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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; 'Antonovka' has long been recognised as a major source of scab (Venturia inaequalis) resistance useful for apple breeding worldwide. Both major gene resistances in the form of the Rvi10 and Rvi17 and quantitative resistance, collectively identified as VA, have been identified in different accessions of 'Antonovka'. Most of the 'Antonovka' scab resistance used in apple-breeding programmes around the world can be traced back to Schmidt 'Antonovka' and predominantly its B VIII progenies 33,25 (PI 172623), 34,6 (PI 172633), 33,8 (PI 172612) and 34,5 (PI 172632). Using genetic profile reconstruction, we have identified "common 'Antonovka' " as the progenitor of the B VIII family, which is consistent with it having been a commercial cultivar in Poland and the single source of scab resistance used by Dr. Martin Schmidt. The major 'Antonovka' scab resistance genes mapped to date are located either very close to Rvi6, or about 20-25 cM above it, but their identities need further elucidation. The presence of the 139 bp allele of the CH-Vf1 microsatellite marker known to be associated with Rvi17 (Va1) in most of the 'Antonovka' germplasm used in breeding suggests that it plays a central role in the resistance. The nature and the genetic relationships of the scab resistance in these accessions as well as a number of apple cultivars derived from 'Antonovka', such as, 'Freedom', 'Burgundy' and 'Angold', are discussed. The parentage of 'Reglindis' is unclear, but the cultivar commercialised as 'Reglindis' was confirmed to be an Rvi6 cultivar.

A reproducible procedure was developed for genetic transformation of Hydrangea macrophylla Ser. cv . Blaumeise by Agrobacterium tumefaciens following the development of an efficient regeneration system using leaf discs excised from 12 to 15 weeks old meristem-derived vitroplants. Explants were cultivated on solid B5 medium complemented with maltose 110 mM, BAP 10 μM and NAA 0.5 μM. A low light regime of 17 μmol m −2 s −1 improved regeneration frequency up to 86%. For transformation, leaf discs were inoculated and co-cultivated with two disarmed A. tumefaciens strains, EHA 101 and LBA 4404, both carrying the binary vector pFAJ3000 which contained the nptII selectable gene and the GUS reporter gene. A pre-culture period of 3 days and a short co-cultivation duration (1 day) improved the efficiency of transformation. Inoculation of only 10 min with agitation including (or not) vacuum infiltration was sufficient. If selection on kanamycin containing medium was applied after a 2 weeks culture period on shoot regeneration medium, the percentage of explants forming kanamycin-resistant shoots increased from 3.3 to 13.3%. Integration and expression of the introduced transgene were confirmed by histochemical GUS assay, PCR and Southern blot analysis. Flowering of transgenic plants in glasshouse occurred 10 months after acclimatization.

International audience; In vitro development of isolated embryos and axillary bud proliferation were studied in Pistacia vera L. Different regulator-free nutrient media were compared to determine the effects of the mineral solution, agar and sucrose concentrations on seedling development from mature embryos. Nutrient-rich MS [Murashige, T., Skoog, F., 1962. A revised medium for rapid growth and bioassays with tabacco tissue cultures. Physiol. Plant. 15,473-479] and DKW [Driver, J.A., Kuniyuki, A.M., 1984. In vitro propagation of Paradox walnut rootstock. HortScience 19, 507-509] mineral solutions were more efficient for the development of aerial parts than nutrient-poor KN [Knop, W., 1884. Bereitung einer concentrierten nahrstofflosung fur pflanzen. Landwersuhssat 30, 292-294] and WT [Withe, P.R., 1936. Plant tissue cultures. Bot. Rev. 2, 419-437] solutions. Reducing the agar concentration enhanced fresh matter production and balanced seedling development. When tested at different concentrations, sucrose was found to orient mature embryo development, with the best results obtained at concentrations of 2-4%, whereas high concentrations (6 and 12%) mainly inhibited elongation of the aerial parts. Plantlets obtained previously from in vitro cultured embryos were propagated by axillary budding. High bud proliferation (six shoots per explant) was achieved when using 17.8 mu M benzyladenine (BA) combined with 0.5 mu M indole-3-butyric acid (IBA). The addition of 2.9 mu M gibberellic acid (GA(3)) to the propagation medium did not improve axillary shoot yields and on average, media with GA(3) produced 2.3-2.6 elongated stems per cultured explant. Shoots were rooted in vitro in half-strength MS medium containing 12.3 mu M IBA

International audience; We formulate a spatially realistic population-genetic model for ascertaining the synergetic effect between genetic and spatial composition of the host population on the pathogen spread reinforced by evolutionary processes. We show that spatial arrangement of host genotypes is crucial to the efficacy of host genetic diversification. In particular, the reductive effect of multigenic resistance on the pathogen density can be produced by a random patterning of monogenic resistances. Random patterns can reduce both density and genetic diversity of the pathogen population and delay invasion promoted by sexual recombination. By contrast, patchy distributions diversify pathogen population and, hence, reduce the efficacy of resistance genes. The proposed approach provides theoretical support for studying fast emergence and spread of novel pathogen genotypes carrying multiple virulence genes. It has a practical applicability to design innovative strategies for the most appropriate deployment of plant resistance genes.

International audience; Fire blight caused by the bacteria Erwinia amylovora is one of the most important diseases of pear. This causal agent produces a siderophore (desferrioxamine), which was identified as one of its virulence factors. This protein enables the bacteria to overcome conditions of iron limitation encountered in host tissues, and may also protect the bacteria against active oxygen species. Previous experiments indicate that the use of an iron chelator protein, encoded by the bovine lactoferrin gene, reduces fire blight symptoms in some transgenic pear clones. The aim of the present work is to test the ability of a more efficient iron chelator, plant ferritin, to reduce fire blight susceptibility in pear. In the literature, ferritin genes have been overexpressed under the control of a constitutive promoter in different transgenic plant species for various purposes. In several cases, the constitutive expression of ferritin produced negative effects such as reduced growth and chlorophyll content. Therefore, we decided to place the exogenous ferritin gene from pea under the control of a pathogen inducible promoter (sgd24) in comparison with a constitutive promoter (CaMV 35S). Two pear varieties, 'Conference' (CF) and 'Passe-Crassane' (PC) were transformed using both constructs. Transformation rates depended on variety and construct. They were respectively of 12 and 4.3% for CF and PC using the sgd24-ferritin construct. Only PC was transformed with the 35S-ferritin construct, with a transformation rate of 2%. First analyses of the transgenic clones by RT-PCR showed the expression of pea ferritin in both constructs and in all clones. The transgenic clones were acclimatized in greenhouse and exhibited normal growth. Quantification of ferritin gene expression, ferritin protein accumulation, and evaluation of fire blight resistance are underway.

UMR DAP équipe AFEF 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; The present study aimed to dissect tree architectural plasticity into genetic, ontogenetic and environmental effects over the first 4 years of growth of an apple F1 progeny by means of quantitative traits loci (QTL) mapping. Both growth and branching processes were phenotyped on the consecutive annual shoots of different axes within a tree. For each studied trait, predicted values (best linear unbiased predictors, BLUPs) of the genotypic (G) effect or its interaction with tree age (G×A) and climatic year (G×Y) were extracted from mixed linear models of repeated data. These BLUPs, which are independent from autocorrelations between repeated measurements, were used for QTL mapping. QTL detection power was improved by this two-step approach. For each architectural process, numerous QTLs were detected and some particularly interesting co-localised in common genomic regions, for internode lengthening, top diameter, and number and percentage of axillary shoots. When several QTLs were detected for a given trait, global models were estimated, which explained a maximum of 40% of the total variance for both internode length and top diameter and 28% for branching. QTLs detected for BLUPs of G×Y effects were interpreted as resulting from the interaction between genetic maximal potential of growth and climatic factors, while those for G×A effects were interpreted in relation to tree ontogeny. Most of the latter ones were found to be concomitant with key development stages during which the trait average started to decrease, but with different magnitudes depending on genotype

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

International audience; Eutypine is a toxin produced by Eutypa lata, the causal agent of eutypa dieback of grapevines. An eutypine detoxifying gene (Vr-ERE) encoding an NADPH-dependent aldehyde reductase converts eutypine into the corresponding alcohol, eutypinol, a non-toxic form of the toxin. A variety of phytotoxic compounds containing an aldehyde group can act as substrates for this enzyme, therefore opening the possibility to use Vr-ERE as an alternative selection system for plant transformation. Our preliminary experiments with apple and pear have demonstrated the following: 1) among the various substrates of VR-ERE, benzaldehyde (BD), a naturally occurring compound, is able to inhibit adventitious bud regeneration from apple as well as pear in vitro leaves, at concentrations ranging from 1 to 1.5 mM; 2) constitutive expression of Vr-ERE in transgenic ‘Greensleeves’ apple does not provoke any abnormal phenotype after one year of growth in greenhouse, but aldehyde reductase activity is reduced in all transgenic clones; 3) production of transgenic pear using BD instead of kanamycin as selection pressure is feasible, but the efficiency of the selection pressure seems low. In conclusion, the efficiency of Vr-ERE as a selective gene to produce apple and pear transgenic plants is not yet proven and further research is needed

FREE ACCESS This is an Open Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License (http://creativecommons.org/licenses/by-nc/2.0/uk/) which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited; International audience; Objective: AppleBreed DataBase (DB) aims to store genotypic and phenotypic data from multiple pedigree verified plant populations (crosses, breeding selections and commercial cultivars) so that they are easily accessible for geneticists and breeders. It will help in elucidating the genetics of economically important traits, in identifying molecular markers associated with agronomic traits, in allele mining and in choosing the best parental cultivars for breeding. It also provides high traceability of data over generations, years and localities. AppleBreed DB could serve as a generic database design for other perennial crops with long economic lifespans, long juvenile periods and clonal propagation. Results: AppleBreed DB is organized as a relational database. The core element is the GENOTYPE entity, which has two sub-classes at the physical level: TREE and DNA-SAMPLE. This approach facilitates all links between plant material, phenotypic and molecular data. The entities TREE, DNA-SAMPLE, PHENOTYPE and MOLECULAR DATA allow multi-annual observations to be stored as individual samples of individual trees, even if the nature of these observations differs greatly (e.g. molecular data on parts of the apple genome, physico-chemical measurements of fruit quality traits, and evaluation of disease resistance). AppleBreed DB also includes synonyms for cultivars and pedigrees. Finally, it can be loaded and explored through the web, and comes with tools to present basic statistical overviews and with validation procedures for phenotypic and marker data to certify data quality. AppleBreed DB was developed initially as a tool for scientists involved in apple genetics within the framework of the European project, ‘High-quality Disease Resistance in Apples for Sustainable Agriculture’ (HiDRAS), but it is also applicable to many other perennial crops

International audience; Several studies have shown that the proanthocyanidins are responsible for some major flavour properties of the red wines such as colour, bitterness and astringency. Nevertheless, their synthesis and properties are still misunderstood. Some studies have suggested that the proanthocyanidin content is set before veraison. This paper describes the setting of the proanthocyanidins, until the veraison stage, and the relations that exist with the environment. The flavan-3-ol composition was monitored from the end of the berry set until mid veraison, on two plots known to induce a different behaviour of the vine. The study considered the value of the parameters for a homogeneous stage of development. The composition in proanthocyanidins in skin was determined by HPLC reversed phase, after fractionation and thiolysis. A further information about the proanthocyanidins was obtained by histochemistry. The sections, 2 [mu]m thick, were stained according to the 4-dimethylaminocinnamaldehyde method. The interest and the limits of these approaches are described and discussed. Moreover, it is shown that the linking of these two analytical techniques which associates quantitative data and the spatializing of the tannins in cellular tissue could improve the understanding of the setting of the proanthocyanidins content. Finally, this original approach pointed out the importance of the period previous ripening in the synthesis of flavan-3-ols in the skin; the heterogeneity among berries was in relation with the stage of development but some other factors, as light exposition could also play a role

International audience

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

Molecular characterization of french local apple cultivars using SSRs. 11.Eucarpia Symposium on Fruit Breeding and Genetics

International audience

Resistance to pests and diseases is a highly desirable objective in fruit breeding but is a long-term endeavor. It requires appropriate tests for early screening and an analysis of host-parasite interactions for durability of resistance. Host resistance is considered durable when it remains effective for a long period despite an intensive exposure to the parasite. Durability has to be the key-point of any fruit breeding programme for resistance. Non-durable resistance is usually associated with a hypersensitive reaction (HR) and major gene resistance which often occur in multiple allelic series and/or complex loci. HR resistance or race-specific resistance is often rendered ineffective because the pathogen population shifts genetically to avoid the resistance gene. Resistance of a more durable nature against fungi and bacteria is often polygenic and based upon the additive or interaction effects of a few to several genes not promoting a hypersensitive reaction. The more complex the genetics of the resistance, the more difficult it is for the pathogen to evolve to a form that can overcome the resistance. Polygenic resistance is usually associated with a quantitative expression that is characterized by a continuous variation between susceptibility and resistance. To approach the number of genes involved in polygenic resistance (QTL) and the relative effect of each of these genes, they have to be mapped on the genome. Marker-assisted selection (MAS) gives the possibility to recombine QTL for resistance; major genes could be associated with QTL. The combining of resistance genes is desired to: acquire a higher level of resistance against a particular pathotype, obtain resistance against a broader range of pathotypes, and combine different mechanisms of resistance. The current situation of fruit crops - monocultivar orchards established for several years - enhances the selection pressure on the pathogen population and results in population shifts which could be dramatic. Guidelines have to be proposed to predict the evolutionary potential of pathogen populations based on analysis of their genetic structure; the most important parameters to consider are reproductive/mating systems and gene/genotype flow. Pathogens that have the greatest risk of breaking down resistance genes are those that possess a mixed reproduction system, with one sexual cycle per growing season and asexual reproduction during the epidemic phase, and a high potential for gene flow. Several examples of different strategies used by fruit breeders will be reported mainly on Coffea (coffee rust), Musa (black leaf streak disease), Prunus (root knot nematodes) and Malus (scab, mildew, fire blight). Fruit breeders can use the lessons learned from deploying resistance genes in annual crops and selecting resistance in forest trees and apply these to their strategies for fruit tree improvement

Sequence conservation among resistance genes (R genes) was exploited to identify 47 R gene analogues (RGAs) from sugar beet (Beta vulgaris L.). Using degenerate primers, 11 RGAs were amplified from genomic DNA and 7 from leaf or beet cDNA. Twenty-nine were selected from an EST sequencing program. Twenty-one RGAs contained structures similar to the nucleotide binding site (NBS) – leucine rich repeat (LRR) domain, a motif commonly found in several R genes. Among the remaining RGAs, 19 revealed similarity to the serine (threonine) protein kinase domain of R genes, 4 showed features related to the LRR region of the rice disease resistance gene Xa21, 1 RGA resembled the sugar beet nematode resistance gene Hs1pro-1, and 2 had homologies to other gene products associated with disease resistance. For 20 EST-derived RGAs, transcript levels were compared in leaf and root tissue revealing organ-specific transcription in 7 cases. Thirty-three RGAs were spread over all nine sugar beet chromosomes, except for a cluster of nine closely linked RGAs on chromosome 7. The analysis of linkage between RGAs and loci for rhizomania and Cercospora resistance identified alleles associated with resistance in both cases.Key words: RGA, Beta vulgaris, NBS–LRR, genetic linkage map, molecular marker.

The contribution of in vitro methods and molecular tools to pear genetic studies and breeding has advanced considerably during the last decade. Haploidization via in situ parthenogenesis induced by irradiated pollen and in vitro rescue of the haploid plantlets has been developed for pear. Techniques of adventitious bud regeneration from in vitro leaves have been developed for several genotypes of European and Asian pear and for quince. So far, applications of these techniques for the induction of somaclonal variation have been very limited. Increased tolerance to fire blight has been obtained in somaclonal variants of the cultivar Durondeau, and selection of quince rootstocks tolerant to iron deficiency has been achieved with adventious regeneration followed by in vitro selection. A major evolution of pear biotechnology is the recent development of Agrobacterium-mediated transformation. Since the first report in 1996, several teams have succeeded to transform about ten different genotypes of varieties and rootstocks. Projects have started in several countries to express various trangenes in pear (lytic peptides, rolC and SAMase genes). Molecular markers developed on pear now include isozymes, RAPDs and RFLPs. They have been used mostly for varietal identification of european as well as asian pears. Markers have been developed recently for a few genes of interest, such as a black-spot resistance gene and an ACC synthase gene. But no genetic map of pear is yet available. Finally, about 20 genes have already been cloned in pear, most of them from P. pyrifolia. They are mostly involved in fruit ripening or quality and self-incompatibility.

International audience

Fire blight, caused by Erwinia amylovora, is a major disease of pear (Pyrus communis L.). Different strategies to enhance fire blight resistance of pear through genetic engineering are currently studied in our group. Among them, transformation of pear with a depolymerase gene from a bacteriophage has been investigated. This gene encodes an enzyme that degrades the capsular exopolysaccharide (EPS) of E. amylovora. Mutant strains without bacterial EPS are known to fail to colonize host plants. Pear cv. ‘Passe Crassane’ was transformed with Agrobacterium tumefaciens EHA105pBinAR-dpo carrying the depolymerase gene under the control of the CaMV35S promoter. Sixteen transgenic diploid clones were obtained and analysed by western blot and RT-PCR. In vitro tests for fire blight resistance were performed by inoculation with a virulent strain of E. amylovora. Two transgenic clones showed a slight reduction of fire blight symptoms (about 15 %) in comparison to non-transformed plants. This partial resistance was correlated with a stronger expression of the transgene at transcriptional and traductional levels. Very low depolymerase activity was detected in most transgenic clones (0.006% of total soluble proteins). Greenhouse evaluation of fire blight resistance of the depolymerase clones is under investigation on acclimatized plants.

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Texture is a major component of consumer preference for eating-quality in apple. A quantitative genetic analysis of traits associated with fruit-flesh firmness was carried out. This was based on segregation in an unselected mapping population replicated at six sites and harvested over 2 years. Different methods of assessment were compared, and a principal components analysis carried out. Instrumental measures used were Magness-Taylor penetrometer readings, stiffness by acoustic resonance, and a range of sensory descriptors assessed by a trained panel. There were good correlations between some measures, although stiffness was poorly correlated. Whilst genotype by environment effects were large, significant effects were attributable to the genotype, and these were used to detect QTLs. Significant QTLs were detected on seven linkage groups, with large effects on linkage groups L01, L10 and L16. Whilst there was a poor correlation between acoustic stiffness and other measures, the significant and suggestive QTL detected for stiffness on linkage group L10 did represent a subset of significant QTLs detected for the penetrometer measure. The use of sensory assessment proved valuable in detecting QTLs representing different attributes of fruit texture. The possibility of interaction between significant QTLs for fruit texture and other strongly selected traits such as scab resistance and fruit acidity is addressed.

The phytopathogenic bacterium Xanthomonas campestris pv. pelargonii causes severe losses in vegetatively propagated Pelargonium cultivars. Symptoms are characterized by wilting of the plant, localized water-soaked lesions and stem rot (1) To overcome this problem horticulturists have to produce Xanthomonas-free planting material in insect-proof greenhouses after indexing the propagating stocks, which is expensive and time consuming. All commercial cultivars of pelargonium are susceptible to Xanthomonas and no source of resistance is known in the compatible Pelargonium germplasm. Thus, attempts were initiated to obtain Xanthomonas resistant plants by a transgenic approach.

Fire blight, caused by Erwinia amylovora, is the most important bacterial disease of pear (Pyrus communis L.). Attacin E, a lytic protein originating from Hyalophora cecropia, has been previously reported to be active against E. amylovora in transgenic apple. Integration of the attacin E gene under control of a derivative of the constitutive promoter CaMV35S was accomplished using a transformation protocol developed for several pear cultivars. The integration was checked by polymerase chain reaction (PCR) for 11 lines of the cultivar Passe Crassane. Expression was determined in in vitro plantlets. The differences in transcription levels among lines revealed by comparative reverse transcription PCR correlated very well with the differences in attacin E accumulation observed by Western blot analysis. Fire blight inoculations were performed in vitro on all transgenic lines. A significant reduction of symptoms was observed for six lines, in comparison with the susceptible control Passe Crassane.

Linkage maps for the apple cultivars ‘Prima’ and ‘Fiesta’ were constructed using RFLP, RAPD, isozyme, AFLP, SCAR and microsatellite markers in a ‘Prima’בFiesta’ progeny of 152 individuals. Seventeen linkage groups, putatively corresponding to the seventeen haploid apple chromosomes, were obtained for each parent. These maps were aligned using 67 multi-allelic markers that were heterozygous in both parents. A large number of duplicate RFLP loci was observed and, in several instances, linked RFLP markers in one linkage group showed corresponding linkage in another linkage group. Distorted segregation was observed mainly in two regions of the genome, especially in the male parent alleles. Map positions were provided for resistance genes to scab and rosy leaf curling aphid (Vf and Sd 1, respectively) for the fruit acidity gene Ma and for the self-incompatibility locus S. The high marker density and large number of mapped codominant RFLPs and some microsatellite markers make this map an ideal reference map for use in other progenies also and a valuable tool for the mapping of quantitative trait loci.

Apple scab, caused by the fungus Venturia inaequalis (Cke.) Wint., is an important disease in commercial apple production. A mapping population of 155 individuals, derived from a cross between the apple varieties ‘Prima’ (resistant)בFiesta’ (susceptible), was scored for response to the disease in replicated field and glasshouse trials throughout Europe. Twenty data sets were selected and cluster analysis was used to form a consensus score for the population fitting a 1 : 1 segregation ratio of resistance:susceptibility. The progeny were scored with molecular markers. A detailed map covering 54 cM of the ‘Prima’ linkage group containing the Vf gene for scab resistance was constructed using 24 molecular markers linked to the resistance gene. One isoenzyme marker (Pgm-1), six RFLP markers and 17 RAPD markers formed a linkage group with the consensus measure of resistance to scab. Four marker bridges were established with the corresponding ‘Fiesta’ linkage group with additional markers (one isozyme, one RFLP, three RAPD and one AFLP). A low chi-square value indicated a good fit of the marker ordering, which was in close agreement with previously reported linkage positions for some of the markers and Vf. Differences were observed in the ability of different scoring methods to resolve susceptible and resistant classes. The results obtained for the consensus classification of resistance to scab for the population may suggest the presence of virulent inocula at some sites, which could overcome the Vf gene for resistance. The consequences of relying on individual scoring occasions for studying Vf scab resistance are discussed in the context of linkage analysis, conventional breeding selection, and marker-assisted selection.

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International audience; We investigated the effects of overstory canopy openness and seedling density on seedling morphology and growth in the mid-successional species Acer nipponicum Hara in a cool-temperate forest. Studied seedlings were 46 seedlings of 30-160 cm height, and their overstory canopy openness ranged between 7.2% and 17.0%. Seedling density, measured as the number of conspecific neighboring seedlings within a 50 cm radius of the target seedling, ranged between 0 and 19. There were no significant correlations between seedling height, canopy openness, and seedling density. Multiple regression analysis showed that crown depth, leaf mass fraction, and leaf area index decreased with decreasing canopy openness and increasing seedling density, while the ratio of trunk-lateral branches mass increased. Overstory canopy openness did not affect crown area, leaf size, or petiole length, all of which decreased with increasing seedling density. Standardized regression coefficients indicated that seedling density affected morphology and growth more than canopy openness did. The morphological responses to canopy openness cannot be considered as adaptive plasticity, as total leaf area and leaf mass fraction decreased with decreasing light levels. In contrast, responses to seedling density indicate adaptive responses to neighborhood competition. The results highlight the importance of seedling density that influenced seedling growth and morphology independently of overstory canopy openness.

In an outbreeding species such as apple, haploid plants may be especially useful in breeding programmes for the production of homozygous material. However, methods must be available to induce chromosome doubling in the haploid plants. Two antimitotic agents, colchicine and oryzalin, were compared as regards their efficiency in inducing chromosome doubling of in vitro haploid apple shoots. Three colchicine levels (0.025, 0.25 and 1.25 mM) and three oryzalin levels (5, 15 and 30 μM) were evaluated. Three techniques were also used and compared. Survival rate and chromosome counts were determined. Differences were observed between the two antimitotic agents and between the three techniques. This study demonstrates that oryzalin could be a better choice than colchicine for chromosome doubling on haploid apple shoots in vitro.

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Human activities are one of the great evolutionary forces on plant pathogen populations. Controlling their impact is currently a great challenge for sustainable agriculture. The aim of this thesis was to evaluate the impact of apple domestication and of use of quantitative resistances in breeding programs, on the pathogenicity of the causal agent of apple scab, Venturia inaequalis. We compared pathogenicity of V. inaequalis isolates from wild and domesticated apples using cross-inoculations. Then, we compared their pathogenicity on domesticated apple pyramiding several quantitative resistance factors (QTL) to evaluate the potential role of wild apple as a source for highly aggressive isolates. Finally we evaluated selective pressures exerted by resistant QTL on a mixed inoculum using quantitative pyrosequencing. We demonstrated that apple domestication was associated with a important change in pathogenicity. Although isolates from wild apple were able to infect domesticated apple pyramiding several QTL, they were not more aggressive than isolates from domesticated apple, suggesting that they did not represent a supplementary threat for apple orchards. Finally, we clearly differentiated the impact of QTL according to their spectrum of action; the broad spectrum QTL did not exert differential selective pressures, which suggests that they have a higher durability., Maîtriser l'impact des modifications liées à l'activité humaine sur les populations de pathogènes des cultures est un des grands enjeux actuels pour la mise en place d'une agriculture durable. L'objectif de cette thèse était d'évaluer l'impact de la domestication du pommier et de l'utilisation de résistances quantitatives dans les variétés en cours de sélection, sur le pouvoir pathogène du champignon Venturia inaequalis, responsable de la tavelure. Par des inoculations croisées sur pommiers sauvages et domestiques, nous avons comparé le pouvoir pathogène de souches isolées de milieux naturels et cultivés. Puis nous avons comparé le pouvoir pathogène de ces souches sur des génotypes de pommier domestique cumulant plusieurs facteurs de résistances quantitatives (QTL) afin d'évaluer si les pommiers sauvages pouvaient héberger des souches plus agressives que les pommiers domestiques. Enfin, nous avons évalué les pressions de sélection exercées par ces QTL de résistance sur un mélange de souches par pyroséquençage quantitatif. Nos résultats montrent que la domestication du pommier se serait accompagnée d'une modification importante du pouvoir pathogène de V. inaequalis. Les souches issues de pommiers sauvages sont capables d'infecter des génotypes cumulant un grand nombre de facteurs de résistance mais elles ne sont pas plus agressives que les souches issues de pommiers domestiques ; elles ne représenteraient donc pas une menace supplémentaire pour les vergers. Enfin nous différencions nettement l'impact de QTL à spectre d'action étroit et large ; ces derniers, n'exerçant pas de pression de sélection différentielle entre souches co-inoculées, seraient plus durables.

National audience; Apple fruit texture is a key quality trait orienting consumer choice and agro-industrial processes. Among the several factors at different scales involved in texture, cell walls and their constitutive hemicelluloses are determinant in controlling the mechanical properties of the cells and tissues. In this report, the dynamic of cell wall composition and hemicelluloses fine structure during fruit development and ripening was studied from different apple genotypes collected over 2 years. Typical loss of cell wall galactose was observed along apple expansion and ripening. The combination of cell wall enzymatic hydrolysis and MALDI-TOF mass spectrometry revealed important hemicelluloses modifications along the fruit construction and maturation affecting both xyloglucan and the manor amount of glucomannans. Similar modifications were observed for individuals collected over the two years. The finetuning of hemicelluloses structure during apple development and ripening appears as an addition mean by which the fruit controls its mechanical properties and resulting texture.

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National audience

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; The genetic variability of apple cell walls polysaccharides chemical composition and structure was assessed in a progeny of 141 individuals harvested over 2 years. The variability of the hemicelluloses oligosaccharides released by glucanase was analyzed by MALDI-TOF MS. The genetic contribution was distinguished from harvest year as well as from parental crossing patterns and scab resistance selection. Results showed that harvest year had a major impact on cell wall polysaccharide composition and structure. Within each harvest, genetic effect impact more significantly cell wall polysaccharide chemistry than does reciprocal crossing or early scab selection. Uronic acids, glucose, galactose and xylose contents as well as some glucomannan and xyloglucan structures have a high heritability. This first cell wall chemotyping of an apple progeny opens the way for future searches of genetic markers for the chemical variability of cell wall polysaccharides.

National audience; Parmi les eucalyptus les plus résistants au froid, le gommier des neiges séduit par son feuillage persistant falciforme, son tronc tortueux bigarré et sa floraison blanche

International audience; A revision of Octoknema Pierre is provided, based on morphological data gathered from a study of herbarium specimens and observations in the field. Fourteen species of Octoknema are recognised including six new species: O. bakossiensis Gosline & Malécot, O. belingensis Gosline & Malécot, O. chailluensis Malécot & Gosline, O. kivuensis Gosline & Malécot, O. mokoko Gosline & Malécot and O. ogoouensis Malécot & Gosline. Data are given for four additional poorly known taxe (Oktoknema species A, B, C and D).

International audience; In order to investigate the genetic bases of the physiological syndrome mealiness that causes abnormal fruit softening and juice loss in apples, an integrative approach was devised, consisting of sensory, instrumental, biochemical, genetic, and genomic methods. High levels of activity of alpha-L-arabinofuranosidase (alpha-AFase), a hydrolase acting on the pectic component of the cell walls, were found in individuals exhibiting the mealiness phenotype in a segregating population. The expression levels of the previously uncharacterized apple AF gene MdAF3 are higher in fruits from plants consistently showing mealiness symptons and high alpha-AFase activity. The transcription of MdAF3 is differentially regulated in distinct genomic contexts and appears to be independent of ethylene. Thus, it is likely to be controlled by endogenous developmental mechanisms associated with fruit ripening. The use of integrative approaches has allowed the identification of a novel contributor to the mealiness phenotype in apple and it has been possible to overcome the problems posed by the unavailability of near-isogenic lines to dissect the genetic bases of a complex physiological trait in woody perennial species.

International audience; Studying grapevine (Vitis vinifera) innate defense mechanisms is a prerequisite to the development of new protection strategies, based on the stimulation of plant signaling pathways to trigger pathogen resistance. Two transcriptional coactivators (VvNPR1.1 and VvNPR1.2) with similarity to Arabidopsis thaliana NPR1 (Non-Expressor of PR genes 1), a well-characterized and key signaling element of the salicylic acid (SA) pathway, were recently isolated in Vitis vinifera. In this study, functional characterization of VvNPR1.1 and VvNPR1.2, including complementation of the Arabidopsis npr1 mutant, revealed that VvNPR1.1 is a functional ortholog of AtNPR1, whereas VvNPR1.2 likely has a different function. Ectopic overexpression of VvNPR1.1 in the Arabidopsis npr1-2 mutant restored plant growth at a high SA concentration, Pathogenesis Related 1 (PR1) gene expression after treatment with SA or bacterial inoculation, and resistance to virulent Pseudomonas syringae pv. maculicola bacteria. Moreover, stable overexpression of VvNPR1.1-GFP in V. vinifera resulted in constitutive nuclear localization of the fusion protein and enhanced PR gene expression in uninfected plants. Furthermore, grapevine plants overexpressing VvNPR1.1-GFP exhibited an enhanced resistance to powdery mildew infection. This work highlights the importance of the conserved SA/NPR1 signaling pathway for resistance to biotrophic pathogens in V. vinifera.

UMR 1334 AGAP : Equipe AFEF ‘Architecture et Fonctionnement des Espèces fruitières’ ; Team AFFS ‘Architecture and Functioning of Fruit Species’ Contact: elisabeth.chevreau@angers.inra.fr 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; The development of alternative selection systems without antibiotic resistance genes is a key issue to produce safer and more acceptable transgenic plants. Eutypine is a toxin produced by Eutypa lata, the causal agent of eutypa dieback of grapevine, which is detoxified in mung bean (Vigna radiata) by the gene Vr-ERE. Many phytotoxic compounds containing an aldehyde group can act as substrates for the Vr-ERE enzyme. The aim of the present work was to evaluate the effects of the overexpression of Vr-ERE in transgenic apple plants, as a first step towards the development of an alternative selection system. Viable transgenic apple clones expressing Vr-ERE were produced from the cultivar Greensleeves under kanamycin selection. Although the Vr-ERE transgene was normally expressed at the RNA and protein levels, the increase in aldehyde reductase activity tested on a range of potential substrates was very low in these clones. None of them revealed a significant increase in tolerance to toxic aldehydes compared to their non-transgenic control. This work with transgenic apple plants overexpressing the detoxifying gene Vr-ERE illustrates some of the difficulties in developing an alternative selection pressure

Mobilisation de sources du XIXe siècle dans les études contemporaines de systématique des plantes cultivées

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; Evaluations of plant resistance to pathogens are rarely made using isolates from wild habitats, although the heterogeneity of such habitats may generate pathogen diversity which could be a source of new virulence in cultivated habitats. The aim of this study was to investigate whether scab resistance factors, identified and characterized in apples using isolates of Venturia inaequalis from a cultivated habitat, remained effective against isolates from a wild habitat. Three V. inaequalis core collections originating from the cultivated apple Malus × domestica and from two wild species, M. sieversii and M. sylvestris, were established to maximize pathogen diversity. For each core collection, 10 isolates were inoculated in mixtures onto 51 genotypes from an apple progeny segregating for two qualitative resistance genes and six quantitative resistance loci (QRL). On each apple genotype, isolates that contributed to the scab symptoms were identified within the mixture using microsatellite markers. The most frequently detected isolates were inoculated singly to compare their aggressiveness according to their host origin. The results showed that isolates from a wild habitat were able to infect the susceptible apple genotypes. However, these isolates were never more aggressive than isolates from the cultivated habitat on the resistance factors tested. It can therefore be concluded that the resistance factors used in this study, identified with V. inaequalis isolates from a cultivated habitat, remained effective against isolates from M. sylvestris and M. sieversii.

National audience; Originaire de la région méditerranéenne, Euphorbia characias se révèle adaptée à une grande partie de la France en situation chaude et bien drainée

Hydrangea macrophylla is a horticultural plant of considerable commercial interest that has been widely studied with the aim to more effectively control the different stages of its development during production. However, although floral transformation is a key factor underlying the commercial quality of the product, it remains difficult to control despite these efforts. The floral transformation sequence consists of three successive phases: floral induction (B1), floral evocation (B2), and floral organogenesis (B3). The first is a phase of vegetative organogenesis without elongation leading to the formation of a bud composed of eight phytomer primordia under inductive climatic conditions. This work shows that climatic conditions favorable to floral transformation must be continuously applied without interruption throughout phase B1 to ensure the formation of the floral bud in Hydrangea macrophylla ‘Leuchtfeuer’. In the opposite case, floral transformation is stopped and vegetative growth begins once again.

Chapter 12; International audience; Presently, about 100–250 species are usually recognized in the genus Rosa. The low levels of DNA sequence divergence found across the genus suggest that it is a young genus with much speciation taking place after the last glaciation. Poor phylogenetic resolution and commonly occurring contradictions between chloroplast and nuclear gene phylogenies suggest that hybridization has been a strong driving force in the evolution of roses, often accompanied by polyploidization. In addition, extensive anthropogenic impact has led to the development of many new semi-wild and/or cultivated rose varieties. Some wild species have become invasive. This chapter describes the taxonomy of roses. It presents examples of interesting traits in wild species that may valuable to broaden the genetic base of cultivated roses, such as thornlessness, winter hardiness, drought resistance, and improved shelf-life. Also fragrance and compounds with possible health effects are discussed. The highest priority in rose breeding research is the development of disease resistant roses. Different resistance mechanisms have been found for black spot and powdery mildew in various wild rose species. We describe how resistance genes are being mapped using crosses between wild, diploid rose species, and how other genes for traits of interest are being identified. We proceed with a discussion of various ways to overcome the taxonomic and ploidy level barriers for introgression of traits into cultivated hybrid rose or garden rose germplasm, including dihaploidization, polyploidization, and transgenics. Even so, breeding programs with wild species are usually time-consuming. For garden roses, the genetic and morphological distance between modern cultivars and the wild species is smaller, which makes the use of wild species easier than in the case of cut roses.

International audience

International audience; The pear breeding programme at INRA Angers-Beaucouze is partly devoted to screen for red skin colour from mutants derived from European pear cultivars. These mutants were selected around 1950 in the USA from 'Doyenne du Comice', 'Williams' and 'Beurre Hardy'. Several combinations involving these red mutants and also cultivars including 'Lombacad' and 'Red Satin' were studied for the inheritance of red colour in the leaves after scoring in the greenhouse. In the field, vigour and tree habit of the seedlings were appreciated and compared to the green seedlings. The hypothesis of a genotype C/c for red leaves is confirmed for 'Red Comice' and 'Royal Red Hardy' as in 'Max Red Bartlett'. Three classes of architecture: standard-spread, erect, and compact, were clearly distinguished according to the genotype at the locus C. The red flesh character of the fruit is also under investigation and the red colour of the hypocotyl is proposed as an early putative marker for this trait.

National audience; Viburnum x hillieri 'Winton' offre une floraison et une fructification remarquables. Sans grande exigence, il s'adapte bien au climat de la plupart des régions de France.

International audience; The apple scab (Venturia inaequalis Malus) pathosystem was one of the first systems for which Flor's concept of gene-for-gene (GfG) relationships between the host plant and the pathogen was demonstrated. There is a rich resource of host resistance genes present in Mains germplasm that could potentially be marshalled to confer durable resistance against this most important apple disease. A comprehensive understanding of the host-pathogen interactions occurring in this pathosystem is a prerequisite for effectively manipulating these host resistance factors. An accurate means of identification of specific resistance and consistent use of gene nomenclature is critical for this process. A set of universally available, differentially resistant hosts is described, which will be followed by a set of defined pathogen races at a later stage. We review pertinent aspects of the history of apple scab research, describe the current status and future directions of this research, and resolve some outstanding issues.

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; Apple (Malus x domestica) leaf phenolics, in particular the three major dihydrochalcones, phloridzin, trilobatin and sieboldin, were studied for their potential contribution to the resistance to Erwinia amylovora, the causative agent of fire blight. The constitutive phenolic compositions of part of a progeny resulting from a cross between two apple genotypes, displaying contrasted susceptibilities to fire blight, were investigated by RP-HPLC-DAD. With regards to the major dihydrochalcones, offsprings were found to display the parent profile: either phloridzin alone or a combination of phloridzin, trilobatin and sieboldin. Despite a strong antioxidant activity that could interfere with the oxidative burst triggered by the bacteria, sieboldin did not segregate with resistance even though it was strongly associated with a high constitutive antioxidant capacity of leaf extracts. Several hydroxycinnamic acids and flavonols were identified in the progeny but profiles were very similar. However, concentrations of several flavonols positively correlated with resistance in the genotypes that possessed trilobatin and sieboldin. The major dihydrochalcones that represent more than 200 mg/g of leaf DW are not directly responsible for resistance to fire blight in this system. In addition, a high constitutive antioxidant activity is unlikely to be sufficient to block bacterial spread

International audience; The possibility of eradicating the pelargonium flower break virus (PFBV) and pelargonium line pattern virus (PLPV) by cryotherapy of axillary shoot apices was investigated using five Pelargonium cultivars. Viruses were detected by DAS-ELISA and their location was determined by immunolocalization. Apex culture did not permit elimination of PFBV and only 15% regenerated plants of 'Stellar Artie' cultivar were ELISA PLPV-negative. Plants regenerated from cryotherapy-treated apices were tested by DAS-ELISA after a 3-month in vitro culture period. Viruses were not detected in 25% and 50% of the plants tested for PFBV and PLPV, respectively. However, immunolocalization carried out on apices originating from cryopreserved shoot tips sampled from DAS-ELISA negative plants showed that they were still virus-infected. Using immunolocalization, PFBV and PLPV could be detected in Pelargoniuni apices, even in the meristematic dome. However, viral particles were more numerous in basal zone cells than in meristematic cells. Our results demonstrate that PFBV and PLPV are present within meristematic cells and that cryopreservation can partly reduce the quantity of these viruses in Pelargonium plants but not eliminate them totally. Additional knowledge on localization and behaviour of viruses during cryopreservation is essential to optimize cryotherapy and plant genetic resource management.