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Optical detection of downy mildew in grapevine leaves: daily kinetics of autofluorescence upon infection.

ABSTRACT: A 15-day survey of autofluorescence has been conducted upon infection by downy mildew [Plasmopara viticola (Berk. & M.A. Curtis) Berl. & de Toni] of leaves of a susceptible grapevine genotype. Different autofluorescence signals were followed from the cellular to the whole-leaf level by using four types of devices for fluorosensing: a macroscope, a spectrofluorimeter, a portable field optical sensor (the Multiplex 3), and a field fluorescence sensor prototype with 335 nm excitation. It was shown for the first time, by the three different techniques and at three different scales, that the stilbene-dependent violet-blue autofluorescence (VBF) had a transitory behaviour, increasing to a maximum 6 days post-inoculation (DPI) and then decreasing to a constant lower level, nevertheless significantly higher than in the control leaf. This behaviour could be sensed from both sides of the leaf. On the abaxial side, VBF could discriminate the presence of infection from 1 DPI, and on the adaxial side from 3 DPI. There was a constant increase in blue-excited green fluorescence starting from 8 DPI, concomitant with a decrease in leaf chlorophyll content sensed by one reflectance and two fluorescence indices available on the Multiplex 3 sensor. These results show that a pre-symptomatic and symptomatic sensing of downy mildew is possible by autofluorescence-based sensors, and this is potentially applicable in the field.

SUBMITTER: Bellow S

PROVIDER: S-EPMC3528039 | biostudies-literature | 2013 Jan

REPOSITORIES: biostudies-literature

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Optical detection of downy mildew in grapevine leaves: daily kinetics of autofluorescence upon infection.

Bellow Sébastien S Latouche Gwendal G Brown Spencer C SC Poutaraud Anne A Cerovic Zoran G ZG

Journal of experimental botany 20121203 1

A 15-day survey of autofluorescence has been conducted upon infection by downy mildew [Plasmopara viticola (Berk. & M.A. Curtis) Berl. & de Toni] of leaves of a susceptible grapevine genotype. Different autofluorescence signals were followed from the cellular to the whole-leaf level by using four types of devices for fluorosensing: a macroscope, a spectrofluorimeter, a portable field optical sensor (the Multiplex 3), and a field fluorescence sensor prototype with 335 nm excitation. It was shown ...[more]

PMID: 23213137

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Project description:BackgroundTo limit the impact of the downy mildew disease of grapevine and reduce the need to recur to chemical treatments, an effective strategy might be recovering adaptive resistance traits in both cultivated and wild V. vinifera germplasm. Considering that stilbenes represent the most important class of phytoalexins in the Vitaceae, the constitutive expression and transcriptional activation of all the functional members of the stilbene synthase gene family were analysed in a group of nine grapevine genotypes following artificial infection with the oomycete Plasmopara viticola, the causal agent of the disease. In addition, in the same genotypes we analyzed the expression of genes encoding for two transcription factors involved in the transcriptional regulation of the stilbene synthase genes, namely VvMYB14 and VvMYB15, and of genes encoding for chalcone synthases.ResultsDowny mildew incidence and severity ranged from nihil to high in the grapevine genotypes considered, being low to moderate in a subgroup of V. vinifera genotypes. The constitutive expression of the stilbene synthase genes as well as the extent of their transcriptional activation following P. viticola inoculation appeared to be inversely related to the proneness to develop disease symptoms upon infection. In a specular manner, following P. viticola inoculation all the chalcone synthase genes were up-regulated in the susceptible grapevine genotypes and down-regulated in the resistant ones. The infection brought by P. viticola appeared to elicit a co-ordinated and sequential transcriptional activation of distinct stilbene synthase genes subsets, each of which may be regulated by a distinct and specific MYB transcription factor.ConclusionsThe present results suggest that the induction of stilbene biosynthesis may contribute to the basal immunity against the downy mildew of grapevine, thus representing an adaptive resistance trait to recover, in both cultivated and wild V. vinifera germplasm. During the early stages of P. viticola infection, an antagonistic interaction between flavonol and stilbene biosynthesis might occur, whose outcome might determine the subsequent extent of disease symptoms. Further studies are needed to decipher the possible regulatory mechanisms involved in the antagonistic crosstalk between these two metabolic pathways in resistant and susceptible genotypes in response to P. viticola.

Project description:Grapevine varieties showing partial resistance to downy mildew, caused by Plasmopara viticola, are a promising alternative to fungicides for disease control. Resistant varieties are obtained through breeding programs aimed at incorporating Rpv loci controlling the quantitative resistance into genotypes characterized by valuable agronomic and wine quality traits by mean of crossing. Traditional phenotyping methods used in these breeding programs are mostly based on the assessment of the resistance level after artificial inoculation of leaf discs in bioassays, by using the visual score proposed in the 2nd Edition of the International Organization of Vine and Wine (OIV) Descriptor List for Grape Varieties and Vitis species (2009). In this work, the OIV score was compared with an alternative approach, not used for the grapevine-downy mildew pathosystem so far, based on the measurement of components of resistance (RCs); 15 grapevine resistant varieties were used in comparison with the susceptible variety 'Merlot'. OIV scores were significantly correlated with P. viticola infection frequency (IFR), the latent period for the downy mildew (DM) lesions to appear (LP50), and the number of sporangia produced per lesion (SPOR), so that when the OIV score increased (i.e., the resistance level increases), IFR and SPOR decreased, while LP50 increased. The relationship was linear for LP50, monomolecular for IFR and hyperbolic for SPOR. No significant correlation was found between OIV score and DM lesion size, sporangia produced per unit area of lesion, length of infectious period, and infection efficiency of the sporangia produced on DM lesions. The correlation between OIV score and area under the disease progress curve (AUDPC) calculated by using the RCs and a simulation model was significant and fit an inverse exponential function. Based on the results of this study, the measurement of the RCs to P. viticola in grapevine varieties by means of monocyclic, leaf disc bioassays, as well as their incorporation into a model able to simulate their effect on the polycyclic development of DM epidemics in vineyards, represents an improved method for phenotyping resistance level.

Project description:The obligatory biotrophic oomycetes Plasmopara viticola is the causal agent of downy mildew, a destructive disease of grapevine worldwide. So far, chemical fungicides are widely employed to limit this pathogen, but their adverse effects are stimulating the quest for environmentally friendly alternative approaches. Here, we report on the search for mycoviruses that might be later developed as biocontrol agents for this pathogen. Symptomatic leaves were collected from various regions in Spain and Italy and mycelia associated to leaf lesions was harvested. Total RNA extractions were depleted of rRNA and metatranscriptomes were generated using a high-throughput sequencing approach. The virome associated to leaf lesions was then characterized through a bioinformatic pipeline relying on blast searches against current viral databases. Here, we present an inventory of 283 new RNA viruses: 222 positive strand RNA viruses, 29 negative strand RNA viruses, 27 double-stranded RNA viruses and 5 ORFan virus RdRP segments, which could not be reliably assigned to any existing group in the Riboviria. In addition to ORFan viruses, we found other surprising new evolutionary trajectories in this wide inventory of viruses. The most represented viruses in our collection are those in phylum Lenarviricota, and, among them, a group of mycovirus segments distantly related to narnaviruses, but characterized by a polymerase palm domain lacking subdomain C, with the putative GDD catalytic triad. We also provided evidence of a strict association between two RNA segments that form a new mycovirus clade of positive strand RNA in the phylum Kitrinoviricota, order Martellivirales. In the phylum Negarnaviricota, we report for the first time in the order Mononegavirales a clade of viruses that is ambisense, a feature that so far was present only in the order Bunyavirales. Furthermore, in the same phylum we detected the widespread occurrence and abundant accumulation in our libraries of a distinct mycovirus clade distantly related to the Muvirales and Goujanvirales orders, which so far include only viruses infecting invertebrates. Possible new oomycetes-specific virus clades are also described in the phylum Duplornaviricota. These data greatly expand the evolutionary history of mycoviruses adding new layers of diversity to the realm Riboviria.

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Optical detection of downy mildew in grapevine leaves: daily kinetics of autofluorescence upon infection.

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Optical detection of downy mildew in grapevine leaves: daily kinetics of autofluorescence upon infection.

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