Identification of race-specific resistance in North American Vitis spp. limiting Erysiphe necator hyphal growth

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.     

Effects of Diffuse Colonization of Grape Berries by Uncinula necator on Bunch Rots, Berry Microflora, and Juice and Wine Quality

ABSTRACT Production of grape (principally cultivars of Vitis vinifera) for high-quality wines requires a high level of suppression of powdery mildew (Uncinula necator syn. Erysiphe necator). Severe infection of either fruit or foliage has well-documented and deleterious effects upon crop and wine quality. We found that berries nearly immune to infection by U. necator due to the development of ontogenic resistance may still support diffuse and inconspicuous mildew colonies when inoculated approximately 3 weeks post-bloom. Fruit with diffuse mildew colonies appear to be healthy and free of powdery mildew in late-season vineyard assessments with the naked eye. Nonetheless, presence of these colonies on berries was associated with (i) elevated populations of spoilage microorganisms; (ii) increased evolution of volatile ethyl acetate, acetic acid, and ethanol; (iii) increased infestation by insects known to be attracted to the aforementioned volatiles; (iv) increased rotting by Botrytis cinerea; and (v) increased frequency of perceived defects in wines prepared from fruit supporting diffuse powdery mildew colonies. Prevention of diffuse infection requires extending fungicidal protection until fruit are fully resistant to infection. Despite a perceived lack of improvement in disease control due to the insidious nature of diffuse powdery mildew, potential deleterious effects upon crop and wine quality thereby would be avoided.     

Detection of Erysiphe necator in Air Samples Using the Polymerase Chain Reaction and Species-Specific Primers

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.     

A single nucleotide polymorphism in the β-tubulin gene distinguishing two genotypes of Erysiphe necator expressing different symptoms on grapevine

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.     

Persistence and spatial autocorrelation of clones of Erysiphe necator overwintering as mycelium in dormant buds in an isolated vineyard in northern Italy

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.     

No indication of strict host associations in a widespread mycoparasite: grapevine powdery mildew (Erysiphe necator) is attacked by phylogenetically distant Ampelomyces strains in the field

Pycnidial fungi belonging to the genus Ampelomyces are common intracellular mycoparasites of powdery mildews worldwide. Some strains have already been developed as commercial biocontrol agents (BCAs) of Erysiphe necator and other powdery mildew species infecting important crops. One of the basic, and still debated, questions concerning the tritrophic relationships between host plants, powdery mildew fungi, and Ampelomyces mycoparasites is whether Ampelomyces strains isolated from certain species of the Erysiphales are narrowly specialized to their original mycohosts or are generalist mycoparasites of many powdery mildew fungi. This is also important for the use of Ampelomyces strains as BCAs. To understand this relationship, the nuclear ribosomal DNA internal transcribed spacer (ITS) and partial actin gene (act1) sequences of 55 Ampelomyces strains from E. necator were analyzed together with those of 47 strains isolated from other powdery mildew species. These phylogenetic analyses distinguished five major clades and strains from E. necator that were present in all but one clade. This work was supplemented with the selection of nine inter-simple sequence repeat (ISSR) markers for strain-specific identification of Ampelomyces mycoparasites to monitor the environmental fate of strains applied as BCAs. The genetic distances among strains calculated based on ISSR patterns have also highlighted the genetic diversity of Ampelomyces mycoparasites naturally occurring in grapevine powdery mildew. Overall, this work showed that Ampelomyces strains isolated from E. necator are genetically diverse and there is no indication of strict mycohost associations in these strains. However, these results cannot rule out a certain degree of quantitative association between at least some of the Ampelomyces lineages identified in this work and their original mycohosts.     

Effects of sunlight exposure on grapevine powdery mildew development

Natural and artificially induced shade increased grapevine powdery mildew (Erysiphe necator) severity in the vineyard, with foliar disease severity 49 to 75% higher relative to leaves in full sun, depending on the level of natural shading experienced and the individual experiment. Cluster disease severities increased by 20 to 40% relative to those on check vines when ultraviolet (UV) radiation was filtered from sunlight reaching vines in artificial shading experiments. Surface temperatures of leaves in full sunlight averaged 5 to 8°C higher than those in natural shade, and in one experiment, filtering 80% of all wavelengths of solar radiation, including longer wavelengths responsible for heating irradiated tissues, increased disease more than filtering UV alone. In controlled environment experiments, UV-B radiation reduced germination of E. necator conidia and inhibited both colony establishment (hyphal formation and elongation) and maturity (latent period). Inhibitory effects of UV-B radiation were significantly greater at 30°C than at 20 or 25°C. Thus, sunlight appears to inhibit powdery mildew development through at least two mechanisms, i.e., (i) UV radiation's damaging effects on exposed conidia and thalli of the pathogen; and (ii) elevating temperatures of irradiated tissues to a level supraoptimal or inhibitory for pathogen development. Furthermore, these effects are synergistic at temperatures near the upper threshold for disease development.     

小麦-粗山羊草双二倍体抗白粉病基因定位及其遗传转移

中国春单体分析表明,四倍体小麦－粗山羊草双二倍体Ａｍ６的抗白粉病基因位于１Ａ染色体上,与２４个已知抗白粉病基因比较说明,Ａｍ６可能为一新的抗白粉病基因。限制性片段长度多态性分析表明,在分布小麦７个部分同源群的３６个探针中未找到与此抗病基因连锁的分子标记。     

Ontogenic resistance to powdery mildew in grape berries

ABSTRACT Berries of Vitis vinifera are reported to be susceptible to infection by Uncinula necator until soluble solids levels (brix) reach 8%, and established colonies are reported to sporulate until brix reach 15%. However, our analysis of disease progress on fruit of selected V. vinifera cultivars indicated that severity became asymptotic several weeks earlier in fruit development. When mildew-free fruit clusters of V. vinifera 'Chardonnay', 'Riesling', 'Gewürztraminer', and 'Pinot Noir' were inoculated at stages ranging from prebloom to 6 weeks postbloom, only fruit inoculated within 2 weeks of bloom developed severe powdery mildew. Substantial ontogenic resistance to infection was expressed in fruit nearly 6 weeks before fruit brix reached 8% and over 2 months before they reached 15%. Rachises of 'Chardonnay' and 'Riesling' fruit clusters developed severe powdery mildew when inoculated at bloom, and disease increased steadily over the next 60 days. The rachis of fruit clusters inoculated 31 days after bloom developed only trace levels of powdery mildew. Berry weight of all four cultivars at harvest was reduced when fruit clusters were inoculated at bloom or 16 days postbloom, primarily by splitting, rotting, and dehydration of mildewed berries, but the weight of later-inoculated berries was not reduced. Inoculation of berries just as ontogenic resistance increased markedly, approximately 3 to 4 weeks postbloom, resulted in the development of inconspicuous, diffuse, non-sporulating mildew colonies on berries, sometimes associated with a network of necrotic epidermal cells. Rather than a protracted and relatively static period of berry susceptibility lasting 3 months, fruit of V. vinifera appear to acquire ontogenic resistance rapidly after fruit set. A refocusing of disease management on this critical period of high fruit susceptibility should greatly improve the efficacy of fungicides directed against powdery mildew.     

Effects of humidity on the development of grapevine powdery mildew

ABSTRACT The effects of humidity on powdery mildew development on grape seedlings and the germination of Uncinula necator conidia in vitro were examined. Studies were conducted at an optimum temperature of 25 +/- 2 degrees C. Disease on foliage was markedly affected by humidity levels in the test range of 39 to 98% relative humidity (RH), corresponding to vapor pressure deficits (VPD) of 1,914 to 61 Pa. Incidence and severity increased with increasing humidity to an optimum near 85% RH, and then appeared to plateau or decrease marginally at higher values. Conidial density and chain length also were proportional to humidity, but were influenced less strongly. There was a strong, positive linear relationship between humidity level and frequency of conidium germination with RH treatments of 相似文献   

Molecular characterization of a powdery mildew resistance gene in wheat cultivar suwon 92

ABSTRACT Powdery mildew, caused by Blumeria graminis f. sp tritici, is an important foliar disease of wheat worldwide. Pyramiding race-specific genes into a single cultivar and combining race-specific resistance genes with durable resistance genes are the preferred strategies to improve the durability of powdery mildew resistance. The objectives of this study were to characterize a powdery mildew resistance gene in Suwon 92 and identify gene-specific or tightly linked molecular markers for marker-assisted selection (MAS). A population of recombinant inbred lines (RILs) was derived by single seed descent from a cross between Suwon 92 and a susceptible cultivar, CI 13227. The RILs were screened for adult-plant infection type of powdery mildew and characterized with amplified fragment length polymorphism (AFLP) and simple sequence repeat (SSR) markers. The linked markers explained 41.3 to 69.2% of the phenotypic variances measured in 2 years. A morphological marker, hairy glume, was also associated with powdery mildew resistance in Suwon 92, and explained 43 to 51% of the phenotypic variance. The powdery mildew resistance gene in Suwon 92 was located on the short arm of chromosome 1A where Pm3 was located. Two gene-specific markers were developed based on the sequence of the cloned Pm3b gene. These two markers, which were mapped at the same locus in the peak region of the LOD score for the RIL population, explained most of the phenotypic variance for powdery mildew resistance in the RIL population. The powdery mildew resistance in Suwon 92 is most likely conditioned by the Pm3 locus. The gene markers developed herein can be directly used for MAS of some of the Pm3 alleles in breeding programs.     

Host Barriers and Responses to Uncinula necator in Developing Grape Berries

ABSTRACT Grape berries are highly susceptible to powdery mildew 1 week after bloom but acquire ontogenic resistance 2 to 3 weeks later. We recently demonstrated that germinating conidia of the grape powdery mildew pathogen (Uncinula necator) cease development before penetration of the cuticle on older resistant berries. The mechanism that halts U. necator at that particular stage was not known. Several previous studies investigated potential host barriers or cell responses to powdery mildew in berries and leaves, but none included observation of the direct effect of these factors on pathogen development. We found that cuticle thickness increased with berry age, but that ingress by the pathogen halted before formation of a visible penetration pore. Cell wall thickness remained unchanged over the first 4 weeks after bloom, the time during which berries progressed from highly susceptible to nearly immune. Autofluorescent polyphenolic compounds accumulated at a higher frequency beneath appressoria on highly susceptible berries than on highly resistant berries; and oxidation of the above phenolics, indicated by cell discoloration, developed at a significantly higher frequency on susceptible berries. Beneath the first-formed appressoria of all germinated conidia, papillae occurred at a significantly higher frequency on 2- to 5-day-old berries than on 30- to 31-day-old fruit. The relatively few papillae observed on older berries were, in most cases (82.8 to 97.3%), found beneath appressoria of conidia that had failed to produce secondary hyphae. This contrasted with the more abundantly produced papillae on younger berries, where only 35.4 to 41.0% were located beneath appressoria of conidia that had failed to produce secondary hyphae. A pathogenesis-related gene (VvPR-1) was much more highly induced in susceptible berries than in resistant berries after inoculation with U. necator. In contrast, a germin-like protein (VvGLP3) was expressed within 16 h of inoculation in resistant, but not in susceptible berries. Our results suggest that several putative barriers to infection, i.e., cuticle and cell wall thickness, antimicrobial phenolics, and two previously described pathogenesis-related proteins, are not principal causes in halting pathogen ingress on ontogenically resistant berries, but rather that infection is halted by one or more of the following: (i) a preformed physical or biochemical barrier near the cuticle surface, or (ii) the rapid synthesis of an antifungal compound in older berries during the first few hours of the infection process.     

Potential and actual ascospore release of Erysiphe necator chasmothecia in Austria

Journal of Plant Diseases and Protection - Ascospores of grape powdery mildew (Erysiphe necator Schw.) play a crucial role in the disease onset in spring in many vine-growing areas. We investigated...     

New tools for studying epidemiology and resistance of grape powdery mildew to DMI fungicides

Using a Random Amplified Polymorphic DNA (RAPD) assay, we investigated the genetic polymorphism existing among 62 European isolates of the grape powdery mildew fungus (Uncinula necator [Schw.] Burr.). Isolates overwintering as mycelium in buds were genetically distinct from isolates overwintering as ascospores, suggesting the existence of two genetically isolated powdery mildew populations, and consequently of two independent sources of inoculum in the vineyard. Isolates resistant to fungicides inhibiting sterol 14α-demethylation (DMIs) were found in both populations, suggesting that resistance to DMIs may arise independently in the two powdery mildew populations. A PCR assay targeting the gene encoding U. necator 14α-demethylase has been developed which will permit an early, specific detection of U. necator infections, and may be useful for spraying programmes. ©1997 SCI     

苦瓜白粉病病原菌和生理小种的鉴定及苦瓜对白粉病的抗性遗传分析

为明确海南省苦瓜白粉病的病原菌、生理小种及苦瓜对白粉病的抗性遗传规律,结合形态学鉴定和分子鉴定解析白粉病菌及生理小种种类,通过显微镜观察白粉病菌侵染过程,并应用主基因+多基因混合遗传模型分析法探讨苦瓜对白粉病的主要抗性遗传规律。结果表明:采集自海南省6个市(县)的苦瓜白粉病病原菌均为单囊壳白粉菌Sphaerotheca fuliginea,属生理小种2F,该菌在侵染苦瓜叶片时有4个关键时期:接种后4 h为分生孢子萌发高峰期,8 h为附着孢形成高峰期,16～24 h为次生菌丝形成高峰期,5 d为分生孢子梗形成高峰期。将其接种于苦瓜抗、感品系,对白粉病的抗性符合2对加性-显性-上位性主基因+加性-显性多基因模型,主基因和多基因共同控制苦瓜对白粉病的抗性,其中以主基因遗传为主,且会受到环境变异的影响。根据苦瓜抗性遗传规律,F2代主基因遗传率最高,受环境影响最小,在苦瓜的白粉病抗性育种中,以早期世代F2代作为有效选择世代。研究表明白粉病菌侵染叶片的前2 d是白粉病防治的最佳时期,所以在白粉病易发的物候期,可将防治时间提前1～2 d。     

小麦新资源对条锈病白粉病的抗性鉴定

１９９１～１９９７年在甘谷田间对远缘小麦新资源进行抗条锈病、白粉病鉴定。结果表明，在供试１６０份材料中，对条锈病免疫或高抗、白粉病免疫的有２５份，其抗性比较稳定，是良好的抗源材料，可供抗病育种利用。     

Powdery mildew resistance in selections from moroccan barley landraces

Thirty barley landraces collected from Morocco in 1985 and 1989, and held in the Polish Gene Bank, IHAR, Radzików, Poland, were screened for resistance to powdery mildew. Fifteen tested landraces (50%) showed powdery mildew resistance reactions and 24 single plant lines were selected. Eighteen lines originating from 13 landraces were tested with 17 isolates of powdery mildew and another six lines originating from six landraces were tested with 23; the isolates were chosen according to their virulence spectra observed on the ‘Pallas’ isolines differential set. Three lines (E 1090-2-2, E 1110-3-2 and E 1077-1-1) showed resistance to all powdery mildew virulence genes prevalent in Europe. In 21 lines, unknown genes alone or in combination with specific ones were detected. Five different resistance alleles(Mlat, Mlal, Mla3, Mlg andMl(CP)) were postulated to be present in the tested lines, alone or in combination:Mlat was postulated to be present in nine (~38%) lines;Mlg andMl(CP) in two lines, andMla1 andMla3 in one tested line each. The use of newly identified sources of resistance in barley breeding as a means of controlling powdery mildew is discussed.     

Field evaluation of mungbeans for resistance to Cercospora leaf spot and powdery mildew

Abstract

Cercospora leaf spot, caused by Cercospora canescens and powdery mildew, caused by Erysiphe polygoni are two of the most important fungal diseases of mungbeans. Nearly 4000 accessions of the global mungbean collection at the Asian Vegetable Research and Development Center were screened for resistance to these two pathogens. Less than 4% and 12% of the accessions showed resistance to Cercospora leaf spot and powdery mildew, respectively. The level of resistance to Cercospora leaf spot varied and highly resistant lines were not found. A few lines were rated highly resistant to powdery mildew over several years but others were rated moderately resistant or susceptible in other years.     

普通小麦“兰考90(6)”品系对白粉病抗性的遗传研究

 普通小麦(Triticum aestivum L.)"兰考90(6)"系列品系是以六倍体小黑麦(X Triticosecale Wittmack;AABBRR)为白粉病抗源培育的新的小麦-黑麦1BL/1RS异易位系。这些品系高抗白粉病。小麦白粉病抗性基因推导试验证明,"豫麦66"携带的抗病基因与大多数已经报道的小麦抗白粉病基因不同。用白粉菌[Blumeria graminis (DC.) E. O. Speer f. sp. tritici]单孢堆分离物进行的遗传分析表明,"兰考90(6)"品系携带一个小种专化的隐性抗白粉病基因。对"中国春"和"兰考90(6)21-12"杂交F₂分离群体进行1RS染色体检测,结果证明该抗白粉病基因不在1RS染色体臂上。本研究为有效利用"兰考90(6)"系列品系中的抗白粉病基因提供了科学依据。