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

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

Heat-induced Maillard reaction of the tripeptide IPP and ribose: Structural characterization and implication on bioactivity

peer-reviewedMaillard reaction products (MRPs) were prepared from aqueous model mixtures containing 60 g L− 1 ribose and 30 g L− 1 of the bioactive tripeptide IPP (Ile-Pro-Pro), heated at 98 °C. MRP and associated reactions with changes in IPP were observed within one hour of heat-treatment. The pH of MRPs decreased significantly during the heat treatment of IPP–ribose mixtures from 9.0 to 7.6 after one hour. The amino group content, IPP and ribose concentration decreased significantly during heat treatment. The fluorescence intensity of the IPP–ribose MRPs reached the maximum within 2 h. Modification of the UV/vis spectra for IPP–ribose MRPs was mainly due to a condensation reaction of IPP with ribose. Compounds with molecular weight between 300 and 650 Da were dominant while compounds smaller than 250 Da were also produced during the reactions, as characterized by size exclusion chromatography. Mass spectrometry revealed that IPP was conjugated to ribose at the N-terminal (m/z of 458.3) upon heat-treatment. The presence of ribose also promoted peptide degradation to dehydrated IP (m/z of 211.1). IPP–ribose MRPs lost the known angiotensin-I-converting enzyme (ACE) inhibitory activity of IPP; however, strong antioxidant properties were detected

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