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Theobroma cacao is the only source that allows the production of chocolate. It is of major economic importance for producing countries such as Ecuador, which is the third-largest cocoa producer in the world. Cocoa is classified into two groups: bulk cocoa and aromatic fine flavour cocoa. In contrast to bulk cocoa, fine flavour cocoa is characterised by fruity and floral notes. One of the characteristics of Nacional cocoa, the emblematic cocoa of Ecuador, is its aromatic ARRIBA flavour. This aroma is mainly composed of floral notes whose genetic and biochemical origin is not well-known. This research objective is to study the genetic and biochemical determinism of the floral aroma of modern Nacional cocoa variety from Ecuador. Genome-Wide Association Study (GWAS) was conducted on a population of 152 genotypes of cocoa trees belonging to the population variety of modern Nacional. Genome-Wide Association Study was conducted by combining SSR and SNP genotyping, assaying biochemical compounds (in roasted and unroasted beans), and sensory evaluations from various tastings. This analysis highlighted different areas of association for all types of traits. In a second step, a search for candidate genes in these association zones was undertaken, which made it possible to find genes potentially involved in the biosynthesis pathway of the biochemical compound identified in associations. Our results show that two biosynthesis pathways seem to be mainly related to the floral note of Nacional cocoa: the monoterpene biosynthesis pathway and the L-phenylalanine degradation pathway. As already suggested, the genetic background would therefore appear as largely explaining the floral note of cocoa.

Theobroma cacao is the only source of cocoa. Cocoa is classified into two types of products: bulk cocoa and fine flavour cocoa. Contrary to bulk cocoa, fine aromatic cocoa is characterized by its floral and fruity aromas [1]. In order to understand the genetic determinism of the formation of these aromas in cocoa beans, a genetic study using the Genetic Wide Association Study (GWAS) method was undertaken. It was carried out on 158 clones belonging to a population of Nacional tree type cultivated in Ecuador, whose volatile compound concentrations and sensory profiles were characterized for its diversity. This study revealed areas of correlation between, on the one hand, the genetic diversity of this population, revealed by molecular marker alleles and the volatile compounds detected in the different clones, and on the other hand, between this same genetic diversity and their sensory profiles. These correlation zones, also called associations, are therefore linked to one of these traits, but also in some cases to both types of traits. Thanks to these associations, which correspond to a restricted area of the cocoa genome, and the knowledge of its complete sequence [2], candidate genes have been brought to light. Some of them are known and identified in biosynthesis pathways of volatile compounds, which are themselves known to have a fruity note. In a preliminary study, a difference in the expression of these genes was identified between four genotypes (two floral and two fruity genotypes) during different stages of development and fermentation of the beans. The results showed that the candidate genes tended to be activated during fermentation and not during the maturation stages of the pods.

The number of grains per panicle is an important yield-related trait in cereals which depends in part on panicle branching complexity. One component of this complexity is the number of secondary branches per panicle. Previously, a GWAS site associated with secondary branch and spikelet numbers per panicle in rice was identified. Here we combined gene capture, bi-parental genetic population analysis, expression profiling and transgenic approaches in order to investigate the functional significance of a cluster of 6 ANK and ANK-TPR genes within the QTL. Four of the ANK and ANK-TPR genes present a differential expression associated with panicle secondary branch number in contrasted accessions. These differential expression patterns correlate in the different alleles of these genes with specific deletions of potential cis-regulatory sequences in their promoters. Two of these genes were confirmed through functional analysis as playing a role in the control of panicle architecture. Our findings indicate that secondary branching diversity in the rice panicle is governed in part by differentially expressed genes within this cluster encoding ANK and ANK-TPR domain proteins that may act as positive or negative regulators of panicle meristem’s identity transition from indeterminate to determinate state., Author summary Grain yield is one of the most important indexes in rice breeding, which is controlled in part by panicle branching complexity. A new QTL with co-location of spikelet number (SpN) and secondary branch number (SBN) traits was identified by genome-wide association study in a Vietnamese rice landrace panel. A set of four Ankyrin and Tetratricopeptide repeat domain-encoding genes was identified from this QTL based on their difference of expression levels between two contrasted haplotypes for the SpN and SBN traits. The differential expression is correlated with deletions in the promoter regions of these genes. Two of the genes act as negative regulators of the panicle meristem’s identity transition from indeterminate to determinate state while the other two act as positive regulators of this meristem fate transition. Based on the different phenotypes between overexpressed and mutant plants, two of these genes were confirmed as playing a role in the control of panicle architecture. These findings can be directly used to assist selection for grain yield improvement.

Theobroma cacao est un arbre originaire des régions tropicales humides d'Amérique latine. Il est cultivé pour ses fèves qui permettent la production de chocolat. Le Nacional, originaire d'Equateur, est une variété de cacao fin connue pour ses arômes floraux et épicées, appelée saveur "Arriba". Les mécanismes de la synthèse de ses arômes restent peu connus. Les travaux de cette thèse ont porté sur l'étude des déterminants génétiques et biochimiques des arômes des cacaos fins équatoriens afin d'initier de contribuer aux connaissances dans ce domaine.Une première partie de l'étude a été réalisée à partir d'une population de cacaoyer de type Nacional moderne, qui est la variété de Nacional actuellement cultivée. Des études d'associations ont été réalisées sur l'ensemble du génome (GWAS) et ont porté sur l'analyse des composés volatils relatifs aux arôme floraux et fruités (fruits frais et fruits secs) contenus dans les fèves, avant et après torréfaction, ainsi que, sur des résultats d'analyses sensorielles de liqueurs. Cette première étude a pu montrer que les arômes floraux du Nacional étaient principalement synthétisés grâce à deux voies de biosynthèse: celle des monoterpènes et la voie de dégradation du L-phénylalanine. Les résultats relatifs aux arômes fruités ont permis de mettre en lumière cinq voies métaboliques majeures: la voie de biosynthèse des monoterpènes, les voies de dégradations du L-phénylalanine, des sucres, des acides gras et des protéines. Des gènes candidats codant pour des enzymes impliqués dans ces voies métaboliques ont été identifiés dans les zones d'associations correspondantes.La variété Nacional moderne est issue de diverses générations de croisements entre des Trinitario (hybrides Amelonado/Criollo) et le Nacional ancestral, mettant ainsi en jeu 3 ancêtres contrastés. L'effet de cette étape de domestication récente sur les arômes de la variété Nacional moderne a été étudié. Grâce aux données de génotypage des trois ancêtres de référence et aux résultats des GWAS portant sur l'ensemble des caractères de qualité (composés volatils et non-volatils, analyses sensorielles), il a été possible de déterminer l'origine des allèles ayant un effet positif sur les arômes dans les différentes zones d'associations. Cette étude a montré que l'ensemble des ancêtres fondateurs ont apporté des allèles favorables à la synthèse d'arômes de qualité (floraux, fruités, …) mais aussi à la synthèse de défauts (amertume, astringence, …). Nous avons pu montrer que les zones d'associations en lien avec les arômes de qualité et celles avec les défauts n'étaient pas liées génétiquement. Il est donc possible de sélectionner les zones d'intérêts pour les arômes tout en contre-sélectionnant les zones apportant des défauts.La deuxième partie de cette étude a porté sur l'analyse des arômes d'une population de cacaoyers natifs d'Amazonie et issus de la zone d'origine de la variété Nacional ancestral. Dans cette étude, des analyses GWAS ont également été effectuées sur l'ensemble des caractères de qualité analysés précédemment. Quatre vingt dix sept gènes candidats sont communs aux deux populations analysées. De nouveaux composés volatils ainsi que de nouvelles zones d'associations ont également été détectées montrant ainsi une plus grande diversité et richesse aromatique de ces nouvelles ressources génétiques et leur intérêt pour la création de nouvelles variétés aromatiques adaptées à l'Amazonie.Enfin, une étude GWAS sur les composés non-volatils, les caractères sensoriels liés à l'amertume et à l'astringence, ainsi que la teneur en matière grasse et en protéines, a également été réalisée sur les deux populations. Des gènes candidats en lien avec la voie de biosynthèse de la caféine et celle des polyphénols, ou en lien avec la voie de biosynthèse des acides gras ont pu être observés dans les zones d'associations.

We explored the Theobroma cacao genome sequence to progress in the knowledge of the T. cacao incompatibility system. Cocoa self-compatibility is an important yield factor and has been described as controlled by a late gameto-sporophytic system involving several locus, and resulting in gametic non-fusion. In this work, we identified two different mechanisms controlling the T. cacao self-incompatibility system at two separate loci, located on chromosome one and four (CH1 and CH4). Both loci are responsible for gametic selection, but only one (the CH4 locus) is involved in the main fruit drop. The CH1 locus acts prior to gamete fusion and independently of CH4 locus. Fine mapping and genome wide association studies focused analyses of restricted regions without recombinant plants where several candidate genes were identified. Their expression analysis showed differential expression during incompatible or compatible reactions for some of them. Highly polymorphic SSR diagnostic markers, designed in the CH4 region identified by fine mapping, allowed the development of efficient diagnostic markers predicting selfcompatibility and fruit setting according to allele or genotype presence. SSR alleles specific to self-compatible Amelonado and Criollo varieties were also identified allowing screening for self-compatible plants in cocoa populations.