Leaf morphological characteristics and stilbene production differently affect downy mildew resistance of Vitis vinifera varieties grown in Italy

The degree of resistance to downy mildew of grape varieties belonging to the oenological tradition of Central Italy was evaluated by the analysis of plant responses to pathogen infections carried out in natural and controlled environments. Leaf morphological traits, such as hair and stornata! density, were determined for each variety, and the percentage of infected stornata and pathogen colonization of host mesophyll at 24, 48, and 72 hours post inoculation were assessed by epifluorescence microscopy. Furthermore, stilbene production at the site of Plasmopara viticola infection was analyzed at 72 hours post inoculation. Results indicate differences in resistance to downy mildew among selected varieties. Different significant values were detected among grapevines in the percentage of infected stornata and average number of successfully penetrated zoospores per stornata and per leaf surface unit. Differences also emerged in the rate of pathogen growth and stilbene production, signifying that defence mechanisms involved or induced during pathogen infection could be differentially effective among grapevine cultivars in limiting disease progression.Peer reviewe

Downy mildew: is resistance linked to inoculum concentration?

Leaves of different Vitis vinifera cultivars, susceptible (‘Chasselas’ and ‘2185’), less susceptible (‘2142’) or resistant to downy mildew ('Solaris’ and ‘2091’), were inoculated with four different concentrations of an aqueous sporangia suspension of Plasmopara viticola (5 × 105, 2 × 105, 6 × 104 and 2 × 104 sporangia·ml-1). The infection rate of these samples was then examined by light microscopy and synthesis of stilbenes was analysed at infection sites. Infection rate increased parallel with inoculum concentration, but there was no correlation between the infection rate and resistance to P. viticola. Moreover, at the lowest inoculum concentration, the infection rate is similar for susceptible and resistant grapevine varieties. Quantification of stilbenes at 72 hpi showed that at the lowest inoculum concentration, the most susceptible grape variety synthesized the largest amount of stilbenes, whose level remained however below the ED50 values defined for each of them. Conversely, at the highest inoculum concentration, the most resistant varieties produced the highest amounts of the most toxic stilbenes against P. viticola. The critical role of the inoculum concentration used for artificial inoculation to evaluate grapevine resistance to downy mildew is discussed.

Leaf morphological characteristics and stilbene production differently affect downy mildew resistance of Vitis vinifera varieties grown in Italy

The degree of resistance to downy mildew of grape varieties belonging to the oenological tradition of Central Italy was evaluated by the analysis of plant responses to pathogen infections carried out in natural and controlled environments. Leaf morphological traits, such as hair and stomatal density, were determined for each variety, and the percentage of infected stomata and pathogen colonization of host mesophyll at 24, 48, and 72 hours post inoculation were assessed by epifluorescence microscopy. Furthermore, stilbene production at the site of Plasmopara viticola infection was analyzed at 72 hours post inoculation. Results indicate differences in resistance to downy mildew among selected varieties. Different significant values were detected among grapevines in the percentage of infected stomata and average number of successfully penetrated zoospores per stomata and per leaf surface unit. Differences also emerged in the rate of pathogen growth and stilbene production, signifying that defence mechanisms involved or induced during pathogen infection could be differentially effective among grapevine cultivars in limiting disease progression.

Effects of resveratrol, viniferins and pterostilbene on Plasmopara viticola zoospore mobility and disease development

The effects of stilbenes (resveratrol, δ- and ε-viniferins, and pterostilbene) on the mobility of zoospores of Plasmopara viticola and on subsequent disease development were studied in vitro. δ-viniferin and pterostilbene are the most toxic stilbenes concerning zoospore mobility (ED50 : 14.6 and 28.3 μM) and disease development (ED50 : 14.7 and 12.7 μM). The analysis of stilbenes in leaf cells of resistant (Solaris) and susceptible (Chasselas) grape cultivars artificially inoculated with P. viticola has shown that very high amounts of stilbenic phytoalexins accumulate at the site of infection of the resistant cultivar compared to the susceptible one

Histological and biochemical criteria for objective and early selection of grapevine cultivars resistant to Plasmopara viticola

Grapevine breeding is the most effective way to create cultivars resistant to downy mildew (Plasmopara viticola), and to reduce the number of fungicide applications. Four criteria, including histological and biochemical analyses, based on the level of different mechanisms of resistance to grapevine downy mildew, were tested on 42 different cultivars. Plantlets were artificially inoculated with downy mildew and the sporangia density was measured spectrophotometrically 6 d after infection. Callose synthesis in stomata and δ- and ε-viniferin levels at the site of infection were recorded 48 h after inoculation. These observations have allowed the 42 cultivars to be divided into 5 groups: very resistant (VR), resistant (R), less susceptible (LS), susceptible (S) and highly susceptible (HS). All 4 criteria have to be applied to assign the resistance level closer to field conditions. This method allows to rapidly evaluate the level of resistance of seedlings to downy mildew thereby leading to a reduction in duration  of the breeding program by several years.

Development of a rapid and highly sensitive direct-PCR assay to detect a single conidium of Botrytis cinerea Pers.:Fr in vitro and quiescent forms in planta

“Direct-PCR” amplifications of Botrytis cinerea-specific genomic sequences, without any DNA purification step or time consuming sample preparation, were developed. A single copy sequence of 0.7 Kb in the Botrytis cinerea genome was amplified in reactions containing no more than 1 x 105 to 1 single conidium. As a demonstrative application, this assay was applied to detect B. cinerea in different parts of immature grape berries (at ‘pea size’), when previously inoculated with conidia at flowering. Using this method we showed the presence of quiescent Botrytis in the receptacle area only. Cloning and sequencing of the fragment confirmed the single sequence gene of B. cinerea. These results demonstrate that the method is easy to apply and of sufficiently high sensitivity to detect the presence of B. cinerea in immature grape berries. Its use for studies on the development of grey mould and improved control of the disease in vineyards is discussed

Evaluation of Hungarian Wines for Resveratrol by Overpressured Layer Chromatography

A method, including solid phase extraction sample preparation, overpressured layer chromatographic separation and subsequent densitometric evaluation, was developed for measurement of total resveratrol (cis- and trans-isomers) content of wine. The amount of resveratrol was determined in wine samples from different winemaking regions of Hungary. The total resveratrol was high in Hungarian red wines (3.6–11 mg/L), and much lower in white ones (0.04–1.5 mg/L)