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International audience; This work introduces a novel online Headspace-Solid Phase Microextraction-Gas Chromatography-Mass Spectrometry-based untargeted metabolomics approach, suggested as an alternative tool to study the environmental fate of volatile xenometabolites in emerging complex biopesticides, e.g. the Myrica gale methanolic extract herbicide containing several unknown metabolites. A “living” microcosm sample was designed for non-destructive analysis by a 35-min HS-SPME automated extraction and a 36-min GC-MS run. A 38-day kinetics study was then applied on two groups of soil samples: control and spiked. Statistical tools were used for the comparative kinetics study. The Principal Component Analysis revealed and explained the evolution and the dissipation of the herbicide volatile xenometabolome over time. The time-series Heatmap and Multivariate Empirical Bayes Analysis of Variance allowed the prioritization of 101 relevant compounds including 22 degradation by-products. Out of them, 96 xenometabolites were putatively identified. They included 63 compounds that are identified as herbicide components for the first time. The Orthogonal Projections to Latent Structures Discriminant Analysis and its Cross-Validation test were used to assess the total dissipation of the herbicide volatile residues and method detection limit. The reproducibility of the method was also assessed. The highest inter-samples (n = 3) Peak Area RSD was 7.75 %. The highest inter-samples (n = 3) and inter-days (n = 8) Retention Time SD were 0.43 sec and 3.44 sec, respectively. The work presents a green, non-laborious and high-throughput approach. It required a small number of environmental samples (6 microcosms) that were analyzed 8 times and were not destroyed during the study.

International audience; Evidence of chemical plant protection products' (PPPs) long-term impact has been found in all environmental compartments. Therefore, other types of PPPs are developed to complement chemical PPPs like PPPs from natural sources, namely biocontrol products (BPs). Little is known about those new BPs, and it is important to assess their potential long-term environmental impact. Recently, the Environmental Metabolic Footprinting (EMF) approach was developed. It permits studying sample's entire meta-metabolome (endometabolome and xenometabolome) through a kinetics tracking of metabolomes of treated and untreated samples. Those metabolomes are compared time-by-time to estimate the “resilience time” of the samples after treatment. The current study aims to investigate BP residues' dissipation on peach fruits (Prunus persica). For that, an untargeted Liquid Chromatography-Mass Spectrometry metabolomics approach based on the EMF was optimised to separate the xenometabolome of the PPP from the endometabolome of the fruits. This “new version” of the EMF approach is able to target the BP treatment residues' (xenometabolome) dissipation exclusively. Thus, it is able to determine the time needed to have no more residues in the studied matrix: the “dissipation interval”. Field experiment was conducted on peach tree orchard against brown rot treated with (i) a plant extract BP (Akivi); (ii) a reference mineral extract BP (Armicarb®); and (iii) a Chemical reference treatment campaign. Formulated Akivi and its by-products' dissipation was monitored, a degradation kinetics appeared but the sampling did not last long enough to allow the determination of the “dissipation interval”. Armicarb® and the Chemical reference's residues and by-products showed a persistence pattern along the sampling kinetics. These results indicate that the EMF approach, formerly developed on soil and sediment, is applicable for fruit matrices and can be used to investigate the fate of complex BP treatment on the matrix through the xenometabolome tracking on treated fruits.

International audience; Sharka disease, caused by Plum pox virus (PPV), induces several changes in Prunus. In leaf tissues, the infection may cause oxidative stress and disrupt the photosynthetic process. Moreover, several defense responses can be activated after PPV infection and have been detected at the phytohormonal, transcriptomic, proteomic, and even translatome levels. As proposed in this review, some responses may be systemic and earlier to the onset of symptoms. Nevertheless, these changes are highly dependent among species, variety, sensitivity, and tissue type. In the case of fruit tissues, PPV infection can modify the ripening process, induced by an alteration of the primary metabolism, including sugars and organic acids, and secondary metabolism, including phenolic compounds. Interestingly, metabolomics is an emerging tool to better understand Prunus–PPV interactions mainly in primary and secondary metabolisms. Moreover, through untargeted metabolomics analyses, specific and early candidate biomarkers of PPV infection can be detected. Nevertheless, these candidate biomarkers need to be validated before being selected for a diagnostic or prognosis by targeted analyses. The development of a new method for early detection of PPV-infected trees would be crucial for better management of the outbreak, especially since there is no curative treatment.

International audience; Salix spp. cortex is a bark extract from the common plant willow including S. purpurea L., S. daphnoides Vill. and S. fragilis L, traditionally used in human medicine (i.e. for acetylsalicylic acid usually called aspirin). It is an approved basic substance with fungicidal properties under agricultural EU pesticide regulation (Regulation (EC) no 1107/2009) since 2015. Currently, its approved uses are for arboriculture and viticulture. An initial use extension to allow nurseries to use it as a plant growth regulator is being evaluated by the EU regulatory assessment organisation. Willow water helps cuttings to root, due to its indole butyric acid content, which is a growth hormone (plant growth regulator) in the auxin family and generally chemically synthesized for this purpose. Recently, fungicide usages for the vegetable and fruit market (tomatoes, berries and kiwis) have been submitted to the EU Commission for evaluation. Initial main field trials to demonstrate its plant protection properties were coordinated in France by the 4P project “Protection des Plantes Par les Plantes” (Plant Protection by Plants) between 2010 and 2013 but additional field trials were collected. We detail in this study the results obtained and all the uses of this extract as a plant protection product, including inducing rooting. In addition, we describe the full approval processes for the basic substance Salix spp. cortex and these extensions of use, covered by the Article 23 of the plant protection regulation in EU.

The emergence of pesticides of natural origin appears as an environmental-friendly alternative to synthetic pesticides for managing weeds. To verify this assumption, leptospermone, a natural β-triketone herbicide, and sulcotrione, a synthetic one, were applied to soil microcosms at 0× (control), 1× or 10× recommended field dose. The fate of these two herbicides (i.e. dissipation and formation of transformation products) was monitored to assess the scenario of exposure of soil microorganisms to natural and synthetic herbicides. Ecotoxicological impact of both herbicides was explored by monitoring soil bacterial diversity and activity using next-generation sequencing of 16S rRNA gene amplicons and soil metabolomics. Both leptospermone and sulcotrione fully dissipated over the incubation period. During their dissipation, transformation products of natural and synthetic β-triketone were detected. Hydroxy-leptospermone was almost completely dissipated by the end of the experiment, while CMBA, the major metabolite of sulcotrione, remained in soil microcosms. After 8 days of exposure, the diversity and structure of the soil bacterial community treated with leptospermone was significantly modified, while less significant changes were observed for sulcotrione. For both herbicides, the diversity of the soil bacterial community was still not completely recovered by the end of the experiment (45 days). The combined use of next-generation sequencing and metabolomic approaches allowed us to assess the ecotoxicological impact of natural and synthetic pesticides on non-target soil microorganisms and to detect potential biomarkers of soil exposure to β-triketones.

The intensive use of synthetic herbicides over the past 50 years has led to a number of deleterous impacts on the terrestrial and aquatic environment and is a serious challenge to the ability to sustain agricultural production into the future. In order to remedy this problem, the use of biocontrols is rapidly accelerating and initiating the transition of the agricultural system into an agroecological system. Microalgae (in its wider sense, eukaryote and cyanobacteria) from marine and freshwater habitats are known to produce a diverse range of toxic or otherwise bioactive allelochemical metabolites for growth, communication and defense. Between the search for innovative active molecules and the development of new production processes at the dawn of biotechnology, microalgae have an immense potential in their contribution to biocontrol products. Microalgal organisms can be grown in mass cultures enabelling their manipulation for optimal production of bioactive substances, giving them a significant advantage over terrestrial plants in exploring their effectiveness. In this review we will present the structural diversity of allelochemical compounds and their role for microalgae, the competitive advantages that these compounds provide to microalgae compared to competing species, as well as their potential as biological control agents and bioherbicides.

Biotic and abiotic stresses strongly affect plant growth, quality of production and crop yield. Respectively biocontrol products and biostimulants have been proposed as agronomic tools to counteract those stresses. Research and development on biocontrol and biostimulant products require the study and characterization of the biotic and abiotic interactions involved in these processes. Plant metabolomics approach plays a key role in the identification of chemical mediators involved in these two types of interactions. Metabolomics is a useful tool for the characterization of volatile organic compounds (VOCs) involved in plant-insect interactions, or for the bioguided identification of active compounds from new raw materials. On the other hand, metabolomics approach can be used for the characterization of metabolic pathways affected by plant priming, biostimulant or elicitor application and/or for mechanistic study of Plant-PGPR (Plant Growth Promoting Rhizobacteria) interaction through biomarker specific metabolic fingerprint study.

International audience; Rationale:Correct biomarker determination in metabolomics is crucial for unbiased conclusions and reliable applications. However, this determination is subject to several drifts, e.g. matrix effects and ion suppression in Liquid Chromatography/Mass Spectrometry (LC/MS)‐based approaches. This phenomenon provokes critical issues for biomarker determination, particularly during comparative studies dealing with samples exhibiting heterogeneous complexities.Methods:Occurrence of the issue was coincidentally noticed when studying the environmental impact of a complex bioinsecticide: Bacillus thuringiensis israelensis. The studied samples comprised insecticide‐spiked sediments and untreated control sediments. QuEChERS extractions followed by LC/ESI‐Q/ToF analyses were performed on sediments after 15 days of incubation. Meta‐metabolomes containing pesticide xenometabolites and sediment endometabolites were analyzed in depth using XCMS‐based computational data preprocessing. Multivariate statistical analyses (PCA, OPLS‐DA) and raw data crosschecks were performed to search for environmental biomarkers.Results:Multivariate analyses and raw data crosschecks led to the selection of nine metabolites as biomarker candidates. However, when exploring the mass spectra, co‐elutions were noticed between seven of these metabolites and multi‐charged macromolecules originating from the pesticide. Provoked false positives were thus suspected due to a potential ion suppression exclusively occurring in the spiked samples. A dilution‐based approach was then applied. It confirmed five metabolites as suppressed ions.Conclusions:Ion suppression should be considered as a critical issue for biomarker determination when comparing heterogeneous metabolic profiles. Raw chromatograms and mass spectra crosschecks are mandatory to reveal potential ion suppressions in such cases. Dilution is a suitable approach to filter reliable biomarker candidates before their identification and absolute quantification.

Chemical composition and antifungal activity of essential oils of Algerian Mentha species were studied. Chemical compositions of different Mentha species oils (Mentha rotundifolia, M. spicata, M. pulegium, and M. piperita) were investigated by capillary GC and GC/MS, and their antifungal activities were evaluated by means of paper disc diffusion method and minimum inhibitory concentration (MIC) assays. In total, 98 components from all Mentha species were identified. All oils were rich in monoterpene-oxygenated components. In addition, we reported fumigant antifungal activity of Algerian Mentha essential oils against four fungi: Botrytis cinerea, Penicillium expansum, Monilinia laxa, and M. fructigena. All oils demonstrated very good inhibition especially against B. cinerea, M. laxa, and M. fructigena. Both Monilinia fungi were extremely sensitive to all Algerian Mentha oils, which suggests that Mentha essential oils have the potential to be used as bio-pesticides to protect fruit trees, such as apple and pear trees, and provides an alternative to chemical pesticides.

Pesticides are regularly used for a variety of applications and are disseminated throughout the environment. These substances may have significant negative impacts. To date, the half-life, t1/2, was often used to study the fate of pesticides in environmental matrices (water, soil, sediment). However, this value gives limited information. First, it does not evaluate the formation of by-products, resulting in the need for additional experiments to be performed to evaluate biodegradation and biotransformation products. T1/2 also fails to consider the chemical's impact on biodiversity. Resilience time, a new and integrative proxy, was recently proposed as an alternative to t1/2, with the potential to evaluate all the post-application effects of the chemical on the environment. The 'Environmental Metabolic Footprinting' (EMF) approach, giving an idea of the resilience time, was used to evaluate the impact of botanicals on soil. The goal is to optimise the EMF to study the impact of a microbial insecticide, the Bacillus thuringiensis israelensis (Bti), on sediment. The difficulty of this work lies in the commercial solution of Bti that is really complex, and this complexity yields chromatograms that are extremely difficult to interpret; t1/2 cannot be used. No methodologies currently exist to monitor the impact of these compounds on the environment. We will test the EMF to determine if it is sensitive enough to tolerate such complex mixtures. A pure chemical insecticide, the α-cypermethrin, will be also studied. The article shows that the EMF is able to distinguish meta-metabolome differences between control and exposed (with Bti) sediments.

Tetranychus cinnabarinus (Boisduval) is one of the most economically important and highly polyphagous herbivorous pests in fields and greenhouses worldwide. We previously reported that 2,4-di-tert-butylphenol (DTBP) and ethyl oleate (EO) showed significantly acaricidal, repellent and oviposition deterrent properties against T. cinnabarinus via an unknown mechanism. In this study, the acaricidal activities of DTBP and EO and their biochemical mechanisms in controlling T. cinnabarinus were investigated at different time points by assessing the associated changes in toxic symptoms, potential target-related enzyme activities and seven neurotransmitters belonging to the biogenic amines (BAs). The results showed that the median lethal times (LT50) for DTBP and EO were 8 and 15 h after treatment, respectively. Using dynamic symptomatology observations, typical neurotoxic symptoms including excitation, convulsion and paralysis were observed in the mites treated with DTBP and EO. Furthermore, the two compounds exerted significant inhibitory activity on monoamine oxidase (MAO) in adult T. cinnabarinus females in vitro and in vivo and had little effect on acetylcholinesterase (AChE) activity. The content levels of the seven BAs analyzed by UPLC-3QMS were higher in the mites treated with DTBP and EO than in the controls, except for phenethylamine (PEA) (for DTBP and EO) and octopamine (OA) (for EO). These results demonstrate that both DTBP and EO exert effects on T. cinnabarinus that are possibly consequences of their preventive effects on the deamination of BAs in the nervous system, most likely through inhibitory effects on MAO or MAO-like enzymes and/or interactions with certain special biogenic amine G protein-coupled receptors.

International audience; This study investigated the environmental fate of myrigalone A, a light absorbing natural herbicide found on leaves and fruits of Myrica gale. Myrigalone A was irradiated in water and as a dry solid deposit to simulate reactions on leaves, alone and in the presence of the terpenes generated by Myrica gale. The phototransformation was fast (t1/2 = 35 min in water). Analyses by liquid chromatography coupled to high resolution orbitrap electrospray mass spectrometry (MS) and gas chromatography-MS revealed the formation of 11 photoproducts in water and solid and 9 in gaseous phase. Some were detected in the leaf glands and oil covering the fruits of Myrica gale, which suggested that photodegradation occurred in the field. Moreover, myrigalone A photoinduced the oxidation of terpenes that in turn protected it against photolysis. This highlights the need for additional research on the effect of terpenes on the photodegradation of pesticides on vegetation.

Chemical composition and herbicidal, antifungal, antibacterial and molluscicidal activities of essential oils from Choukzerk, Eryngium triquetrum, and Alexander, Smyrnium olusatrum, from western Algeria were characterized. Capillary GC-FID and GC/MS were used to investigate chemical composition of both essential oils, and the antifungal, antibacterial, molluscicidal and herbicidal activities were determined by % inhibition. Collective essential oil of E. triquetrum was dominated by falcarinol (74.8%) and octane (5.6%). The collective essential oil of S. olusatrum was dominated by furanoeremophilone (31.5%), furanodiene+curzurene (19.3%) and (E)-β-caryophyllene (11%). The E. triquetrum oil was tested and a pure falcarinol (99%) showed virtuous herbicidal and antibacterial activities against potato blackleg disease, Pectobacterium atrosepticum, and Gram-negative soil bacterium, Pseudomonas cichorii (85 and 100% inhibition, respectively), and high ecotoxic activity against brine shrimp, Artemia salina, and the freshwater snail, Biomphalaria glabrata, with an IC50 of 0.35 µg/mL and 0.61 µg/mL, respectively. Essential oil of S. olusatrum showed interesting antibacterial and ecotoxic activity and good herbicidal activity against watercress seeds, Lepidium sativum (74% inhibition of photosynthesis, 80% mortality on growth test on model watercress), while the furanoeremophilone isolated from the oil (99% pure) showed moderate herbicidal activity. Both oils showed excellent antifungal activity against Fusarium. Both oils and especially falcarinol demonstrated good potential as new biocontrol agents in organic crop protection.

Wind observations are important for a wide range of domains including among others meteorology, agriculture and extreme wind engineering. To ensure the provision of high quality surface wind data over Belgium, a new semi-automated data quality control (QC) has been developed and applied to wind observations from the automated weather stations operated by the Royal Meteorological Institute of Belgium. This new QC applies to 10 m 10 min averaged wind speed and direction, 10 m gust speed and direction, 2 m 10 min averaged wind speed and 30 m 10 min averaged wind speed records. After an existence test, automated procedures check the data for limits consistency, internal consistency, temporal consistency and spatial consistency. At the end of the automated QC, a decision algorithm attributes a flag to each particular data point. Each day, the QC staff analyzes the preceding day's observations in the light of the assigned quality flags.

3th biennal congress on natural products and biocontrol, Perpignan, september 2016 EA BIOME; International audience; Bioherbicides appear as an ecofriendly alternative to synthetic herbicides, generally used for weed management, because they are supposed to have low side on human health and ecosystems. In this context, our work aims to study abiotic (i.e., photolysis) and biotic (i.e,. biodegradation) processes involved in the fate of leptospermone, a natural β-triketone herbicide, by combining chemical and microbiological approaches. Under controlled conditions, the photolysis of leptospermone was sensitive to pH. Leptospermone has a half-life of 72 h under simulated solar light irradiations. Several transformation products, including hydroxy-leptospermone, were identified. For the first time, a bacterial strain able to degrade leptospermone was isolated from an arable soil. Based on its 16S ribosomal RNA (rRNA) gene sequence, it was affiliated to the Methylophilus group and was accordingly named as Methylophilus sp. LS1. Interestingly, we report that the abundance of OTUs, similar to the 16S rRNA gene sequence of Methylophilus sp. LS1, was strongly increased in soil treated with leptospermone. The leptospermone was completely dissipated by this bacteria, with a half-life time of 6 days, allowing concomitantly its growth. Hydroxy-leptospermone was identified in the bacterial culture as a major transformation product, allowing us to propose a pathway of transformation of leptospermone including both abiotic and biotic processes.

International audience

Five plant extracts traditionally used in organic and biodynamic farming for pest control and antifungal (downy mildew) disease management were selected after a farmer survey and analyzed for their chemical composition in LC-PDA-MS-MS and using adapted analytical method from food chemistry for determination of class of component (e.g., protein, sugar, lipids…). Their antifungal activity against Penicillium expansum, Botrytis cinerea, Botrytis allii, brown rot causing agents (Monilinia laxa and Monilinia fructigena), and grape downy mildew (Plasmopara viticola) was examined in vitro. White willow (Salix alba) and absinthe (Artemisia absinthium) ethanolic extracts were found to be the most effective in particular against Plasmopara viticola, with a total inhibition of spores germination when applied at 1000 mg/L. These extracts also showed a relatively low toxicity during preliminary ecotoxicological assays on Daphnia pulex. Extract from the bark of white willow contained some flavonoids, especially flavanones (eriodyctiol and derivates) and flavanols (catechins and derivates), as major compounds, whereas absinthe extract was rich in O-methylated flavanols and hydroxycinnamic acids. Thujone content in this extract was also determined by external calibration in GC-MS analysis, and its value was 0.004% dry extract.

International audience; Elionurus muticus is a native grass from Brazil that has been shown to have large variability in thechemical constituents of its essential oil. However, few studies have evaluated this species. Theaims of this study were to chemically characterize four wild populations of E. muticus that werecollected in southern Brazil as well as to evaluate the allelopathic effect of the essential oil, which isrich in citral. Chemical analyses were performed using gas chromatography and mass spectrometry.The phytotoxic activity was evaluated with onion and lettuce seeds. The results showed that thestudied populations have chemical variability based on their geographic origin, which indicatesthe possibility of chemotypes. The results obtained with E. muticus essential oil, reveals phytotoxicactivity on the germination and growth of lettuce and onion.

BACKGROUND New herbicidal formulations were designed by combining wheat gluten (WG), two montmorillonites (MMTs) (unmodified and organically modified) and a model pesticide (ethofumesate), and their performances were assessed through an integrative study conducted in soil using an experimental methodology with data modelling. RESULTS All the WG formulations tested were effective in decreasing the apparent diffusivity of ethofumesate in soil in comparison with the non-formulated active substance. The slow-release effect was significantly more pronounced in the presence of the organically modified MMT, confirming the importance of sorption mechanisms to reduce ethofumesate diffusion. The bioassays undertaken on watercress to evaluate herbicidal antigerminating performances showed that all the WG formulations (with or without MMT) were more effective than both the commercial formulation and the non-formulated ethofumesate, whatever the concentration tested. To explain such results, it was proposed that WG formulations would enable ethofumesate to be more available and thus more effective in inhibiting seed germination, as they would be less prone to be leached by water transport due to watering and also less subject to photodegradation. CONCLUSION The use of pesticide formulations based on wheat gluten and nanoclays appeared to be a promising strategy both to reduce the mobility of pesticides in soil and to protect UV-photosensitive pesticides from photodegradation.

This study presents a novel approach for assessing the risk of agrochemicals in soil microcosms through the use of non-targeted metabolomics. The metabolome of treated soils was extracted and tested through LCMS profiling in order to generate an "Environmental Metabolic Footprint" (EMF). A dynamic characterization of pollution biomarkers was obtained through a multivariate statistical analysis of EMF data, where our results show the possible evolution towards a state of resilience. The EMF methodology was applied to two β-triketone herbicides in soil microcosms: one natural, leptospermone, and one synthetic, sulcotrione. In spite of a four-fold higher application dose, leptospermone exhibited a lower resilience time than did sulcotrione (ca. 30 days vs ca. 45 days respectively).

The ecotoxicological impact of leptospermone, a β-triketone bioherbicide, on the bacterial community of two arable soils was investigated. Soil microcosms were exposed or not (control, 0×) to leptospermone applied at 1× or 10× recommended dose. Leptospermone was moderately adsorbed to both soils (i.e.: Kfa  1.2 and Koc  140 ml.g-1). Dissipation of leptospermone was lower in sterilized than in unsterilized soils suggesting that it was mainly influenced by biotic factors. Within 45 days, leptospermone disappeared almost entirely from one of the two soils (i.e. DT50 < 10 days), while 25% of the leptospermone remained in the other. The composition of the microbial community assessed by qPCR targeting 11 microbial groups was found to be significantly modified in soil microcosms exposed to leptospermone. Pyrosequencing of 16S rRNA gene amplicons showed a shift in the bacterial community structure and a significant impact of leptospermone on the diversity of the soil bacterial community. Changes in the composition as well as in the α- and β-diversity of microbial community were transient in the soil able to fully dissipate the leptospermone. They were persistent in the soil where leptospermone remained. To conclude the bacterial community of the two soils was sensitive to leptospermone and its resilience was observed only when leptospermone was fully dissipated.

*† ‡ SUMMARY In Arabidopsis, four homeotic gene classes, A, B, C and E, are required for the patterning of floral organs. However, very little is known about how the activity of these master genes is translated into regulatory processes leading to specific growth patterns and the formation of organs with specific shapes and sizes. Previously we showed that the transcript variant BPEp encodes a bHLH transcription factor that is involved in limiting petal size by controlling post-mitotic cell expansion. Here we show that the phytohormone jasmonate is required for control of BPEp expression. Expression of BPEp was negatively regulated in opr3 mutant flowers that are deficient in jasmonate synthesis. Moreover, the expression of BPEp was restored in opr3 flowers following exogenous jasmonate treatments. Expression of the second transcript variant BPEub, which originates from the same gene as BPEp via an alternative splicing event, was not affected, indicating that BPEp accumulation triggered by jasmonate occurs at the post-transcriptional level. Consistent with these data, opr3 exhibited an increase in petal size as a result of increased cell size, as well as a modified vein pattern, phenotypes that are similar to those of thebpe-1 mutant. Furthermore, exogenous treatments with jasmonate rescued petal phenotypes associated with loss of function of OPR3. Our data demonstrate that jasmonate signaling downstream of OPR3 is involved in the control of cell expansion and in limiting petal size, and that BPEp is a downstream target that functions as a component mediating jasmonate signaling during petal growth.

International audience; Baptistonia is endemic to the Brazilian Atlantic Forest and is made up of 23 species, the phylogenetic relationships of which are analysed here. Three plastid regions ( psbA-trnH , rpoB-trnC and trnS-G ) were sequenced. Using seven inter-simple sequence repeat (ISSR) markers, we scored 183 loci of ISSR dominant markers. Trees were obtained by successively using (a) maximum parsimony and Bayesian analysis of individual and combined sequence data sets, (b) neighbour joining (NJ) and Dollo parsimony of the ISSR data and (c) NJ analysis of the floral oil composition. These approaches, as well as the results of a previous morphological analysis, were combined using a supertree method, yielding clearer relationships within the genus. Various results of our study indicate that the Baptistonia species are recently radiated. B. colorata and B. venusta (previously known as Oncidium trulliferum ) are confirmed as being sister to the remaining species of the genus. Four taxonomic groups are clearly inferred. The evolution of some morphological traits and floral oil composition proved to be strongly linked to the phylogeny.

International audience; Azospirillum is a plant growth-promoting rhizobacterium (PGPR) able to enhance growth and yield of cereals such as rice, maize and wheat. The growth-promoting ability of some Azospirillum strains appears to be highly specific to certain plant species and cultivars. In order to ascertain the specificity of the associative symbiosis between rice and Azospirillum, the physiological response of two rice cultivars, Nipponbare and Cigalon, inoculated with two rice-associated Azospirillum was analyzed at two levels: plant growth response and plant secondary metabolic response. Each strain of Azospirillum (Azospirillum lipoferum 4B isolated from Cigalon and Azospirillum sp. B510 isolated from Nipponbare) preferentially increased growth of the cultivar from which it was isolated. This specific effect is not related to a defect in colonization of host cultivar as each strain colonizes effectively both rice cultivars, either at the rhizoplane (for 4B and B510) and inside the roots (for B510). The metabolic profiling approach showed that, in response to PGPR inoculation, profiles of rice secondary metabolites were modified, with phenolic compounds such as flavonoids and hydroxycinnamic derivatives being the main metabolites affected. Moreover, plant metabolic changes differed according to Azospirillum strain \texttimes cultivar combinations; indeed, 4B induced major secondary metabolic profile modifications only on Cigalon roots, while B510, probably due to its endophytic feature, induced metabolic variations on shoots and roots of both cultivars, triggering a systemic response. Plant secondary metabolite profiling thereby evidences the specific interaction between an Azospirillum strain and its original host cultivar.

The volatile components from Croton campestris root bark were localized by an anatomical study and analyzed by gas chromatography-mass spectrometry for the first time. The roots of this plant showed secretory cells. These volatile constituents, isolated from the dichloromethane extract by hydrodistillation, were analyzed by gas chromatography-mass spectrometry. We found 69 components. They were characterized, and the major constituents of crude oil root barks were spathulenol (23.3%) and borneol (18.7%). Growth inhibitory activity of the active compounds in solution was evaluated by measuring minimal inhibitory concentrations using a broth micromethod. The minimal inhibitory concentration of root bark volatile constituents was 1.56 μg/mL for Staphylococcus aureus, 3.125 μg/mL for Candida albicans, and 6.25 μg/mL for Aspergillusniger.

International audience; Abstract: GC-FID and GC-MS analysis of essential oil from oregano leaves (Origanum compactum) resulted in the identification of 46 compounds, representing more than 98% of the total composition. Carvacrol was the predominant compound (36.46%), followed by thymol (29.74%) and p-cymene (24.31%). Serial extractions with petroleum ether, ethyl acetate, ethanol, and water were performed on aerials parts of Origanum compactum. In these extracts, different chemical families were characterized: polyphenols (gallic acid equivalent 21.2 to 858.3 g/kg), tannins (catechin equivalent 12.4 to 510.3 g/kg), anthocyanins (cyanidin equivalent 0.38 to 5.63 mg/kg), and flavonoids (quercetin equivalent 14.5 to 54.7 g/kg). The samples (essential oil and extracts) were subjected to a screening for antioxidant (DPPH and ABTS assays) and antimalarial activities and against human breast cancer cells. The essential oil showed a higher antioxidant activity with an IC50= 2 \textpm 0.1 mg/L. Among the extracts, the aqueous extract had the highest antioxidant activity with an IC50= 4.8 \textpm 0.2 mg/L (DPPH assay). Concerning antimalarial activity, Origanum compactum essential oil and ethyl acetate extract showed the best results with an IC50 of 34 and 33 mg/mL, respectively. In addition, ethyl acetate extract (30 mg/L) and ethanol extract (56 mg/L) showed activity against human breast cancer cells (MCF7). The oregano essential oil was considered to be nontoxic.

International audience; We report the identification of the allelochemical 3-(1-oxo-3-phenylpropyl)- 1,1,5-trimethylcyclo-hexane-2,4,6-trione, known as myrigalone A, from the fruits and leaves of Myrica gale. The structure of the compound was confirmed by high-resolution techniques (UV, MS and NMR analysis). The compound is phytotoxic towards classical plant species used for allelochemical assays and also against Fallopia x bohemica, a highly invasive plant. Application of either powdered dry leaves or dry fruits of M. gale also showed in vitro phytotoxic activity. We hypothesize that M. gale could be used as a green allelopathic shield to control Fallopia x bohemica invasion, in addition to its potential use as an environmentally friendly herbicide.

International audience; • Most Azospirillum plant growth-promoting rhizobacteria (PGPR) benefit plant growth through source effects related to free nitrogen fixation and ⁄ or phytohormone production, but little is known about their potential effects on plant physiology. These effects were assessed by comparing the early impacts of three Azospirillum inoculant strains on secondary metabolite profiles of two different maize (Zea mays) cultivars. • After 10 d of growth in nonsterile soil, maize methanolic extracts were analyzed by reverse-phase high-performance liquid chromatography (RP-HPLC) and secondary metabolites identified by liquid chromatography ⁄ mass spectrometry (LC ⁄ MS) and nuclear magnetic resonance (NMR). • Seed inoculation resulted in increased shoot biomass (and also root biomass with one strain) of hybrid PR37Y15 but had no stimulatory effect on hybrid DK315. In parallel, Azospirillum inoculation led to major qualitative and quantitative modifications of the contents of secondary metabolites, especially benzoxazinoids, in the maize plants. These modifications depended on the PGPR strain · plant cultivar combination. • Thus, Azospirillum inoculation resulted in early, strain-dependent modifications in the biosynthetic pathways of benzoxazine derivatives in maize in compatible interactions. This is the first study documenting a PGPR effect on plant secondary metabolite profiles, and suggests the establishment of complex interactions between Azospirillum PGPR and maize.

International audience

Two plant species belonging to the genus Brunfelsia, commonly known as ‘‘Yesterday-Today-and-Tomorrow'', are closely related, however, differ by their flower fragrance. Flowers of Brunfelsia australis present a pleasant fragrance, whereas flowers of Brunfelsia pauciflora are scentless. SPME/GC/MS analysis on flower samples of both species of Brunfelsia indicated that flowers of B. australis emitted a fresh flowery fragrance, essentially comprising two monoterpenic compounds (linalool and (E)-ocimene). B. pauciflora, on the other hand, produced only a few sesquiterpenoids. These results are considered in an ecological and evolutionary context.