Delineation of a scab resistance gene cluster on linkage group 2 of apple

National audienc

Genome mapping of three major resistance genes to woolly apple aphid (Eriosoma lanigerum Hausm.)

International audienceWoolly apple aphid (WAA; Eriosoma lanigerum Hausm.) can be a major economic problem to apple growers in most parts of the world, and resistance breeding provides a sustainable means to control this pest. We report molecular markers for three genes conferring WAA resistance and placing them on two linkage groups (LG) on the genetic map of apple. The Er1 and Er2 genes derived from 'Northern Spy' and 'Robusta 5',respectively, are the two major genes that breeders have used to date to improve the resistance of apple rootstocks to this pest. The gene Er3, from 'Aotea 1' (an accession classified as Malus sieboldii), is a new major gene for WAA resistance. Genetic markers linked to the Er1 and Er3 genes were identified by screening random amplification of polymorphic deoxyribonucleic acid (DNA; RAPD) markers across DNA bulks from resistant and susceptible plants from populations segregating for these genes. The closest RAPD markers were converted into sequence-characterized amplified region markers and the genome location of these two genes was assigned to LG 08 by aligning the maps around the genes with a reference map of 'Discovery' using microsatellite markers. The Er2 gene was located on LG 17 of 'Robusta 5' using a genetic map developed in a M.9 x 'Robusta 5' progeny. Markers for each of the genes were validated for their usefulness for marker-assisted selection in separate populations. The potential use of the genetic markers for these genes in the breeding of apple cultivars with durable resistance to WAA is discusse

A proposal for the nomenclature of Venturia inaequalis races

International audienceThe 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

Development and test of 21 multiplex PCRs composed of SSRs spanning most of the apple genome

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 project

Analyses of Expressed Sequence Tags from Apple

The domestic apple (Malus domestica; also known as Malus pumila Mill.) has become a model fruit crop in which to study commercial traits such as disease and pest resistance, grafting, and flavor and health compound biosynthesis. To speed the discovery of genes involved in these traits, develop markers to map genes, and breed new cultivars, we have produced a substantial expressed sequence tag collection from various tissues of apple, focusing on fruit tissues of the cultivar Royal Gala. Over 150,000 expressed sequence tags have been collected from 43 different cDNA libraries representing 34 different tissues and treatments. Clustering of these sequences results in a set of 42,938 nonredundant sequences comprising 17,460 tentative contigs and 25,478 singletons, together representing what we predict are approximately one-half the expressed genes from apple. Many potential molecular markers are abundant in the apple transcripts. Dinucleotide repeats are found in 4,018 nonredundant sequences, mainly in the 5′-untranslated region of the gene, with a bias toward one repeat type (containing AG, 88%) and against another (repeats containing CG, 0.1%). Trinucleotide repeats are most common in the predicted coding regions and do not show a similar degree of sequence bias in their representation. Bi-allelic single-nucleotide polymorphisms are highly abundant with one found, on average, every 706 bp of transcribed DNA. Predictions of the numbers of representatives from protein families indicate the presence of many genes involved in disease resistance and the biosynthesis of flavor and health-associated compounds. Comparisons of some of these gene families with Arabidopsis (Arabidopsis thaliana) suggest instances where there have been duplications in the lineages leading to apple of biosynthetic and regulatory genes that are expressed in fruit. This resource paves the way for a concerted functional genomics effort in this important temperate fruit crop

Analysis of expressed sequence tags from Actinidia : Applications of a cross species EST database for gene discovery in the areas of flavor, health, color and ripening

Background Kiwifruit (Actinidia spp.) are a relatively new, but economically important crop grown in many different parts of the world. Commercial success is driven by the development of new cultivars with novel consumer traits including flavor, appearance, healthful components and convenience. To increase our understanding of the genetic diversity and gene-based control of these key traits in Actinidia, we have produced a collection of 132,577 expressed sequence tags (ESTs). Results The ESTs were derived mainly from four Actinidia species (A. chinensis, A. deliciosa, A. arguta and A. eriantha) and fell into 41,858 non redundant clusters (18,070 tentative consensus sequences and 23,788 EST singletons). Analysis of flavor and fragrance-related gene families (acyltransferases and carboxylesterases) and pathways (terpenoid biosynthesis) is presented in comparison with a chemical analysis of the compounds present in Actinidia including esters, acids, alcohols and terpenes. ESTs are identified for most genes in color pathways controlling chlorophyll degradation and carotenoid biosynthesis. In the health area, data are presented on the ESTs involved in ascorbic acid and quinic acid biosynthesis showing not only that genes for many of the steps in these pathways are represented in the database, but that genes encoding some critical steps are absent. In the convenience area, genes related to different stages of fruit softening are identified. Conclusion This large EST resource will allow researchers to undertake the tremendous challenge of understanding the molecular basis of genetic diversity in the Actinidia genus as well as provide an EST resource for comparative fruit genomics. The various bioinformatics analyses we have undertaken demonstrates the extent of coverage of ESTs for genes encoding different biochemical pathways in Actinidia

Development of a set of apple SSRs markers spanning the apple genome, genotyping of HiDRAS plant material and validation of genotyping data

International audienceThe Pedigree Genotyping concept (van de Weg et al., 2004) forms the base of the EU-project HiDRAS, aimed at the identification of molecular markers for fruit quality and disease resistance. The concept allows the exploitation of breeding material in the assessment of marker-trait associations and in allele mining by using multiple pedigreed plant populations, which can be any combination of crosses, cultivars, and breeding selections. The Pedigree Genotyping model makes use of directed genotyping and the so-called Identity By Descent (IBD) concept. Prerequisites for this innovative technique are a good coverage of the apple genome with highly polymorphic simple sequence repeats (SSR) markers, and consistent molecular data throughout the whole pedigree. At the beginning of the HiDRAS project, about 160 apple SSRs were available and mapped. However the coverage of the apple genome was not sufficient as there were several regions of the genome with no SSRs available, or the level of polymorphism was low. To fill these gaps with new SSRs, two approaches were used: (i) development and mapping of SSRs from SSR-enriched genomic libraries and available apple EST sequences containing repeated motifs; and (ii) making use of the synteny existing between apple and pear, by using SSR markers developed in pear. These strategies enriched the apple reference map, 'Fiesta' * 'Discovery', with 156 new SSRs. A subset of 88 reliable SSR markers, highly polymorphic and evenly distributed across the apple genome, was selected. Eighty-three out of 88 SSR markers were organized in 21 multiplex PCR reactions and used to genotype the HiDRAS plant material (around 2000 DNA samples representing over 1750 genotypes). After the generation of raw data, each single data point was verified for consistency throughout the whole pedigree by the use of newly developed software packages. This allowed the identification of not only scoring errors, but also mistakes in the reported pedigrees and the "(not) true-to-typeness" of some genotypes (wrong plants). The complete procedure from SSR development to data validation is presented

The Vh8 locus of a new gene-for-gene interaction between Venturia inaequalis and the wild applie Malus sieversii is closely linked to the Vh2 locus in Malus pumila R12740-7A

The wild apple (Malus sieversii) is a large-fruited species from Central Asia, which is used as a source of scab resistance in cultivar breeding. Phytopathological tests with races of Venturia inaequalis were performed to differentiate scab-resistance genes in Malus as well as an avirulence gene in the pathogen. A novel gene-for-gene interaction between V. inaequalis and Malus was identified. The locus of the scab-resistance gene Vh8 is linked with, or possibly allelic to, that of the Vh2 gene in Malus pumila Russian apple R12740-7A, at the lower end of linkage group 2 of Malus. Race 8 isolate NZ188B.2 is compatible with Vh8, suggesting the loss or modification of the complementary AvrVh8 gene, while isolate 1639 overcomes both Vh2 and Vh8, but is incompatible with at least one other gene not detected by any of the other race isolates tested. Our research is the first to differentiate scab-resistance genes in a putative gene cluster in apple with the aid of races of V. inaequalis