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1.
ABSTRACT Erysiphe necator overwinters as ascospores in cleistothecia and mycelium in dormant buds of grapevines. Shoots developing from infected buds early in the growing season are covered with dense mycelium and are known as "flag shoots". Combining epidemiological and genetic analyses, the objective of this study was to analyze the spatial and genetic structure of a flag shoot subpopulation of E. necator as a way to assess the contribution of flag shoots as primary inoculum, and to determine if flag shoot subpopulations are clonal with only one mating type. One vineyard in Tuscany, Italy was surveyed intensively for flag shoots for 8 years; isolations of E. necator were made from flag shoots for 5 years. We observed distinct disease foci developing around flag shoots early in epidemics, demonstrating a steep dispersal gradient of conidia and the importance of flag shoots as primary inoculum sources. Flag shoots were spatially aggregated within and between years, most likely as a result of short-distance dispersal of conidia from flags early in the season when dormant buds for the next year's shoots are formed and are susceptible to infection. The two mating types were found in 1:1 ratios in this flag shoot subpopulation. Genotypic diversity, based on inter-simple sequence repeat markers, was high in all years with only two haplotypes occurring twice, and subpopulations were genetically differentiated between years. Similarities between haplotypes were not spatially autocorrelated. One multilocus analysis of population structure is consistent with the hypothesis of random mating but another is not. These results are not consistent with expectations for a strictly clonal or strictly randomly mating flag shoot subpopulation. Instead, the hypothesis that the flag shoot subpopulation of E. necator may reproduce clonally and sexually needs further testing. 相似文献
2.
The population structure of the grape powdery mildew fungus, Erysiphe necator (formerly Uncinula necator), has been hypothesized to vary from being clonal to highly diverse and recombining. We report here on the structure of an E. necator population sampled during a 4-year period from an isolated vineyard in northern Italy (Voghera, Pavia Province). We obtained 54 isolates of E. necator that overwintered asexually as mycelium in grapevine buds and caused severe symptoms on the emerging shoots, known as flag shoots. All isolates were genotyped for mating type, four multilocus polymerase chain reaction (PCR)-based markers (a total of 64 loci were scored), and two single-copy loci designed to identify genetic subgroups in E. necator. All isolates had the same mating type and single-locus alleles that correlate to isolates from flag shoots in other areas. Only 2 of the 64 loci scored from multilocus markers were polymorphic; 46 of the 54 isolates had the same multilocus haplotype. Seven isolates had a second haplotype that was recovered over 3 years, and only a single isolate was found with a third haplotype. Both variant haplotypes differed from the main clonal haplotype by single loci. Spatial autocorrelation analyses showed that vines with flag shoots were not aggregated within years, but they were aggregated between consecutive years. These results demonstrate that this subpopulation of E. necator on flag shoots is composed of a single clonal lineage that has persisted for at least 4 years. We speculate that the lack of diversity in the flag shoot subpopulation in this vineyard is the result of restricted immigration from surrounding areas and genetic drift operating through founder effects and periodic bottlenecks. We propose a model that integrates epidemiology and population genetics to explain the variation observed in genetic structure of E. necator flag shoot subpopulations from different vineyards or viticultural regions. 相似文献
3.
Erysiphe necator, the causative agent of powdery mildew in grapevine, was introduced into Europe from North America during the middle of the
19th century. Our objective was to analyze the genetic variation and the population structure of the fungus in southern France.
The sample comprised 101 isolates and was mainly of flag shoot origin, i.e., infection of sprouting shoots after overwintering
of mycelium in buds. RAPD analysis identified different haplotypes that clustered in two genetic groups (A and B). The most
frequent haplotypes of each group were found in several different locations in two areas separated by 100 km and throughout
the 3 year period. Several haplotypes of both groups originated from flag shoots and were recovered over successive years
indicating that there is no correlation between genetic group and overwintering mode. All isolates of group A were of mating
type +, but those in group B could be either + or −. Lower genotypic diversity was detected within group A than within group
B. These results were consistent with the hypothesis that group A reproduces only asexually. 相似文献
4.
A. Rügner J. Rumbolz † B. Huber G. Bleyer U. Gisi H.-H. Kassemeyer R. Guggenheim 《Plant pathology》2002,51(3):322-330
Developing shoots of grapevine ( Vitis vinifera cv. Kerner) were inoculated with conidia of the powdery mildew, Uncinula necator , at well-defined phenological stages of the host to provoke the development of flag shoots in the field and to investigate these shoots as early as possible in the following growing season for the presence of the pathogen. The disease progress was monitored and fungal growth and development on samples from a field trial were analysed by means of low-temperature scanning electron microscopy (LT–SEM). Mycelium was detected on the surface and in the interior parts of buds. The suitability of the field plot to analyse flag shoots was proven by the occurrence of such shoots in the following spring. Furthermore, early stages of cleistothecia development on berries were described for the first time. Establishment of U. necator in dormant buds of grapevine, giving rise to flag shoots in the following spring, is considered to play a significant role in overwintering of the pathogen in the vineyards of southern Germany. 相似文献
5.
The biotrophic fungus Erysiphe necator (formerly Uncinula necator ), for which two genetic groups have been described in European vineyards, is the causal agent of grapevine powdery mildew. By analysing the pathogen population with respect to polymorphism in the sequence of the β-tubulin gene, which distinguishes two groups of isolates, a new tool was developed for epidemiological and population studies and tested in the vineyard. As in many ascomycetes, the β-tubulin gene of E. necator ( Entub ) includes six introns and seven exons and encodes a 447-amino-acid protein. A single nucleotide polymorphism (SNP) in the intron-3 region of the Entub gene distinguished two genetic groups (A and B). This method was used to examine differences in the ratio of the two groups from a total of 289 grape powdery mildew samples collected at the beginning of the growing season from either flag shoots or leaves with sparse-spot symptoms in four different vineyards. The SNP in the intron-3 region of the β-tubulin gene, similar to SNPs in the CYP51 gene, was associated with genotypes A and B of E. necator and confirmed the existence of two sympatric populations of the pathogen in the French vineyards. Differences in the relative proportions of each group varied with the presence or absence of flag-shoot symptoms and with the region in which isolates had been collected. 相似文献
6.
ABSTRACT Ninety isolates of grape powdery mildew (Uncinula necator) from Europe (sixty-two) and India (twenty-eight) were collected. Ten of the sixty-two European isolates originated from mycelium overwintering in dormant buds ("flagshoots"). Mating types were determined, and genetic variation was assessed by random amplified polymorphic DNA (RAPD). Forty-one European isolates, including all "flagshoot" isolates, were mating type +, and twenty-one were mating type -. All Indian isolates were mating type -. Phenetic analysis based on 414 amplicons revealed three main groups. Most European isolates (53) clustered together. Nine flagshoot isolates clustered in a second distinct group. These isolates, which coexisted with other isolates in the field, may represent a genetically isolated biotype of U. necator. Indian isolates clustered into two groups. The first group (15 isolates) was a subgroup of the group containing European nonflagshoot isolates. The second group (12 isolates) was distinct from the other groups. These two groups of Indian isolates may represent genetically isolated populations with different climatic tolerances. A polymerase chain reaction primer pair, derived from a RAPD fragment specific to the Indian isolates, proved to be suitable for field studies. 相似文献
7.
Falacy JS Grove GG Mahaffee WF Galloway H Glawe DA Larsen RC Vandemark GJ 《Phytopathology》2007,97(10):1290-1297
ABSTRACT A polymerase chain reaction (PCR) assay employing species-specific primers was developed to differentiate Erysiphe necator from other powdery mildews common in the northwest United States. DNA was extracted from mycelia, conidia, and/or chasmothecia that were collected from grape leaves with a Burkard cyclonic surface sampler. To differentiate E. necator from other erysiphaeceous fungi, primer pairs Uncin144 and Uncin511 were developed to select unique sequences of the internal transcribed spacer regions of E. necator. Using these primers in PCR amplifications, a 367-bp amplicon specific to E. necator was generated, but no amplicons were generated from other erysiphaceous species collected from 48 disparate hosts representing 26 vascular plant families. The PCR limit of detection was one to five conidia of E. necator placed directly into reaction mixtures or 100 to 250 conidia placed on glass rods coated with silicon grease. During field studies, this PCR assay facilitated the detection of E. necator inoculum in air samples within hours of sample rod collection and prior to disease onset. Amplification of E. necator DNA did not occur when the PCR assay was conducted on vineyard air samples collected while grapes were dormant or during periods when vine growth occurred but E. necator remained dormant. The initial PCR detection of E. necator of the season occurred during seasonal ascospore releases caused by precipitation events between bud burst and the prebloom period during the 3 years of the study. Detection ceased for 7 to 11 days following ascospore release and then resumed several days prior to the observance of microscopic symptoms and signs of powdery mildew in the field. Results of this study represent the initial step toward the goal of incorporating an inoculum availability component into current and future grapevine powdery mildew risk assessment models. 相似文献
8.
Bud colonization and perennation of powdery mildew ( Erysiphe necator ) was studied by inoculating shoots of grapevine ( Vitis vinifera cv. Carignane) at different phenological stages. Disease incidence and severity assessments indicated that buds were most susceptible at the three- to six-unfolded-leaf stage. Incidence of powdery mildew colonies on the surface of buds collected from these shoots 7 weeks postinoculation was highest at these stages (68 and 62%, respectively), which indicates that colonization of the bud interior via the infected bud surface is likely to occur within this period. Histological analyses of buds revealed hyphae with haustoria, conidiophores and conidia on all parts of the bud interior except for the meristems. In particular, trichomes were frequently parasitized by haustoria. In total, 13·2% of all buds analysed, and 32·3% of all buds originating from shoots inoculated at the three-unfolded-leaf stage, were infected by E. necator . In the spring of the following year, buds from inoculated shoots yielded 18 flag shoots (1·6% of all emerging shoots). These primary infections caused an epidemic 28 days after the appearance of the first flag shoot. A linear regression analysis on the frequency of infections of the bud exterior, bud interior and flag shoots revealed that incidence of external bud infection in the first season is strongly correlated with flag shoot incidence in the following season ( R 2 = 0·94). Hence predictions of flag shoot incidence may be reliably based on the incidence of infection on the outer bud scales in the preceding season. 相似文献
9.
ABSTRACT Population genetic and epidemiological studies have resulted in different hypotheses about the predominant source of primary inoculum in the Phaeosphaeria nodorum-wheat pathosystem (i.e., sexually derived, windborne ascospores versus asexual or seedborne inoculum). We examined the genetic structure of seedborne populations of P. nodorum as a further step toward evaluating the hypothesis that seedborne inoculum is an important contributor to foliar epidemics in New York's rotational wheat fields. In all, 330 seedborne isolates from seven field populations were genotyped at 155 amplified fragment length polymorphism loci. Seedborne populations possessed high levels of genotypic diversity, with virtually every isolate (326/330) having a unique haplotype. As in previous population genetic studies of P. nodorum, we found low levels of gametic disequilibrium, although we could reject the null hypothesis of random mating with the index of association test for two populations. Thus, genotypically diverse and seemingly panmictic populations of P. nodorum that have been observed in wheat foliage could be derived from seedborne primary inoculum. Although sexual reproduction and recombination may contribute to the diversity of foliar populations of P. nodorum, population genetic data do not rule out seed as a source of primary inoculum. Further experimentation will be needed to determine definitively the relative importance of windborne ascospores and seed-borne asexual inoculum in epidemics of Stagonospora nodorum blotch in New York. 相似文献
10.
In this study, 18 microsatellite loci were used to examine the genetic structure and mode of reproduction of Phaeomoniella chlamydospora, the fungus that causes Petri disease of grapevine and is involved in another grapevine disease, esca. A total of 60 southeastern Australian isolates were tested and compared with 64 isolates from southern France. The French population possessed relatively high genotypic diversity (G = 29·2) whilst the Australian population showed low genotypic diversity (G = 11·1), consistent with a limited founder population. Haplotypes from four different grapevine cultivars were not significantly differentiated (Φpt values effectively zero). Likewise, genetic differentiation of haplotypes from different regions within each country was not significant, although small but significant genetic differentiation (9%) was identified between Australia and France. Based on bootstrapped cluster analysis, there did not appear to be any genetic groups within the overall sample of isolates. Significant linkage disequilibrium identified in both countries, together with overrepresented, widespread identical haplotypes, indicated limited genetic recombination and a largely clonal structure consistent with the absence of an observed sexual cycle in this species. 相似文献
11.
Ramming DW Gabler F Smilanick JL Margosan DA Cadle-Davidson M Barba P Mahanil S Frenkel O Milgroom MG Cadle-Davidson L 《Phytopathology》2012,102(1):83-93
Race-specific resistance against powdery mildews is well documented in small grains but, in other crops such as grapevine, controlled analysis of host-pathogen interactions on resistant plants is uncommon. In the current study, we attempted to confirm powdery mildew resistance phenotypes through vineyard, greenhouse, and in vitro inoculations for test cross-mapping populations for two resistance sources: (i) a complex hybrid breeding line, 'Bloodworth 81-107-11', of at least Vitis rotundifolia, V. vinifera, V. berlandieri, V. rupestris, V. labrusca, and V. aestivalis background; and (ii) Vitis hybrid 'Tamiami' of V. aestivalis and V. vinifera origin. Statistical analysis of vineyard resistance data suggested the segregation of two and three race-specific resistance genes from the two sources, respectively. However, in each population, some resistant progeny were susceptible in greenhouse or in vitro screens, which suggested the presence of Erysiphe necator isolates virulent on progeny segregating for one or more resistance genes. Controlled inoculation of resistant and susceptible progeny with a diverse set of E. necator isolates clearly demonstrated the presence of fungal races differentially interacting with race-specific resistance genes, providing proof of race specificity in the grape powdery mildew pathosystem. Consistent with known race-specific resistance mechanisms, both resistance sources were characterized by programmed cell death of host epidermal cells under appressoria, which arrested or slowed hyphal growth; this response was also accompanied by collapse of conidia, germ tubes, appressoria, and secondary hyphae. The observation of prevalent isolates virulent on progeny with multiple race-specific resistance genes before resistance gene deployment has implications for grape breeding strategies. We suggest that grape breeders should characterize the mechanisms of resistance and pyramid multiple resistance genes with different mechanisms for improved durability. 相似文献
12.
Osmotin and thaumatin from grape: a putative general defense mechanism against pathogenic fungi 总被引：1，自引：0，他引：1
Monteiro S Barakat M Piçarra-Pereira MA Teixeira AR Ferreira RB 《Phytopathology》2003,93(12):1505-1512
ABSTRACT Little information is available concerning the expression of pathogenesis-related (PR) proteins in grapevine (Vitis vinifera) and their effect properties on the major fungal pathogens of grape. A systematic study was performed on the effect of total or individual grape proteins on mycelial growth, spore germination, and germ tube growth of Uncinula necator, Phomopsis viticola, and Botrytis cinerea. Two proteins, identified as PR proteins by immunological methods and by N-terminal sequencing as osmotin and thaumatin-like protein, exhibited strong antifungal activities in vitro, blocking the growth of Phomopsis viticola and Botrytis cinerea mycelia. In addition, they inhibited spore germination and germ tube growth of U. necator, Phomopsis viticola, and Botrytis cinerea. The presence of both proteins displayed a synergistic effect. The expression of osmotin and thaumatin-like protein was induced in grapevine leaves and berries infected with U. necator and Phomopsis viticola. Thaumatin previously was thought to occur exclusively in berries. Immunoblot analyses revealed the accumulation of the two PR proteins in infected leaves and berries, supporting a role in vivo in increasing the resistance of grapevine to fungal attack. 相似文献
13.
Spatial aggregation in Fusarium pseudograminearum populations from the Australian grain belt 总被引：1，自引：0，他引：1
A. R. Bentley † M. G. Milgroom J. F. Leslie B. A. Summerell L. W. Burgess 《Plant pathology》2009,58(1):23-32
Previous studies have evaluated the overall structure of populations of Fusarium pseudograminearum (teleomorph, Gibberella coronicola ), causal agent of cereal crown rot, but there is no information available on spatial relationships of genetic variation in field populations. Three 1-m-row sections in crown-rot-affected wheat fields in the Australian grain belt were intensively sampled to estimate population genetic parameters and the spatial aggregation, or clustering, of disease aggregates and genotypes. Estimates of population genetic parameters based on amplified fragment length polymorphisms (AFLPs) indicated that the genetic diversity in isolates from the 1-m-row populations described a significant portion of the diversity recorded for corresponding field and regional populations. In point pattern analysis, there was physical clustering and aggregation of F. pseudograminearum isolates from two of the three sites. Analysis of the spatial distribution of clonal haplotypes (DICE similarity ≥ 97%) indicated significant aggregation of clones in all three 1-m-row populations. Based on matrix comparison tests, both mating types and genetic distances had significant spatial aggregation for at least two of the three 1-m-row populations. This is consistent with the presence of non-random spatial genetic structure due to clonal aggregation. High levels of genetic diversity and spatial structuring of disease and genotypes in at least two of the three 1-m-row populations is consistent with the hypothesis that stubble is a primary inoculum source in no-tillage farming systems, resulting in aggregated patterns of disease and allowing for haplotypes to be maintained in the field over a number of annual cropping cycles. 相似文献
14.
ABSTRACT Genetic variability and population structure of Cercospora sorghi from wild and cultivated sorghum were investigated to gain insight into their potential impact on epidemics of gray leaf spot of sorghum in Africa. Population structure was examined using data derived from amplified fragment length polymorphism (AFLP) of C. sorghi by Nei's test for population differentiation, G(ST), and analysis of molecular variation (AMOVA). Two ecological populations of C. sorghi in Uganda were devoid of population structure (G(ST) = 0.03, small ef, CyrillicF(ST) = 0.01, P = 0.291). AMOVA revealed that genetic variability was due mainly to variations within (99%) rather than between (0.35%) populations, and Nei's genetic distance between the two populations was 0.014. Phenetic analysis based on AFLP data and polymerase chain reaction-restriction fragment length polymorphism analyses of the internal transcribed spacer regions of rDNA and mitochondrial small subunit rDNA separated Cercospora cereal pathogens from dicot pathogens but did not differentiate among C. sorghi isolates from wild and cultivated sorghum. Our results indicate that Ugandan populations of C. sorghi compose one epidemiological unit and suggest that wild sorghum, while not affecting genetic variability of the pathogen population, provides an alternative host for generating additional inoculum. 相似文献
15.
Grapevine leafroll-associated virus 3 (GLRaV-3; Ampelovirus, Closteroviridae), associated with grapevine leafroll disease, is an important pathogen found across all major grape-growing regions of the world. The genetic diversity of GLRaV-3 in Napa Valley, CA, was studied by sequencing 4.7 kb in the 3' terminal region of 50 isolates obtained from Vitis vinifera 'Merlot'. GLRaV-3 isolates were subdivided into four distinct phylogenetic clades. No evidence of positive selection was observed in the data set, although neutral selection (ratio of nonsynonymous to synonymous substitution rates = 1.1) was observed in one open reading frame (ORF 11, p4). Additionally, the four clades had variable degrees of overall nucleotide diversity. Moreover, no geographical structure among isolates was observed, and isolates belonging to different phylogenetic clades were found in distinct vineyards, with one exception. Considered with the evidence of purifying selection (i.e., against deleterious mutations), these data indicate that the population of GLRaV-3 in Napa Valley is not expanding and its effective population size is not increasing. Furthermore, research on the biological characterization of GLRaV-3 strains might provide valuable insights on the biology of this species that may have epidemiological relevance. 相似文献
16.
Ruijiao Kang Yanfeng Hu Limin Wang Shunpei Xie Yonghui Li Hongxia Yuan Min Wang Linlin Chen Shengli Ding Honglian Li 《Plant pathology》2021,70(1):87-99
Wheat root rot, caused by Bipolaris sorokiniana, has led to severe losses of wheat products worldwide. To evaluate the pathogenicity and genetic variation of B. sorokiniana, diseased wheat samples were collected from 97 locations in the Huanghuai floodplain of China in 2014 and 2015 for analysis. A total of 673 isolates were obtained, 262 of which were identified as B. sorokiniana. Pathogenicity analysis of the isolates revealed variation in pathogenicity, which was not directly correlated with geographic region. Large variations in pathogenicity were also found within geographic groups. To determine the genetic structure of the populations, PCR was performed with universal rice primers (URP). Cluster analysis based on amplification patterns showed that the classified groups were correlated with geographical regions. Thus, analysis of the genetic diversity of the population indicated a negative correlation with geographic origin, that is, the greater the distance between sites, the lower the genetic variation similarity coefficient. Identification of wheat germplasm resistance showed that resistant cultivars accounted for a low percentage, while susceptible and highly susceptible cultivars were in the majority. Overall, these results are meaningful for developing strategies to prevent and control wheat root rot. 相似文献
17.
ABSTRACT The population structure of Heterobasidion annosum in the Pacific Northwest (PNW) Christmas tree plantations was estimated at two spatial scales to assess the relative importance of primary and secondary infection, colonization, and spread of the pathogen. Ninety-three isolates from single trees in 27 discrete mortality pockets and 104 isolates from 12 individual root systems of noble and Fraser fir trees were sampled near Mossyrock, Washington. Isolates were genotyped using somatic compatibility assays and microsatellite markers to determine the spatial scale at which dispersal of single genotypes (genets) was occurring. All isolates sampled from different trees in discrete mortality pockets had distinct genotypes, whereas the root systems of single trees were dominated by one or two genotypes. These results suggest that infection of PNW Christmas trees results from frequent primary infection events of adjacent stumps and localized secondary spread within root systems rather than clonal spread of the pathogen between adjacent trees. We hypothesize that mortality pockets may be due to availability of infection courts and/or variation in inoculum levels during selective harvesting of patches of mature trees. 相似文献
18.
19.
Variation of Diplodia seriata, a fungal species associated with botryosphaeria dieback of grapevine, was investigated with respect to its genetic, phenotypic and pathogenic characteristics. The inter‐simple sequence repeat (ISSR) technique was used to investigate the genetic diversity of 83 isolates of D. seriata. Five ISSR primers were able to provide reproducible and polymorphic DNA fingerprint patterns, thus showing a relevant genetic variability in the species. Analyses of ISSR data by different clustering methods grouped the isolates into two distinct clusters through the Bayesian and DAPC analyses. No relationships between either geographic or host origin of isolates and genetic clusters were observed. Several representative isolates from each genetic cluster were chosen for studying their conidial dimensions, in vitro mycelial growth, vegetative and mating compatibility, and pathogenicity on detached grapevine canes and potted vines. No significant differences in conidial dimensions were detected among the groups. Vegetative compatibility reactions were observed among isolates but this was not related with the genetic clustering. Production of sexual fruiting bodies in vegetative compatible crossings was not observed under the experimental conditions used in the study. All 14 isolates tested for pathogenicity were confirmed to be pathogenic according to the length of the necrotic lesions that they caused and their reisolation frequencies from the infected plant tissues. Differences in the length of necrosis were detected among isolates, thus revealing the existence of different virulence levels in the species. 相似文献
20.
The relative importance of conidia and ascospores as primary inoculum of Venturia inaequalis in a southeast England orchard 
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下载免费PDF全文 Apple scab, caused by Venturia inaequalis, can lead to large losses of marketable fruit if left uncontrolled. The disease appears in orchards during spring as lesions on leaves. These primary lesions are caused by spores released at bud burst from overwintering sources; these spores can be sexually produced ascospores from the leaf litter or asexual conidia from mycelium in wood scab or within buds. The relative importance of conidia and ascospores as primary inoculum were investigated in an orchard in southeast England, UK. Potted trees not previously exposed to apple scab were placed next to (c. 1 m) orchard trees to trap air‐dispersed ascospores. Number and position of scab lesions were assessed on the leaves of shoots from both the potted trees (infection by airborne ascospores) and neighbouring orchard trees (infection by both ascospores and splash‐dispersed, overwintered conidia). The distribution and population similarity of scab lesions were compared in the two tree types by molecular analysis and through modelling of scab incidence and count data. Molecular analysis was inconclusive. Statistical modelling of results suggested that conidia may have contributed approximately 20–50% of the primary inoculum in early spring within this orchard: incidence was estimated to be reduced by 20% on potted trees, and lesion number by 50%. These results indicate that, although conidia are still a minority contributor to primary inoculum, their contribution in this orchard is sufficient to require current management to be reviewed. This might also be true of other orchards with a similar climate. 相似文献