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     

Breeding for resistance: breeding for resistance to Plum pox virus in Italy

Plum pox virus (PPV) resistance in apricot and peach in Italy has been researched in several Italian institutions, including the Institutes for Fruit Tree Culture and Plant Pathology in Rome, which are assessing resistance, and the Universities of Bari, Bologna and Milan, examining breeding techniques, both by cross-breeding and genetic transformation approaches. A reliable protocol for resistance assessment has been set up, in which biological indexing, ELISA and PCR techniques are used. No source of resistance has yet been found in peach. Hybridization with Prunus davidiana could be investigated given the promising results obtained within the INRA programme in France. Almost 6000 apricot seedlings from controlled crosses between resistant parents have been investigated in search of possible molecular markers, but no positive results have been obtained so far, probably due the fact that resistance to both D and M strains is sought. The transformation of cultivars has proven difficult in apricot where the regeneration step is particularly critical, while in peach a reliable regeneration protocol still remains to be set up.     

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.     

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.     

Components of quantitative resistance to powdery mildew (Erysiphe pisi) in pea (Pisum sativum)

Components of quantitative resistance in pea ( Pisum sativum ) to Erysiphe pisi , the pathogen causing powdery mildew, were investigated. Conidium germination, infection efficiency, latent period and conidium production dynamics on cv. Quantum (quantitatively resistant) were compared with those on Pania and Bolero (susceptible). There was an additional comparison in conidium germination experiments with the resistant cv. Resal. Quantitative resistance in Quantum did not affect conidium germination, but infection efficiency of conidia on this cultivar was 34% less than on the susceptible Pania. More conidia germinated on 5-day-old leaflets than on 15-day-old leaflets but the age of the plant did not affect percentage germination or infection efficiency. The length of the latent period did not differ between cultivars. Total conidium production (AUC) per unit leaflet area on Quantum was 25% less than on Pania. The maximum conidium production per day (CMAX) per unit leaflet area on Quantum was 33% less than on Pania. The time to maximum conidium production per day (TMAX) was 10% longer on Quantum than on Pania. The cv. Bolero, reported to be susceptible, also showed some degree of quantitative resistance, but this differed from that of Quantum. Total conidium production was less on Bolero than on Quantum, but the conidia on Bolero were produced sooner, and for a shorter period, than on Quantum. The stability of these responses was tested by analysing components in three different temperature regimes and testing for interactions with temperature, and with leaflet age. Temperature affected all conidium production variables. AUC per leaflet area was nearly seven times as great and CMAX nearly 15 times greater at 23°C than at 13°C. TMAX increased by 1.5 times when temperature increased from 13°C to 18°C or 23°C. Several interactions occurred and these are described.     

Genetics of resistance to downy mildew in Brassica oleracea and breeding towards durable disease control for UK vegetable production

Downy mildew resistance was previously identified from screening a Brassica oleracea collection against two standard UK isolates of Hyaloperonospora parasitica. Sources of resistance were chosen from this material and developed further in this study by generating doubled haploid (DH) and inbred lines. Seedlings from the new lines were tested for resistance to a larger collection of H. parasitica isolates collected in 2001–2002 and 2007–2008 from the main broccoli and cauliflower production regions of the UK. Three lines (derived from borecole or summer cabbage) were broadly resistant to the pathogen isolates. Three of the remaining lines exhibited strong isolate‐specific resistance; several examples of weak or basal level of resistance to some isolates were observed. A new H. parasitica variant collected in 2008 was virulent in the broadly resistant lines, but was avirulent in a line with narrow specificity of resistance. The F₂ and BC₁ seedlings derived from outcrossing each of the three broadly resistant lines to susceptible broccoli and cauliflower lines segregated in a manner indicating that the resistance was controlled by a single dominant gene. No susceptibility was observed amongst F₂ seedlings derived from intercrossing the three resistant lines, indicating that they all share the same or closely linked broad‐spectrum resistance gene(s). DH lines were produced from F₁ plants, and resistant plants were further backcrossed to produce broccoli and cauliflower‐like lines that could be useful pre‐breeding material. A combination of resistance from lines with broad and narrow specificity is recommended for controlling downy mildew in UK brassica production.     

New genes for resistance to downy mildew (Peronospora parasitica) in oilseed rape (Brassica napus ssp. oleifera)

The response of cotyledons of oilseed rape ( Brassica napus ssp. oleifera ) accessions to infection by isolates of Peronospora parasitica under controlled conditions was assessed on a 0–7 scale (disease reaction). In interactions scored 0–3, 4–5 and 6–7, the host was considered resistant, partially resistant and susceptible, respectively. Accession RES-26, selected from the spring oilseed rape cultivar Janetzkis, was partially resistant to isolate R1 and resistant to isolate P003 of P. parasitica , which distinguishes it from three previously described differential response groups ('A', 'B' and 'C') of accessions in B. napus . The resistance of RES-26 to isolate P003 seemed to be conditioned by a single, partially dominant gene and the resistance of RES-02, which belongs to group 'A' (resistant to R1 and P003), by two independent partially dominant genes. The gene for resistance to P003 in RES-26 is either closely linked, allelic or identical to one of the two genes for resistance in RES-02. Resistance of RES-02 to R1 is conditioned by a single, incompletely dominant gene. The genes for resistance to isolates R1 and P003 in RES-02 are either closely linked, allelic or identical. The cotyledonary leaves of each seedling responded independently when inoculated simultaneously each with a different isolate of the pathogen.     

Adult plant resistance to powdery mildew (Erysiphe graminis)in three barley cultivars

The mechanism(s) of adult plant resistance (a.p.r.) to Erysiphe graminis f. sp. hordei were examined in two spring barley cultivars, Athos and Porthos, which possess similar identified mildew resistance genes and seedling resistance, but differential field resistance. These cultivars were compared with Golden Promise, a universally susceptible cultivar. Differential a.p.r. to a compatible race was detected as a reduction in the number and sporulation of colonies on later-formed Athos leaves. Golden Promise exhibited some a.p.r., but much less than either Athos or Porthos. Differential a.p.r. was expressed at several infection stages, viz. appressorial formation, penetration and sporulation. Epidermal hypersensitivity occurred infrequently in all leaf position/cultivar interactions. Papillae were associated with >90% of all penetration failures. The incidence of papillae in successful penetrations at 96 h was 20% on later-formed leaves of Athos compared to c. 60% on all other leaves.     

Further characterization of field resistance in lettuce to Bremia lactucae (downy mildew)

The development of five Bremia lactucae isolates on the field-resistant crisphead lettuce cultivar Iceberg was compared under field conditions with that on the more susceptible crisphead cultivar Ithaca. With each of five isolates, the epidemic developed more rapidly on Ithaca than Iceberg; lesions on Ithaca were more numerous, larger and sporulated more profusely than on Iceberg. There was no evidence for adaptation of isolates to Iceberg, which provides some additional evidence that the resistance of this cultivar could be non-differential (horizontal). There was, however, evidence of non-specific differences between isolates in their quantitative virulence.     

50个小麦生产及后备品种(系)的抗白粉病基因推导

为明确我国小麦品种(系)中抗白粉病基因的组成,利用25个不同毒性的小麦白粉菌菌株对50个小麦生产及后备品种(系)进行抗白粉病基因推导,结果表明,参试的50个小麦品种(系)中有8个小麦品种(系)对供试的25个菌株全部感病,5个品种含有抗病基因Pm8,2个品种含有Pm4a,9个品种含有Pm2+6,4个品种含有Pm2,22个品种(系)可能含有供试基因之外的其他抗性基因或新基因。此研究结果可为小麦抗病育种以及品种利用提供依据。     

Occurrence in Italy and characteristics of lettuce downy mildew (Bremia lactucae) resistant to phenylamide fungicides

Control of lettuce downy mildew (Bremia lactucae) with phenylamide fungicides has failed in some intensive lettuce-producing areas in Northern Italy since Spring 1993. Before then, these chemicals and particularly metalaxyl, provided the best disease control. The sensitivity of Bremia lactucae isolates collected in such areas to metalaxyl was evaluated in the laboratory. These strains grew and sporulated profusely on lettuce seedlings treated with 100 and 200 ppm of metalaxyl, whereas sensitive control strains were completely inhibited when treated with fungicide concentrations ranging from 0.5–1 ppm. Thus in Italy occurrence of resistance to phenylamide fungicides in Bremia lactucae has also been demonstrated, as in almost all the countries where these chemicals were previously authorised. Subsequently, a demonstration of occurrence of resistance was made and the virulence pattern of several strains (resistant and sensitive to metalaxyl) was characterised using differential NL series containing the 13 DM resistant genes or R factors. The results suggest the occurrence of a new pathotype in Italy different from all the 16 NL Bremia lactucae races studied.     

Virulence phenotypes in powdery mildew (Blumeria graminis) populations and resistance genes in triticale (x Triticosecale)

Triticale (xTriticosecale) is a new host for powdery mildew (Blumeria graminis) being not infected by this pathogen in Germany before 2001. To evaluate population structure of the pathogen and race-specific resistances in the host, 694 isolates were collected in 12 states of Germany in the years 2007 to 2009 on triticale. They were tested by a newly developed initial differential set of 20 triticale cultivars. Corresponding virulences were found for all differentials except for cultivar Grenado. In total, 272 different virulence phenotypes (=pathotypes) were detected. The virulence complexity of the isolates ranged from 6 to 19 of 20 possible virulences with a mean of 15. In all years, a high level of diversity of the powdery mildew populations was observed with Simpson indices in the range 0.95 to 0.97. The distribution of the pathotypes was even across Germany with an Evenness index in the range 0.82 to 0.88. A set of 19 isolates with different virulence pathotypes and 10 cultivars were selected to be used to identify race-specific resistances of triticale cultivars and breeding lines. Some cultivars susceptible to most of the isolates in seedling stage were moderately resistant in adult-plant stage. The high diversity and complexity of the pathotypes found in German powdery mildew populations as well as an increasing acreage of only a few dominating triticale cultivars accelerate the adaptation of the pathogen to race-specific host resistances suggesting restricted durability only. More durable resistance might be achieved by combining new effective race-specific (qualitative) resistance genes with race-nonspecific (quantitative) resistances effective in the adult-plant stage that are already available.     

Occurrence and inheritance of resistance to powdery mildew (Oidium lycopersici) in Lycopersicon species

Among 146 accessions of Lycopersicon pimpinellifolium , 132 of L. esculentum var. cerasiforme and 53 of L. peruvianum screened for resistance to powdery mildew, caused by Oidium lycopersici , a wide variation in reactions was found. Two plants of L. esculentum var. cerasiforme accession LA-1230 were resistant. One resistant symptomless plant of accession LA-1230, designated LC-95, produced homozygous resistant progenies. LC-95 was crossed with cv. Marmande (susceptible parent) and F₁, F₂ and backcrosses to the resistant and the susceptible parents were derived. These genotypes were grown in glasshouses at 23°C and 95–100% RH and inoculated with O. lycopersici . The F₁ plants were susceptible. F₂ and backcross segregations fitted the hypothesis of a single recessive gene which is here designated ol-2 .     

Riboflavin (Vitamin B2) induces defence responses and resistance to Plasmopara viticola in grapevine

Grapevine (Vitis vinifera) is susceptible to a variety of pathogenic fungi that affect yield and wine quality. Their control is generally achieved by widespread application of fungicides in the vineyards. The economic costs and negative environmental impact associated with these applications has led to a quest for alternative strategies, focusing on manipulation of host defence mechanisms. Here, we evaluated the ability of riboflavin (i.e. Vitamin B₂) to induce resistance against downy mildew in grapevine. Our results showed that 2 mM riboflavin applied 1–3 days before pathogen inoculation provided a disease reduction efficiency of 86 %. A microscopic analysis of the time course of P. viticola colonization in riboflavin-treated leaf discs suggests early inhibition of hyphae spreading in the intercellular space. This resistance does not result from a direct fungitoxic effect of riboflavin. However, this vitamin activates host-defence responses including H₂O₂ generation, upregulation of an array of defence-related genes and synthesis of callose in stomata cells, while stilbene synthesis is not affected. Using a pharmacological approach, we deduced that both jasmonic acid and callose biosynthesis pathways are significantly involved in riboflavin-induced resistance against downy mildew in grapevine.     

Spatial and temporal spread of powdery mildew (Erysiphe pisi) in peas (Pisum sativum) varying in quantitative resistance

A field experiment was conducted to assess the progress in time and spread in space of powdery mildew (caused by Erysiphe pisi ) in pea ( Pisum sativum ) cultivars differing in resistance to the disease. Disease severity (proportion of leaf area infected) was measured in 19 × 23 m plots of cultivars Pania and Bolero (both susceptible) and Quantum (quantitatively resistant). Inoculum on infected plants was introduced into the centre of each plot. Leaves (nodes) were divided into three groups within the canopy (lower, middle, upper) at each assessment because of the large range in disease severity vertically within the plants. Disease severity on leaves at upper nodes was less than 4% until the final assessment 35 days after inoculation. Exponential disease progress curves were fitted to disease severity data from leaves at middle nodes. The mean disease relative growth rate was greater on Quantum than on Pania or Bolero, but it was delayed, resulting in an overall lower disease severity on Quantum. Gompertz growth curves were fitted to disease progress on leaves at lower nodes. Disease progress on Quantum was delayed compared with Pania and Bolero. The average daily rates of increase in disease severity from Gompertz curves did not differ between the cultivars on these leaves. Disease gradients in the plots from the inoculum focus to 12 m were detected at early stages of the epidemic, but the effects of background inoculum inputs and the rate of disease progress meant that these gradients decreased with time as the disease epidemic intensified. Spread was rapid, and there were no statistically significant differences between cultivar isopathic rates (Pania 2.2, Quantum 2.9 and Bolero 4.0 m d⁻¹).     

小麦品种(系)抗白粉病基因推导及分子标记鉴定

利用基因推导法和分子标记对我国主要麦区的小麦品种(系)进行了抗白粉病基因的鉴定。结果表明,南30-10等15个品种(系)含有Pm8,新麦2号等9个品种(系)含有Pm4,中植4号等9个品种(系)含有Pm21,郑麦113含有Pm4b+5b,杨09-111和新紫1号含有Pm2+mld。研究发现,基因推导和分子标记相结合,可大大提高小麦品种(系)抗白粉病基因鉴定结果的准确性,鉴定结果可为抗病育种和品种布局及白粉病的防治提供依据。