部成树的叶片组织中测定了δ ¹³C含量。与此同时,在试验果园进行了为期7年的3个水平的灌溉试验。在试验结束时,测定了叶片、树枝、树干和根的生长参数和δ ¹³C含量。研究结果表明,叶片组织中δ ¹³C含量约为−27‰,其同位素贫化程度远超过生长在地球上最干燥和最热环境中的沙漠树种的预期值。在不同的相思树种和不同器官中,所有灌溉水平处理中的δ ¹³C含量并没有富集(−28‰到ca. −27‰),证实了在成熟相思树中的测定结果。在不同器官中,叶片δ ¹³C含量与树枝和根的δ ¹³C含量异常相似,甚至比树干的δ ¹³C含量负值更少。高度贫化的叶片δ ¹³C表明,尽管这些树木生长在极端干燥的生境中,但其气孔气体交换较高。非光合组织中缺乏δ ¹³C富集可能与叶片和异养组织生长的季节耦合有关。     

Removal of GABA(A) receptor γ2 subunits from parvalbumin neurons causes wide-ranging behavioral alterations

We investigated the behavioral significance of fast synaptic inhibition by αβγ2-type GABA(A) receptors on parvalbumin (Pv) cells. The GABA(A) receptor γ2 subunit gene was selectively inactivated in Pv-positive neurons by Cre/loxP recombination. The resulting Pv-Δγ2 mice were relatively healthy in the first postnatal weeks; but then as Cre started to be expressed, the mice progressively developed wide-ranging phenotypic alterations including low body weight, motor deficits and tremor, decreased anxiety levels, decreased pain sensitivity and deficient prepulse inhibition of the acoustic startle reflex and impaired spatial learning. Nevertheless, the deletion was not lethal, and mice did not show increased mortality even after one year. Autoradiography with t-butylbicyclophosphoro[(35)S]thionate suggested an increased amount of GABA(A) receptors with only α and β subunits in central nervous system regions that contained high levels of parvalbumin neurons. Using BAC-transgenesis, we reduced some of the Pv-Δγ2 phenotype by selectively re-expressing the wild-type γ2 subunit back into some Pv cells (reticular thalamic neurons and cerebellar Pv-positive neurons). This produced less severe impairments of motor skills and spatial learning compared with Pv-Δγ2 mice, but all other deficits remained. Our results reveal the widespread significance of fast GABAergic inhibition onto Pv-positive neurons for diverse behavioral modalities, such as motor coordination, sensorimotor integration, emotional behavior and nociception.     

Woodland strawberry axillary bud fate is dictated by a crosstalk of environmental and endogenous factors

Plant architecture is defined by fates and positions of meristematic tissues and has direct consequences on yield potential and environmental adaptation of the plant. In strawberries (Fragaria vesca L. and F. × ananassa Duch.), shoot apical meristems can remain vegetative or differentiate into a terminal inflorescence meristem. Strawberry axillary buds (AXBs) are located in leaf axils and can either remain dormant or follow one of the two possible developmental fates. AXBs can either develop into stolons needed for clonal reproduction or into branch crowns (BCs) that can bear their own terminal inflorescences under favorable conditions. Although AXB fate has direct consequences on yield potential and vegetative propagation of strawberries, the regulation of AXB fate has so far remained obscure. We subjected a number of woodland strawberry (F. vesca L.) natural accessions and transgenic genotypes to different environmental conditions and growth regulator treatments to demonstrate that strawberry AXB fate is regulated either by environmental or endogenous factors, depending on the AXB position on the plant. We confirm that the F. vesca GIBBERELLIN20-oxidase4 (FvGA20ox4) gene is indispensable for stolon development and under tight environmental regulation. Moreover, our data show that apical dominance inhibits the outgrowth of the youngest AXB as BCs, although the effect of apical dominance can be overrun by the activity of FvGA20ox4. Finally, we demonstrate that the FvGA20ox4 is photoperiodically regulated via FvSOC1 (F. vesca SUPPRESSOR OF OVEREXPRESSION OF CONSTANS1) at 18°C, but at higher temperature of 22°C an unidentified FvSOC1-independent pathway promotes stolon development.

Environmental conditions and apical dominance dictate woodland strawberry plant architecture by regulating axillary bud fate.     

Endotoxin depletion of recombinant protein preparations through their preferential binding to histidine tags

The presence of endotoxins in preparations of recombinantly produced therapeutic proteins poses serious problems for patients. Endotoxins can cause fever, respiratory distress syndromes, intravascular coagulation, or endotoxic shock. A number of methods have been devised to remove endotoxins from protein preparations using separation procedures based on molecular mass or charge properties. Most of the methods are limited in their endotoxin removal capacities and lack general applicability. We are describing a biotechnological approach for endotoxin removal. This strategy exploits the observation that endotoxins form micelles that expose negative charges on their surface, leading to preferential binding of endotoxins to cationic surfaces, allowing the separation from their resident protein. Endotoxins exhibit high affinity to stretches of histidines, which are widely used tools to facilitate the purification of recombinant proteins. They bind to nickel ions and are the basis for protein purification from cellular extracts by immobilized metal affinity chromatography. We show that the thrombin-mediated cleavage of two histidine tags from the purified recombinant protein and the adsorption of these histidine tags and their associated endotoxins to a nickel affinity column result in an appreciable depletion of the endotoxins in the purified protein fraction.     

Signals of Climate Change in Butterfly Communities in a Mediterranean Protected Area

The European protected-area network will cease to be efficient for biodiversity conservation, particularly in the Mediterranean region, if species are driven out of protected areas by climate warming. Yet, no empirical evidence of how climate change influences ecological communities in Mediterranean nature reserves really exists. Here, we examine long-term (1998–2011/2012) and short-term (2011–2012) changes in the butterfly fauna of Dadia National Park (Greece) by revisiting 21 and 18 transects in 2011 and 2012 respectively, that were initially surveyed in 1998. We evaluate the temperature trend for the study area for a 22-year-period (1990–2012) in which all three butterfly surveys are included. We also assess changes in community composition and species richness in butterfly communities using information on (a) species’ elevational distributions in Greece and (b) Community Temperature Index (calculated from the average temperature of species'' geographical ranges in Europe, weighted by species'' abundance per transect and year). Despite the protected status of Dadia NP and the subsequent stability of land use regimes, we found a marked change in butterfly community composition over a 13 year period, concomitant with an increase of annual average temperature of 0.95°C. Our analysis gave no evidence of significant year-to-year (2011–2012) variability in butterfly community composition, suggesting that the community composition change we recorded is likely the consequence of long-term environmental change, such as climate warming. We observe an increased abundance of low-elevation species whereas species mainly occurring at higher elevations in the region declined. The Community Temperature Index was found to increase in all habitats except agricultural areas. If equivalent changes occur in other protected areas and taxonomic groups across Mediterranean Europe, new conservation options and approaches for increasing species’ resilience may have to be devised.     

Butterfly and grasshopper diversity patterns in humid Mediterranean grasslands: the roles of disturbance and environmental factors

The present paper studies butterfly, grasshopper and vascular plant communities in ten seasonally flooded grasslands with different anthropogenic disturbance regimes (NW Greece). Disturbance intensity was assessed on the basis of disturbance frequency and type (grazing, mowing, trampling, constructions). The distribution patterns of butterflies are regulated by humidity and elevation (Redundancy Analysis). Elevation, flower-heads abundance, low disturbance intensity and plant species richness predict grasshopper species richness well, while the latter together with humidity predict plant species richness (Generalized Linear Models). Chorthippus lacustris, a critically endangered endemic grasshopper species, is positively associated with humid microhabitats with high flower-heads abundance. An indicator value procedure reveals four butterfly species as being typical species for habitats with a pronounced character of hedgerows and tree lines. Conservation management of grassland butterflies should focus on the maintenance of the humid character of the humid grasslands as well as on the maintenance of hedgerows and tree lines. The reduction of human-induced disturbance towards occasional grazing and mowing seems to benefit both butterfly and grasshopper communities. Finally, we suggest the use of grasshoppers as surrogates for vascular plants and vice versa, given their congruent species richness patterns.     

Rational design of a drug for Alzheimer's disease with cholinesterase inhibitory and neuroprotective activity

The rate and duration of inhibition of recombinant human acetylcholinesterase (AChE) and human butyrylcholinesterase (BuChE) by nine N-methyl,N-alkyl derivatives of (R)-3-prop-2-ynylamino-indan, designed as potential treatment of Alzheimer's disease, was obtained from measurement of the carbamylation k(i) and decarbamylation k(3) rate constants. This also provided information about the rate of formation of the leaving group, 6-OH-(R)-3-prop-2-ynylamino-indan, designed as an MAO-B inhibitor with neuroprotective activity. The N-dimethyl derivative had the highest k(i) of the alkyl derivatives. Substitution of one N-methyl by N-ethyl resulted in a 14-fold decrease in k(i) and 28-fold decrease in k(3). A progressive increase in k(i) occurred as the length of the alkyl chain progressed from propyl to n-hexyl and cyclo-hexyl, with relatively little or no increase in k(3). Higher k(i) values than that of the dimethyl analogue were obtained with the N-aryl substitutes, N-phenyl and N-methoxy-phenyl. Six of the compounds had much higher k(i) values for BuChE than AChE, but the N-cyclo-hexyl and N-methoxy-phenyl compounds were inactive. However, an inverse relation was found between k(i) and the degree of brain AChE inhibition ex vivo after parenteral administration of the compounds in rats. This could have resulted from more rapid hydrolysis of the compounds with high k(i) values by esterases in blood and liver. Only the N-ethyl and N-propyl derivatives showed AChE and BuChE inhibitory activity in vivo of a suitably slow onset and long duration, together with MAO-B inhibition.     

Tobacco mosaic virus movement protein functions as a structural microtubule-associated protein

The cell-to-cell spread of Tobacco mosaic virus infection depends on virus-encoded movement protein (MP), which is believed to form a ribonucleoprotein complex with viral RNA (vRNA) and to participate in the intercellular spread of infectious particles through plasmodesmata. Previous studies in our laboratory have provided evidence that the vRNA movement process is correlated with the ability of the MP to interact with microtubules, although the exact role of this interaction during infection is not known. Here, we have used a variety of in vivo and in vitro assays to determine that the MP functions as a genuine microtubule-associated protein that binds microtubules directly and modulates microtubule stability. We demonstrate that, unlike MP in whole-cell extract, microtubule-associated MP is not ubiquitinated, which strongly argues against the hypothesis that microtubules target the MP for degradation. In addition, we found that MP interferes with kinesin motor activity in vitro, suggesting that microtubule-associated MP may interfere with kinesin-driven transport processes during infection.     

Towards the co‐ordination of terrestrial ecosystem protocols across European research infrastructures

The study of ecosystem processes over multiple scales of space and time is often best achieved using comparable data from multiple sites. Yet, long‐term ecological observatories have often developed their own data collection protocols. Here, we address this problem by proposing a set of ecological protocols suitable for widespread adoption by the ecological community. Scientists from the European ecological research community prioritized terrestrial ecosystem parameters that could benefit from a more consistent approach to data collection within the resources available at most long‐term ecological observatories. Parameters for which standard methods are in widespread use, or for which methods are evolving rapidly, were not selected. Protocols were developed by domain experts, building on existing methods where possible, and refined through a process of field testing and training. They address above‐ground plant biomass; decomposition; land use and management; leaf area index; soil mesofaunal diversity; soil C and N stocks, and greenhouse gas emissions from soils. These complement existing methods to provide a complete assessment of ecological integrity. These protocols offer integrated approaches to ecological data collection that are low cost and are starting to be used across the European Long Term Ecological Research community.