High-Resolution Melting (HRM) Analysis Reveals Genotypic Differentiation of Venturia inaequalis Populations in Greece (original) (raw)
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Molecular Ecology Notes, 2004
Twenty-one new polymorphic microsatellite markers were isolated in the phytopathogenic fungus Venturia inaequalis , the causal agent of apple scab. An enrichment protocol was used to isolate microsatellite loci and the level of polymorphism was assessed on 44 European isolates. All loci were polymorphic with an average of 9.1 alleles per locus (range 2 -24). Tests of cross-species amplifications suggest that at least some of these microsatellites could be used in different species, mainly Spilocaea pyracanthae and S. eriobotryae .
TURKISH JOURNAL OF AGRICULTURE AND FORESTRY, 2016
Introduction Apple scab is a major problem in both Turkish and global apple markets. This disease is responsible for a decrease in the total market value of the fruit by 70%, resulting in production losses ranging from 30% to 60% (Türkoğlu, 1978; Agrios, 1997). On average, apple producers in Turkey apply fungicides 20-30 times per season in order to control the incidence of the causal pathogen, Venturia inaequalis, which is attributed to excess environmental pollutants (Boyraz et al., 2005; Soriano et al., 2009). V. inaequalis has a high degree of genetic variability due to recombination, thus leading to selective pressures on the fungal pathogen to overcome host resistance. An example of this selective pressure leading to host plant susceptibility was observed in Golden Delicious apple varieties. During the 1900s, Golden Delicious apples were highly resistant to apple scab, whereas today this variety is considered one of the most susceptible apple varieties (Gessler et al., 2006).
Tarım Bilimleri Dergisi, 2017
Apple scab (Venturia inaequalis (Cke) Wint) is the main disease of apple trees not only in the world but also in Turkey. The disease can cause severe yield losses in all Turkish apple orchards. Genetic diversity and relatedness of apple scab (Venturia inaequalis (Cke) Wint) isolates collected from Turkey and Europe were investigated in this study through molecular markers. RAPD, ISSR, SSR and SRAP markers were used in the molecular studies. Results obtained by marker system revealed that isolates were quite separated from each other and it is appeared to exist a variation between them. Genetic relatedness between the isolates are very close to each other, and difference among the groups is not significant due to host specificity and geographical location. The groups are consisted of more number of isolates when SSR and SRAP markers are used compared with ISSR and RAPD markers. Using SSR and SRAP markers are preferable to provide more informative outcomes because of ease of use, repeatability and specificity.
PLOS ONE
Apple scab caused by Venturia inaequalis Cooke (Wint.) is one the important diseases of trade and industrial significance in apple. In present study variability studies in pathogen isolates were studied, which is one of the most important factors for devising management studies of scab disease in apple. Genetic diversity of 30 Venturia inaequalis isolates from 12 districts of two geographical distinct regions of Jammu and Kashmir was calculated based on the allele frequencies of 28 SSR markers and the internal transcribed spacer (ITS) region of the ribosomal DNA. The ITS based characterized sequences were submitted to NCBI Gen-Bank and accession numbers were sanctioned. Dendrogram showed that all the accessions formed 2 main clusters with various degree of sub clustering within the clusters. Analysis based on SSR study reveals that the heterozygosity ranged from 0.0 and 0.5, with an average value of 0.39. The expected heterozygosis or gene diversity (He) ranged from 0.0 to 0.50 with an average of 0.40. The F st value ranges from 0 to 0.6 with an average of 0.194. Diversity within each population (HS) values ranging from 0.26 to 0.33. Average differentiation among populations (GST) was 0.11 and populations were isolated by significant distance (r 2 = 0.50, P < 0.01). From the AMOVA analysis, 25% of variation was observed among population, 9% among individuals and 66% within individuals observed in the population. Structure analysis grouped isolates into two populations. Principle coordinate analysis explained variation of 36.6% in population 1, 14.30% in population 2 and 13.10% in population 3(Admixture) with 64.07% as overall cumulative percentage of variation. This indicates that extensive short-distance gene flow occurs in Kashmir region that dispersal over longer distances also appears to occur frequently enough to prevent differentiation due to genetic drift. Also it is evident that Jammu and Kashmir most likely has V. inaequalis subpopulations linked to diverse climatic conditions of the Jammu region compared to the mountainous inland Kashmir region. The results of present study would help to understand the genetic diversity of V. inaequalis from Jammu and Kashmir that would lead in the development of more effective management strategies and development of new resistant cultivars through marker-assisted selection.
Phytopathology, 2004
The recent breakdown of Vf, a major resistance gene to apple scab, provided an opportunity to analyze a population genetic process within the matching virulent subpopulation of the fungus Venturia inaequalis. We utilized the amplified fragment length polymorphism technique and allelic variation at four microsatellite loci to assess genetic structure of 133 isolates of V. inaequalis from a single commercial apple orchard sampled from one cultivar carrying the Vf gene (Judeline) and three cultivars devoid of the Vf gene. Both analyses indicated a strong decrease of the genetic diversity among isolates from the Vf cultivar compared with the high level of diversity among isolates from the three other cultivars. This leads to a high genetic differentiation between virVf and avrVf groups (F(ST) > 0.17). Analyses of the genetic distance between AFLP patterns based on the Jaccard index indicate that all virVf isolates could be assigned to a single clonal lineage. These results lead us to...
A genetic linkage map of Venturia inaequalis, the causal agent of apple scab
BMC Research Notes, 2009
Background: Venturia inaequalis is an economically-important disease of apple causing annual epidemics of scab worldwide. The pathogen is a heterothallic ascomycete with an annual cycle of sexual reproduction on infected apple leaf litter, followed by several cycles of asexual reproduction during the apple growing season. Current disease control is achieved mainly through scheduled applications of fungicides. Genetic linkage maps are essential for studying genome structure and organisation, and are a valuable tool for identifying the location of genes controlling important traits of interest such as avirulence, host specificity and mating type in V. inaequalis. In this study, we performed a wide cross under in vitro conditions between an isolate of V. inaequalis from China and one from the UK to obtain a genetically diverse mapping population of ascospore progeny isolates and produced a map using AFLP and microsatellite (SSR) markers. Findings: Eighty-three progeny were obtained from the cross between isolates C0154 (China) × 01/213 (UK). The progeny was screened with 18 AFLP primer combinations and 31 SSRs, and scored for the mating type locus MAT. A linkage map was constructed consisting of 294 markers (283 AFLPs, ten SSRs and the MAT locus), spanning eleven linkage groups and with a total map length of 1106 cM. The length of individual linkage groups ranged from 30.4 cM (Vi-11) to 166 cM (Vi-1). The number of molecular markers per linkage group ranged from 7 on Vi-11 to 48 on Vi-3; the average distance between two loci within each group varied from 2.4 cM (Vi-4) to 7.5 cM (Vi-9). The maximum map length between two markers within a linkage group was 15.8 cM. The MAT locus was mapped to a small linkage group and was tightly linked to two AFLP markers. The map presented is over four times longer than the previously published map of V. inaequalis which had a total genetic distance of just 270 cM. Conclusion: A genetic linkage map is an important tool for investigating the genetics of important traits in V. inaequalis such as virulence factors, aggressiveness and mating type. The linkage map presented here represents a significant improvement over currently published maps for studying genome structure and organisation, and for mapping genes of economic importance on the V. inaequalis genome.
Population Genetic Structure of Apple Scab (Venturia inaequalis (Cooke) G. Winter) in Iran
PLOS ONE, 2016
The population genetic structure of 278 Venturia inaequalis isolates, collected from different apple cultivars of eighteen different provinces in Iran, was investigated using 22 polymorphic microsatellite markers. Analysis of molecular variation, Bayesian clustering and Nei's genetic distance analyses based on 88 microsatellite alleles indicated substantial levels of gene flow among the collection sites. Ninety three percent of the variation was observed among the individuals within the populations and only 7% variation was observed among the populations. Structure analysis grouped the isolates into two populations. Maximum number of pathogen genotypes (44) was observed in the North of Iran that grows various different apple cultivars. Investigation on the variation of the pathogen on different cultivars in the North of Iran suggested a significant differentiation of the pathogen populations between wild apple and commercial cultivars. During sampling, varying ranges of scab infection were observed on various apple cultivars in forests, monoculture and mix orchards. Wild type apple (Malus orientalis) along the Caspian Sea Coast had the most infection in comparison with the Iranian endemic and commercial cultivars. Based on the genetic analysis and host tracking scenario of the pathogen, it was presumed that Iran could potentially be the center of origin of V. inaequalis, which requires further detailed studies with isolates collected from different parts of central Asia and world for confirmation.
International Journal of Molecular Sciences, 2012
The goals of this investigation were to identify and evaluate the use of polymorphic microsatellite marker (PMM) analysis for molecular typing of seventeen plant pathogenic fungi. Primers for di-, tri-, and tetranucleotide loci were designed directly from the recently published genomic sequence of Mycospherlla graminicola and Fusarium graminearum. A total of 20 new microsatellite primers as easy-to-score markers were developed. Microsatellite primer PCR (MP-PCR) yielded highly reproducible and complex genomic fingerprints, with several bands ranging in size from 200 to 3000 bp. Of the 20 primers tested, only (TAGG)4, (TCC)5 and (CA)7T produced a high number of polymorphic bands from either F. graminearum or F. culmorum. (ATG)5 led to successful amplifications in M. graminicola isolates collected from Germany. Percentage of polymorphic bands among Fusarium species ranged from 9 to 100%. Cluster analysis of OPEN ACCESS banding patterns of the isolates corresponded well to the established species delineations based on morphology and other methods of phylogenetic analysis. The current research demonstrates that the newly designed microsatellite primers are reliable, sensitive and technically simple tools for assaying genetic variability in plant pathogenic fungi.
Molecular Ecology, 2010
Evaluating the impact of plant domestication on the population structure of the associated pathogens provides an opportunity to increase our understanding of how and why diseases emerge. Here, we investigated the evolution of the population structure of the apple scab fungus Venturia inaequalis in response to the domestication of its host. Inferences were drawn from multilocus microsatellite data obtained from samples collected on (i) the Central Asian Malus sieversii, the main progenitor of apple, (ii) the European crabapple, Malus sylvestris, a secondary progenitor of apple, and (iii) the cultivated apple, Malus · domestica, in orchards from Europe and Central Asia. Using clustering methods, we identified three distinct populations: (i) a large European population on domesticated and wild apples, (ii) a large Central Asian population on domesticated and wild apples in urban and agricultural areas, and (iii) a more geographically restricted population in M. sieversii forests growing in the eastern mountains of Kazakhstan. Unique allele richness and divergence time estimates supported a host-tracking co-evolutionary scenario in which this latter population represents a relict of the ancestral populations from which current populations found in human-managed habitats were derived. Our analyses indicated that the domestication of apple induced a significant change in the genetic differentiation of populations of V. inaequalis in its centre of origin, but had little impact on its population dynamics and mating system. We discuss how the structure of the apple-based agrosystem may have restricted changes in the population structure of the fungus in response to the domestication of its host.
2003
This study was initiated to determine whether differences in genotypic diversity among populations of Venturia inaequalis (Cke.) Wint., as detected using neutral genetic markers, were related to the ecological conditions in which apples are grown in Israel. Since sexual reproduction in this fungal pathogen has an obligate requirement for sustained low winter temperatures, and since these requirements in Israel are met only on the Golan Heights, we were interested in whether lower elevation populations of this pathogen might be comprised of asexual clonal lineages. Unlike temperate apple growing regions, where the primary spring inoculum is ascosporic derived from overwintered pseudothecia, Israeli apple orchards at lower elevations in the Hula Valley and along the coastal plain rarely if ever experience low winter temperatures and pseudothecia have never been recovered. Two orchards were sampled from the Golan Heights (El Rom and Ortal, n = 38) and three orchards from the Hula Valley and coastal plain (Sede Eliezer, Ginaton and Be'er Tuvia, n = 40). Microsatellite primers were used to analyze population structure and the resulting binary data analyzed by both cluster and parsimony analysis. Populations from the coastal plain were genetically uniform within each of the orchards sampled, whereas populations from the Golan Heights showed levels of genotypic diversity ten times as high. The data support field observations that this pathogen does not reproduce sexually in regions characterized by the absence of low winter temperatures and is instead composed of clonal lineages. This may have bearing on control strategies for the disease in Israel.