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Many ornamental crops are polyploid or even exist at different ploidy levels. Polyploid QTL analysis tools have been developed in recent years, yet they are limited in the population types they accept. Biparental populations are nowadays being regarded as a limited tool for QTL discovery, as only a limited number of QTLs occurs in an experimental cross and their effects might not be stable across genetic backgrounds. Genome-Wide Association Studies include more genetic diversity but suffer from (hidden) genetic structure and low frequency of QTL alleles. Both factors influence QTL detection and effect estimation, decreasing the sensitivity of QTL analysis. Alternatively, multiparental populations (MPP) can be used, potentially combining multiple QTLs and QTL alleles with known population structure and balanced allele frequencies. Breeding populations of interconnected crosses also constitute a form of MPP and QTLs identified in them might be more applicable to commercial cultivars. To perform QTL analysis in polyploids, mixed models or Bayesian approaches that consider pedigree information are recommended. During the analysis, QTL effects are ideally estimated using Identity by Descent (IBD) alleles (genomic regions that originate from the same ancestor) which can be obtained through haplotype estimation. Although MPPs could thus be a powerful set-up to estimate polyploid haplotypes, a software gap was identified as no current polyploid haplotyping tools are able to utilize MPP pedigree information to obtain haplotypes across an MPP. In order to utilize MPPs to their full extent and expand polyploid QTL analyses to encompass typical breeding populations, new haplotyping tools must be developed.

Thanks to recent advances in high-throughput genotyping tools and software for genetic analyses, it is now possible to perform QTL mapping and genome-wide association analysis in tetraploid roses and to establish the effect of favorable SNP marker haplotype(s) (“plus-alleles”). The challenge is how to use these tools and technologies for rose breeding in a cost-effective way. At least four points need to be addressed: (1) which steps of the breeding process may benefit from marker information; (2) what is the value of the identified plus-alleles for the trait; (3) where in the breeding germplasm do these plus-alleles occur, and (4) how can plants carrying these plus-alleles be selected efficiently. Here we discuss for which purposes markers may be applied.

Black spot disease (Diplocarpon rosae) is the most important leaf disease of garden roses in warm humid areas. Although partial resistance to black spot has been shown to be moderately heritable, the responsible quantitative trait loci (QTL) remain unidentified. Because of the interspecific nature and high heterozygosity in commercial roses, as well as the relatively small research input compared to row crops, the genomic resources available for rose are limited. To effectively identify markers associated with QTL controlling black spot resistance, abundant markers across the genome and careful phenotyping are required. Fifteen inter-related diploid rose populations with black spot resistant cultivar R. wichuraiana ‘Basye’s Thornless’ in the genetic background were assessed based on the percent of total foliage covered with lesions in June, October and November of 2016 in College Station. Broad sense heritability was estimated at 0.51 which indicates black spot partial resistance is a moderately heritable trait. Genotyping-by-sequencing technology was used to generate SNP markers for linkage map construction. Previous anchor SSR markers were used to designate the linkage group number. The final consensus map used for black spot QTL detection contained 791 SNP covering 430 cM with the biggest gap being 6.6 cM on LG4. One major black spot QTL was discovered on LG3 explaining ~13% of the total phenotypic variance (equaling approximately 26% of the genetic variation) using pedigree-based analysis among all fifteen populations.

Acidity is a critical component of the apple fruit consumption experience. In previous biparental family studies, two large-effect acidity QTLs were reported using freshly harvested fruit. Objectives of this study were to determine the number and location of QTLs for acidity variation in a large apple breeding program and ascertain the quantitative effects and breeding relevance of QTL allelic combinations at harvest and after commercially relevant periods of cold storage. Pedigree-connected germplasm of 16 full-sib families representing nine important breeding parents, genotyped for the 8K SNP array, was assessed for titratable acidity at harvest and after 10- and 20-week storage treatments, for three successive seasons. Using pedigree-based QTL mapping software, FlexQTL™, evidence was found for only two QTLs, on linkage groups 16 (the reported Ma locus) and LG 8 (here called Ma3) that jointly explained 66 ± 5% of the phenotypic variation. An additive allele dosage model for the two QTLs effectively explained most acidity variation, with an average of + 1.8 mg/L at harvest per high-acidity allele. The more high-acidity alleles, the faster the depletion with storage, with all combinations appearing to eventually converge to a common baseline. All parent cultivars and selections had one or two of the four possible high-acidity alleles. Each QTL had a rare second high-acidity allele with stronger or reduced effect. Diagnostic SNP markers were identified for QTL alleles derived from distinct sources. Combined QTL effects highlighted utility of the DNA-based information in new cultivar development for targeting desired fruit acidity levels before or after storage.

International audience; Although many peach QTLs responsible for the variability of traits have been identified and published so far, the number of molecular markers currently used in breeding is still limited. One of the reasons is the large QTL intervals produced, in part, by the limited progeny size. Here we report a QTL mapping approach that enlarges the progeny size by analyzing jointly multiple progenies. Our analysis included 1467 individuals from eighteen Prunus progenies, 13 from intra-specific crosses and five inter-specific between peach and closely related species. The progenies were grown in five locations, with no duplication between orchards. Data from phenology, tree, flower and fruit traits, fruit quality and yield measured in different locations and years were subjected to different standardization methods and integrated in a single data file. The populations were genotyped with the 9K SNP Illumina array, which increased considerably the marker density compared to previous studies. The QTL analysis was conducted with FlexQTLTM software. Here we describe and discuss the preliminary QTLs obtained for some of the traits analyzed (maturity date, percentage of red skin color and soluble solid content). The identification of donors of favorable alleles will represent an important tool for marker-assisted breeding. This study has been conducted in the frame of the Fruit Breedomics European project.

Horticultural crop improvement would benefit from a standardized, systematic, and statistically robust procedure for validating quantitative trait loci (QTLs) in germplasm relevant to breeding programs. Here, we describe and demonstrate a strategy for developing reference germplasm sets of perennial, clonally propagated crops, especially those with long juvenile periods. Germplasm is chosen to efficiently represent important members of larger pedigree-connected genepools. To facilitate validation of multiple QTLs, genome-wide representation of alleles is optimized for designated important breeding parents (IBPs) by estimating average allelic representation in relatives. The strategy and arising principles were demonstrated in a simulated germplasm set. Strong statistical power can be achieved with a carefully chosen germplasm set composed of IBPs, their numerous unselected progenies and close relatives, and all available founders and intermediate ancestors. Crop Reference Sets were developed in the marker-assisted breeding (MAB)-enabling “RosBREED” project as a base resource for QTL validation in US breeding germplasm of apple (Malus × domestica), peach (Prunus persica), and sweet cherry (Prunus avium) consisting of 467, 452, and 268 individuals, respectively. These sets adequately represent the most designated IBPs, have distinct advantages for QTL validation over other germplasm arrangements of equal size, and are recommended as a base resource for QTL validation by breeders of these US crops. The strategy described here can be used to develop efficient reference germplasm sets suiting other breeding genepools or to calculate the statistical power for QTL validation of germplasm sets already established.

Fruit ripening can be considered as a complex set of biochemical and physiological changes occurring at the end of the developmental stage. Ripe fruit texture notably affects overall quality and consumer appreciation. Excessive softening limits shelf-life and storability, thereby increasing disease susceptibility and economic loss. Fruit softening is a process due to the depolymerisation of different polysaccharide classes, an event controlled by a synergic and coordinated action of several enzymes among which expansins play a fundamental role. To date, six expansin genes are known to be expressed during apple fruit ontogeny, from full bloom up to fruit ripening. We identified a novel expansin apple homolog (Md-Exp7) sharing high sequence similarity with specific-ripening expansin genes of other crops. A functional marker (Md-Exp7SSR) based on an SSR motif located within the untranslated region of the gene was developed and mapped on Linkage Group 1 of the apple and pear genomes in a region where one major apple QTL for fruit firmness had been previously identified. The allelic composition of 31 apple varieties for the SSR marker was associated with differences in fruit softening.

A Bayesian approach to analyze complex traits is presented that can help plant eneticists and breeders in exploiting the marker and phenotypic data on pedigreed populations as available from ongoing breeding programs. The statistical model for the quantitative trait may include non-genetic and genetic components. The latter component can be divided into QTL on known marker linkage groups, major genes and a polygenic component. The full probability model, prior assumptions on model variables are presented and criterion for model selection and posterior inferences are given. Simulated data on a known pedigreed population structure of the EU project HiDRAS was used to illustrate the use of the Bayesian approach to analyze complex traits. It was shown that estimates for QTL parameters were more accurate when non-genetic factors were included in the model and when a polygenic component was included when not all linkage groups were analyzed simultaneously. The Bayesian approach has been implemented into the software package FlexQTL and allows plant breeders explore their pedigreed populations for segregating QTL alleles that are relevant in their breeding program.

Publication Inra prise en compte dans l'analyse bibliométrique des publications scientifiques mondiales sur les Fruits, les Légumes et la Pomme de terre. Période 2000-2012. http://prodinra.inra.fr/record/256699; International audience; An integrated consensus genetic map for apple was constructed on the basis of segregation data from four genetically connected crosses (C1 = Discovery x TN10-8, C2 = Fiesta x Discovery, C3 = Discovery x Prima, C4 = Durello di Forli x Fiesta) with a total of 676 individuals using CarthaGene(R) software. First, integrated female-male maps were built for each population using common female-male simple sequence repeat markers (SSRs). Then, common SSRs over populations were used for the consensus map integration. The integrated consensus map consists of 1,046 markers, of which 159 are SSR markers, distributed over 17 linkage groups reflecting the basic chromosome number of apple. The total length of the integrated consensus map was 1,032 cM with a mean distance between adjacent loci of 1.1 cM. Markers were proportionally distributed over the 17 linkage groups (chi(2) = 16.53, df = 16, p = 0.41). A non-uniform marker distribution was observed within all of the linkage groups (LGs). Clustering of markers at the same position (within a 1-cM window) was observed throughout LGs and consisted predominantly of only two to three linked markers. The four integrated female-male maps showed a very good colinearity in marker order for their common markers, except for only two (CH01h01, CH05g03) and three (CH05a02z, NZ02b01, Lap-1) markers on LG17 and LG15, respectively. This integrated consensus map provides a framework for performing quantitative trait locus (QTL) detection in a multi-population design and evaluating the genetic background effect on QTL expression

In apple breeding fruit quality is a crucial aspect to guarantee commercial success of new varieties. Among the several quality attributes, individual sugars and acid components, as well as texture, are considered important features due to their significant contribution to sensory quality and consumer's acceptance. In traditional breeding, however, selection for these particular traits is a time consuming process, due to the long unproductive juvenile phase of apple seedlings. This step can be assisted to date by the use of molecular markers, an important diagnostic tool useful to anticipate and assist the identification of novel apple varieties characterized by superior fruit quality behaviour. However, the routinely implementation of molecular markers in breeding programs is still limited. In this study a new approach named Pedigree Based Analysis will be used on six full-sib progenies and their pedigree (composed by 15 varieties, represented by founders and ancestors), to target the main set of quantitative trait loci (QTLs). This strategy will allow the identification of the most valuable alleles present in the breeding activities of both Italian institutes. The ultimate goal of the Laimburg - FEM joint project is the identification and validation of molecular markers as a suitable tool for Marker Assisted Breeding in the two on-going apple breeding programs.

To date, molecular markers have been made available for many economically important traits. Unfortunately, lack of knowledge of their allelic variation hampers their full exploitation in commercial breeding programs. These markers have usually been identified in one single cross. Consequently, only one or two favourable alleles of the related QTL are identified and may be exploited for marker-assisted breeding (MAB), while a breeding program may include several alleles. Selection for only these alleles means that many favourable genotypes are ignored, which decreases efficiency and leads to genetic erosion. A new approach, called pedigree genotyping, allows the identification and exploitation of the majority of alleles present in an ongoing breeding program. This is achieved by including breeding material itself in QTL detection, so covering multiple generations and linking many crosses through their common ancestors in the pedigree. The principle of Identity by Descent (IBD) is utilised to express the identity of an allele of a modern selection in terms of alleles of founding cultivars. These founder alleles are used as factors in statistical analysis. Co-dominant markers, like SSR (= microsatellite) markers, are essential in this approach since they are able to connect cultivars, breeding selections and progenies at the molecular marker level by monitoring specific chromosomal segments along family trees. Additional advantages of the use of breeding genetic material are (1) a major reduction in experimental costs since plant material is already available and phenotyped by default (2) continuity over generations within breeding programs with regard to marker research (3) the testing of QTL-alleles against a wide range of genetic backgrounds, making results generally applicable (4) possibility to explore intra- as well as inter-QTL interactions. Fruit firmness in apple is used as an example to illustrate the principles of this powerful approach to detect QTLs and estimate their allelic variation. Prospects for strawberry are also indicated

Fire blight, caused by the bacterial pathogen Erwinia amylovora, is one of the most destructive diseases of apple (Malus x domestica). In order to analyse the genetic determinism of resistance to fire blight in apple, a quantitative trait analysis (QTL) approach was used. A F1 progeny of 164 individuals derived from a cross between the apple cultivars `Prima¿ and `Fiesta¿ was inoculated in greenhouse conditions. Seven copies per genotype were used. The length of the necrosis observed on shoots was scored 7 and 14 days after inoculation. The MapQTL software was used for QTL analyses, using two previously built maps of the parents, and the symptoms scored on shoots. Digenic interactions between all pairwise combinations of genetic markers were tested using a two-way ANOVA model with the SAS software. QTL were detected at the same locations both 7 and 14 days after inoculation. Two weak effect QTL deriving from `Prima¿ were detected on linkage groups (LG) 3 and LG16. One strong effect QTL deriving from `Fiesta¿ was detected on LG7 that explained 46.6% of the phenotypic variation observed in the progeny. Two additional significant (P

To date, molecular markers are available for many economically important traits. Unfortunately, lack of knowledge of the allelic variation of the related genes hampers their full exploitation in commercial breeding programs. These markers have usually been identified in one single cross. Consequently, only one or two favourable alleles of the related QTL are identified and exploitable for marker-assisted breeding (MAB), whereas a breeding program may include several alleles. Selection for just these alleles means that many favourable genotypes are ignored, which decreases efficiency and leads to genetic erosion. A new approach, called Pedigree Genotyping, allows the identification and exploitation of most alleles present in an ongoing breeding program. This is achieved by including breeding material itself in QTL detection, thus covering multiple generations and linking many crosses through their common ancestors in the pedigree. The principle of Identity by Descent (IBD) is utilised to express the identity of an allele of a modern selection in terms of alleles of founding cultivars. These founder alleles are used as factors in statistical analyses. Co-dominant markers like SSR (microsatellite) markers are essential in this approach since they are able to connect cultivars, breeding selections and progenies at the molecular marker level by monitoring specific chromosomal segments along family trees. Additional advantages of the use of breeding genetic material are (1) a major reduction in experimental costs since plant material is already available and phenotyped by default (2) continuity over generations within breeding programs with regard to marker research (3) the testing of QTL-alleles against a wide range of genetic backgrounds, making results generally applicable, (4) intra- as well as inter-QTL interactions can be explored. Fruit firmness in apple will be used as an example to illustrate the principles of this powerful approach to detect QTLs and estimate their allelic variation

International audience; The Venturia inaequalis-Malus pathosystem was one of the first for which gene-for-gene relationships were demonstrated following the discovery of such relationships between Melampsora lini and flax by Flor in the 1950s. An understanding of these relationships forms the basis for monitoring pathotypes of V. inaequalis at the population level and is employed to assess the usefulness of resistance genes for breeding durable resistance to scab. These pathotypes are difficult to accommodate in the current system of nomenclature for V. inaequalis races where each new combination of avirulence alleles is assigned a simple numerical descriptor as its name. We propose a system that is better suited to the increasing complexities of combinations of genes involved in both race-specific and race-nonspecific recognition by the host, while at the same time updating the name of scab resistance loci (Rvik) and QTL loci (Qvik) to international standards. For the race-specific interactions, the basic premise is that each Rvik-AvrRvik and Qvik-AvrQvik relationship should be represented by a differential host (k), abbreviated to h(k), carrying only the specific Rvik or (major) Qvik resistance allele and an isolate of the pathogen having lost only the complementary allele at the AvrRvik or AvrQvik locus, race (k), with k representing the number of the specific interaction. Races lacking more than one avirulence gene at different loci will be identified as race (k,l,m,...) and apple hosts carrying multiple scab resistance genes as host (k,l,m,...). The proposed system has some continuity with the current system, but should simplify the presentation and interpretation of studies on avirulence alleles in V. inaequalis at the population level. Gene-for-gene relationships reported to date for this pathosystem are reviewed, some inconsistencies clarified, and several new interactions added. The gene-for-gene relationships of European isolates collected in the last decade have been studied in order to establish a new set of reference isolates that represent various V.inaequalis races.

To date, molecular markers are available for many economically important traits in grapevine (Vitis vinifera L.) as well as in other fruit crops. Unfortunately, lack of knowledge of the allelic variation of the related genes hampers their full exploitation in commercial breeding programs. These markers have usually been identified in a single cross. Consequently, only one or two favourable alleles of the related quantitative trait loci (QTL) are identified and exploitable for marker-assisted breeding (MAB), whereas a breeding program may include several alleles. Our novel employed approach, namely pedigree-based analysis (PBA), allows for the identification and exploitation of most alleles present in an ongoing breeding program. This is achieved by including breeding material itself in QTL detection, thus covering multiple generations and linking many crosses through their common ancestors in the pedigree. In this study we focus on the genetic characterization of numerous Vitis hybrids, donors of resistance to downy and/or powdery mildew, in order to identify selection signatures. In particular, 120 hybrids of American origin and 100 European hybrids, along with 44 related V. vinifera cultivars, have been genotyped at 190 SSR loci. These markers have been chosen based on their physical distance in order to have 10 SSRs well-scattered along each chromosome. Based on the SSR profiles, the historical pedigree records have been verified. Following the trueness-to-type validation for most of the studied hybrids, the identity-by-descent (IBD) analysis has been performed, tracing the allelic flow through the successive generations. Here we report results about the identification of non-vinifera genomic intervals retained into the vinifera background and the anchoring of the discovered breeding signatures to the grapevine genome. In addition, we report on the PBA-based QTL results for segregating populations coming from pedigree-supported parental genotypes donors of resistance to downy mildew in north-eastern Italy.

International audience; Key message: Proof of concept of Bayesian integrated QTL analyses across pedigree-related families from breeding programs of an outbreeding species. Results include QTL confidence intervals, individuals' genotype probabilities and genomic breeding values. Abstract: Bayesian QTL linkage mapping approaches offer the flexibility to study multiple full sib families with known pedigrees simultaneously. Such a joint analysis increases the probability of detecting these quantitative trait loci (QTL) and provide insight of the magnitude of QTL across different genetic backgrounds. Here, we present an improved Bayesian multi-QTL pedigree-based approach on an outcrossing species using progenies with different (complex) genetic relationships. Different modeling assumptions were studied in the QTL analyses, i.e., the a priori expected number of QTL varied and polygenic effects were considered. The inferences include number of QTL, additive QTL effect sizes and supporting credible intervals, posterior probabilities of QTL genotypes for all individuals in the dataset, and QTL-based as well as genome-wide breeding values. All these features have been implemented in the FlexQTL (TM) software. We analyzed fruit firmness in a large apple dataset that comprised 1,347 individuals forming 27 full sib families and their known ancestral pedigrees, with genotypes for 87 SSR markers on 17 chromosomes. We report strong or positive evidence for 14 QTL for fruit firmness on eight chromosomes, validating our approach as several of these QTL were reported previously, though dispersed over a series of studies based on single mapping populations. Interpretation of linked QTL was possible via individuals' QTL genotypes. The correlation between the genomic breeding values and phenotypes was on average 90 %, but varied with the number of detected QTL in a family. The detailed posterior knowledge on QTL of potential parents is critical for the efficiency of marker-assisted breeding.

Apple (Malus x domestica Borkh.) is one of the most important fruit trees grown in Europe and around the world for human consumption, and therefore plant breeders aim at producing new apple varieties with high fruit quality. The availability of molecular markers suitable for marker-assisted selection could greatly increase the efficiency and power of breeding. The recent release of the cv. Golden Delicious genome sequence contributed to an exponential increase in the number of Malus sequences publicly available, while the successful achievement of two apple expressed sequence tag (EST) projects permitted the development of molecular markers from coding sequences. Here we present the setting up and mapping of new EST-based markers specific for fruit development and fruit quality traits. Since a large proportion of the ESTs used in our work were transcriptionally characterized and some of them co-localize within quantitative trait locus regions controlling fruit quality traits, the data reported will be effective in the identification of candidate genes. Due to the high level of polymorphism present in the Malus genome, 70% of the entire set of ESTs analyzed were polymorphic and 80% of them were successfully located using the conventional map-based approach. Fifty new EST-based markers were placed on the apple reference genetic map Fiesta x Discovery, thus enhancing the saturation of some regions, and 17 on Prima x Fiesta. Finally, another 17 markers were located on the Golden Delicious genome sequence using an in-silico approach.

International audience; A series of 21 multiplex (MP) polymerase chain reactions containing simple sequence repeat (SSR) markers spanning most of the apple genome has been developed. Eighty-eight SSR markers, well distributed over all 17 linkage groups (LGs), have been selected. Eighty-four of them were included in 21 different MPs while four could not be included in any MPs. The 21 MPs were then used to genotype approximately 2,000 DNA samples from the European High-quality Disease-Resistant Apples for a Sustainable agriculture project. Two SSRs (CH01d03 and NZAL08) were discarded at an early stage as they did not produce stable amplifications in the MPs, while the scoring of the multilocus (ML) SSR Hi07d11 and CN44794 was too complex for large-scale genotyping. The testing of the remaining 80 SSRs over a large number of different genotypes allowed: (1) a better estimation of their level of polymorphism; as well as of (2) the size range of the alleles amplified; (3) the identification of additional unmapped loci of some ML SSRs; (4) the development of methods to assign alleles to the different loci of ML SSRs and (5) conditions at which an SSR previously described as ML would amplify alleles of a single locus to be determined. These data resulted in the selection of 75 SSRs out of the 80 that are well suited and recommended for large genotyping projects

A set of test crosses of diploid potatoes was used to identify QTLs for foliage resistance against Phytophthora infestans and QTLs for foliage maturity type, and to assess their genetic relationship. The most important locus for both traits was found on chromosome 5 near marker GP21: the allele of marker GP21 that is associated with resistance to late blight is also associated with late foliage maturity. An additional QTL with a small effect on foliage maturity type was identified on chromosome 3, and additional QTLs for late blight resistance were found on chromosomes 3 and 10. Another QTL was detected on chromosome 7 when resistance was adjusted for the effect of foliage maturity type. The additional QTLs for resistance against P. infestans on chromosomes 3 and 10 seem to be independent of foliage maturity type and are not affected by epistatic effects of the locus on chromosome 5. The effects of the additional QTLs for resistance are small, but early maturing genotypes that necessarily have the allele for susceptibility for late blight on chromosome 5 may benefit from the resistance that is provided by these QTLs on chromosomes 3 and 10.

Although fire blight, caused by the bacterium Erwinia amylovora, is one of the most destructive dis- eases of apple (Malus · domestica) worldwide, no ma- jor, qualitative gene for resistance to this disease has been identified to date in apple. We conducted a quan- titative trait locus (QTL) analysis in two F1 progenies derived from crosses between the cultivars Fiesta and either Discovery or Prima. Both progenies were inocu- lated in the greenhouse with the same strain of E. am- ylovora, and the length of necrosis was scored 7 days and 14 days after inoculation. Additive QTLs were identified using the MAPQTL software, and digenic epi- static interactions, which are an indication of putative epistatic QTLs, were detected by two-way analyses of variance. A major QTL explaining 34.3-46.6% of the phenotypic variation was identified on linkage group (LG) 7 of Fiesta in both progenies at the same genetic position. Four minor QTLs were also identified on LGs 3, 12 and 13. In addition, several significant digenic interactions were identified in both progenies. These results confirm the complex polygenic nature of resis- tance to fire blight in the progenies studied and also reveal the existence of a major QTL on LG7 that is stable in two distinct genetic backgrounds. This QTL could be a valuable target in marker-assisted selection to obtain new, fire blight-resistant apple cultivars and forms a starting point for discovering the function of the genes underlying such QTLs involved in fire blight control.

In the frame of the D.A.R.E. project, five mapping populations have been studied for partial scab resistance against several races of Venturia inaequalis. A main objective was to identify QTLs (quantitative traits loci) with broad spectrum of resistance towards a wide range of strains of the fungus. Genetic markers (mainly SSR and AFLP) were tested on each population and genetic maps were constructed for both parents of each population. Meanwhile, pathological tests with several isolates of different races of V. inaequalis were performed. Four major genomic regions appear to be involved in scab resistance: they are located on linkage groups (LG) LG-1, LG-2, LG-11, and LG-17. Some other linkage groups carry either QTLs or major resistance genes that are isolate specific: a QTL on LG-5, Vd on LG-10, and Vg on LG-12. The QTL region located on LG-17 clearly exhibited the widest spectrum of resistance

With the advent of genetic maps for apple that carry common transferable markers, it is possible to investigate genomic relationships between genes present in different accessions. Co-dominant markers, such as microsatellites, are particularly useful for this purpose. In recent years, genetic markers have been developed for a number of resistance genes for apple scab (Venturia inaequalis). In this paper, we present the discovery of a new scab resistance gene (Vh8) that maps to linkage group 2 (LG2). We then bring together the findings from different research groups on other scab resistance genes that also map to LG2 in an attempt to delineate their arrangement. These other genes comprise major genes (Vh2, Vr, Vbj and Vr2), as well as several race specific quantitative trait loci (QTLs) from `Discovery? and `TN10-8?. Results indicate that Vh2 and Vr are probably identical, and that Vh2/Vr, Vh8, Vbj and at least one of the QTLs are closely linked to each other on the lower half of LG2. A putative map of this gene cluster is presented. The fourth major gene, Vr2, maps at a significant distance from this gene cluster at the top end of LG2. We discuss the consequences of resistance gene cluster arrangements on breeding strategies for durable resistance to apple scab and the use of marker-assisted selection

We used a new method called nucleotide-binding site (NBS) profiling to identify and map resistance gene analogues (RGAs) in apple. This method simultaneously allows the amplification and the mapping of genetic markers anchored in the conserved NBS-encoding domain of plant disease resistance genes. Ninety-four individuals belonging to an F1 progeny derived from a cross between the apple cultivars ‘Discovery’ and ‘TN10-8’ were studied. Two degenerate primers designed from the highly conserved P-loop motif within the NBS domain were used together with adapter primers. Forty-three markers generated with NBS profiling could be mapped in this progeny. After sequencing, 23 markers were identified as RGAs, based on their homologies with known resistance genes or NBS/leucine-rich-repeat-like genes. Markers were mapped on 10 of the 17 linkage groups of the apple genetic map used. Most of these markers were organized in clusters. Twenty-five markers mapped close to major genes or quantitative trait loci for resistance to scab and mildew previously identified in different apple progenies. Several markers could become efficient tools for marker-assisted selection once converted into breeder-friendly markers. This study demonstrates the efficiency of the NBS-profiling method for generating RGA markers for resistance loci in apple.

The ABC model for floral development was proposed more than 10 years ago and since then many studies have been performed on model species, such as Arabidopsis thaliana, Antirrhinum majus, and many other species in order to confirm this hypothesis. This led to additional information on flower development and to more complex molecular models. AGAMOUS (AG) is the only C type gene in Arabidopsis and it is responsible for stamen and carpel development as well as floral determinacy. LLAG1, an AG homologue from lily (Lilium longiflorum Thunb.) was isolated by screening a cDNA library derived from developing floral buds. The deduced amino acid sequence revealed the MIKC structure and a high homology in the MADS-box among AG and other orthologues. Phylogenetic analysis indicated a close relationship between LLAG1 and AG orthologues from monocot species. Spatial expression data showed LLAG1 transcripts exclusively in stamens and carpels, constituting the C domain of the ABC model. Functional analysis was carried out in Arabidopsis by overexpression of LLAG1 driven by the CaMV35S promoter. Transformed plants showed homeotic changes in the two outer floral whorls with some plants presenting the second whorl completely converted into stamens. Altogether, these data strongly indicated the functional homology between LLAG1 and AG.

Although varying with context, quality of fresh fruits includes several properties such as color, texture, flavor and health promoting compounds. This thesis focused on two important quality aspects, namely texture and aroma in apple, and defining the genomic regions involved in the control of these two features. The genetic control of texture and VOCs production have been investigated using two marker-trait association analysis approaches: Pedigree Based Analysis (PBA) and Genome Wide Association Study (GWAS). In chapter 2, ASSIsT (Automatic SNP ScorIng Tool), a software dedicated for the efficient calling and filtering of SNPs from Illumina InfiniumÒ arrays is presented. ASSIsT builds on GenomeStudio® derived data and identifies markers showing reliable genotype calls (bi-allelic segregation pattern). In addition, ASSIsT identifies and re-edits SNP calls of markers showing additional alleles (null alleles or additional SNPs in the probe annealing site). Chapter 3 aimed to dissect the genetic control of fruit firmness in apple during storage through PBA and employing 24 bi-parental families (1216 individuals) connected by a common pedigree structure. Ten QTLs were identified encompassing eight linkage groups, which unravelled a QTL dynamics over storage shedding light on the specific genetic control at each time-point. Chapter 4: aimed to comprehensively decipher the genetic control of fruit texture. Two complementing QTL mapping approaches were employed together with a novel and high sophisticated phenotyping device for fruit texture. The PBA was carried out on six full-sib pedigreed families (416 individuals), while the GWAS was performed on a collection of 233 apple accessions. The texture analyser employed (TAXT-AED texture analyser) allowed the measurement of both the mechanical properties (firmness) and the acoustic properties (crispness) of fruit texture. The QTL results indicated chromosome 10 being associated in changes of the mechanical properties of fruit texture, while chromosomes 2 and 14 were more associated to the acoustic response. In Chapter 5 the interplay between texture and volatile organic compounds (VOCs) was investigated in 162 apple accessions. The array of volatile compounds phenotyped was implemented into a GWAS identifying seven chromosomes harbouring important candidate genes for aroma, such as MdAAT1 and MdIGS. Next, volatilome and fruit texture data were integrated revealing a negative correlation between these two features.