Expression of cell wall related genes in basal and ear internodes of silking brown-midrib-3, caffeic acid O-methyltransferase (COMT) down-regulated, and normal maize plants

<p>Abstract</p> <p>Background</p> <p>Silage maize is a major forage and energy resource for cattle feeding, and several studies have shown that lignin content and structure are the determining factors in forage maize feeding value. In maize, four natural <it>brown-midrib </it>mutants have modified lignin content, lignin structure and cell wall digestibility. The greatest lignin reduction and the highest cell wall digestibility were observed in the <it>brown-midrib-3 </it>(<it>bm3</it>) mutant, which is disrupted in the caffeic acid <it>O</it>-methyltransferase (COMT) gene.</p> <p>Results</p> <p>Expression of cell wall related genes was investigated in basal and ear internodes of normal, COMT antisens (AS225), and <it>bm3 </it>maize plants of the INRA F2 line. A cell wall macro-array was developed with 651 gene specific tags of genes specifically involved in cell wall biogenesis. When comparing basal (older lignifying) and ear (younger lignifying) internodes of the normal line, all genes known to be involved in constitutive monolignol biosynthesis had a higher expression in younger ear internodes. The expression of the COMT gene was heavily reduced, especially in the younger lignifying tissues of the ear internode. Despite the fact that AS225 transgene expression was driven only in sclerenchyma tissues, COMT expression was also heavily reduced in AS225 ear and basal internodes. COMT disruption or down-regulation led to differential expressions of a few lignin pathway genes, which were all over-expressed, except for a phenylalanine ammonia-lyase gene. More unexpectedly, several transcription factor genes, cell signaling genes, transport and detoxification genes, genes involved in cell wall carbohydrate metabolism and genes encoding cell wall proteins, were differentially expressed, and mostly over-expressed, in COMT-deficient plants.</p> <p>Conclusion</p> <p>Differential gene expressions in COMT-deficient plants highlighted a probable disturbance in cell wall assembly. In addition, the gene expressions suggested modified chronology of the different events leading to cell expansion and lignification with consequences far beyond the phenylpropanoid metabolism. The reduced availability of monolignols and S units in <it>bm3 </it>or AS225 plants led to plants also differing in cell wall carbohydrate, and probably protein, composition. Thus, the deficiency in a key-enzyme of the lignin pathway had correlative effects on the whole cell wall metabolism. Furthermore, the observed differential expression between <it>bm3 </it>and normal plants indicated the possible involvement in the maize lignin pathway of genes which up until now have not been considered to play this role.</p

Investigating the unusually high cell wall digestibility of the old INRA early flint F4 maize inbred line

The old INRA flint early line F4, which belongs to the northern flint group, is typified by its high cell wall digest- ibility which reaches values as high as those observed in several brown-midrib bm3 mutant lines. The F4 line thus appeared as a model that could contribute to the understanding of genetic mechanisms involved in variation of secondary wall traits. Different strategies and results were thus gathered including especially cell wall biochemical and digestibility investigations, expression approaches, QTL investigations, and colocalizations between QTLs and candidate genes. Lignin content was lower in F4 than in other lines, with a tendency to lower p-coumarate content. The Syringyl/Guaiacyl lignin unit ratio was similar in F4 as in other lines, but this ratio was nearly not reduced in F4bm3, conversely to what is observed in bm3 mutants. In comparison with the INRA F2 control line, expressions of three PAL genes including the ZmPAL, of the ZmF5H1 and the ZmCOMT genes were significantly reduced in F4 lignifying ear internodes at early silking stage. In the F7025 x F4 RIL progeny, seven QTLs were shown with favorable alleles (increasing cell wall digestibility) originating from F4. Two strong QTLs were located in bins 1.03 and 2.03 colocalizing with the ZmMYB019 and ZmSWN6 transcription factors, respectively. Orthologs of ZmMYB019 have been shown to be involved in lignin biosynthesis, and the PpMYB8 ortholog was shown to regu- late PAL gene expression in maritime pine. The ZmSWN6 NAC transcription factor is an upstream master regulator of the secondary wall biosynthetic programs. At the other QTL positions, colocalizations were also shown with other secondary wall related ZmMYB, but also with BAHD genes involved in arabinoxylan feruloylation, and with the position of the bm6 mutation. Three QTL positions were shown with favorable alleles originating from F7025, which colocalized with ZmMYB and ZmNAC transcriptions factors. As a tentative conclusion, the F4 unusually high cell wall digestibility is likely greatly related to the altered working of at least two major transcription factors regulating cell wall biosynthesis and assembly

Nucleotide diversity of the ZmPox3 maize peroxidase gene: Relationships between a MITE insertion in exon 2 and variation in forage maize digestibility

BACKGROUND: Polymorphisms were investigated within the ZmPox3 maize peroxidase gene, possibly involved in lignin biosynthesis because of its colocalization with a cluster of QTL related to lignin content and cell wall digestibility. The purpose of this study was to identify, on the basis of 37 maize lines chosen for their varying degrees of cell wall digestibility and representative of temperate regions germplasm, ZmPox3 haplotypes or individual polymorphisms possibly associated with digestibility. RESULTS: Numerous haplotypes with high diversity were identified. Frequency of nucleotide changes was high with on average one SNP every 57 bp. Nucleotide diversity was not equally distributed among site categories: the estimated π was on average eight times higher for silent sites than for non-synonymous sites. Numerous sites were in linkage disequilibrium that decayed with increasing physical distance. A zmPox3 mutant allele, carrying an insertion of a transposable element in the second exon, was found in lines derived from the early flint inbred line, F7. This element possesses many structural features of miniature inverted-repeat transposable elements (MITE). The mutant allele encodes a truncated protein lacking important functional sites. An ANOVA performed with a subset of 31 maize lines indicated that the transposable element was significantly associated with cell wall digestibility. This association was confirmed using an additional set of 25 flint lines related to F7. Moreover, RT-PCR experiments revealed a decreased amount of corresponding mRNA in plants with the MITE insertion. CONCLUSION: These results showed that ZmPox3 could possibly be involved in monolignol polymerisation, and that a deficiency in ZmPox3 peroxidase activity seemingly has a negative effect on cell wall digestibility. Also, genetic diversity analyses of ZmPox3 indicated that this peroxidase could be a relevant target for grass digestibility improvement using specific allele introgressions

Characterization of a cinnamoyl-CoA reductase 1 (CCR1) mutant in maize: effects on lignification, fibre development, and global gene expression

Cinnamoyl-CoA reductase (CCR), which catalyses the first committed step of the lignin-specific branch of monolignol biosynthesis, has been extensively characterized in dicot species, but few data are available in monocots. By screening a Mu insertional mutant collection in maize, a mutant in the CCR1 gene was isolated named Zmccr1–. In this mutant, CCR1 gene expression is reduced to 31% of the residual wild-type level. Zmccr1– exhibited enhanced digestibility without compromising plant growth and development. Lignin analysis revealed a slight decrease in lignin content and significant changes in lignin structure. p-Hydroxyphenyl units were strongly decreased and the syringyl/guaiacyl ratio was slightly increased. At the cellular level, alterations in lignin deposition were mainly observed in the walls of the sclerenchymatic fibre cells surrounding the vascular bundles. These cell walls showed little to no staining with phloroglucinol. These histochemical changes were accompanied by an increase in sclerenchyma surface area and an alteration in cell shape. In keeping with this cell type-specific phenotype, transcriptomics performed at an early stage of plant development revealed the down-regulation of genes specifically associated with fibre wall formation. To the present authors’ knowledge, this is the first functional characterization of CCR1 in a grass species

Haploid induction in plants

International audienceIntroduce the technique of haploid induction in plant breeding

Increased genomic prediction accuracy in wheat breeding using a large Australian panel

Key message: Genomic prediction accuracy within a large panel was found to be substantially higher than that previously observed in smaller populations, and also higher than QTL-based prediction.Abstract: In recent years, genomic selection for wheat breeding has been widely studied, but this has typically been restricted to population sizes under 1000 individuals. To assess its efficacy in germplasm representative of commercial breeding programmes, we used a panel of 10,375 Australian wheat breeding lines to investigate the accuracy of genomic prediction for grain yield, physical grain quality and other physiological traits. To achieve this, the complete panel was phenotyped in a dedicated field trial and genotyped using a custom Axiom TM Affymetrix SNP array. A high-quality consensus map was also constructed, allowing the linkage disequilibrium present in the germplasm to be investigated. Using the complete SNP array, genomic prediction accuracies were found to be substantially higher than those previously observed in smaller populations and also more accurate compared to prediction approaches using a finite number of selected quantitative trait loci. Multi-trait genetic correlations were also assessed at an additive and residual genetic level, identifying a negative genetic correlation between grain yield and protein as well as a positive genetic correlation between grain size and test weight

POLYNUCLEOTIDE RESPONSIBLE OF HAPLOID INDUCTION IN MAIZE PLANTS AND RELATED PROCESSES

The present invention concerns an isolated polynucleotide responsible of haploid induction in maize plants and related processes. Additionally, the invention relates to plants that have been genetically transformed with the polynucleotide of the invention. The invention also relates to a process for screening a mutant plant population for enhanced haploid induction by using said isolated polynucleotide. The invention further relates to molecular markers associated with haploid induction in maize plants and their use in quality control for inducer lines

Increased genomic prediction accuracy in wheat breeding using a large Australian panel

Key message: Genomic prediction accuracy within a large panel was found to be substantially higher than that previously observed in smaller populations, and also higher than QTL-based prediction.Abstract: In recent years, genomic selection for wheat breeding has been widely studied, but this has typically been restricted to population sizes under 1000 individuals. To assess its efficacy in germplasm representative of commercial breeding programmes, we used a panel of 10,375 Australian wheat breeding lines to investigate the accuracy of genomic prediction for grain yield, physical grain quality and other physiological traits. To achieve this, the complete panel was phenotyped in a dedicated field trial and genotyped using a custom Axiom TM Affymetrix SNP array. A high-quality consensus map was also constructed, allowing the linkage disequilibrium present in the germplasm to be investigated. Using the complete SNP array, genomic prediction accuracies were found to be substantially higher than those previously observed in smaller populations and also more accurate compared to prediction approaches using a finite number of selected quantitative trait loci. Multi-trait genetic correlations were also assessed at an additive and residual genetic level, identifying a negative genetic correlation between grain yield and protein as well as a positive genetic correlation between grain size and test weight

Maize In Planta Haploid Inducer Lines: A Cornerstone for Doubled Haploid Technology

International audienceDoubled haploid (DH) technology produces strictly homozygous fertile plant thanks to doubling the chromosomes of a haploid embryo/seedling. Haploid embryos are derived from either male or female germ line cells and hold only half the number of chromosomes found in somatic plant tissues, albeit in a recombinant form due to meiotic genetic shuffling. DH production allows to rapidly fix these recombinant haploid genomes in the form of perfectly homozygous plants (inbred lines), which are produced in two rather than six or more generations. Thus, DH breeding enables fast evaluation of phenotypic traits on homogenous progeny. While for most crops haploid embryos are produced by costly and often genotype-dependent in vitro methods, for maize, two unique in planta systems are available to induce haploid embryos directly in the seed. Two "haploid inducer lines", identified from spontaneous maize mutants, are able to induce embryos of paternal or maternal origin. Although effortless crosses with lines of interest are sufficient to trigger haploid embryos, substantial improvements were necessary to bring DH technology to large scale production. They include the development of modern haploid inducer lines with high induction rates (8-12%), and methods to sort kernels with haploid embryos from the normal ones. Chromosome doubling represents also a crucial step in the DH process. Recent identification of genomic loci involved in spontaneous doubling opens up perspectives for a fully in planta DH pipeline in maize. Although discovered more than 60 years ago, maize haploid inducer lines still make headlines thanks to novel applications and findings. Indeed, maternal haploid induction was elegantly diverted to deliver genome editing machinery in germplasm recalcitrant to transformation techniques. The recent discovery of two molecular players controlling haploid induction allowed to revisit the mechanistic basis of maize maternal haploid induction and to successfully translate haploid induction ability to other crops

Héros militaire, culture et société (XIXe-XXe siècles)

À travers des exemples emblématiques et variés pris dans l’histoire contemporaine française, les communications de ce cahier montrent la récurrence d’un modèle d’héroïsation guerrière construit autour de la figure du héros sacrificiel inspiré des récits homériques, des chansons de geste et de l’hagiographie chrétienne. Tout récit d’événement militaire se décline en hommage – voire en sacralisation – de héros, individualisés ou collectifs, qui vise à la fois à donner une identité quasi généalogique au groupe combattant et à servir de modèle en son sein et, au-delà, dans l’ensemble de la société, pour galvaniser l’ardeur au combat ou l’acceptation de la mort individuelle au nom d’un idéal sacré. À l’image des vicissitudes politiques et militaires du pays, l’hommage aux héros amène une réécriture épique de pages sombres de l’histoire. La logique du récit héroïque aboutit ainsi à une sorte de sublimation grandiose où s’effacent les responsabilités et les culpabilités. Ce processus d’héroïsation témoigne à la fois d’un rapport au temps propre à l’époque contemporaine et de la spécificité de la culture militaire au sein de la société. À travers la mise en avant de ces personnages, ce sont de véritables pédagogies souvent concurrentes et évolutives de la nation qui sont mises en œuvre. L’étude du héros militaire s’inscrit ainsi dans la veine de l’analyse des usages politiques du passé le plus contemporain qui se développe depuis les années 1980 jusqu’à aujourd’hui