Non-targeted metabolic profiling of BW312 Hordeum vulgare semi dwarf mutant using UHPLC coupled to QTOF high resolution mass spectrometry

Abstract Barley (Hordeum vulgare) is the fourth crop cultivated in the world for human consumption and animal feed, making it important to breed healthy and productive plants. Among the threats for barley are lodging, diseases, and pathogens. To avoid lodging, dwarf and semi-dwarf mutants have been selected through breeding processes. Most of these mutants are affected on hormonal biosynthesis or signalling. Here, we present the metabolic characterization of a brassinosteroid insensitive semi-dwarf mutant, BW312. The hormone profile was determined through a targeted metabolomics analysis by UHPLC-triple quadrupole-MS/MS, showing an induction of gibberellic acid and jasmonic acid in the semi-dwarf mutant. A non-targeted metabolomics analysis by UHPLC-QTOF-MS/MS revealed a differential metabolic profile, with 16 and 9 metabolites showing higher intensities in the mutant and wild-type plants respectively. Among these metabolites, azelaic acid was identified. Gibberellic acid, jasmonic acid, and azelaic acid are involved in pathogen resistance, showing that this semi-dwarf line has an enhanced basal pathogen resistance in absence of pathogens, and therefore is of interest in breeding programs to fight against lodging, but also probably to increase pathogen resistance

Correlated production and consumption of chloromethane in the Arabidopsis thaliana phyllosphere

Chloromethane (CH3Cl) is a toxic gas mainly produced naturally, in particular by plants, and its emissions contribute to ozone destruction in the stratosphere. Conversely, CH3Cl can be degraded and used as the sole carbon and energy source by specialised methylotrophic bacteria, isolated from a variety of environments including the phyllosphere, i.e. the aerial parts of vegetation. The potential role of phyllospheric CH3Cl-degrading bacteria as a filter for plant emissions of CH3Cl was investigated using variants of Arabidopsis thaliana with low, wild-type and high expression of HOL1 methyltransferase previously shown to be responsible for most of CH3Cl emissions by A. thaliana. Presence and expression of the bacterial chloromethane dehalogenase cmuA gene in the A. thaliana phyllosphere correlated with HOL1 genotype, as shown by qPCR and RT-qPCR. Production of CH3Cl by A. thaliana paralleled HOL1 expression, as assessed by a fluorescence-based bioreporter. The relation between plant production of CH3Cl and relative abundance of CH3Cl-degrading bacteria in the phyllosphere suggests that CH3Cl-degrading bacteria co-determine the extent of plant emissions of CH3Cl to the atmosphere

Mutations in UDP-Glucose:Sterol Glucosyltransferase in Arabidopsis Cause Transparent Testa Phenotype and Suberization Defect in Seeds

In higher plants, the most abundant sterol derivatives are steryl glycosides (SGs) and acyl SGs. Arabidopsis (Arabidopsis thaliana) contains two genes, UGT80A2 and UGT80B1, that encode UDP-Glc:sterol glycosyltransferases, enzymes that catalyze the synthesis of SGs. Lines having mutations in UGT80A2, UGT80B1, or both UGT80A2 and UGT8B1 were identified and characterized. The ugt80A2 lines were viable and exhibited relatively minor effects on plant growth. Conversely, ugt80B1 mutants displayed an array of phenotypes that were pronounced in the embryo and seed. Most notable was the finding that ugt80B1 was allelic to transparent testa15 and displayed a transparent testa phenotype and a reduction in seed size. In addition to the role of UGT80B1 in the deposition of flavanoids, a loss of suberization of the seed was apparent in ugt80B1 by the lack of autofluorescence at the hilum region. Moreover, in ugt80B1, scanning and transmission electron microscopy reveals that the outer integument of the seed coat lost the electron-dense cuticle layer at its surface and displayed altered cell morphology. Gas chromatography coupled with mass spectrometry of lipid polyester monomers confirmed a drastic decrease in aliphatic suberin and cutin-like polymers that was associated with an inability to limit tetrazolium salt uptake. The findings suggest a membrane function for SGs and acyl SGs in trafficking of lipid polyester precursors. An ancillary observation was that cellulose biosynthesis was unaffected in the double mutant, inconsistent with a predicted role for SGs in priming cellulose synthesis

Vitamin D5 in Arabidopsis thaliana

Abstract Vitamin D3 is a secosterol hormone critical for bone growth and calcium homeostasis, produced in vertebrate skin by photolytic conversion of the cholesterol biosynthetic intermediate provitamin D3. Insufficient levels of vitamin D3 especially in the case of low solar UV-B irradiation is often compensated by an intake of a dietary source of vitamin D3 of animal origin. Small amounts of vitamin D3 were described in a few plant species and considered as a peculiar feature of their phytochemical diversity. In this report we show the presence of vitamin D5 in the model plant Arabidopsis thaliana. This plant secosterol is a UV-B mediated derivative of provitamin D5, the precursor of sitosterol. The present work will allow a further survey of vitamin D distribution in plant species

Fundamental parameters of Be stars located in the seismology fields of COROT

In preparation for the COROT space mission, we determined the fundamental parameters (spectral type, temperature, gravity, vsini) of the Be stars observable by COROT in its seismology fields (64 Be stars). We applied a careful and detailed modeling of the stellar spectra, taking into account the veiling caused by the envelope, as well as the gravitational darkening and stellar flattening due to rapid rotation. Evolutionary tracks for fast rotators were used to derive stellar masses and ages. The derived parameters will be used to select Be stars as secondary targets (i.e. observed for 5 consecutive months) and short-run targets of the COROT mission. Furthermore, we note that the main part of our stellar sample is falling in the second half of the main sequence life time, and that in most cases the luminosity class of Be stars is inaccurate in characterizing their evolutionary status.Comment: 25 pages, 9 figures, Accepted for publication in A&

Early-type objects in NGC6611 and Eagle Nebula

An important question about Be stars is whether Be stars are born as Be stars or not. It is necessary to observe young clusters to answer this question. Observations of stars in NGC6611 and the star-formation region of Eagle Nebula have been carried out with the ESO-WFI in slitless spectroscopic mode and at the VLT-GIRAFFE. The targets for the GIRAFFE observations were pre-selected from the literature and our catalogue of emission-line stars (ELS) based on the WFI study. GIRAFFE observations allowed us to study accurately the population of the early-type stars with and without emission lines. For this study, we determined the fundamental parameters of OBA stars. We also studied the status of the objects (main sequence or pre-main sequence stars) by using IR data, membership probabilities, and location in HR diagrams. The nature of the early-type ELS in M16 is derived. The slitless observations with the WFI clearly indicate a small number of ELS in M16. We observed with GIRAFFE 101 OBA stars, among them 9 are ELS with circumstellar emission in Halpha. We found that: W080 could be a new He-strong star, like W601. W301 is a possible classical Be star, W503 is a mass-transfer eclipsing binary with an accretion disk, and the other ones are possible Herbig Ae/Be stars. We also found that the rotational velocities of main sequence B stars are 18% lower than those of pre-main sequence B stars, in good agreement with theory about the evolution of rotational velocities. Combining different indications and technics, we found that 27% of the B-type stars are binaries. We also redetermined the age of NGC6611 found equal to 1.2--1.8 Myears in good agreement with the most recent determinations.Comment: Accepted by A&A, english not yet correcte

On the evolutionary status of Be stars. I. Field Be stars near the Sun

A sample of 97 galactic field Be stars were studied by taking into account the effects induced by the fast rotation on their fundamental parameters. All program stars were observed in the BCD spectrophotometric system in order to minimize the perturbations produced by the circumstellar environment on the spectral photospheric signatures. This is one of the first attempts at determining stellar masses and ages by simultaneously using model atmospheres and evolutionary tracks, both calculated for rotating objects. The stellar ages ($\tau$) normalized to the respective inferred time that each rotating star can spend in the main sequence phase ($\tau\_{\rm MS}$) reveal a mass-dependent trend. This trend shows that: a) there are Be stars spread over the whole interval 0 \la \tau/\tau\_{\rm MS} \la 1 of the main sequence evolutionary phase; b) the distribution of points in the ($\tau/\tau\_{\rm MS},M/M\_{\odot}$) diagram indicates that in massive stars (M \ga 12M\_{\odot}) the Be phenomenon is present at smaller $\tau/\tau\_{\rm MS}$ age ratios than for less massive stars (M \la 12M\_{\odot}). This distribution can be due to: $i$) higher mass-loss rates in massive objets, which can act to reduce the surface fast rotation; $ii$) circulation time scales to transport angular momentum from the core to the surface, which are longer the lower the stellar mass.Comment: 18 pages, 6 figures, A&A, in pres

Effects of gravitational darkening on the determination of fundamental parameters in fast rotating B-type stars

In this paper we develop a calculation code to account for the effects carried by fast rotation on the observed spectra of early-type stars. Stars are assumed to be in rigid rotation and the grid of plane-parallel model atmospheres used to represent the gravitational darkening are calculated by means of a non-LTE approach. Attention is paid on the relation between the apparent and parent non-rotating counterpart stellar fundamental parameters and apparent and true vsini parameters as a function of the rotation rate Omega/Omega_c, stellar mass and inclination angle. It is shown that omission of gravitational darkening in the analysis of chemical abundances of CNO elements can produce systematic overestimation or underestimation, depending on the lines used, rotational rate and inclination angle. The proximity of Be stars to the critical rotation is re-discussed by correcting not only the vsini of 130 Be stars, but also their effective temperature and gravity to account for stellar rotationally induced geometrical distortion and for the concomitant gravitational darkening effect. We concluded that the increase of the vsini estimate is accompanied by an even higher value of the stellar equatorial critical velocity, so that the most probable average rate of angular velocity of Be stars attains Omega/Omega_c ~ 0.88.Comment: 20 pages, 16 figures. Submitted for publication in A&

Triterpenoids in Echinoderms: Fundamental Differences in Diversity and Biosynthetic Pathways

Echinoderms form a remarkable phylum of marine invertebrates that present specific chemical signatures unique in the animal kingdom. It is particularly the case for essential triterpenoids that evolved separately in each of the five echinoderm classes. Indeed, while most animals have ∆5-sterols, sea cucumbers (Holothuroidea) and sea stars (Asteroidea) also possess ∆7 and ∆9(11)-sterols, a characteristic not shared with brittle stars (Ophiuroidea), sea urchins (Echinoidea), and crinoids (Crinoidea). These particular ∆7 and ∆9(11) sterols emerged as a self-protection against membranolytic saponins that only sea cucumbers and sea stars produce as a defense mechanism. The diversity of saponins is large; several hundred molecules have been described in the two classes of these saponins (i.e., triterpenoid or steroid saponins). This review aims to highlight the diversity of triterpenoids in echinoderms by focusing on sterols and triterpenoid glycosides, but more importantly to provide an updated view of the biosynthesis of these molecules in echinoderms

Antiferromagnetic interlayer exchange coupling across an amorphous metallic spacer layer

By means of magneto-optical Kerr effect we observe for the first time antiferromagnetic coupling between ferromagnetic layers across an amorphous metallic spacer layer. Biquadratic coupling occurs at the transition from a ferromagnetically to an antiferromagnetically coupled region. Scanning tunneling microscopy images of all involved layers are used to extract thickness fluctuations and to verify the amorphous state of the spacer. The observed antiferromagnetic coupling behavior is explained by RKKY interaction taking into account the amorphous structure of the spacer material.Comment: Typset using RevTex, 4 pages with 4 figures (.eps