The reliability of leaf bioassays for predicting disease resistance on fruit: a case study on grapevine resistance to downy and powdery mildew

This study was designed to assess the reliability of grapevine leaf bioassays for predicting disease resistance on fruit in the field. The efficacy of various grapevine quantitative trait loci (QTLs) for conferring resistance to downy and powdery mildew was evaluated in bioassays and in a 2‐year field experiment for downy mildew. The resistance genes studied were inherited from Muscadinia rotundifolia (Rpv1 and Run1) and from American Vitis species through cv. Regent (QTLRgP and QTLRgD). In bioassays, genotypes carrying Run1 blocked powdery mildew development at early stages. Genotypes combining Run1 with QTLRgP displayed no greater level of resistance. For downy mildew, genotypes carrying Rpv1 and/or QTLRgD were more resistant than the susceptible cv. Merlot, and showed a high level of leaf resistance in the field (<10% severity). Disease levels on bunches were much higher than those on leaves, with a high variability between Rpv1 genotypes (1–48%). A Bayesian decision theory framework predicted that an OIV‐452 threshold of 5 in leaf bioassays allowed accurate selection of grapevine genotypes (P = 0·83) with satisfactory disease severity on bunches. Therefore, this study validates that the use of early bioassays on leaves, as currently performed by grapevine breeders, ensures a satisfactory level of resistance to downy mildew of bunches in the field.     

The role of the abaxial leaf surface waxes of Lolium spp. in resistance to Erysiphe graminis

Field and glasshouse observations of Lolium spp. grasses indicated that the lower, abaxial, leaf surface was rarely infected by powdery mildew ( Erysiphe graminis ) even when the upper, adaxial, surface was densely colonized. Experiments showed that conidia of two strains of E. graminis , one from Lolium and one from Avena , germinated equally well on both surfaces of Lolium and Avena leaves, but that the subsequent growth and development of germlings was impaired on the lower surface of Lolium leaves, so that most formed only multiple short germ tubes or an abnormal long tube, and only c. 25% or fewer formed infection structures. This contributes to the apparent resistance of the lower Lolium leaf surface to powdery mildew and may help to explain why the disease is relatively unimportant in UK ryegrass crops, since infection structures develop at a high frequency on only 50% of the leaf area, i.e. the upper surface. Scanning electron microscopy showed that the epicuticular waxes on the lower Lolium leaf surface form amorphous sheets. This contrasts with the crystalline plate waxes seen on the upper surface of Lolium leaves and on both surfaces of oat leaves. However, when the lower Lolium leaf surface was washed with chloroform to remove epicuticular wax, normal germling and infection structure development was obtained on the wax-free surface. This suggests that the sheet waxes prevent the pathogen gaining access to features of the cuticular membrane which trigger normal germling development.     

Downy mildew resistance evaluation in 28 grapevine hybrids promising for breeding programs in Trentino region (Italy)

Downy mildew is a major grapevine disease caused by the biotrophic oomycete, Plasmopara viticola. Numerous disease resistance studies of diverse Vitis germplasm have been previously carried out to identify downy mildew resistance sources; however, ratings were mainly reported using leaf disc in vitro testing and foliage field assessment, or upon leaf and cluster field evaluations. In the current study, 28 grapevine hybrid cultivars were screened using leaf disc bioassay, for disease resistance characterization of both existing and wild-collected materials. 16 hybrids were identified as highly resistant or resistant, and will serve as relevant resistance donors in future pre-breeding and breeding programs. All grapevine hybrids were evaluated for foliar and cluster downy mildew resistance in an untreated field trial over three successive years. This study showed that the leaf disc bioassay provided some information on the resistance level of the genotypes under scrutiny, but it was a weak predictor of their resistance level under field conditions on leaves and even more on bunches. These findings are relevant to future applications in both traditional and marker-assisted breeding programs which promote sustainable viticulture.     

A standardised method for the quantitative analysis of resistance to grapevine powdery mildew

Powdery mildew caused by Erysiphe necator is one of the most important diseases affecting grapevine (Vitis vinifera, L.). Control of this pathogen is based on the use of fungicides, which cause environmental damage and increase production costs. A cost-effective and environmentally friendly alternative to control the disease relies on using resistant varieties. While most V. vinifera cultivars are susceptible to powdery mildew, several species belonging to the Vitaceae have been described as resistant. Several loci for resistance to grapevine powdery mildew have been identified through genetic analysis of segregating populations derived from different resistance sources. Identifying quantitative trait loci (QTL) with minor effects on the resistance may prove valuable in a strategy of pyramiding, which aims at increasing the durability of the resistance. However, current methods for evaluation of resistance either do not take into account quantitative variations, or, if they do, are not adapted to large sample sets. Here we develop a method for the analysis of quantitative resistance to grapevine powdery mildew in large populations. We devised a semi-quantitative resistance scale and confirmed the usefulness of a cell counter to quantify sporulation. We compared three inoculation methods and identified dry inoculation using a settling tower as the one giving the best infection. Finally, we confirmed the value of the method by applying it to a set of plants segregating for resistance to E. necator. Using the method described here for the quantitative analysis of the resistance to powdery mildew will prove valuable for breeding for durable resistance.     

Breakdown of Pl2, a major gene of resistance to apple powdery mildew, in a French experimental orchard

Pl2 , a major resistance gene to powdery mildew originating from Malus zumi , is mainly used in apple-breeding programmes. In an experimental orchard in France, an increase of susceptibility to powdery mildew was observed on apple genotypes carrying Pl2 . This increase of susceptibility could not be explained by an effect of the age of the trees, or by an effect of the amount of inoculum on expression of the resistance Pl2 . It was demonstrated by tests of pathogenicity in controlled conditions that isolates of Podosphaera leucotricha sampled in this orchard were virulent to Pl2 . The appearance of virulent isolates within 6 years after planting apple genotypes selected for the presence of Pl2 resulted in a breakdown of the resistance. Ten years after planting, the percentage of genotypes that were still resistant to powdery mildew varied between 2 and 56%, depending on the progenies. This suggested that additional quantitative resistance genes may influence the durability of the resistance Pl2 .     

云南蔷薇属部分种质资源对白粉病的抗性鉴定

对14个云南蔷薇属种质资源（包括7个栽培品种和7个野生种）进行白粉病抗性的离体鉴定和田间鉴定,其中免疫品种（种）2个,中抗品种（种）8个,中感品种1个,高感品种（种）3个。白粉病离体鉴定与田间鉴定两种方法的抗性评价结果完全一致,表明该方法可作为白粉病抗性快速鉴定的方法。     

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.     

Variation in adult plant resistance to powdery mildew in spring oats under field and laboratory conditions

Ten oat genotypes were assessed for adult plant resistance (APR) to powdery mildew ( Erysiphe graminis f.sp. avenae ) in small field plots and on detached leaf segments in the laboratory using glasshouse-grown plants. Significant genotype differences were observed in the field and on leaf segments for latent period, infection frequency and spore production, components used to assess APR. The genotype OM1711, which was selected because it expressed a higher level of mildew resistance than its most resistant parent, cv. Maldwyn, showed this transgressive resistance in both field and laboratory tests. By contrast, the genotype OM1387, also selected for high mildew resistance, only showed this enhancement in the field. It is concluded that quantitative assessment of mildew in detached leaves is not sufficiently precise to be used alone in selecting segregants for high levels of APR, and that field data are also needed.     

Correlations of Morphological, Anatomical, and Chemical Features of Grape Berries with Resistance to Botrytis cinerea

ABSTRACT Resistance of mature berries of grapevine cultivars and selections to postharvest infection by Botrytis cinerea was assessed. Little or no resistance existed in most popular table grape Vitis vinifera cultivars, except in moderately resistant 'Emperor' and 'Autumn Black'. Highly resistant grapes were V. rotundifolia, V. labrusca, or other complex hybrids. Morphological, anatomical, and chemical characteristics of 42 genetically diverse cultivars and selections with various levels of resistance to B. cinerea were examined to determine which features were associated with resistance. We quantified the (i) density of berries within a cluster; (ii) number of pores and lenticels on the berry surface; (iii) thickness and number of cell layers in the epidermis and external hypodermis; (iv) amount of cuticle and wax; (v) berry skin protein content; (vi) total phenolic content of the skin before and after B. cinerea inoculation; and (vii) catechin and trans- and cis-resveratrol contents of the skin before and after inoculation. The number of pores was negatively correlated with resistance. Highly resistant cultivars had few or no pores in the berry surface. The number and thickness of epidermal and hypodermal cell layers and cuticle and wax contents were positively correlated with resistance. Other characteristics evaluated were not associated with resistance. trans-Resveratrol and cis-resveratrol were induced by B. cinerea inoculation only in sensitive and moderately resistant cultivars and selections.     

Role of stilbenes in the resistance of grapevine to powdery mildew

Stilbene phytoalexins are identified as defence response in pathogen–grapevine interactions, but little information is available on the role of stilbenes on Erysiphe necator, causal agent of grapevine powdery mildew. Analysis of stilbenes in artificially infected leaf discs from susceptible to highly resistant cultivars was performed and compared to the development of the pathogen. Results indicate that stilbene synthesis is confined in infected cells, penetrated by an appressorium–peg. Stilbene amounts expressed by infection site allow discriminating susceptible and resistant cultivars. Highest viniferins concentrations on resistant cultivars are in correspondence with the observed inhibition of the pathogen growth. The analysis of stilbenes at the infection site and viniferins accumulation in grapevine defence reaction is discussed.     

Wax matters: absence of very‐long‐chain aldehydes from the leaf cuticular wax of the glossy11 mutant of maize compromises the prepenetration processes of Blumeria graminis

Conidial germination and differentiation, the so‐called prepenetration processes, of the barley powdery mildew fungus (Blumeria graminis f.sp. hordei) are triggered in vitro by very‐long‐chain aldehydes, minor constituents of barley leaf wax. However, until now it has not been demonstrated that these cuticle‐derived molecules also play a significant role in the initiation and promotion of the fungal prepenetration processes in vivo, on the surface of a living plant leaf. In the maize (Zea mays) wax mutant glossy11, which is completely devoid of cuticular very‐long‐chain aldehydes, germination and appressorial differentiation of B. graminis were strongly impeded. Spraying the mutant leaf surface with aldehyde‐containing wild‐type wax or pure n‐hexacosanal (C₂₆‐aldehyde) fully restored fungal prepenetration, whereas maize wild‐type leaf surfaces coated with n‐docosanoic acid exhibited reduced conidial germination rates of 23%, and only 5% of the conidia differentiated infection structures. In vitro studies were performed to further corroborate the extensive prevention of fungal germination and differentiation in response to artificial surfaces coated with aldehyde‐deficient maize wax. Because of its phenotype affecting the B. graminis prepenetration processes, the glossy11 mutation of maize may become a valuable molecular target and genetic tool that could provide a means of developing basal powdery mildew resistance in the globally important crops wheat and barley.     

The epidemiology of powdery mildew on concord grapes

ABSTRACT Vitis labruscana 'Concord' is a grape cultivar widely grown in the United States for processing into juice and other grape products. Concord grapes are sporadically but sometimes severely damaged by the grape powdery mildew pathogen, Uncinula necator. Although the foliage is often reported to be moderately resistant to powdery mildew, severe fruit infection occurs in some years. We observed the seasonal development of powdery mildew on leaves, rachises, and berries of unsprayed Concord grapevines. Inoculations of flower and fruit clusters revealed a brief period of berry susceptibility and a protracted period of rachis susceptibility. The rachis remained highly susceptible to infection, and the severity of rachis infection increased throughout the growing season until the rachis formed a periderm shortly before harvest. In contrast, berries were nearly immune to infection within 2 weeks after fruit set. Rachis and berry infections were detected before the disease was observed on foliage, and the incidence of rachis and berry infection often exceeded disease incidence observed on foliage until after fruit acquired substantial ontogenic resistance. Excellent control of fruit infection, and adequate control of leaf infection, was achieved by two fungicide applications targeted at the peak period of fruit susceptibility. Although Concord is thought to be moderately resistant to powdery mildew, the rachis is highly susceptible, and may be the avenue by which prebloom infections make their way onto the developing fruit. Late-season infection of the rachis neither spread to the fruit, nor did it cause fruit to drop prematurely, and may be of little economic consequence on fruit destined for processing. Although fruit of V. vinifera cultivars have been reported to remain susceptible to infection until berry sugar levels reach 8 to 15%, Concord fruit become nearly immune to infection nearly 6 weeks before this stage of development. Because powdery mildew does not become conspicuous on foliage until late summer, it is generally regarded as a late-season problem on Concord grapes, and previous management programs have reflected this belief. However, the greatest contribution to control of fruit infection is due to fungicides applied during the peak period of fruit susceptibility, from bloom until shortly after fruit set, long before the disease is observed on foliage.     

Factors affecting the activities of nitrophenol fungicides IV. The influence of hydrogen bonding

The control of powdery mildew on marrows (caused by Sphaerotheca fuliginea) achieved with 4-(1-substituted alkyl)-2,6-dinitrophenols, 2-(1-substituted alkyl)-4,6-dinitrophenols, 4-(1-substituted alkyl)-2,6-dinitrophenyl crotonates, cycloalkyldinitrophenols, 2-cyclohexyl-halogeno-nitrophenols and 4-cyclohexyl-2-substituted-6-nitrophenols is discussed. Certain combinations of partition and intra- and intermolecular hydrogen bonding capacities lead to vapour phase and/or zonal activity or to penetration of the leaf surface followed by possible translocation, phytotoxicity or translaminar activity. The capacity of the phenol to hydrogen bond to the leaf cuticular waxes has a considerable influence upon the fate of these materials when used as foliar fungicides.     

黄瓜白粉病菌在不同抗性黄瓜材料上的侵染过程

白粉病是黄瓜生产中发生普遍,危害严重的主要病害之一。pm5.1和PM5.2是黄瓜上的2个白粉病抗性位点,本文对7份不同抗病基因型的黄瓜自交系进行了黄瓜白粉病抗性鉴定,并开展了黄瓜白粉病菌侵染过程的研究,对侵染后12、24、72 h的萌发率、菌丝形成率及菌落形成率等进行了分析。结果表明,当基因型为PM5.1PM5.1 pm5.2pm5.2时,黄瓜病情指数最高,表现为高感白粉病;当基因型为PM5.1PM5.1PM5.2 PM5.2和pm5.1pm5.1pm5.2pm5.2时,表现为中感白粉病;当基因型为pm5.1pm5.1PM5.2PM5.2时黄瓜自交系病情指数最低,表现为抗白粉病。分生孢子在抗、感黄瓜自交系植株叶片上均能萌发,但只能在感病黄瓜材料上完成整个无性生长周期,产生分生孢子。此外,黄瓜白粉病菌分生孢子在感病材料上的萌发率、菌丝形成率及菌落形成率均高于抗病材料。     

Comparative analysis of powdery mildew development on leaves, seedlings and flower panicles of different genotypes of cashew

Resistance of cashew ( Anacardium occidentale ) genotypes to powdery mildew ( Oidium anacardii ) was assessed on detached leaves, seedlings and flower panicles. Leaves were more susceptible before the development of a waxy cuticle, which confers a dull appearance. Differences in susceptibility/resistance were observed on detached leaves inoculated in the laboratory. Some genotypes supported dense growth of mildew, whereas on others only slight infection was observed under comparable conditions. Mature leaves were immune to colonization. In the field the rate of mildew infection, as estimated from disease progress curves on both seedlings and flower panicles, varied significantly between genotypes allowing their categorization into susceptible and partially resistant clones. A similar rank order for degree of susceptibility/resistance was determined from studies on detached leaves, seedlings and flower panicles. Brazilian introductions of A. occidentale and the AZA2 and AC6 genotypes proved to be partially resistant, while AM6, ATA19 and AIN62 were highly susceptible. The relationship between leaf and flower mildew infection showed positive and highly significant correlations. It is concluded that each of the three approaches to the assessment of disease susceptibility can be used to screen genotypes against mildew attack. Screening in the laboratory provides a rapid indication of potential resistance providing that leaves are compared at the same stages of maturity.     

Differential expression of resistance to powdery mildew incited by race 1 or 2 ofsphaerotheca fuliginea inCucumis melo genotypes at various stages of plant development

Thirty-nine genotypes ofCucumis melo (plant introduction entries, open-pollinated cultivars and F₁ hybrids) were evaluated for resistance to powdery mildew under either natural field conditions or artificial inoculation in growth chambers at the cotyledonary stage and the 2-true-leaf stage. Results confirmed that susceptibility in cotyledons was not necessarily associated with susceptibility in either true leaves in growth chambers or adult plants in the field. However, resistance at the 2-true-leaf stage in growth chambers was highly correlated with resistance of field-grown plants. Results also showed that 20 muskmelon genotypes resistant to race 1 at the cotyledonary stage were also resistant at the 2-leaf-stage and as adult plants in the field. The same was true for ten genotypes with race 2 inoculations. Because muskmelon genotypes expressing resistance in cotyledons were also resistant in true leaves in growth chambers or the field, the use of plants at the cotyledonary stage is recommended for screening for powdery mildew resistance caused by race 1 or race 2 ofS. fuliginea. When cotyledons are susceptible, screening should be done at the 2-true-leaf stage.     

Genetics of resistance to wheat leaf rust, stem rust, and powdery mildew in Aegilops sharonensis

Aegilops sharonensis (Sharon goatgrass) is a wild relative of wheat and a rich source of genetic diversity for disease resistance. The objectives of this study were to determine the genetic basis of leaf rust, stem rust, and powdery mildew resistance in A. sharonensis and also the allelic relationships between genes controlling resistance to each disease. Progeny from crosses between resistant and susceptible accessions were evaluated for their disease reaction at the seedling and/or adult plant stage to determine the number and action of genes conferring resistance. Two different genes conferring resistance to leaf rust races THBJ and BBBB were identified in accessions 1644 and 603. For stem rust, the same single gene was found to confer resistance to race TTTT in accessions 1644 and 2229. Resistance to stem rust race TPMK was conferred by two genes in accessions 1644 and 603. A contingency test revealed no association between genes conferring resistance to leaf rust race THBJ and stem rust race TTTT or between genes conferring resistance to stem rust race TTTT and powdery mildew isolate UM06-01, indicating that the respective resistance genes are not linked. Three accessions (1644, 2229, and 1193) were found to carry a single gene for resistance to powdery mildew. Allelism tests revealed that the resistance gene in accession 1644 is different from the respective single genes present in either 2229 or 1193. The simple inheritance of leaf rust, stem rust, and powdery mildew resistance in A. sharonensis should simplify the transfer of resistance to wheat in wide crosses.     

A single dominant locus, ren4, confers rapid non-race-specific resistance to grapevine powdery mildew

In the present study we screened the progeny of Vitis vinifera × V. romanetii populations segregating for resistance to powdery mildew and determined the presence of a single, dominant locus, Ren4, conferring rapid and extreme resistance to the grapevine powdery mildew fungus Erysiphe necator. In each of nine Ren4 pseudo-backcross 2 (pBC(2)) and pBC(3) populations (1,030 progeny), resistance fit a 1:1 segregation ratio and overall segregated as 543 resistant progeny to 487 susceptible. In full-sib progeny, microscopic observations revealed the reduction of penetration success rate (as indicated by the emergence of secondary hyphae) from 86% in susceptible progeny to below 10% in resistant progeny. Similarly, extreme differences were seen macroscopically. Ratings for Ren4 pBC(2) population 03-3004 screened using natural infection in a California vineyard and greenhouse and using artificial inoculation of an aggressive New York isolate were fully consistent among all three pathogen sources and environments. From 2006 to 2010, Ren4 pBC(2) and pBC(3) vines were continuously screened in California and New York (in the center of diversity for E. necator), and no sporulating colonies were observed. For population 03-3004, severity ratings on leaves, shoots, berries, and rachises were highly correlated (R(2) = 0.875 to 0.996) in the vineyard. Together, these data document a powdery mildew resistance mechanism not previously described in the Vitaceae or elsewhere, in which a dominantly inherited resistance prevents hyphal emergence and is non-race-specific and tissue-independent. In addition to its role in breeding for durable resistance, Ren4 may provide mechanistic insights into the early events that enable powdery mildew infection.     

2002年部分麦区小麦白粉病菌对三唑酮的抗药性监测及苯氧菌酯敏感基线的建立

 采用拌种离体叶段法测定了2002年采自北京、江苏、山东、山西、河北、新疆和四川7省市部分麦区的109个小麦白粉病菌菌株对三唑酮和苯氧菌酯的敏感性,结果表明小麦白粉病菌群体对三唑酮敏感性EC₅₀平均值是63.70 μg/mL,平均抗性水平为30.48倍,最高抗性水平达142.97倍。其中江苏、山东、四川三地的菌株抗药性水平明显高于河北、北京、山西和新疆。同时测得病菌群体对苯氧菌酯的敏感基线EC₅₀值为85.82μg/mL,且白粉病菌对甲氧基丙烯酸酯类杀菌剂苯氧菌酯和三唑类杀菌剂三唑酮之间无交互抗性。该结果可为这两类杀菌剂在生产上的合理应用和抗药性治理提供依据。     

Resistance to powdery mildew (Blumeria graminis f.sp. avenae) in oat seedlings and adult plants

In this work, 165 Avena sativa and Avena byzantina accessions were screened for resistance to powdery mildew caused by Blumeria graminis f.sp. avenae and the defence mechanisms of resistant plants were further characterized. Ten resistant and moderately resistant accessions were selected according to macroscopic assessment. A detailed histological study of selected genotypes showed a range of defence mechanisms, acting alone or in combination, that impeded fungal development at different stages. Since the resistance observed in the collection was scarce, a study of adult plant resistance was carried out in 45 genotypes selected from field trials. Nine oat landraces and two commercial varieties showed very high levels of adult plant resistance. A detailed study of the components of the adult plant resistance revealed a high increase of penetration and post‐haustorial resistance in the fifth compared to the first leaves. Identification of the resistance sources and characterization of underlying defence mechanisms will be useful for future breeding programmes and for further cellular and molecular studies to unravel the genetic basis of resistance, in this species in particular and in cereal–powdery mildew interactions in general.