Dispersive and non-dispersive waves through plants: implications for arthropod vibratory communication

Vibratory communication in arthropods is a widespread phenomenon. Arthropods living on plants have been reported to use only dispersive bending waves in the context of prey-predator, competition, social and sexual interactions. Differences in signal structure have also been postulated to work as species recognition mechanisms and speciation agents. Using two identical laser Doppler vibrometers and a wavelet analysis, we quantified the wave propagation modes in rush stems (Juncus effusus) over the whole range of frequencies used by arthropods. A non-dimensionalized analysis shows that mechanical waves propagate not only as dispersive bending waves, but also as non-dispersive waves. Our analysis implies that an arthropod can communicate through non-dispersive bending waves by either producing signals of high frequencies or by choosing large stems, two widely different options tapping into the physiological and the behavioural repertoires, respectively. Non-dispersive waves, unreported so far in insect vibratory communication in plants, present serious advantages over dispersive bending waves in terms of signal integrity and may well be much more widely used than anticipated, in particular for species recognition.     

Ecotourism disturbance to wildfowl in protected areas: historical, empirical and experimental approaches in the Camargue, Southern France

Ecotourism is becoming very popular, especially in protected areas where wildlife concentrate and is easier to observe, but the consequences of associated disturbance have seldom be quantified other than in the short-term, making the sustainability of this activity untested. We combined a historical, an empirical and an experimental approach to assess the long-, medium- and short-term consequences of disturbance to wintering wildfowl (Anatidae) in a wetland of international importance in the Camargue, Southern France. In the short-term, disturbance made teal (Anas crecca) move away temporarily from observation blinds without leaving the waterbody. Wildfowl fed more after disturbance, disrupting their normal resting activities. In the medium-term, waterbodies with more tourists did not host fewer birds: conversely the most heavily disturbed one hosted the highest wildfowl density. In the long term, wildfowl numbers were not related with the number of visitors. When practiced with appropriate guiding of people, and where appropriate facilities are provided to limit human disturbance as done here, ecotourism may not affect wintering wildfowl other than reversibly in the very short term. The legitimate demand of the public for access, even in fragile protected areas, may therefore be sustainable under some conditions.     

Effect of hydrophobic modification on rheological and swelling features during chemical gelation of aqueous polysaccharides

Rheological characteristics during chemical gelation with the cross-linker ethylene glycol diglycidyl ether (EGDE) of semidilute aqueous solutions of hydroxyethylcellulose (HEC) and of two hydrophobically modified analogues (HM-1-HEC and HM-2-HEC) are reported. In addition, rheological features of gelling samples (dextran and its hydrophobically modified analogue (HM-dextran)) of a different structure have been examined. Some swelling experiments on these gels in the postgel region are also reported. The gelation time of the hydroxyethylcellulose systems decreased with increasing cross-linker concentration, and incorporation of hydrophobic units of HEC resulted in a slower gelation. The time of gelation for the dextran system was only slightly affected by the incorporation of hydrophobic groups (HM-dextran). At the gel point, a power law frequency dependence of the dynamic storage modulus (G' proportional to omegan') and loss modulus (G' proportional to omegan') was observed for all gelling systems with n' = n' = n. The attachment of hydrophobic moieties on the dextran chains had virtually no impact on the value of n (n = 0.77), and the percolation model describes the incipient dextran gels. By increasing the number of hydrophobic groups of the HEC polymer, the value of n for the corresponding incipient gel drops significantly, and the value of the gel strength parameter increases strongly. Incorporation of hydrophobic units in the HEC chains promotes the formation of stronger incipient gels because of the contribution from the hydrophobic association effect. The frequency dependence of the complex viscosity reveals that all the investigated gels become more solidlike in the postgel domain. Far into the postgel region, the hydrophobicity of HEC plays a minor role for the strength of the gel network, whereas the values of the complex viscosity are significantly higher for HM-dextran than for the corresponding dextran gel. The swelling experiments on HEC, HM-1-HEC, and HM-2-HEC systems disclose that the degree of swelling of the postgels in water is quite different, depending on the relative distance from the gel point at which the cross-linker reaction is quenched. At a given distance from the gel point, the swelling of the HEC gel is less pronounced than for the corresponding hydrophobically modified samples. At this stage, the swelling of the HM-dextran gel is stronger than for the dextran gel.     

Evidence for ACD5 ceramide kinase activity involvement in Arabidopsis response to cold stress

Although sphingolipids emerged as important signals for plant response to low temperature, investigations have been limited so far to the function of long‐chain base intermediates. The formation and function of ceramide phosphates (Cer‐Ps) in chilled Arabidopsis were explored. Cer‐Ps were analysed by thin layer chromatography (TLC) following in vivo metabolic radiolabelling. Ceramide kinase activity, gene expression and growth phenotype were determined in unstressed and cold‐stressed wild type (WT) and Arabidopsis ceramide kinase mutant acd5. A rapid and transient formation of Cer‐P occurs in cold‐stressed WT Arabidopsis plantlets and cultured cells, which is strongly impaired in acd5 mutant. Although concomitant, Cer‐P formation is independent of long‐chain base phosphate (LCB‐P) formation. No variation of ceramide kinase activity was measured in vitro in WT plantlets upon cold stress but the activity in acd5 mutant was further reduced by cold stress. At the seedling stage, acd5 response to cold was similar to that of WT. Nevertheless, acd5 seed germination was hypersensitive to cold and abscisic acid (ABA), and ABA‐dependent gene expression was modified in acd5 seeds when germinated at low temperature. Our data involve for the first time Cer‐P and ACD5 in low temperature response and further underline the complexity of sphingolipid signalling operating during cold stress.     

Probing the Acceptor Active Site Organization of the Human Recombinant β1,4-Galactosyltransferase 7 and Design of Xyloside-based Inhibitors

Among glycosaminoglycan (GAG) biosynthetic enzymes, the human β1,4-galactosyltransferase 7 (hβ4GalT7) is characterized by its unique capacity to take over xyloside derivatives linked to a hydrophobic aglycone as substrates and/or inhibitors. This glycosyltransferase is thus a prime target for the development of regulators of GAG synthesis in therapeutics. Here, we report the structure-guided design of hβ4GalT7 inhibitors. By combining molecular modeling, in vitro mutagenesis, and kinetic measurements, and in cellulo analysis of GAG anabolism and decorin glycosylation, we mapped the organization of the acceptor binding pocket, in complex with 4-methylumbelliferone-xylopyranoside as prototype substrate. We show that its organization is governed, on one side, by three tyrosine residues, Tyr¹⁹⁴, Tyr¹⁹⁶, and Tyr¹⁹⁹, which create a hydrophobic environment and provide stacking interactions with both xylopyranoside and aglycone rings. On the opposite side, a hydrogen-bond network is established between the charged amino acids Asp²²⁸, Asp²²⁹, and Arg²²⁶, and the hydroxyl groups of xylose. We identified two key structural features, i.e. the strategic position of Tyr¹⁹⁴ forming stacking interactions with the aglycone, and the hydrogen bond between the His¹⁹⁵ nitrogen backbone and the carbonyl group of the coumarinyl molecule to develop a tight binder of hβ4GalT7. This led to the synthesis of 4-deoxy-4-fluoroxylose linked to 4-methylumbelliferone that inhibited hβ4GalT7 activity in vitro with a K_i 10 times lower than the K_m value and efficiently impaired GAG synthesis in a cell assay. This study provides a valuable probe for the investigation of GAG biology and opens avenues toward the development of bioactive compounds to correct GAG synthesis disorders implicated in different types of malignancies.     

Disruption of Nucleotide Homeostasis by the Antiproliferative Drug 5-Aminoimidazole-4-carboxamide-1-β-d-ribofuranoside Monophosphate (AICAR)

5-Aminoimidazole-4-carboxamide-1-β-d-ribofuranoside monophosphate (AICAR) is a natural metabolite with potent anti-proliferative and low energy mimetic properties. At high concentration, AICAR is toxic for yeast and mammalian cells, but the molecular basis of this toxicity is poorly understood. Here, we report the identification of yeast purine salvage pathway mutants that are synthetically lethal with AICAR accumulation. Genetic suppression revealed that this synthetic lethality is in part due to low expression of adenine phosphoribosyl transferase under high AICAR conditions. In addition, metabolite profiling points to the AICAR/NTP balance as crucial for optimal utilization of glucose as a carbon source. Indeed, we found that AICAR toxicity in yeast and human cells is alleviated when glucose is replaced by an alternative carbon source. Together, our metabolic analyses unveil the AICAR/NTP balance as a major factor of AICAR antiproliferative effects.     

Involvement of Arabidopsis Hexokinase1 in Cell Death Mediated by Myo-Inositol Accumulation

Programmed cell death (PCD) is essential for several aspects of plant life, including development and stress responses. We recently identified the mips1 mutant of Arabidopsis thaliana, which is deficient for the enzyme catalyzing the limiting step of myo-inositol (MI) synthesis. One of the most striking features of mips1 is the light-dependent formation of lesions on leaves due to salicylic acid (SA)-dependent PCD. Here, we identified a suppressor of PCD by screening for mutations that abolish the mips1 cell death phenotype. Our screen identified the hxk1 mutant, mutated in the gene encoding the hexokinase1 (HXK1) enzyme that catalyzes sugar phosphorylation and acts as a genuine glucose sensor. We show that HXK1 is required for lesion formation in mips1 due to alterations in MI content, via SA-dependant signaling. Using two catalytically inactive HXK1 mutants, we also show that hexokinase catalytic activity is necessary for the establishment of lesions in mips1. Gas chromatography-mass spectrometry analyses revealed a restoration of the MI content in mips1 hxk1 that it is due to the activity of the MIPS2 isoform, while MIPS3 is not involved. Our work defines a pathway of HXK1-mediated cell death in plants and demonstrates that two MIPS enzymes act cooperatively under a particular metabolic status, highlighting a novel checkpoint of MI homeostasis in plants.