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Brachypodium distachyon has emerged as the model species for important temperate grass crops such as wheat and barley and the genome of the B. distachyon community inbred line Bd21 has been sequenced. Methods for tissue culture and Agrobacterium-mediated transformation have been developed for this model grass as a resource for reverse genetics and functional genomic analyses. In order to obtain a high quantity and quality of compact embryogenic callus (CEC) in B. distachyon, it is important to examine and optimize the optimal concentration of the auxin 2,4-D (dichlorophenoxyacetic acid) to use in both callus induction and callus proliferation media. Here, we investigated the effects of different concentrations of 2,4-D on callus induction and callus proliferation of B. distachyon Bd21. Our results showed that 2.5 mg l−1 2,4-D is an optimal concentration to use for both callus induction and proliferation, although 5.0 mg l−1 may also be used for callus proliferation. Additionally, the suitability of hygromycin or bialaphos as selectable markers was examined and results indicated that hygromycin is significantly more efficient than bialaphos when using the Agrobacterium-mediated transformation system.

Fusarium head blight (FHB) is one of the most devastating diseases of wheat (Triticum aestivum), barley (Hordeum vulgare) and other small grain cereals grown in warm and humid regions worldwide. In addition to yield loss, the disease compromises the quality of infected grain as a result of contamination with a range of Fusarium mycotoxins that are harmful to human and animal health. Deoxynivalenol (DON) is the most prevalent trichothecene mycotoxin found in Fusarium-infected grains. DON acts as a virulence factor for Fusarium, facilitating disease spread within wheat heads. Resistance to DON is an innate component of FHB resistance. Here we review FHB as a globally important disease, with a specific focus on the role of DON in disease development, the importance of its’ resistance in plant defence against Fusarium and the current knowledge regarding the genes activated as part of the cereal defence against the toxin.

Deoxynivalenol (DON) is a mycotoxin produced by phytopathogenic Fusarium fungi in cereal grain and plays a role as a disease virulence factor. TaFROG (Triticum aestivum Fusarium Resistance Orphan Gene) enhances wheat resistance to DON and it interacts with a sucrose non-fermenting-1 (SNF1)-related protein kinase 1 catalytic subunit α (SnRK1α). This protein kinase family is central integrator of stress and energy signalling, regulating plant metabolism and growth. Little is known regarding the role of SnRK1α in the biotic stress response, especially in wheat. In this study, 15 wheat (Triticum aestivum) SnRK1α genes (TaSnRK1αs) belonging to four homoeologous groups were identified in the wheat genome. TaSnRK1αs are expressed ubiquitously in all organs and developmental stages apart from two members predominantly detected in grain. While DON treatment had either no effect or downregulated the transcription of TaSnRK1αs, it increased both the kinase activity associated with SnRK1α and the level of active (phosphorylated) SnRK1α. Down-regulation of two TaSnRK1αs homoeolog groups using virus induced gene silencing (VIGS) increased the DON-induced damage of wheat spikelets. Thus, we demonstrate that TaSnRK1αs contribute positively to wheat tolerance of DON and conclude that this gene family may provide useful tools for the improvement of crop biotic stress resistance. Science Foundation Ireland Irish Department of Agriculture Biotechnological and Biological Sciences Research Council of the United Kingdom Designing Future Wheat (DFW) strategic programme update copyright and citation details during checkdate report - AC

The cyclization of aza-β3-tetrapeptides gives access to new CTP (cyclotetrapeptide) analogues. These stereocontrolled templates are assembled without any asymmetric synthesis. X-ray crystallographic structure and NMR analysis show that the macrocyclic scaffold is characterized by a fully cooperative intramolecular H-bond network, in sharp contrast with the nanotubular assemblies observed for β3-cyclotetrapeptides. This folding property reduces considerably the polarity of aza-β3-tetrapeptides and should be useful in addressing intracellular targets.

Highlight Genes that enhance resistance to the Fusarium virulence factor deoxynivalenol (DON) are targets for disease resistance breeding. This study provides direct evidence that a wheat ABCC3 enhances resistance to DON., The mycotoxin deoxynivalenol (DON) acts as a disease virulence factor for Fusarium fungi, and tolerance of DON enhances wheat resistance to Fusarium head blight (FHB) disease. Two variants of an ATP-binding cassette (ABC) family C transporter gene were cloned from DON-treated wheat mRNA, namely TaABCC3.1 and TaABCC3.2. These represent two of three putative genes identified on chromosomes 3A, 3B, and 3D of the wheat genome sequence. Variant TaABCC3.1 represents the DON-responsive transcript previously associated with DON resistance in wheat. PCR-based mapping and in silico sequence analyses located TaABCC3.1 to the short arm of wheat chromosome 3B (not within the FHB resistance quantitative trait locus Fhb1). In silico analyses of microarray data indicated that TaABCC3 genes are expressed in reproductive tissue and roots, and in response to the DON producer Fusarium graminearum. Gene expression studies showed that TaABCC3.1 is activated as part of the early host response to DON and in response to the FHB defence hormone jasmonic acid. Virus-induced gene silencing (VIGS) confirmed that TaABCC3 genes contributed to DON tolerance. VIGS was performed using two independent viral construct applications: one specifically targeted TaABCC3.1 for silencing, while the other targeted this gene and the chromosome 3A homeologue. In both instances, VIGS resulted in more toxin-induced discoloration of spikelets, compared with the DON effects in non-silenced spikelets at 14 d after toxin treatment (≥2.2-fold increase, P

International audience; A novel radical gold dithiolene complex exhibits a hexagonal columnar mesophase, as confirmed by optical microscopy , DSC analysis and X-ray diffraction . The extent of delocalization of the spin density in such a complex was analyzed by EPR . Temperature dependent magnetization measurements reveal that the global magnetic moment is remarkably affected at the liquid-crystalline phase transition with a marked hysteresis signature, rare behavior among the few described paramagnetic discotic phases. In addition, these molecules were found to strongly aggregate in solution into one-dimensional fibers with a mean diameter of 60 nm extending over micrometres, leading to the formation of gel-like structures. These fibers are stable and can be isolated on surfaces. The gelation of the system can also be detected by temperature-dependent magnetic measurements.

Calmodulin (CaM) is a primary calcium sensor in all eukaryotes. It binds calcium and regulates the activity of a wide range of effector proteins in response to calcium signals. The list of CaM targets includes plant-specific proteins whose functions are progressively being elucidated. Plants also possess numerous calmodulin-like proteins (CMLs) that appear to have evolved unique functions. Functional studies of CaM and CMLs in plants highlight the importance of this protein family in the regulation of plant development and stress responses by converting calcium signals into transcriptional responses, protein phosphorylation or metabolic changes. This review summarizes some of the significant progress made by biochemical and genetic studies in identifying the properties and physiological functions of plant CaMs and CMLs. We discuss emerging paradigms in the field and highlight the areas that need further investigation.

Through steady-state and time-resolved fluorescence experiments, the polarity of the bilayers of egg phosphatidylcholine vesicles was studied by means of the solvatochromic 2-anthroyl fluorophore which we have recently introduced for investigating the environmental micropolarity of membranes and which was incorporated synthetically in phosphatidylcholine molecules (anthroyl-PC) in the form of 8-(2-anthroyl)octanoic acid. Fluorescence quenching experiments carried out with N,N-dimethylaniline and 12-doxylstearic acid as quenchers showed that the 2-anthroyl chromophore was located in depth in the hydrophobic region of the lipid bilayer corresponding to the C9-C16 segment of the acyl chains. Steady-state fluorescence spectroscopy revealed a nonstructured and red-shifted (lambda em(max) = 464 nm) spectrum for the probe in egg-PC bilayers, which greatly differed from the structured and blue (lambda em(max) = 404 nm) spectrum the fluorophore was shown to display in n-hexane. While the fluorescence decays of the fluorophore in organic solvents were monoexponential, three exponentials were required to account for the fluorescence decays of anthroyl-PC in egg-PC vesicles, with average characteristic times of 1.5 ns, 5.5 ns, and 20 ns. These lifetime values were independent of the emission wavelength used. Addition of cholesterol to the lipid did not alter these tau values. One just observed an increase in the fractional population of the 1.5-ns short-living species detrimental to the population of the 20-ns long-living ones. These observations enabled time-resolved fluorescence spectroscopy measurements to be achieved in the case of the 1/1 (mol/mol) egg-PC/cholesterol mixture. Three distinct decay associated spectra (DAS) were recorded, with maximum emission wavelengths, respectively, of 410 nm, 440 nm, and 477 nm for the 1.5-ns, 6-ns, and 20-ns lifetimes found in this system. On account of the properties and the polarity scale previously established for the 2-anthroyl chromophore in organic solvents, these data strongly suggest the occurrence of three distinct excited states for anthroyl-PC in egg-PC bilayers, corresponding to three environments for the 2-anthroyl chromophore, differing in polarity. The lifetime of 1.5 ns and the corresponding structured and blue (lambda em(max) = 410 nm) DAS account for a hydrophobic environment, with an apparent dielectric constant of 2, which is that expected for the hydrophobic core of the lipid bilayer.(ABSTRACT TRUNCATED AT 400 WORDS)

10 pages; International audience; The sulfiding of benzils with P4S10 in 1,3-dimethyl 2-imidazolidinone (DMI) as solvent allows for a direct synthesis of neutral radical, gold dithiolene complexes based on 1,2-bis-(4-alkoxyphenyl) ethylene-1,2-dithiolate ligands with n-butyl, n-octyl and n-dodecyl chains. The three neutral and soluble complexes Au-OC4, Au-OC8 and Au-OC12 exhibit a near infrared (NIR) absorption band around 1.5 mm and EPR characteristics which confirm a strong delocalization of the spin density on the electron-rich dithiolene ligands. X-Ray crystal structures of Au-OC4 and Au-OC12 are compared with those of the corresponding nickel complexes. They are characterised by segregation of the alkyl chains into layered structures with a stacking of the radical complexes into alternated spin chains, confirmed by the temperature dependence of the magnetic susceptibility which attests for antiferromagnetic interactions and a singlet ground state. Observations under polarising microscope and DSC experiments do not reveal a thermotropic behaviour for Au-OC12

Reaction of racemic (dl)- or enantiopure (D)-camphorquinone with P4S10 in dimethylimidazolidinone followed by addition of NiCl2·6H2O or KAuCl4 affords the corresponding bornylenedithiolato metal complexes [(dl-bordt)2Ni], [(D-bordt)2Ni], [(dl-bordt)2Au], and [(D-bordt)2Au] in their neutral form. Cyclic voltammetry and UV/Vis/NIR spectrometry show that the bornylenedithiolato ligand acts as a highly electron-rich dithiolene ligand. Complete assignment of the 1H and 13C NMR signals of the diamagnetic enantiopure [(D-bordt)2Ni] is reported thanks to a combination of 2D NMR spectroscopy experiments. X-ray crystal structure determinations were performed for the four complexes, showing that the two diastereomeric mixtures, [(dl-bordt)2Ni] and [(dl-bordt)2Au], are isomorphous (space group P21/c), as are also the two enantiopure complexes, [(D-bordt)2Ni] and [(D-bordt)2Au] (space group P21). Positional disorder of the two (D)- and (L)-bornylenedithiolate enantiomers on the same position is observed in the structure of the diastereomeric mixtures, with the complex located on the inversion center. The temperature dependence of the magnetic susceptibility of the paramagnetic gold complexes [(dl-bordt)2Au] and [(D-bordt)2Au] demonstrates the presence of antiferromagnetic interactions, to a larger extent in the diastereoisomeric mixture [(dl-bordt)2Au] than in the enantiopure complex [(D-bordt)2Au], as a consequence of stronger lateral S···S intermolecular interactions in the former. (© Wiley-VCH Verlag GmbH & Co. KGaA, 69451 Weinheim, Germany, 2009)

RNA-d2 is a user-friendly program developed for interactively generating aesthetic and non-overlapping drawings of RNA secondary structures. It designed so that the drawings can be edited in a very natural and intuitive way, in order to emphasize structural homologies between several molecules, as well as the foldings themselves to update the base-pair sets according to new data. The program automatically produces a polygonal display in which the unpaired nucleotides are regularly positioned on circles and the stems harmoniously distributed on their periphery. RNA secondary structures can be encoded via the keyboard, but the program also automatically draws output files from thermodynamic prediction programs. The user interacts directly on different screen displays according to the editing functions. Rotation/translation of any subdomain and deletion of stems are performed on a coloured backbone view to make easier the identification of structural features, whereas addition of new base-pairings and numbering manipulation are realized on a complete polygonal view. Each modification is displayed in real time on the screen. When the display is obscured by numerous overlaps despite the colour code of the backbone view, an automatic function progressively straightens the subdomains which are highly compacted by very dissymmetric internal loops. RNA-d2 allows easy untangling and editing of RNA molecules > 1000 nucleotides long.

Mismatch between the hydrophobic thicknesses of transmembrane proteins and the supporting lipid bilayer and its consequences on the lateral organization of lipids have been investigated with bacteriorhodopsin and phosphatidylcholine species with a variety of acyl-chain lengths. The purple membrane, from the bacterium Halobacterium halobium, was used and reconstituted with dilauroyl-(Lau2GroPCho), dimyristoyl- (Myr2GroPCho), dipalmitoyl- (Pam2GroPCho) and distearoyl- (Ste2-GroPCho) glycerophosphocholine. The phase behaviour of the lipids was investigated at different temperatures and different protein/lipid molar ratios, by analyzing the fluorescence excitation spectra of the 1-acyl-2-[8-(2-anthroyl)-octanoyl]-sn-glycero-3-phosphocholine probe, and by measuring the fluorescence depolarization of the 1,6-diphenyl-1,3,5-hexatriene probe. Data obtained with 1-acyl-2-[8-(2-anthroyl)-octanoyl]-sn-glycero-3-phosphocholine shows that bacteriorhodopsin produced positive or negative shifts in the phase transition temperature of the host lipids depending on the strength and sign of the mismatch between the lipid and protein hydrophobic thicknesses and also on the protein concentration and aggregation state in the lipid bilayer. In the region of high protein concentration (bacteriorhodopsin/phosphatidylcholine molar ratios ∼ 1:50) and despite the presence of the endogenous lipids, bacteriorhodopsin (hydrophobic length dp∼ 3.0–3.1 nm) brought about a large upward shift in the phase-transition temperature of Lau2GroPCho (ΔT∼ 40 K, mean hydrophobic thickness d∼ 2.4 nm), and to a lesser extent of Myr2GroPCho (ΔT∼ 23 K, d∼ 2.8 nm), accounting for a strong rigidifying effect of the protein on these short-chain lipids. Bacteriorhodopsin had no influence on the phase properties of Pam2GroPCho (ΔT∼ 0 K, d∼ 3.2 nm), a lipid whose mean hydrophobic thickness is similar to that of the protein. In contrast, the transition temperature of Ste2GroPCho was decreased (ΔT∼−13 K, d∼ 3.7 nm), indicating a fluidifying effect of the protein on this long-chain lipid. Similar effects on the lipid acyl-chain order were observed in the region of high-protein dilution (bacteriorhodopsin/phosphatidylcholine molar ratios

Fluorescence excitation and emission spectra, relative fluorescence quantum yield φ r and fluorescence lifetime τ of methyl 8-(2-anthroyl)-octanoate have been studied in a set of organic solvents covering a large scale of polarity and in the presence of water. In this probe, the 2-anthroyl chromophore exhibits quite remarkable and unique fluorescence properties. Thus, when going from n -hexane to methanol, the maximum emission wavelength λ em max shifts from 404 nm to 492 nm while φ r and τ increase from 1 to 17.7 and from 0.91 ns to 13.5 ns, respectively. These increments are still more accentuated in the presence of water with estimated values of 526 nm for λ em max , 27 for φ r and 20 ns for τ in this solvent. Because of the presence of a keto group which is a hydrogen bond acceptor and which can conjugate with the aromatic ring so as to provide the chromophore with a high dipole moment, the fluorescence properties of the probe strongly depend on the polarity of the surrounding medium. They can be accounted for in terms of general solvent effects (dipolar solute/solvent interactions) in the presence of aprotic solvents and in terms of specific solvent effects (hydrogen bonding) in protic solvents. Such properties of solvatochromism make the 2-anthroyl chromophore, after 8-(2-anthroyl)octanoic acid has been attached to phospholipids (E. Perochon and J.F. Tocanne (1991) Chem. Phys. Lipids 58, 7–17) a potential tool for studying microenvironmental polarity in biological membranes.