A host-pathogen simulation model: powdery mildew of grapevine

An epidemiological model simulating the growth of a single grapevine stock coupled to the dispersal and disease dynamics of the airborne conidia of the powdery mildew pathogen Erysiphe necator was developed. The model input variables were either climatic (temperature, wind speed and direction) or related to the pathogen (location and onset of primary infection). The environmental input variables dictated plant growth and pathogen spread (latent period, infection, lesion growth, conidial spore production and release). Input parameters characterized the crop production system, the growth conditions and the epidemiological characteristics of the pathogen. Output described, at each time step, number, age and pattern of healthy and infected organs, infected and infectious leaf area and aerial density of spores released. Validation of the model was achieved by comparing model output with experimental data for epidemics initiated at different times of host growth. Epidemic behaviour for two contrasting years of crop development and 7 phenological stages at the time of primary infection (PI) was examined. For PI occurring at day 115 a vine with late budbreak (1998) showed 58% of primary leaves diseased at flowering compared with only 19% for a vine with early budbreak (2003). Depending on the phenological stage at PI (1–4 leaves), the proportion of diseased primary leaves decreased from 42% to 6% at flowering. Simulations suggested that differences resulted from the interplay between the timing of the first sporulation event, the phenological stage at the time of initial infection, and the age structure and spatial distribution of the leaf population.     

Chitosan oligomers and copper sulfate induce grapevine defense reactions and resistance to gray mold and downy mildew

ABSTRACT Chitosan (CHN), a deacetylated derivative of chitin, was shown to be efficient in promoting plant defense reactions. CHN oligomers of different molecular weight (MW) and degree of acetylation (DA) triggered an accumulation of phytoalexins, trans- and cis-resveratrol and their derivatives epsilon-viniferin and piceid, in grapevine leaves. Highest phytoalexin production was achieved within 48 h of incubation with CHN at 200 mug/ml with an MW of 1,500 and a DA of 20% (CHN1.5/20), while oligomers with greater MW were less efficient, indicating that a specific MW threshold could be required for phytoalexin response. Treatment of grapevine leaves by highly active CHN1.5/20 also led to marked induction of chitinase and beta-1,3-glucanase activities. CHN1.5/20 applied together with copper sulfate (CuSO(4)) strongly induced phytoalexin accumulation. CuSO(4) alone, especially at low concentrations also elicited a substantial production of phytoalexins in grapevine leaves. Evidence is also provided that CHN1.5/20 significantly reduced the infection of grapevine leaves by Botrytis cinerea and Plasmopara viticola, and in combination with CuSO(4) conferred protection against both pathogens.     

Resistance assessment of grapevine leaves to downy mildew with sporulation area scoring

The use of resistant individuals obtained through a breeding program, aimed at decreasing chemical treatments, is one of the most promising strategies for control of downy mildew caused by Plasmopara viticola in grapevine. In this study, 869 hybrid grape individuals, obtained by crossing the resistant Regent grape cultivar with sensitive Alphonse Lavallée, were tested after artificial inoculation of detached leaves by drops of the pathogen inoculum. Sporulation severity, as visual evaluation of sporulation density including necrosis, and sporulation incidence were scored to assess resistance level, then the sporulation area (mm²) was measured for each individual. The criterium of sporulation incidence could not describe all resistance levels. Sporulation area made it possible to measure the entire area in detail for how the pathogen spread at the inoculation site. Sporulation area was not fully consistent with sporulation severity. In this context, a new scale for sporulation area was developed to determine resistance level to the pathogen by taking into account the distribution of sporulation area values within those obtained from sporulation severity scales. Six categories ranging from 0 to 0.05 mm² (extremely resistant-ER) to ≥15.1 mm² (extremely sensitive-ES) were defined. 43 of the 869 individuals tested through the three evaluation criteria, in which no sporulation was observed, and 18 additional individuals with sporulation area of <0.05 mm² were scored in the ER group. Microscopic analysis confirmed findings for the ER group and other resistance levels. The authors suggest that the scale of sporulation area will be useful for assessment of resistance to downy mildew on grapevine leaves.

     

Endophytic Beauveria bassiana activates expression of defence genes in grapevine and prevents infections by grapevine downy mildew Plasmopara viticola

Fungal entomopathogens like Beauveria bassiana (Ascomycota: Hypocreales) are known as antagonists of insects with multiple functional and ecological roles, and have attracted increased attention as biocontrol agents in integrated pest management programmes. For some crop plants, it has been proven that endophytic B. bassiana, besides its entomopathogenic habit, can provide protection against plant pathogens or limit their damaging effects. However, for grapevine, limited knowledge is available on the influence of endophytic B. bassiana on fungal pathogens and about the mechanisms underlying putative protection effects. This study assessed the protective potential of endophytic B. bassiana against grapevine downy mildew Plasmopara viticola in greenhouse experiments. Three and seven days after a B. bassiana treatment, potted grapevine plants were inoculated with P. viticola and the evolving disease severity was assessed. Disease severity was significantly reduced in B. bassiana-treated plants compared to control plants, depending on the age of leaves. Furthermore, a microarray and an RT-qPCR analysis were performed to work out fundamental aspects of genes involved in the interaction between grapevine and B. bassiana. The results indicate an up-regulation of diverse defence-related genes in grapevine as a response to endophytic establishment of B. bassiana. Thus, endophytic establishment of an entomopathogenic fungus such as B. bassiana in grapevine plants would represent an alternative and sustainable plant protection strategy, with the potential for reducing pesticide applications in viticulture.     

Modelling the effect of partial resistance on epidemics of downy mildew of grapevine

European Journal of Plant Pathology - The cultivation of grape varieties with partial resistance to disease may become an important component for disease management in the future. The impact of...     

Importance of secondary inoculum of Plasmopara viticola to epidemics of grapevine downy mildew

To quantify the magnitude and the spatial spread of grapevine downy mildew secondary sporangia, 4685 Plasmopara viticola single lesion samples were collected from 18 plots spread across central Europe. Disease symptoms were collected on two to 22 sampling dates per plot between 2000 and 2002. Four multiallelic microsatellite markers were used for genotypic identification of pathogen samples. Genetic analysis showed more than 2300 site-specific P. viticola genotypes, indicating that populations are genetically rich demographic units. Approximately 70% of the genotypes were sampled once and 14% were sampled twice throughout the various epidemics. In the 18 populations only seven genotypes (0.3%) were identified more than 50 times. Three genotypes particularly successful in causing disease through secondary cycles showed mainly a clustered distribution. The distance of sporangial migration per secondary cycle was less than 20 m and their plot colonization rate was calculated at around 1–2 m² day⁻¹. Downy mildew epidemics of grapevine are therefore the result of the interaction of a multitude of genotypes, each causing limited (or a few) lesions, and of a dominant genotype able to spread stepwise at plot-scale. These findings contrast with current theories about grapevine downy mildew epidemiology, which postulate that there is massive vineyard colonization by one genotype and long-distance migration of sporangia.     

不同葡萄品种对霜霉病的田间苗期抗性评价

对36个葡萄品种(品系)苗期对霜霉病的田间抗病性进行了评价, 结果表明, 不同葡萄品种(品系) 对葡萄霜霉病的抗性存在明显差异, 绝大多数欧美杂交种的抗性要强于欧亚种葡萄, 供试的10个欧美杂种中, ‘瑞峰无核’对葡萄霜霉病表现为高抗; ‘DEMIR’、‘龙宝’、‘红富士’、‘黑奥林’、‘峰后’5个品种表现为中抗。     

Overexpression of phytoglobin in barley alters both compatible and incompatible interactions with the mildew pathogen Blumeria graminis

This study showed how barley plants can be shifted in their response to isolates of the mildew pathogen Blumeria graminis with different host adaptation by overexpression of the barley phytoglobin gene HvHb1. At early infection stages, plants overexpressing phytoglobin (GPHb1) showed less papilla formation and more hypersensitive response against both virulent and avirulent pathogen isolates compared to the wildtype (WT) plants. The shift was most pronounced in a wheat-adapted isolate (B. graminis f. sp. tritici). At later infection stages, GPHb1 plants infected with a virulent pathogen isolate (A6) showed less leaf chlorosis compared to the WT plants, indicating delayed senescence. The chlorophyll level was significantly higher in A6-infected GPHb1 plants 9 days after inoculation (dai) and the senescence indicators sphingosine-1-phosphate:ceramide ratio and phytol content confirmed delayed senescence. At 14 dai the percentage of fungal DNA was significantly higher on the GPHb1 plants than on WT plants, probably as a result of the delayed senescence. The results show that overexpression of phytoglobin (previously known as plant haemoglobin) can be an important tool to understand disease-related stress effects in plants of agronomic importance and for understanding basic resistance mechanisms. Studying this process in more detail may provide insights into how to alleviate stress-related senescence in plants.     

Evidence for sporangial dispersal leading to a single infection event and a sudden high incidence of grapevine downy mildew

In a German vineyard (PRO) between 10 and 13 June 2004, the incidence of grapevine downy mildew ( Plasmopara viticola ) increased abruptly from 0 to 99%. Infected vines bore on average between 45 and 60 lesions each, corresponding to about 220 000 lesions ha⁻¹ in a non-aggregated distribution. A second vineyard (FUT), approximately 50 m distant from PRO, had been inoculated 3 weeks before the abrupt increase in incidence of disease in PRO. Using microsatellites to ascertain the sources of inoculum and likelihood and extent of interplot spread from FUT to PRO, 555 samples were collected and 20 unique genotypes were identified, of which one caused 80% of the sampled lesions in both vineyards. Three genotypes responsible for 95% of the lesions in FUT and PRO were identified as the genotypes originally established through earlier inoculations in FUT. This is the first report of definitive and quantitative evidence of sporangial migration up to 130 m in a single infection event. The utility of molecular tools to address practical epidemiological issues in this pathosystem is illustrated. The results of this study provide an example of how P. viticola was able to rapidly colonize European vineyards after the pathogen was introduced from North America in 1878.     

Transboundary system for grapevine downy mildew risk evaluation on a territorial scale (P. Rada project)*

A wide and homogenous viticultural area traverses the border between Friuli Venezia Giulia (Italy) and the western part of Slovenia. Despite similar agricultural, climatic and geographical conditions on each side of the border, the management of agricultural activities and, in particular, the organization of phytosanitary campaigns is carried out separately and differently between the two countries incurring a waste of money, time and energy. The project P. Rada (EU Interreg IIIA Italy Slovenia 2000–06) was set up with the aim of establishing a common expert agrometeorological system to evaluate the development of Plasmopara viticola on a territorial scale. The required agrometeorological data are collected by a network of weather stations and spatially interpolated in a grid with a resolution of 500 × 500 m. The spatial dataset obtained is integrated with rainfall images processed by a polarimetric radar located in Fossalon di Grado (Gorizia, Italy). Once the territorial dataset has been completed, data are used to feed two agrometeorological models: SWEB, for the estimation of leaf wetness, and PLASMO for the simulation of grapevine downy mildew. The main output of the system is represented by meteorological (temperature, rainfall, leaf wetness) and epidemiological (total and current number of infections, days for the outbreak of the current infection) maps capable of supplying end users with detailed information useful for planning and management of grapevine protection activity.     

Contribution of molecular studies to botanical epidemiology and disease modelling: grapevine downy mildew as a case-study

After being accidentally introduced from the USA at the end of the 19^th century, downy mildew caused by Plasmopara viticola (Berk. et Curt.) Berlese et De Toni became one of the most damaging diseases affecting Vitis vinifera in Europe. Downy mildew causes both direct and indirect losses and can lead to severe reduction of yield. Our understanding of the life cycle and epidemiology of P. viticola has been recently altered by molecular studies that revealed that the overwintering inoculum (i.e., the oospores) does more than initiate disease, as was previously thought. A mechanistic model was developed for predicting the entire chain of processes leading to primary infections, and this primary infection model was linked to other models of secondary infection cycles. The model for primary infections defines the length of the primary inoculum season and a seasonal oospore dose consisting of several cohorts of oospores that progressively mature. The model was evaluated by means of Bayesian analysis in both Italy and eastern Canada, and showed high sensitivity, specificity, and accuracy both for potted plants and vineyards. Fungicide applications are necessary to control downy mildew because preventive agronomic practices are not very effective, including host resistance. The use of warning systems based on weather-driven models leads to a reduction in the use and cost of chemicals and a reduction in their environmental impact.     

Involvement of phenolic compounds in the resistance of grapevine callus to downy mildew (Plasmopara viticola)

Callus tissue of different grapevines (Vitis spp.) was inoculated withPlasmopara viticola. Short, highly-branched hyphae with necrosis, and long hyphae with heavy sporulation were observed on resistant and susceptible callus respectively. Thin-layer chromatography and spectrophotometric analysis showed that resistant callus contained greater quantities of gallocatechin derivatives than susceptible callus. Regression analysis between the field disease rating of each variety and its gallocatechin derivatives content indicated 92.2% correlation. Histochemical studies showed that, after infection withP. viticola, flavonoids appeared in the superficial cell walls of the callus, to a lesser degree on susceptible callus than on resistant callus. At a late stage of infection, the superficial cells of resistant callus were suberized, which did not occur in susceptible callus. This study showed that the preformed gallocatechin derivatives, the induced flavonoids and suberized superficial cells might play a role in the resistance of grapevine callus tissue to this fungus.Abbreviations CallusI Callus ofV. riparia var. Gloire de Montpellier - CallusV Callus ofV. vinifera var. Grenache - TLC Thin Layer Chromatography - var. variety - GAD Gallic acid derivatives - GD gallocatechin derivatives - RC resistant callus - SC susceptible callus     

Controlling downy mildew (Plasmopara viticola) in field-grown grapevine with β-aminobutyric acid (BABA)

Two foliar sprays of BABA (DL-3-amino-n-butanoic acid, DL-β-aminobutyric acid), or a mixture of BABA and different fungicides at reduced rates, effectively controlled (>90%) downy mildew, caused byPlasmopara viticola, in the foliage of field-grown grapevines. In five field trials BABA sprays resulted in a significant reduction of infectious leaf area and fungal sporulation, and of necrosis of oilspots. The level of disease control for BABA in four trials was similar to that achieved by metalaxyl-Cu or ‘Acrobat Plus’ (dimethomorph + mancozeb). Two-way tank-mixtures of BABA + fosetyl-AI, BABA + folpet, or BABA + Bion (benzothiadiazole), each at half the recommended rate, provided an additive effect againstP. viticola, performing as well as the full rate of each fungicide alone. BABA was not phytotoxic and did not affect pH, total titratable acids, or °Brix of the juice, as determined by commercial fungicidal standards. The results indicate that foliar applications of BABA can efficiently be integrated into a downy mildew control program in vineyards.     

Advances in sunflower downy mildew research

This review summarises the progress in research on sunflower downy mildew as reported in publications of the past 10 to 15 years, the period since the last comprehensive review on Plasmopara halstedii. Particular attention is paid to subjects that showed much progress and may be of particular interest to sunflower pathologists, mycologists or molecular biologists. Accordingly, single sections are devoted to the problems of taxonomic and phylogenetic aspects, host specificity, the host—pathogen interaction including resistance phenomena, as well as epidemiology and disease management. Reflecting the progress achieved in some fields and illuminating the deficits in others should stimulate the reader's interest in this very significant pathosystem.     

Characterization of resistance mechanisms activated by Trichoderma harzianum T39 and benzothiadiazole to downy mildew in different grapevine cultivars

Downy mildew, caused by Plasmopara viticola, is one of the most destructive diseases of grapevine and is controlled with intense application of chemical fungicides. Treatment with Trichoderma harzianum T39 (T39) or benzothiadiazole‐7‐carbothioic acid S‐methyl ester (BTH) has been previously shown to activate grapevine resistance to downy mildew and reduce disease symptoms in the Pinot noir cultivar. However, enhancement of plant resistance can be affected by several factors, including plant genotype. In order to further extend the use of resistance inducers against downy mildew, the physiological and molecular properties of T39‐ and BTH‐activated resistance in different cultivars of table and wine grapes were characterized under greenhouse conditions. T39 treatment reduced downy mildew symptoms, but the degree of efficacy differed significantly among grapevine cultivars. However, efficacy of BTH‐activated resistance was consistently high in the different cultivars. Expression profiles of defence‐related genes differed among cultivars in response to resistance inducers and to pathogen inoculation. T39 treatment enhanced the expression of defence‐related genes in the responsive cultivars, before and after P. viticola inoculation. A positive correlation between the efficacy of T39 and the expression level of defence‐related genes was found in Primitivo and Pinot noir plants, while different genes or more complex processes were probably activated in Sugraone and Negroamaro. The data reported here suggest that the use of a responsive cultivar is particularly important to maximize the efficacy of resistance inducers and new natural inducers should be explored for the less responsive cultivars.     

Glycosylation and oxidative dimerization of resveratrol are respectively associated to sensitivity and resistance of grapevine cultivars to downy mildew

Levels of resistance to Plasmopara viticola, from susceptible to highly resistant, of different grapevine cultivars were observed in vineyards and confirmed by the symptoms developed after inoculations. On the abaxial surface of infected leaves, P. viticola developed abundant sporangiophores on susceptible cultivars (Chasselas, Gamay, Gamaret and Pinot Noir), whereas on less susceptible cultivars (Seyval Blanc and Johanniter) the parasite produced few sporangiophores and some necrotic spots at the site of infection. On resistant cultivars (Bronner, Solaris, IRAC 2091), P. viticola induced a hypersensitive response and only necrotic spots were visible and the disease ceased to develop. Stilbenes were analyzed 4, 7, 24, 48 h post-infection (hpi) on small leaf samples cut from the site of infection. Large differences were observed between the cultivars at 24 and 48 hpi. Susceptible grapevines produced resveratrol and its glycoside, piceide. In contrast, resistant plants produced high concentration of ε- and δ-viniferin. Resveratrol and piceide have little or no toxicity activity against P. viticola, whereas δ-viniferin is highly toxic and can be considered an important marker for resistance of grapevine to downy mildew. The importance of oxidative dimerization of resveratrol in comparison to the extent of its glycosylation in defense reaction of grapevines against P. viticola is discussed.     

Efficiency of resistance elicitors in the management of grapevine downy mildew Plasmopara viticola: epidemiological, biochemical and economic aspects

This paper aimed to evaluate the efficiency of resistance elicitors in the management of grapevine downy mildew (Plasmopara viticola), identify the action of the elicitors on host metabolism, and determine their economic viability. The experiments were performed in a commercial vineyard variety ‘Isabel’ (Vitis labrusca) at Vale do Sirijí [Natuba, Paraiba State in Brazil, in the period of September 2009 to January 2010 (first season) and February to June 2010 (second season)]. The statistical design of randomized blocks consisted of eight treatments (untreated control, fungicide (pyraclostrobin+metiram), potassium phosphite, Agro-Mos?, Fungicide+potassium phosphite, Fungicide+Agro-Mos?, potassium phosphite+AgroMós? and Fungicide+potassium phosphite+Agro-Mos?) with four replications, with the experimental unit consisting of 45 leaves. Applications were made every 7?days, starting 20?days after pruning (DAP) with a total of 12 sprays. The evaluations were carried out biweekly, analyzing the following variables: incubation period, disease incidence and severity, area under the disease progress curve, and efficiency of control. The enzymatic determination was performed using pulp extracts from three fruits harvested at 45, 60, 90 and 120 DAP for each treatment. The resistance elicitors were able to reduce the disease incidence under different climatic conditions, indicating their viability as an alternative for the management of Plasmopara viticola.     

新疆不同地区葡萄霜霉病菌致病性分化及遗传多样性的分析

为了解新疆不同地区葡萄霜霉病菌Plasmopara viticola的遗传特征,分别采用叶盘接菌法及SSR分子标记技术对采自新疆吐鲁番、阿克苏、石河子等11个不同产区的葡萄霜霉病菌菌株的致病性进行测定,并分析各菌株的遗传多样性。结果显示,来自新疆不同地区的菌株种内存在着致病性分化现象,依据其在鉴别寄主上致病性的差异,将供试菌株划分为强、中、弱3类,其中强致病性菌株为优势菌株,且种间致病性分化与菌株的地理来源无关;SSR标记结果表明,供试菌株之间存在遗传变异现象,并且菌株之间亲缘关系都较近,在相似系数为0.93时,48株菌株聚为4大类,且遗传分化与地理分布具有一定的相关性。研究表明新疆不同地区的葡萄霜霉病菌菌株之间存在致病性分化与遗传变异现象。