Subdivision and genetic structure of four populations of Venturia inaequalis in Switzerland

Analyses of four populations of Venturia inaequalis in Switzerland were performed to obtain information about migration and to predict the probable speed of the spread of new pathotypes able to overcome resistance, e.g. Vf-resistance, of new cultivars. Genetic and haplotype diversity was calculated based on allele frequencies of random amplified polymorphic DNA (RAPD) markers and the internal transcribed spacer (ITS)-region of ribosomal DNA, which are regarded to be neutral, and the β-tubulin locus which may be under selection pressure. Within-population diversity was found to be quite similar over all four populations. Normalised haplotype diversity based on RAPD and ITS data was very high with a mean of 0.95. Diversity among populations (GST) was consistent over all neutral loci with a low mean of 0.04, but reached the high value of 0.26 for the selected β-tubulin locus. Low GST based on neutral loci may suggest a high level of gene flow. Considering these results, new pathotypes would be expected soon outside their place of identification. But actual gene flow is easily overestimated because of effects of gene flow in the past. However, naturally occurring gene flow could be increased by human activity. Therefore, it is very difficult to predict durability of the Vf-resitance in Switzerlan

GEOBASE: a simple geographical information system on a personal computer

Spatially distributed data are often encountered in the biological sciences. Representation and analysis of such data requires specific tools. A simple geographical information system is presented, which allows representation and elementary analysis of geographically coded information. The system handles two kinds of data: maps and facts, where map data describe the basis on which the fact data are located. Maps consist of objects described through a set of coordinates, while for facts a coordinate pair is associated with an unlimited number of data records containing five fields: a date, an element from a list, a two-character code, an integer number and a real number. The input data can be displayed interactively on screen by logically combining selection criteria for each field. The facts corresponding to the selected criteria are either displayed as such, or are clustered and displayed as polygons or pies. A short example showing a possible application of the program is presented and advantages as well as limitations are discusse

Vf scab resistance of Malus

The apple production in temperate regions with spring rains, the Scab caused by the fungus Venturia inaequalis is the most important constraint. To produce spotless apples and avoid damage that develops during storage, growers apply fungicide on a regular or weather-determined basis. All major apple cultivars are highly susceptible to this disease. To limit the need for fungicide applications, apple breeders are currently introgressing disease resistance from wild Malus accessions into commercial lines. The first attempts to do this were made 100years ago. As apples are self-incompatible, pseudo-backcrossing is used to eliminate unwanted traits from wild Malus and select new cultivars that are attractive to both producers and consumers. This process, from the first cross of a commercial cultivar with a wild, disease-resistant Malus, is extremely long due to apple's long juvenile phase, the need for more than seven backcross steps and the high heterozygosity of this genus. Therefore, most of today's scab-resistant cultivars rely on a single introduction of scab resistance from Malus floribunda 821, referred to as Vf. In this paper, we trace the history of Vf from its initial identification through its use in breeding and commercial production. We sum up the literature describing how and where Vf resistance has been overcome by new pathotypes of V. inaequalis. Finally, we describe the current knowledge of the genes behind Vf resistance, its mode of action and the use of Vf genes in gene technolog

Genetic Dissection of Plasmopara viticola Population from a Greek Vineyard in Two Consecutive Years

Oilspots of the Plasmopara viticola population in a Greek vineyard over 2 years were collected and the pathogen genotyped with the use of four microsatellite loci. In 2001, five samplings occurred and 327 lesions were collected, which were classified to 23 genotypes. In 2002, the 426 lesions collected in four samplings belonged to 54 genotypes. A reproducible pattern of the evolution of the epidemic was exhibited that can be described as 'predominance-of-one-clone'. In 2001, the predominant clone covered 72-92% of each sample, while in 2002 the incidence of the predominant clone was 38-90%. The remaining genotypes showed low clonal reproduction and dispersal. Oosporic infections occurred throughout the season. The severity and the diversity in 2002 were, however, higher than in 2001 and this was not in accordance with the climatic conditions, suggesting that other factors, like the epidemic of the previous year, also influence the epidemic of the disease. These results produce new concepts about the epidemiology of the pathoge

Genetic Diversity of Venturia inaequalis Across Europe

The genetic diversity of eleven populations of Venturia inaequalis from five European countries was calculated based on the allele frequencies of 18 random amplified polymorphic DNA markers and the internal transcribed spacer region of the ribosomal DNA. Diversity within each population (HS) was high with values ranging from 0.26 to 0.33. Average differentiation among populations (GST) was 0.11 and populations were isolated by distance (r2=0.50, P<0.01). This indicates that extensive short-distance gene flow occurs in Europe and that dispersal over longer distances also appears to occur frequently enough to prevent differentiation due to genetic drift. We had expected more pronounced differentiation between populations north and south of the Alps, because V. inaequalis is primarily dispersed by rain splashing and wind distribution of leaf litter, and because the Alps might form a barrier to dispersal. However, our results indicate that human-mediated gene flow due to transport of spores or infected plant material has probably occurre

Identification of Microsatellite Markers for Plasmopara viticola and Establishment of High throughput Method for SSR Analysis

The Oomycete Plasmopara viticola is the causal organism of downy mildew on grapevine (Vitis spp.). In order to set up the techniques for investigating downy mildew disease dynamics and genetic structure, co-dominant, neutral, highly reproducible and polymorphic microsatellite markers for P. viticola were developed. Five markers, two with a (TC)n repeat (loci BER and ISA), two with a (TC)n(AC)n repeat (loci CES and REX) and one with a (CT)n(CTAT)n repeat (locus GOB), were selected. Simple sequence repeat (SSR) markers revealed different degrees of polymorphism within 190 oil spots (disease symptoms) collected from an infected Italian vineyard. The most polymorphic SSR marker GOB showed 43 alleles (Nei's expected gene diversity He = 0.89) while CES, ISA, BER and REX showed 14 (He = 0.71), 4 (He = 0.57), 3 (He = 0.24) and 1 allele (He = 0), respectively. A high throughput DNA extraction method, that allowed molecular analysis of this obligate pathogen directly in the host without any isolation procedure, was developed. The quality and quantity of oil spots did not influence the SSR analysis. Amplified SSR loci were separated by electrophoresis on a Beckman-Coulter 2000XL sequencer and automatically analysed. The objective of this study was to develop molecular biological tools and methods that allow high throughput analysis of the downy mildew population

Microsatellite based population structure of Plasmopara viticola at single vine scale

The genetic structure of a Plasmopara viticola population was characterized on five single vines, one for each cultivar Regent, Merlot, Isabella, Müller-Thurgau and Solaris, using four neutral specific polymorphic microsatellite markers. Five-hundred and seventy samples were collected at four dates in the period between the 10th of July and the 23rd of August 2006. On average over all five cultivars, 67% of the genotypes present on the single selected vines derived from primary infections and caused 37% of the lesions genotyped. Fifty-three percent of these genotypes occurred only once on the vine throughout the survey period, while 14% were able to asexually reproduce on the selected single vine throughout the survey period, causing 23% of the lesions. Thirty-three percent of the genotypes on the single vine derived from other vines, 28% from vines of other cultivars in the other rows, and 5% from vines of the same cultivar in the same row. New primary infections appear all along the sampling dates. The overwhelmingly quantitative role of primary infections at vineyard scale was known, however here we observed the phenomenon also at the single vine scale and the reduced contribution of secondary lesions to the populations present on more resistant cultivars compared to the susceptible cultivars. As the sampling extended almost to defoliation, the results are judged to be representative of a typical P. viticola epidemi

Co-inoculated Plasmopara viticola genotypes compete for the infection of the host independently from the aggressiveness components

During Plasmopara viticola epidemics only few genotypes produce most of the secondary lesions and dominate in the population. Selection of dominant genotypes is hypothesized to be linked to environmental conditions and can occur rapidly, particularly if there is also difference between genotypes in terms of fitness and aggressiveness. Measurements of aggressiveness components can largely determine the rate of epidemic development, although the components of aggressiveness do not take into account potential direct competition between genotypes. Differences in aggressiveness have been also reported to be greater under non-optimal conditions suggesting for genotype adaptation to different conditions. To evaluate differences in latency at non-optimal conditions, we characterized genotypes deriving from different climatic regions at three different temperatures (15, 25 and 35°C) and we found no differences. To investigate whether other factors may impact on competition between P. viticola genotypes, we evaluated polycyclic infections of P. viticola by co-inoculating three genotypes with similar aggressiveness components in two different co-inoculation experiments and an increasing prevalence of one of the two genotypes was observed. Competition was not related to the origin of the genotype and we hypothesize that competitive selection is modulated by differences in the secretion of effector molecules which can contribute to the establishment of dominant genotypes over an epidemic seaso

QTL mapping of fire blight resistance in apple

Fire blight caused by the bacterium Erwinia amylovora is a severe threat to apple and pear orchards worldwide. Apple varieties exhibit a wide range of relative susceptibility/tolerance to fire blight. Although, no monogenic resistance against fire blight has been identified yet, recent evidence indicates the existence of quantitative resistance. Potential sources of fire blight resistance include several wild Malus species and some apple cultivars. F1 progenies of ‘Fiesta'בDiscovery' were inoculated with the Swiss strain Ea 610 and studied under controlled conditions to identify quantitative trait loci (QTLs) for fire blight resistance. Disease was evaluated at four time points after inoculation. Shoot lesion length and the area under disease progress curve (AUDPC) values were used for QTL analysis. One significant (LOD score of 7.5-8.1, p<0.001) QTL was identified on the linkage group 7 of ‘Fiesta' (F7). The F7 QTL explained about 37.5-38.6% of the phenotypic variatio

Comparison between volatile emissions from transgenic apples and from two representative classically bred apple cultivars

While most risk assessments contrast a transgenic resistant to its isogenic line, an additional comparison between the transgenic line and a classically bred cultivar with the same resistance gene would be highly desirable. Our approach was to compare headspace volatiles of transgenic scab resistant apple plants with two representative cultivars (the isogenic ‘Gala' and the scab resistance gene-containing ‘Florina'). As modifications in volatile profiles have been shown to alter plant relationships with non-target insects, we analysed headspace volatiles from apple plants subjected to different infection types by gas chromatography-mass spectrometry. Marked differences were found between healthy and leafminer (Phyllonorycter blancardella) infested genotypes, where emissions between the transgenic scab resistant line and the two cultivars differed quantitatively in four terpenes and an aromatic compound. However, these modified odour emissions were in the range of variability of the emissions recorded for the two standard cultivars that proved to be crucial reference