A clarified position for solanum lycopersicum var. cerasiforme in the evolutionary history of tomatoes (solanaceae)

The natural phenotypic variability present in the germplasm of cultivated plants can be linked to molecular polymorphisms using association genetics. However it is necessary to consider the genetic structure of the germplasm used to avoid false association. The knowledge of genetic structure of plant populations can help in inferring plant evolutionary history. In this context, we genotyped 360 wild, feral and cultivated accessions with 20 simple sequence repeat markers and investigated the extent and structure of the genetic variation. The study focused on the red fruited tomato clade involved in the domestication of tomato and confirmed the admixture status of cherry tomatoes (Solanum lycopersicum var. cerasiforme). We used a nested sample strategy to set-up core collection maximizing the genetic diversity with a minimum of individuals. Results Molecular diversity was considerably lower in S. lycopersicum i.e. the domesticated form. Model-based analysis showed that the 144 S. lycopersicum var. cerasiforme accessions were structured into two groups: one close to the domesticated group and one resulting from the admixture of the S. lycopersicum and S. pimpinellifolium genomes. SSR genotyping also indicates that domesticated and wild tomatoes have evolved as a species complex with intensive level of hybridization. We compiled genotypic and phenotypic data to identify sub-samples of 8, 24, 32 and 64 cherry tomato accessions that captured most of the genetic and morphological diversity present in the entire S. lycopersicum var. cerasiforme collection. Conclusion The extent and structure of allelic variation is discussed in relation to historical events like domestication and modern selection. The potential use of the admixed group of S. lycopersicum var. cerasiforme for association genetics studies is also discussed. Nested core collections sampled to represent tomato diversity will be useful in diversity studies. Molecular and phenotypic variability of these core collections is defined. These collections are available for the scientific community and can be used as standardized panels for coordinating efforts on identifying novel interesting genes and on examining the domestication process in more detail

Exploration du polymorphisme moléculaire et protéique de la tomate pour l'identification de QTL de qualité du fruit

L amélioration de la qualité du fruit de tomate dépend largement de la variation génétique. A la suite de la domestication et de la sélection moderne, la diversité moléculaire chez la tomate a été profondément réduite, limitant les possibilités d amélioration. De nouveaux marqueurs moléculaires révélant les polymorphismes nucléotidiques (SNP) constituent des outils précieux pour saturer les cartes génétiques et identifier des QTL (quantitative trait loci) et des associations chez une espèce peu polymorphe comme la tomate. Les objectifs de cette étude étaient de caractériser la diversité génétique de la tomate au niveau moléculaire et de tenter d identifier des QTL, des gènes et des protéines responsables de la variation de caractères de qualité du fruit. Pour cela, trois études indépendantes ont conduit à (1) la découverte de nouveaux marqueurs SNP, (2) leur utilisation en génétique d association et (3) l analyse de la diversité du protéome en relation avec des caractères physiologiques du fruit.Dans la première étude, nous avons comparé deux plateformes de reséquençage pour reséquencer des zones ciblées couvrant environ 0.2% du génome de deux accessions contrastées. Plus de 3000 SNP sont été identifiés. Nous avons ensuite validé 64 SNPs en développant des marqueurs CAPS. Nous avons ainsi montré l intérêt des techniques de reséquençage pour la découverte de SNP chez la tomate et produit des marqueurs simples qui peuvent être utiles pour caractériser de nouvelles ressources.Nous avons ensuite développé un ensemble de 192 SNPs et génotypé une collection de 188 accessions comportant des accessions cultivées, des types cerise et des formes sauvages apparentées et recherché des associations avec 10 caractères de qualité du fruit. Une quarantaine d associations a été détectée dans des régions où des QTL avaient été préalablement identifiés. D autres associations ont été identifiées dans de nouvelles régions. Nous avons ainsi confirmé le potentiel de la génétique d association pour la découverte de QTL chez la tomate. Finalement une approche combinant l analyse du protéome, du métabolome et de traits phénotypiques a été mise en oeuvre pour étudier la variabilité naturelle de la qualité du fruit de huit lignées contrastées et de quatre de leurs hybrides, à deux stades de développement (expansion cellulaire et orange-rouge). Nous avons identifié 424 spots protéiques variables en combinant électrophorèse bidimensionnelle et nano LC MS/MS et construit une carte de référence du protéome de fruit de tomate. En parallèle, nous avons mesuré la variation de teneurs en 34 métabolites, les activités de 26 enzymes et cinq caractères phénotypiques. La variabilité génétique et les modes d hérédité ont été décrits. L intégration des données a été réalisée par construction de réseaux de corrélations et régression sPLS. Plusieurs associations ont été détectées intra et inter niveau d expression, permettant une meilleure compréhension de la variation de la qualité des fruits de tomateFruit quality in tomato is highly dependent on genetic variation. Following domestication and modernbreeding, molecular diversity of tomato has been strongly reduced, limiting the possibility to improvetraits of interest. New molecular markers such as single nucleotide polymorphisms (SNP) constituteprecious tools to saturate tomato genetic maps and identify quantitative trait loci (QTL) andassociations in a poorly polymorphic species like tomato. The objectives of this study were tocharacterize tomato genetic diversity at the molecular levels and to try to identify QTLs, genes andproteins responsible for fruit quality traits in tomato. For this purpose, three independent studies wereconducted leading to the discovery of new SNP markers, their use for association study and finally theanalysis of proteome diversity in relation to physiological phenotypes. We first used two nextgenrationsequencing platforms (GA2 Illumina and 454 Roche) to re-sequence targeted sequencescovering about 0.2% of the tomato genome from two contrasted accessions. More than 3000 SNPswere identified between the two accessions. We then validated 64 SNPs by developing CAPS markers.We thus showed the value of NGS for the discovery of SNPs in tomato and we produced low costCAPS markers which could be used to characterize other tomato collections. A SNPlexTM arraycarrying 192 SNPs was then developed and used to genotype a broad collection of 188 accessionsincluding cultivated, cherry type and wild tomato species and to associate these polymorphisms to tenfruit quality traits using association mapping approach. A total of 40 associations were detected andco-localized with previously mapped QTLs. Some other associations were identified in new regions.We showed the potential of using association genetics in tomato. Finally, a new analytical approachcombining proteome, metabolome and phenotypic profiling were applied to study natural geneticvariation of fruit quality traits in eight diverse accessions and their four corresponding F1s at cellexpansion and orange-red stages. We identified 424 variable spots by combining 2-DE and nano LCMS/MS and built the first comprehensive proteome reference map of the tomato fruit pericarp at twodevelopmental stages from the 12 genotypes. In parallel, we measured the variation of 34 metabolites,26 enzyme activities and five phenotypic traits. A large range of variability and several inheritancemodes were described in the four groups of traits. Data integration was achieved through sPLS andcorrelation networks. Many significant associations were detected within level and between levels ofexpression. This systems biology approach provides better understanding of networks of elements(proteins, enzymes, metabolites and phenotypic traits) in tomato fruitsAVIGNON-Bib. numérique (840079901) / SudocSudocFranceF

Increase in Tomato Locule Number Is Controlled by Two Single-Nucleotide Polymorphisms Located

In tomato (Solanum lycopersicum) fruit, the number of locules (cavities containing seeds that are derived from carpels) varies from two to up to 10 or more. Locule number affects fruit shape and size and is controlled by several quantitative trait loci(QTLs). The large majority of the phenotypic variation is explained by two of these QTLs, fasciated (fas) and locule number (lc), that interact epistatically with one another. FAS has been cloned, and mutations in the gene are described as key factors leading to the increase in fruit size in modern varieties. Here, we report the map-based cloning of lc. The lc QTL includes a 1,600-bp region that is located 1,080 bp from the 3# end of WUSCHEL, which encodes a homeodomain protein that regulates stem cell fate in plants. The molecular evolution of lc showed a reduction of diversity in cultivated accessions with the exception of two single-nucleotide polymorphisms. These two single-nucleotide polymorphisms were shown to be responsible for the increase in locule number. An evolutionary model of locule number is proposed herein, suggesting that the fas mutation appeared after the mutation in the lc locus to confer the extreme high-locule-number phenotype

Metabolic characterization of loci affecting sensory attributes in tomato allows an assessment of the influence of the levels of primary metabolites and volatile organic contents

Numerous studies have revealed the extent of genetic and phenotypic variation between both species and cultivars of tomato. Using a series of tomato lines resulting from crosses between a cherry tomato and three independent large fruit cultivar (Levovil, VilB, and VilD), extensive profiling of both central primary metabolism and volatile organic components of the fruit was performed. In this study, it was possible to define a number of quantitative trait loci (QTLs) which determined the levels of primary metabolites and/or volatile organic components and to evaluate their co-location with previously defined organoleptic QTLs. Correlation analyses between either the primary metabolites or the volatile organic compounds and organoleptic properties revealed a number of interesting associations, including pharmaceutical aroma–guaiacol and sourness–alanine, across the data set. Considerable correlation within the levels of primary metabolites or volatile organic compounds, respectively, were also observed. However, there was relatively little association between the levels of primary metabolites and volatile organic compounds, implying that they are not tightly linked to one another. A notable exception to this was the strong association between the levels of sucrose and those of a number of volatile organic compounds. The combined data presented here are thus discussed both with respect to those obtained recently from wide interspecific crosses of tomato and within the framework of current understanding of the chemical basis of fruit taste

Breeding Tomato Hybrids for Flavour: Comparison of GWAS Results Obtained on Lines and F1 Hybrids

[EN] Tomato flavour is an important goal for breeders. Volatile organic compounds (VOCs) are major determinants of tomato flavour. Although most tomato varieties for fresh market are F1 hybrids, most studies on the genetic control of flavour-related traits are performed on lines. We quantified 46 VOCs in a panel of 121 small fruited lines and in a test cross panel of 165 hybrids (the previous panel plus 44 elite cherry tomato lines crossed with a common line). High and consistent heritabilities were assessed for most VOCs in the two panels, and 65% of VOC contents were strongly correlated between lines and hybrids. Additivity was observed for most VOCs. We performed genome wide association studies (GWAS) on the two panels separately, along with a third GWAS on the test cross subset carrying only F1 hybrids corresponding to the line panel. We identified 205, 183 and 138 associations, respectively. We identified numerous overlapping associations for VOCs belonging to the same metabolic pathway within each panel; we focused on seven chromosome regions with clusters of associations simultaneously involved in several key VOCs for tomato aroma. The study highlighted the benefit of testcross panels to create tasty F1 hybrid varieties.This research was funded by the CIFRE project Qualhytom, grant number 2018/1239, the ANR project TomEpiSet, grant number ANR-16-CE20-0014 and European Union's Horizon 2020 research and innovation programme, HARNESSTOM, grant number No. 101000716.Bineau, E.; Rambla Nebot, JL.; Priego-Cubero, S.; Hereil, A.; Bitton, F.; Plissonneau, C.; Granell Richart, A.... (2021). Breeding Tomato Hybrids for Flavour: Comparison of GWAS Results Obtained on Lines and F1 Hybrids. Genes. 12(9):1-20. https://doi.org/10.3390/genes12091443S12012

Identification of growth processes involved in QTLs for tomato fruit size and composition

Many quantitative trait loci (QTLs) for quality traits have been located on the tomato genetic map, but introgression of favourable wild alleles into large fruited species is hampered by co-localizations of QTLs with antagonist effects. The aim of this study was to assess the growth processes controlled by the main QTLs for fruit size and composition. Four nearly isogenic lines (NILs) derived from an intraspecific cross between a tasty cherry tomato (Cervil) and a normal-tasting large fruit tomato (Levovil) were studied. The lines carried one (L2, L4, and L9) or five (Lx) introgressions from Cervil on chromosomes 1, 2, 4, and 9. QTLs for fruit size could be mainly associated with cell division processes in L2 and L9, whereas cell expansion was rather homogeneous among the genotypes, except Cervil for which the low expansion rate was attributed to low cell plasticity. The link between endoreduplication and fruit size remained unclear, as cell or fruit sizes were positively correlated with the cell DNA content, but not with the endoreduplication factor. QTLs for fruit composition reflected differences in water accumulation rather than in sugar accumulation, except in L9 for which the up-regulation of sucrose unloading and hexose transport and/or starch synthesis was suggested. This may explain the increased amount of carbon allocated to cell structures in L9, which could be related to a QTL for fruit texture. In Lx, these effects were attenuated, except on fruit size and cell division. Finally, the region on top of chromosome 9 may control size and composition attributes in tomato, by a combination of QTL effects on cell division, cell wall synthesis, and carbon import and metabolism

Combining ecophysiological modelling and quantitative trait locus analysis to identify key elementary processes underlying tomato fruit sugar concentration

A mechanistic model predicting the accumulation of tomato fruit sugars was developed in order (i) to dissect the relative influence of three underlying processes: assimilate supply (S), metabolic transformation of sugars into other compounds (M), and dilution by water uptake (D); and (ii) to estimate the genetic variability of S, M, and D. The latter was estimated in a population of 20 introgression lines derived from the introgression of a wild tomato species (Solanum chmielewskii) into S. lycopersicum, grown under two contrasted fruit load conditions. Low load systematically decreased D in the whole population, while S and M were targets of genotype×fruit load interactions. The sugar concentration positively correlated to S and D when the variation was due to genetic introgressions, while it positively correlated to S and M when the variation was due to changes in fruit load. Co-localizations between quantitative trait loci (QTLs) for sugar concentration and QTLs for S, M, and D allowed hypotheses to be proposed on the processes putatively involved at the QTLs. Among the five QTLs for sugar concentration, four co-localized with QTLs for S, M, and D with similar allele effects. Moreover, the processes underlying QTLs for sugar accumulation changed according to the fruit load condition. Finally, for some genotypes, the processes underlying sugar concentration compensated in such a way that they did not modify the sugar concentration. By uncoupling genetic from physiological relationships between processes, these results provide new insights into further understanding of tomato fruit sugar accumulation