Metabolite profiling of Triterpene Saponins in medicago truncatula hairy roots by liquid chromatography fourier transform Ion Cyclotron resonance mass spectrometry

Triterpenes are one of the largest classes of plant natural products, with an enormous variety in structure and bioactivities. Here, triterpene saponins from hairy roots of the model legume Medicago truncatula were profiled with reversed-phase liquid chromatography coupled to negative-ion electrospray ionization Fourier transform ion cyclotron resonance mass spectrometry (LC ESI FT-ICR MS). Owing to the accuracy of the FT-ICR MS, reliable molecular formulas of the detected compounds could be predicted, which, together with the generated MS" spectra, allowed the tentative identification of 79 different saponins, of which 61 had not been detected previously in M. truncatula. Upon collision-induced dissociation of saponins that contain a uronic acid residue in the sugar chain, fragment ions resulting from cross-ring cleavages of the uronic acid residues were observed. The identified saponins are glycosides of 10 different sapogenins, of which three were not detected before in M. truncatula. Zanhic acid glycosides, which are prevalent in the aerial parts of M. truncatula, were absent in the hairy root extract's. This metabolite compendium will facilitate future functional genomic studies of triterpene saponin biosynthesis in M. truncptula

The bHLH transcription factors TSAR1 and TSAR2 regulate triterpene saponin biosynthesis in Medicago truncatula

Plants respond to stresses by producing a broad spectrum of bioactive specialized metabolites. Hormonal elicitors, such as jasmonates, trigger a complex signaling circuit leading to the concerted activation of specific metabolic pathways. However, for many specialized metabolic pathways, the transcription factors involved remain unknown. Here, we report on two homologous jasmonate-inducible transcription factors of the basic helix-loop-helix family, TRITERPENE SAPONIN BIOSYNTHESIS ACTIVATING REGULATOR1 (TSAR1) and TSAR2, which direct triterpene saponin biosynthesis in Medicago truncatula. TSAR1 and TSAR2 are coregulated with and transactivate the genes encoding 3-HYDROXY-3-METHYLGLUTARYL-COENZYME A REDUCTASE1 (HMGR1) and MAKIBISHI1, the rate-limiting enzyme for triterpene biosynthesis and an E3 ubiquitin ligase that controls HMGR1 levels, respectively. Transactivation is mediated by direct binding of TSARs to the N-box in the promoter of HMGR1. In transient expression assays in tobacco (Nicotiana tabacum) protoplasts, TSAR1 and TSAR2 exhibit different patterns of transactivation of downstream triterpene saponin biosynthetic genes, hinting at distinct functionalities within the regulation of the pathway. Correspondingly, overexpression of TSAR1 or TSAR2 in M. truncatula hairy roots resulted in elevated transcript levels of known triterpene saponin biosynthetic genes and strongly increased the accumulation of triterpene saponins. TSAR2 overexpression specifically boosted hemolytic saponin biosynthesis, whereas TSAR1 overexpression primarily stimulated nonhemolytic soyasaponin biosynthesis. Both TSARs also activated all genes of the precursor mevalonate pathway but did not affect sterol biosynthetic genes, pointing to their specific role as regulators of specialized triterpene metabolism in M. truncatula

The TriForC database : a comprehensive up-to-date resource of plant triterpene biosynthesis

Triterpenes constitute a large and important class of plant natural products with diverse structures and functions. Their biological roles range from membrane structural components over plant hormones to specialized plant defence compounds. Furthermore, triterpenes have great potential for a variety of commercial applications such as vaccine adjuvants, anti-cancer drugs, food supplements and agronomic agents. Their biosynthesis is carried out through complicated, branched pathways bymultiple enzyme types that include oxidosqualene cyclases, cytochrome P450s, and UDP-glycosyltransferases. Given that the number of characterized triterpene biosynthesis enzymes has been growing fast recently, the need for a database specifically focusing on triterpene enzymology became eminent. Here, we present the TriForC database (http://bioinformatics. psb. ugent. be/triforc/), encompassing a comprehensive catalogue of triterpene biosynthesis enzymes. This highly interlinked database serves as a user-friendly access point to versatile data sets of enzyme and compound features, enabling the scanning of a complete catalogue of experimentally validated triterpene enzymes, their substrates and products, as well as the pathways they constitute in various plant species. The database can be accessed by direct browsing or through convenient search tools including keyword, BLAST, plant species and substructure options. This database will facilitate gene mining and creating genetic toolboxes for triterpene synthetic biology

‘Good-genes' and ‘compatible-genes' effects in an Alpine whitefish and the information content of breeding tubercles over the course of the spawning season

Some models of sexual selection predict that individuals vary in their genetic quality and reveal some of this variation in their secondary sexual characteristics. Alpine whitefish (Coregonus sp.) develop breeding tubercles shortly before their spawning season. These tubercles are epidermal structures that are distributed regularly along the body sides of both males and females. There is still much unexplained variation in the size of breeding tubercles within both sexes and with much overlap between the sexes. It has been suggested that breeding tubercles function to maintain body contact between the mating partners during spawning, act as weapons for defence of spawning territories, or are sexual signals that reveal aspects of genetic quality. We took two samples of whitefish from their spawning place, one at the beginning and one around the peak of spawning season. We found that females have on average smaller breeding tubercles than males, and that tubercle size partly reveals the stage of gonad maturation. Two independent full-factorial breeding experiments revealed that embryo mortality was significantly influenced by male and female effects. This finding demonstrates that the males differed in their genetic quality (because offspring get nothing but genes from their fathers). Tubercle size was negatively linked to some aspects of embryo mortality in the first breeding experiment but not significantly so in the second. This lack of consistency adds to inconsistent results that were reported before and suggests that (i) some aspects of genetic quality are not revealed in breeding tubercles while others are, or (ii) individuals vary in their signaling strategies and the information content of breeding tubercles is not always reliable. Moreover, the fact that female whitefish have breeding tubercles of significant size while males seem to have few reasons to be choosy suggests that the tubercles might also serve some functions that are not linked to sexual signalin

Stochastic Approximation with Biased MCMC for Expectation Maximization

The expectation maximization (EM) algorithm is a widespread method for empirical Bayesian inference, but its expectation step (E-step) is often intractable. Employing a stochastic approximation scheme with Markov chain Monte Carlo (MCMC) can circumvent this issue, resulting in an algorithm known as MCMC-SAEM. While theoretical guarantees for MCMC-SAEM have previously been established, these results are restricted to the case where asymptotically unbiased MCMC algorithms are used. In practice, MCMC-SAEM is often run with asymptotically biased MCMC, for which the consequences are theoretically less understood. In this work, we fill this gap by analyzing the asymptotics and non-asymptotics of SAEM with biased MCMC steps, particularly the effect of bias. We also provide numerical experiments comparing the Metropolis-adjusted Langevin algorithm (MALA), which is asymptotically unbiased, and the unadjusted Langevin algorithm (ULA), which is asymptotically biased, on synthetic and real datasets. Experimental results show that ULA is more stable with respect to the choice of Langevin stepsize and can sometimes result in faster convergence.Comment: Accepted to AISTATS'2

A seed-specific regulator of triterpene saponin biosynthesis in Medicago truncatula

Plants produce a vast array of defense compounds to protect themselves from pathogen attack or herbivore predation. Saponins are a specific class of defense compounds comprising bioactive glycosides with a steroidal or triterpenoid aglycone backbone. The model legume Medicago truncatula synthesizes two types of saponins, hemolytic saponins and nonhemolytic soyasaponins, which accumulate as specific blends in different plant organs. Here, we report the identification of the seed-specific transcription factor TRITERPENE SAPONIN ACTIVATION REGULATOR3 (TSAR3), which controls hemolytic saponin biosynthesis in developing M. truncatula seeds. Analysis of genes that are coexpressed with TSAR3 in transcriptome data sets from developing M. truncatula seeds led to the identification of CYP88A13, a cytochrome P450 that catalyzes the C-16α hydroxylation of medicagenic acid toward zanhic acid, the final oxidation step of the hemolytic saponin biosynthesis branch in M. truncatula. In addition, two uridine diphosphate glycosyltransferases, UGT73F18 and UGT73F19, which glucosylate hemolytic sapogenins at the C-3 position, were identified. The genes encoding the identified biosynthetic enzymes are present in clusters of duplicated genes in the M. truncatula genome. This appears to be a common theme among saponin biosynthesis genes, especially glycosyltransferases, and may be the driving force of the metabolic evolution of saponins

3-D Magnetotelluric Investigations for geothermal exploration in Martinique (Lesser Antilles). Characteristic Deep Resistivity Structures, and Shallow Resistivity Distribution Matching Heliborne TEM Results

Within the framework of a global French program oriented towards the development of renewable energies, Martinique Island (Lesser Antilles, France) has been extensively investigated (from 2012 to 2013) through an integrated multi-methods approach, with the aim to define precisely the potential geothermal ressources, previously highlighted (Sanjuan et al., 2003). Amongst the common investigation methods deployed, we carried out three magnetotelluric (MT) surveys located above three of the most promising geothermal fields of Martinique, namely the Anses d'Arlet, the Montagne Pel{\'e}e and the Pitons du Carbet prospects. A total of about 100 MT stations were acquired showing single or multi-dimensional behaviors and static shift effects. After processing data with remote reference, 3-D MT inversions of the four complex elements of MT impedance tensor without pre-static-shift correction, have been performed for each sector, providing three 3-D resistivity models down to about 12 to 30 km depth. The sea coast effect has been taken into account in the 3-D inversion through generation of a 3-D resistivity model including the bathymetry around Martinique from the coast up to a distance of 200 km. The forward response of the model is used to calculate coast effect coefficients that are applied to the calculated MT response during the 3-D inversion process for comparison with the observed data. 3-D resistivity models of each sector, which are inherited from different geological history, show 3-D resistivity distribution and specificities related to its volcanological history. In particular, the geothermal field related to the Montagne Pel{\'e}e strato-volcano, is characterized by a quasi ubiquitous conductive layer and quite monotonic typical resistivity distribution making interpretation difficult in terms of geothermal targets. At the opposite, the resistivity distribution of Anse d'Arlet area is radically different and geothermal target is thought to be connected to a not so deep resistive intrusion elongated along a main structural axis. Beside these interesting deep structures, we demonstrate, after analyzing the results of the recent heliborne TEM survey covering the whole Martinique, that surface resistivity distribution obtained from 3-D inversion reproduce faithfully the resistivity distribution observed by TEM. In spite of a very different sampling scale, this comparison illustrates the ability of 3-D MT inversion to take into account and reproduce static shift effects in the sub-surface resistivity distribution.Comment: Wordl Geothermal Congress 2015, Apr 2015, Melbourne, Australi

Male dominance linked to size and age, but not to 'good genes' in brown trout (Salmo trutta)

BACKGROUND: Males that are successful in intra-sexual competition are often assumed to be of superior quality. In the mating system of most salmonid species, intensive dominance fights are common and the winners monopolise most mates and sire most offspring. We drew a random sample of mature male brown trout (Salmo trutta) from two wild populations and determined their dominance hierarchy or traits linked to dominance. The fish were then stripped and their sperm was used for in vitro fertilisations in two full-factorial breeding designs. We recorded embryo viability until hatching in both experiments, and juvenile survival during 20 months after release into a natural streamlet in the second experiment. Since offspring of brown trout get only genes from their fathers, we used offspring survival as a quality measure to test (i) whether males differ in their genetic quality, and if so, (ii) whether dominance or traits linked to dominance reveal 'good genes'. RESULTS: We found significant additive genetic variance on embryo survival, i.e. males differed in their genetic quality. Older, heavier and larger males were more successful in intra-sexual selection. However, neither dominance nor dominance indicators like body length, weight or age were significantly linked to genetic quality measured as embryo or juvenile survival. CONCLUSION: We found no evidence that females can improve their offspring's genetic viability by mating with large and dominant males. If there still were advantages of mating with dominant males, they may be linked to non-genetic benefits or to genetic advantages that are context dependent and therefore possibly not revealed under our experimental conditions - even if we found significant additive genetic variation for embryo viability under such conditions

Analyse du Consentement à Payer pour l’Assurance Agricole Indicielle des Producteurs du Sud-Borgou au Bénin

L’objectif de l’étude est d’analyser le consentement à payer des producteurs pour l’assurance agricole indicielle. A cet effet, une enquête s’est déroulée auprès de 200 producteurs assurés ou non dans les villages d’intervention de l’assurance agricole indicielle dans le sud du département du Borgou au Bénin. Grâce aux méthodes de la statistique descriptive et économétrique, les données collectées ont été analysées à l’aide du logicielle Stata15. Les résultats issus de nos analyses montrent que 72,5% des producteurs enquêtés ont connaissance de l’assurance agricole indicielle, 81% sont disposés à payer pour l’assurance agricole indicielle et seulement 49% de l’échantillon ont réellement estimé un montant à payer pour l’assurance agricole indicielle relativement au prime payé qui s’élève à 10000 FCFA (17 USD). Ces producteurs estiment payer en moyenne 6132 FCFA soit 10,5 USD pour adopter l’assurance agricole indicielle. L’analyse des facteurs déterminants le consentement à payer pour l’assurance agricole indicielle par la méthode de double hurdle a montré que les variables appartenance à une association agricole, revenu du maïs et la superficie cultivée du maïs sont les variables qui influencent positivement le consentement du producteur à accepter payer pour l’assurance agricole indicielle au seuil de 5%.  Dans le second modèle, les variables revenues du maïs et appartenance à un groupement ont une influence positive au seuil de 5%. Les variables alphabétisation, niveau d’instruction, héritage et la location des terres ont une influence négative au seuil de 5%. L’accès au crédit agricole a une influence positive sur le montant à payer des producteurs au seuil de 10%. Ainsi, le développement de l’assurance agricole indicielle dans ces communes doit tenir compte de ces variables pour sensibiliser les producteurs sur l’assurance agricole indicielle et établir des conditions pouvant permettre aux producteurs d’avoir un accès facile aux crédits afin d’améliorer le revenu de leurs exploitations agricoles. Cela favoriserait son adoption en masse par les producteurs.   The objective of the study is to analyze producers' willingness to pay for insurance agricultural index. To this end, a survey was carried out among 200 insured producers or no in the villages where index agricultural insurance operates in the south of the department from Borgou to Benin. Thanks to the methods of descriptive and econometric statistics, Data collected was analyzed using Stata15 software. The results from our analyzes show that 72.5% of the producers surveyed are aware of agricultural insurance index, 81% are willing to pay for index agricultural insurance and only 49% of the sample actually estimated an amount to pay for index agricultural insurance in relation to the premium paid which amounts to 10,000 FCFA (17 USD). These producers believe they pay on average 6132 FCFA or 10.5 USD to adopt index agricultural insurance. The analysis of factors determining willingness to pay for index agricultural insurance double hurdle showed that the variables belonging to an agricultural association, income from the method by corn and the cultivated area of corn are the variables that positively influence the consent of the producer to accept paying for index agricultural insurance at the threshold of 5%.  In the second model, the variables corn income and membership in a group have a positive influence at the 5% threshold. The variables literacy, level of education, inheritance and land rental has a negative influence at the 5% threshold. Access to agricultural credit has a positive influence on the amount to be paid by producers at the 10% threshold. Thus, the development of index agricultural insurance in these municipalities must take these variables into account to raise awareness among producers about index agricultural insurance and establish conditions that can allow producers to have easy access to credits in order to improve the income of their agricultural exploitations. This would encourage its mass adoption by producers