Genome-wide association mapping of septoria nodorum blotch resistance in Nordic winter and spring wheat collections.

KEY MESSAGE: A new QTL for SNB, QSnb.nmbu-2AS, was found in both winter and spring wheat panels that can greatly advance SNB resistance breeding Septoria nodorum blotch (SNB), caused by the necrotrophic fungal pathogen Parastagonospora nodorum, is the dominant leaf blotch pathogen of wheat in Norway. Resistance/susceptibility to SNB is a quantitatively inherited trait, which can be partly explained by the interactions between wheat sensitivity loci (Snn) and corresponding P. nodorum necrotrophic effectors (NEs). Two Nordic wheat association mapping panels were assessed for SNB resistance in the field over three to four years: a spring wheat and a winter wheat panel (n = 296 and 102, respectively). Genome-wide association studies found consistent SNB resistance associated with quantitative trait loci (QTL) on eleven wheat chromosomes, and ten of those QTL were common in the spring and winter wheat panels. One robust QTL on the short arm of chromosome 2A, QSnb.nmbu-2AS, was significantly detected in both the winter and spring wheat panels. For winter wheat, using the four years of SNB field severity data in combination with five years of historical data, the effect of QSnb.nmbu-2AS was confirmed in seven of the nine years, while for spring wheat, the effect was confirmed for all tested years including the historical data from 2014 to 2015. However, lines containing the resistant haplotype are rare in both Nordic spring (4.0%) and winter wheat cultivars (15.7%), indicating the potential of integrating this QTL in SNB resistance breeding programs. In addition, clear and significant additive effects were observed by stacking resistant alleles of the detected QTL, suggesting that marker-assisted selection can greatly facilitate SNB resistance breeding.

Interaction between the Bird Cherry-Oat Aphid (Rhopalosiphum padi) and Stagonospora Nodorum Blotch (Parastagonospora nodorum) on Wheat.

Wheat plants are under constant attack by multiple pests and diseases. Until now, there are no studies on the interaction between the aphid Rhopalosiphum padi and the plant pathogenic fungus Parastagonospora nodorum causal agent of septoria nodorum blotch (SNB) on wheat. Controlled experiments were conducted to determine: (i) The preference and reproduction of aphids on P. nodorum inoculated and non-inoculated wheat plants and (ii) the effect of prior aphid infestation of wheat plants on SNB development. The preference and reproduction of aphids was determined by releasing female aphids on P. nodorum inoculated (SNB+) and non-inoculated (SNB-) wheat leaves. The effect of prior aphid infestation of wheat plants on SNB development was determined by inoculating P. nodorum on aphid-infested (Aphid+) and aphid free (Aphid-) wheat plants. Higher numbers of aphids moved to and settled on the healthy (SNB-) leaves than inoculated (SNB+) leaves, and reproduction was significantly higher on SNB- leaves than on SNB+ leaves. Aphid infestation of wheat plants predisposed the plants to P. nodorum infection and colonization. These results are important to understand the interactions between multiple pests in wheat and hence how to develop new strategies in future integrated pest management (IPM).

Identification and cross-validation of genetic loci conferring resistance to Septoria nodorum blotch using a German multi-founder winter wheat population.

KEY MESSAGE: We identified allelic variation at two major loci, QSnb.nmbu-2A.1 and QSnb.nmbu-5A.1, showing consistent and additive effects on SNB field resistance. Validation of QSnb.nmbu-2A.1 across genetic backgrounds further highlights its usefulness for marker-assisted selection. Septoria nodorum blotch (SNB) is a disease of wheat (Triticum aestivum and T. durum) caused by the necrotrophic fungal pathogen Parastagonospora nodorum. SNB resistance is a typical quantitative trait, controlled by multiple quantitative trait loci (QTL) of minor effect. To achieve increased plant resistance, selection for resistance alleles and/or selection against susceptibility alleles must be undertaken. Here, we performed genetic analysis of SNB resistance using an eight-founder German Multiparent Advanced Generation Inter-Cross (MAGIC) population, termed BMWpop. Field trials and greenhouse testing were conducted over three seasons in Norway, with genetic analysis identifying ten SNB resistance QTL. Of these, two QTL were identified over two seasons: QSnb.nmbu-2A.1 on chromosome 2A and QSnb.nmbu-5A.1 on chromosome 5A. The chromosome 2A BMWpop QTL co-located with a robust SNB resistance QTL recently identified in an independent eight-founder MAGIC population constructed using varieties released in the United Kingdom (UK). The validation of this SNB resistance QTL in two independent multi-founder mapping populations, regardless of the differences in genetic background and agricultural environment, highlights the value of this locus in SNB resistance breeding. The second robust QTL identified in the BMWpop, QSnb.nmbu-5A.1, was not identified in the UK MAGIC population. Combining resistance alleles at both loci resulted in additive effects on SNB resistance. Therefore, using marker assisted selection to combine resistance alleles is a promising strategy for improving SNB resistance in wheat breeding. Indeed, the multi-locus haplotypes determined in this study provide markers for efficient tracking of these beneficial alleles in future wheat genetics and breeding activities.

Volatile Organic Compound Profiles From Wheat Diseases Are Pathogen-Specific and Can Be Exploited for Disease Classification.

Plants and fungi emit volatile organic compounds (VOCs) that are either constitutively produced or are produced in response to changes in their physico-chemical status. We hypothesized that these chemical signals could be utilized as diagnostic tools for plant diseases. VOCs from several common wheat pathogens in pure culture (Fusarium graminearum, Fusarium culmorum, Fusarium avenaceum, Fusarium poae, and Parastagonospora nodorum) were collected and compared among isolates of the same fungus, between pathogens from different species, and between pathogens causing different disease groups [Fusarium head blight (FHB) and Septoria nodorum blotch (SNB)]. In addition, we inoculated two wheat varieties with either F. graminearum or P. nodorum, while one variety was also inoculated with Blumeria graminis f.sp. tritici (powdery mildew, PM). VOCs were collected 7, 14, and 21 days after inoculation. Each fungal species in pure culture emitted a different VOC blend, and each isolate could be classified into its respective disease group based on VOCs with an accuracy of 71.4 and 84.2% for FHB and SNB, respectively. When all collection times were combined, the classification of the tested diseases was correct in 84 and 86% of all cases evaluated. Germacrene D and sativene, which were associated with FHB infection, and mellein and heptadecanone, which were associated with SNB infection, were consistently emitted by both wheat varieties. Wheat plants infected with PM emitted significant amounts of 1-octen-3-ol and 3,5,5-trimethyl-2-hexene. Our study suggests that VOC blends could be used to classify wheat diseases. This is the first step toward a real-time disease detection in the field based on chemical signatures of wheat diseases.

Septoria Nodorum Blotch of Wheat: Disease Management and Resistance Breeding in the Face of Shifting Disease Dynamics and a Changing Environment.

The fungus Parastagonospora nodorum is a narrow host range necrotrophic fungal pathogen that causes Septoria nodorum blotch (SNB) of cereals, most notably wheat (Triticum aestivum). Although commonly observed on wheat seedlings, P. nodorum infection has the greatest effect on the adult crop. It results in leaf blotch, which limits photosynthesis and thus crop growth and yield. It can also affect the wheat ear, resulting in glume blotch, which directly affects grain quality. Reports of P. nodorum fungicide resistance, the increasing use of reduced tillage agronomic practices, and high evolutionary potential of the pathogen, combined with changes in climate and agricultural environments, mean that genetic resistance to SNB remains a high priority in many regions of wheat cultivation. In this review, we summarize current information on P. nodorum population structure and its implication for improved SNB management. We then review recent advances in the genetics of host resistance to P. nodorum and the necrotrophic effectors it secretes during infection, integrating the genomic positions of these genetic loci by using the recently released wheat reference genome assembly. Finally, we discuss the genetic and genomic tools now available for SNB resistance breeding and consider future opportunities and challenges in crop health management by using the wheat-P. nodorum interaction as a model.

Genetic Structure of the Norwegian Parastagonospora nodorum Population.

The necrotrophic fungal pathogen Parastagonospora nodorum causes Septoria nodorum blotch (SNB), which is one of the dominating leaf blotch diseases of wheat in Norway. A total of 165 P. nodorum isolates were collected from three wheat growing regions in Norway from 2015 to 2017. These isolates, as well as nine isolates from other countries, were analyzed for genetic variation using 20 simple sequence repeat (SSR) markers. Genetic analysis of the isolate collection indicated that the P. nodorum pathogen population infecting Norwegian spring and winter wheat underwent regular sexual reproduction and exhibited a high level of genetic diversity, with no genetic subdivisions between sampled locations, years or host cultivars. A high frequency of the presence of necrotrophic effector (NE) gene SnToxA was found in Norwegian P. nodorum isolates compared to other parts of Europe, and we hypothesize that the SnToxA gene is the major virulence factor among the three known P. nodorum NE genes (SnToxA, SnTox1, and SnTox3) in the Norwegian pathogen population. While the importance of SNB has declined in much of Europe, Norway has remained as a P. nodorum hotspot, likely due at least in part to local adaptation of the pathogen population to ToxA sensitive Norwegian spring wheat cultivars.

Genetic mapping using a wheat multi-founder population reveals a locus on chromosome 2A controlling resistance to both leaf and glume blotch caused by the necrotrophic fungal pathogen Parastagonospora nodorum.

KEY MESSAGE: A locus on wheat chromosome 2A was found to control field resistance to both leaf and glume blotch caused by the necrotrophic fungal pathogen Parastagonospora nodorum. The necrotrophic fungal pathogen Parastagonospora nodorum is the causal agent of Septoria nodorum leaf blotch and glume blotch, which are common wheat (Triticum aestivum L.) diseases in humid and temperate areas. Susceptibility to Septoria nodorum leaf blotch can partly be explained by sensitivity to corresponding P. nodorum necrotrophic effectors (NEs). Susceptibility to glume blotch is also quantitative; however, the underlying genetics have not been studied in detail. Here, we genetically map resistance/susceptibility loci to leaf and glume blotch using an eight-founder wheat multiparent advanced generation intercross population. The population was assessed in six field trials across two sites and 4 years. Seedling infiltration and inoculation assays using three P. nodorum isolates were also carried out, in order to compare quantitative trait loci (QTL) identified under controlled conditions with those identified in the field. Three significant field resistance QTL were identified on chromosomes 2A and 6A, while four significant seedling resistance QTL were detected on chromosomes 2D, 5B and 7D. Among these, QSnb.niab-2A.3 for field resistance to both leaf blotch and glume blotch was detected in Norway and the UK. Colocation with a QTL for seedling reactions against culture filtrate from a Norwegian P. nodorum isolate indicated the QTL could be caused by a novel NE sensitivity. The consistency of this QTL for leaf blotch at the seedling and adult plant stages and culture filtrate infiltration was confirmed by haplotype analysis. However, opposite effects for the leaf blotch and glume blotch reactions suggest that different genetic mechanisms may be involved.

Genome-Wide Association Mapping of Resistance to Septoria Nodorum Leaf Blotch in a Nordic Spring Wheat Collection.

CORE IDEAS: First genome-wide association mapping of adult plant Septoria nodorum blotch resistance. Some adult plant resistance loci were shared with seedling resistance loci. Other adult plant resistance loci were significant across environments. Resistant haplotypes were identified, which can be used for breeding. Parastagonospora nodorum is the causal agent of Septoria nodorum leaf blotch (SNB) in wheat (Triticum aestivum L.). It is the most important leaf blotch pathogen in Norwegian spring wheat. Several quantitative trait loci (QTL) for SNB susceptibility have been identified. Some of these QTL are the result of underlying gene-for-gene interactions involving necrotrophic effectors (NEs) and corresponding sensitivity (Snn) genes. A collection of diverse spring wheat lines was evaluated for SNB resistance and susceptibility over seven growing seasons in the field. In addition, wheat seedlings were inoculated and infiltrated with culture filtrates (CFs) from four single spore isolates and infiltrated with semipurified NEs (SnToxA, SnTox1, and SnTox3) under greenhouse conditions. In adult plants, the most stable SNB resistance QTL were located on chromosomes 2B, 2D, 4A, 4B, 5A, 6B, 7A, and 7B. The QTL on chromosome 2D was effective most years in the field. At the seedling stage, the most significant QTL after inoculation were located on chromosomes 1A, 1B, 3A, 4B, 5B, 6B, 7A, and 7B. The QTL on chromosomes 3A and 6B were significant both after inoculation and CF infiltration, indicating the presence of novel NE-Snn interactions. The QTL on chromosomes 4B and 7A were significant in both seedlings and adult plants. Correlations between SnToxA sensitivity and disease severity in the field were significant. To our knowledge, this is the first genome-wide association mapping study (GWAS) to investigate SNB resistance at the adult plant stage under field conditions.

Understanding Yield Loss and Pathogen Biology to Improve Disease Management: Septoria Nodorum Blotch - A Case Study in Wheat.

The estimated potential yield losses caused by plant pathogens is up to 16% globally and most research in plant pathology aims to reduce yield loss in our crops directly or indirectly. Yield losses caused by a certain disease depend not only on disease severity, but also on the weather factors, the pathogen's aggressiveness, and the ability of the crop to compensate for reduced photosynthetic area. The yield loss-disease relationship in a certain host-pathogen system might therefore change from year to year, making predictions for yield loss very difficult at the regional or even at the farmer's level. However, estimating yield losses is essential to determine disease management thresholds at which acute control measures such as fungicide applications, or strategic measures such as crop rotation or use of resistant cultivars are economically and environmentally sensible. Legislation in many countries enforces implementation of integrated pest management (IPM), based on economic thresholds at which the costs due to a disease justify the costs for its management. Without a better understanding of the relationship between disease epidemiology and yield loss, we remain insufficiently equipped to design adequate IPM strategies that will be widely adapted in agriculture. Crop loss studies are resource demanding and difficult to interpret for one particular disease, as crops are usually not invaded by only one pest or pathogen at a time. Combining our knowledge on disease epidemiology, crop physiology, yield development, damage mechanisms involved, and the effect of management practices can help us to increase our understanding of the disease-crop loss relationship. The main aim of this paper is to review and analyze the literature on a representative host-pathogen relationship in an important staple food crop to identify knowledge gaps and research areas to better assess yield loss and design management strategies based on economic thresholds.

Identification of quantitative trait loci associated with resistance to net form net blotch in a collection of Nordic barley germplasm.

KEY MESSAGE: Association mapping of resistance to Pyrenophora teres f. teres in a collection of Nordic barley germplasm at different developmental stages revealed 13 quantitative loci with mostly small effects. Net blotch, caused by the necrotrophic fungus Pyrenophora teres, is one of the major diseases in barley in Norway causing quantitative and qualitative yield losses. Resistance in Norwegian cultivars and germplasm is generally insufficient and resistance sources have not been extensively explored yet. In this study, we mapped quantitative trait loci (QTL) associated with resistance to net blotch in Nordic germplasm. We evaluated a collection of 209 mostly Nordic spring barley lines for reactions to net form net blotch (NFNB; Pyrenophora teres f. teres) in inoculations with three single conidia isolates at the seedling stage and in inoculated field trials at the adult stage in 4 years. Using 5669 SNP markers genotyped with the Illumina iSelect 9k Barley SNP Chip and a mixed linear model accounting for population structure and kinship, we found a total of 35 significant marker-trait associations for net blotch resistance, corresponding to 13 QTL, on all chromosomes. Out of these QTL, seven conferred resistance only in adult plants and four were only detectable in seedlings. Two QTL on chromosomes 3H and 6H were significant during both seedling inoculations and adult stage field trials. These are promising candidates for breeding programs using marker-assisted selection strategies. The results elucidate the genetic background of NFNB resistance in Nordic germplasm and suggest that NB resistance is conferred by a number of genes each with small-to-moderate effects, making it necessary to pyramid these genes to achieve sufficient levels of resistance.

Mapping of quantitative trait loci associated with resistance to net form net blotch (Pyrenophora teres f. teres) in a doubled haploid Norwegian barley population.

Barley net blotch caused by the necrotrophic fungus Pyrenophora teres is a major barley disease in Norway. It can cause grain shriveling and yield losses, and resistance in currently grown cultivars is insufficient. In this study, a set of 589 polymorphic SNP markers was used to map resistance loci in a population of 109 doubled haploid lines from a cross between the closely related Norwegian cultivars Arve (moderately susceptible) and Lavrans (moderately resistant). Resistance to three net form net blotch (P. teres f. teres) single spore isolates was evaluated at the seedling stage in the greenhouse and at the adult plant stage under field conditions during three years. Days to heading and plant height were scored to assess their influence on disease severity. At the seedling stage, three to four quantitative trait loci (QTL) associated with resistance were found per isolate used. A major, putatively novel QTL was identified on chromosome 5H, accounting for 23-48% of the genetic variation. Additional QTL explaining between 12 and 16.5% were found on chromosomes 4H, 5H, 6H and 7H, with the one on 6H being race-specific. The major QTL on 5H was also found in adult plants under field conditions in three years (explaining up to 55%) and the 7H QTL was found in field trials in one year. Additional adult plant resistance QTL on 3H, 6H and 7H were significant in single years. The resistance on chromosomes 3H, 5H, 6H and 7H originates from the more resistant parent Lavrans, while the resistance on 4H is conferred by Arve. The genetic markers associated with the QTL found in this study will benefit marker-assisted selection for resistance against net blotch.

Role of the cyclic lipopeptide massetolide A in biological control of Phytophthora infestans and in colonization of tomato plants by Pseudomonas fluorescens.

Pseudomonas strains have shown promising results in biological control of late blight caused by Phytophthora infestans. However, the mechanism(s) and metabolites involved are in many cases poorly understood. Here, the role of the cyclic lipopeptide massetolide A of Pseudomonas fluorescens SS101 in biocontrol of tomato late blight was examined. Pseudomonas fluorescens SS101 was effective in preventing infection of tomato (Lycopersicon esculentum) leaves by P. infestans and significantly reduced the expansion of existing late blight lesions. Massetolide A was an important component of the activity of P. fluorescens SS101, since the massA-mutant was significantly less effective in biocontrol, and purified massetolide A provided significant control of P. infestans, both locally and systemically via induced resistance. Assays with nahG transgenic plants indicated that the systemic resistance response induced by SS101 or massetolide A was independent of salicylic acid signalling. Strain SS101 colonized the roots of tomato seedlings significantly better than its massA-mutant, indicating that massetolide A was an important trait in plant colonization. This study shows that the cyclic lipopeptide surfactant massetolide A is a metabolite with versatile functions in the ecology of P. fluorescens SS101 and in interactions with tomato plants and the late blight pathogen P. infestans.

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.

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.

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 Ontogenic Resistance upon Establishment and Growth of Uncinula necator on Grape Berries.

ABSTRACT Grape berries become resistant to powdery mildew early in development and are nearly immune to infection within 4 weeks after bloom. In this study, ontogenic resistance did not reduce attachment, germination, or appressorium formation of Uncinula necator on 3- to 4-week-old berries of Vitis vinifera 'Chardonnay' or 3-week-old berries of V. labruscana 'Concord'. Pathogen ingress halted at the cuticle before formation of a penetration pore. As berries aged, hyphal elongation and colony growth slowed until finally no secondary hyphae formed on fully resistant berries. More appressoria formed per unit of hyphal length as berries aged, indicating that failure to penetrate older berries led to increased attempts to penetrate resistant fruit. Additionally, hyphae within the colonies began to die as berries aged. Finally, the number of degree-hours between germination and sporulation of the colony (latent period) increased and sporophore density decreased with berry age at time of inoculation. Thus, ontogenic resistance both slows, and eventually halts disease development on grape berries, and limits the likelihood of spread by reducing absolute supply of conidia and delaying their formation. It furthermore has a consistent, stable, and predictable impact on grape powdery mildew and operates in a similar fashion and to a similar degree in both V. labruscana and V. vinifera, although at a slightly earlier phenological stage in V. labruscana.

Ontogenic resistance and plant disease management: a case study of grape powdery mildew.

ABSTRACT A fundamental principle of integrated pest management is that actions taken to manage disease should be commensurate with the risk of infection and loss. One of the less-studied factors that determines this risk is ontogenic, or age-related resistance of the host. Ontogenic resistance may operate at the whole plant level or in specific organs or tissues. Until recently, grape berries were thought to remain susceptible to powdery mildew (Uncinula necator) until late in their development. However, the development of ontogenic resistance is actually quite rapid in berries, and fruit become nearly immune to infection within 4 weeks after fruit set. Our objective was to determine how and at what stage the pathogen was halted in the infection process on ontogenically resistant berries. Adhesion of conidia, germination, and appressorium formation were not impeded on older berries. However, once berries were approximately 3 weeks old and older, few germlings were able to form secondary hyphae. Ontogenically resistant berries responded rapidly to infection by synthesis of a germin-like protein that had been previously shown to play a role in host defense against barley powdery mildew. On susceptible berries, cell discoloration around penetration sites indicated the oxidation of phenolic compounds; a process that was followed by localized cell death. However, the pathogen was still able to infect such cells prior to their death, continue secondary growth, and thereby colonize young berries. Formation of papillae was not involved in the differential resistance mechanism of older berries. In susceptible berries, papillae formed frequently at infection sites but did not always contain the pathogen, whereas in resistant berries, the pathogen was always halted prior to the formation of papillae. The host defense, which conditions ontogenic resistance, operates in the earliest stages of the infection process, in the absence of gross anatomical barriers, prior to the formation of a functional haustorium and prior to the development of a conspicuous penetration pore. We also found that diffuse powdery mildew colonies that were not visible in the field predisposed berries to bunch rot by Botrytis cinerea, increased the levels of infestation by spoilage microorganisms, and substantially degraded wine quality. Our improved understanding of the nature, causes, and stability of ontogenic resistance in the grapevine/ powdery mildew system has supported substantial changes in how fungicides are used to control the disease. Present applications are more focused on the period of maximum fruit susceptibility instead of following a calendar-based schedule. This has improved control, reduced losses, and in many cases reduced the number of fungicide applications required to suppress the disease. Particularly where fungicides are deployed in a programmatic fashion and ontogenic resistance is dynamic, there may be equivalent improvements to be made in other hostpathogen systems through studies of how host susceptibility changes through time.