Metabolic engineering of astaxanthin biosynthesis in maize endosperm and characterization of a prototype high oil hybrid

Maize was genetically engineered for the biosynthesis of the high value carotenoid astaxanthin in the kernel endosperm. Introduction of a β-carotene hydroxylase and a β-carotene ketolase into a white maize genetic background extended the carotenoid pathway to astaxanthin. Simultaneously, phytoene synthase, the controlling enzyme of carotenogenesis, was over-expressed for enhanced carotenoid production and lycopene ε-cyclase was knocked-down to direct more precursors into the β-branch of the extended ketocarotenoid pathway which ends with astaxanthin. This astaxanthin-accumulating transgenic line was crossed into a high oil- maize genotype in order to increase the storage capacity for lipophilic astaxanthin. The high oil astaxanthin hybrid was compared to its astaxanthin producing parent. We report an in depth metabolomic and proteomic analysis which revealed major up- or down- regulation of genes involved in primary metabolism. Specifically, amino acid biosynthesis and the citric acid cycle which compete with the synthesis or utilization of pyruvate and glyceraldehyde 3-phosphate, the precursors for carotenogenesis, were down-regulated. Nevertheless, principal component analysis demonstrated that this compositional change is within the range of the two wild type parents used to generate the high oil producing astaxanthin hybrid

Finding Single Copy Genes Out of Sequenced Genomes for Multilocus Phylogenetics in Non-Model Fungi

Historically, fungal multigene phylogenies have been reconstructed based on a small number of commonly used genes. The availability of complete fungal genomes has given rise to a new wave of model organisms that provide large number of genes potentially useful for building robust gene genealogies. Unfortunately, cross-utilization of these resources to study phylogenetic relationships in the vast majority of non-model fungi (i.e. “orphan” species) remains an unexamined question. To address this problem, we developed a method coupled with a program named “PHYLORPH” (PHYLogenetic markers for ORPHans). The method screens fungal genomic databases (107 fungal genomes fully sequenced) for single copy genes that might be easily transferable and well suited for studies at low taxonomic levels (for example, in species complexes) in non-model fungal species. To maximize the chance to target genes with informative regions, PHYLORPH displays a graphical evaluation system based on the estimation of nucleotide divergence relative to substitution type. The usefulness of this approach was tested by developing markers in four non-model groups of fungal pathogens. For each pathogen considered, 7 to 40% of the 10–15 best candidate genes proposed by PHYLORPH yielded sequencing success. Levels of polymorphism of these genes were compared with those obtained for some genes traditionally used to build fungal phylogenies (e.g. nuclear rDNA, β-tubulin, γ-actin, Elongation factor EF-1α). These genes were ranked among the best-performing ones and resolved accurately taxa relationships in each of the four non-model groups of fungi considered. We envision that PHYLORPH will constitute a useful tool for obtaining new and accurate phylogenetic markers to resolve relationships between closely related non-model fungal species

High-quality SNPs from genic regions highlight introgression patterns among European white oaks (Quercus petraea and Q. robur)

International audienceIn the post-genomics era, non-model species like most Fagaceae still lack operational diversity resources for population genomics studies. Sequence data were produced from over 800 gene fragments covering ~530 kb across the genic partition of European oaks, in a discovery panel of 25 individuals from western and central Europe (11 Quercus petraea, 13 Q. robur, one Q. ilex as an outgroup). Regions targeted represented broad functional categories potentially involved in species ecological preferences, and a random set of genes. Using a high-quality dedicated pipeline, we provide a detailed characterization of these genic regions, which included over 14500 polymorphisms, with ~12500 SNPs −218 being triallelic-, over 1500 insertion-deletions, and ~200 novel di- and tri-nucleotide SSR loci. This catalog also provides various summary statistics within and among species, gene ontology information, and standard formats to assist loci choice for genotyping projects. The distribution of nucleotide diversity (θπ) and differentiation (FST) across genic regions are also described for the first time in those species, with a mean n θπ close to ~0.0049 in Q. petraea and to ~0.0045 in Q. robur across random regions, and a mean FST ~0.13 across SNPs. The magnitude of diversity across genes is within the range estimated for long-term perennial outcrossers, and can be considered relatively high in the plant kingdom, with an estimate across the genome of 41 to 51 million SNPs expected in both species. Individuals with typical species morphology were more easily assigned to their corresponding genetic cluster for Q. robur than for Q. petraea, revealing higher or more recent introgression in Q. petraea and a stronger species integration in Q. robur in this particular discovery panel. We also observed robust patterns of a slightly but significantly higher diversity in Q. petraea, across a random gene set and in the abiotic stress functional category, and a heterogeneous landscape of both diversity and differentiation. To explain these patterns, we discuss an alternative and non-exclusive hypothesis of stronger selective constraints in Q. robur, the most pioneering species in oak forest stand dynamics, additionally to the recognized and documented introgression history in both species despite their strong reproductive barriers. The quality of the data provided here and their representativity in terms of species genomic diversity make them useful for possible applications in medium-scale landscape and molecular ecology projects. Moreover, they can serve as reference resources for validation purposes in larger-scale resequencing projects. This type of project is preferentially recommended in oaks in contrast to SNP array development, given the large nucleotide variation and the low levels of linkage disequilibrium revealed

QUBIC: A Fizeau Interferometer Targeting Primordial B-Modes

Q and U Bolometric Interferometer for Cosmology (QUBIC) is a Fizeau interferometer sensitive to linear polarisation, to be deployed at the Antarctic station of Dome C. This experiment in its final configuration will be operated at 97, 150 and 220 GHz and is intended to target CMB primordial B-modes in a multipole window 20<ℓ<150. A sensitivity of r=0.05 (95 % CL) can be reached by the first module alone, after 2 years of operation. Here we review in particular its working principles, and we show how the QUBIC interferometric configuration can be considered equivalent to a pupil-plane filtered imaging system. In this context, we show how our instrument can be self-calibrated. Finally, we conclude by showing an overview of the first dual-band module (150/220 GHz), which will serve also as a demonstrator for the subsequent units, and review the technological choices we made for each subsystem, with particular emphasis on the detection system

LEKIDs as mm-Wave Polarisation Analysers: Fabrication, Test Bench and Early Results

International audienceWe have demonstrated in an earlier paper that LEKIDs can be used in a polarisation selective way in a filled array configuration. A polarised response can be achieved by means of thick Nb polarising grids lithographed on the rear side of a 300 microns silicon wafer, on which Al resonators have been previously patterned. In the most interesting scheme that we have investigated, a unit cell formed by 4 pixels (2 by 2) responds simultaneously to two orthogonal (cartesian) polarisation states. To assess the effectiveness of this detection scheme, we have fabricated a first generation of devices (9 small arrays, 20–25 pixels each, on a 4$^{\prime \prime }$ Silicon wafer) by using a double-sided mask aligner suitable for a precise positioning of the individual grids in correspondence of each resonator’s meander, for the different LEKID geometries. We describe here the realisation of these first devices. The construction of a dedicated polarimetric test bench is also described in this contribution, together with the first characterisation results. We consider this activity as a first and necessary step to evaluate the polarisation purity attainable with polarisation-sensitive pixels whose size is comparable to the wavelength. This is a fundamental information to drive further studies

Impact of protein, lipid and carbohydrate on the headspace delivery of volatile compounds from hydrating powders

The release of volatile compounds, such as aroma, from a food material during hydration is of wide relevance to the food industry. To this end, dry powders of varying chemical composition were hydrated in a controlled system to investigate the impact of varying composition (protein, lipid and carbohydrate) on the delivery rate of volatile compounds to the headspace. Additional lipid and carbohydrate reduced the concentration of volatile compounds in the headspace and accelerated their rate of delivery to the headspace. Protein had no measurable impact. Of the volatile compounds measured, 2,3 butanedione and acetaldehyde were shown to be released slowly into the headspace, and pyrrol, methyl acetate and pyridine were released rapidly; this differential release rate was strongly correlated with hydrophobicity and would indicate that during hydration there is a temporal dimension to the relative abundance of volatile compounds in the headspace

Complementary Measurement of Thermal Architecture of NbSi TES with Alpha Particle and Complex Impedance

As shown by the Planck mission (Planck Collaboration. Astronomy and astrophysics. arXiv1303.5071P, 2013), background limited bolometers in a space environment are very sensitive to high energy particles. In order to not degrade their sensitivity, it is necessary to reduce this unwanted signal. To achieve this goal, a good understanding of the detector's thermal architecture is mandatory. To investigate this question, we used an particle source in front of our niobium silicon (NbSi) alloy Transition edge sensors (TES). The number of time constants required to fit the data and the way these time constants behave as we change the bias power gave us a good insight on the TES thermal architecture. Indeed we expect a decrease of the detector time constant due to the electro-thermal feedback properties. We will first present some standard characterizations of NbSi TES using a simple thermal model and how they indicate the presence of multiple thermal decouplings. Then we will show the results of the particles measurements and how we used them to build our thermal model for Complex Impedance fitting. All this work has been done for the QUBIC experiment, a B-modes instrument

Role of egg-laying behavior, virulence and local adaptation in a parasitoid’s chances of reproducing in a new host

International audienceUnderstanding the ability of parasitoid insects to succeed in new host populations is a relevant question for biological control and adaptive mechanisms. Cotesia typhae is an African parasitoid specialized on the moth Sesamiae nonagrioides, also called the Mediterranean corn borer. Two Kenyan strains of C. typhae differ in their virulence against a new host population from France. We explored behavioral and physiological hypotheses about this differentiation. Cotesia genus belongs to a group of Hymenoptera in which females inject a domesticated virus in their host to overcome its resistance. Since viral particles are injected along with eggs and since the strain with the higher virulence injects more eggs, we hypothesized that virulence could be explained by the quantity of virus injected. To test this assumption, we measured the injected quantities of eggs and viral particles (estimated by viral DNA circles) of each parasitoid strain along several ovipositions, to vary these quantities. Unexpectedly, results showed that virulence against the French host was not correlated to the injected quantities of eggs or viral circles, indicating that virulence differentiation is explained by other causes. The virulence against the respective natural hosts of the two C. typhae strains was also measured, and results suggest that local adaptation to a more resistant natural host may explain the pre-adaptation of one strain to the new host population. We also identified a differentiation of oviposition strategy and subsequent offspring number between the parasitoid strains, which is important in a biocontrol perspective