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Flavescence dorée (FD) is a European quarantine grapevine disease transmitted by the Deltocephalinae leafhopper Scaphoideus titanus. Whereas this vector had been introduced from North America, the possible European origin of FD phytoplasma needed to be challenged and correlated with ecological and genetic drivers of FD emergence. For that purpose, a survey of genetic diversity of these phytoplasmas in grapevines, S. titanus, black alders, alder leafhoppers and clematis were conducted in five European countries. Out of 132 map genotypes, only 11 were associated to FD outbreaks, three were detected in clematis, whereas 127 were detected in alder trees, alder leafhoppers or in grapevines out of FD outbreaks. Most of the alder trees were found infected, including 8% with FD genotypes M6, M38 and M50, also present in alders neighboring FD-free vineyards and vineyard-free areas. The Macropsinae Oncopsis alni could transmit genotypes unable to achieve transmission by S. titanus, while the Deltocephalinae Allygus spp. and Orientus ishidae transmitted M38 and M50 that proved to be compatible with S. titanus. Variability of vmpA and vmpB adhesin-like genes clearly discriminated 3 genetic clusters. Cluster Vmp-I grouped genotypes only transmitted by O. alni, while clusters Vmp-II and -III grouped genotypes transmitted by Deltocephalinae leafhoppers. Interestingly, adhesin repeated domains evolved independently in cluster Vmp-I, whereas in clusters Vmp-II and–III showed recent duplications. Latex beads coated with various ratio of VmpA of clusters II and I, showed that cluster II VmpA promoted enhanced adhesion to the Deltocephalinae Euscelidius variegatus epithelial cells and were better retained in both E. variegatus and S. titanus midguts. Our data demonstrate that most FD phytoplasmas are endemic to European alders. Their emergence as grapevine epidemic pathogens appeared restricted to some genetic variants pre-existing in alders, whose compatibility to S. titanus correlates with different vmp gene sequences and VmpA binding properties., Author summary Since the first outbreaks, Flavescence dorée epidemics had been associated to the introduction of the North American leafhopper vector Scaphoideus titanus. We hereby show that the associated phytoplasma originated from European alders and that epidemics in grapevine are restricted to some phytoplasma genetic variants pre-existing in this wild asymptomatic plant host. The compatibility of this phytoplasma to the introduced S. titanus insect vector resulted from the adaptation of phytoplasma variable membrane proteins Vmps to leafhoppers of the same subfamily living on alders. Vmps organization is similar to adhesion related proteins (ARP) and seems to allow the duplication of pre-adapted repeated domains. This suggests a key role of the Vmp adhesins in the life-style of phytoplasmas infecting woody hosts that rely on the adaptation to new insect vectors to expand their plant-host range.

International audience

UMR BFP - Equipe Mollicutes; When a Palearctic bacterium meets a Nearctic insect vector: genetic and ecological insights into the emergence of the grapevine ‘[i]Candidatus Phytoplasma vitis[/i]’ and ‘[i]Ca. P. vitialni[/i]’ epidemics in Europe. 25. International Congress of Entomology (ICE 2016)

International audience; Impact of "bois noir" disease (BN) on performance of 'Chardonnay' grown in Eger (Hungary) is reported from plant (vegetative and reproductive parameters, fruit composition) to wine (wine quality, sensory analysis). BN-associated chlorophyll degradation and leaf rolling resulted in a loss in functioning surface of the canopy. Lack of lignification and non-viable buds evoked unpredictable crop production. Severity and extent of reduction in production varied between years, but the crop loss was always above 53%. In the case of BN-affected grapevines the reduced quantity resulted in a low quality of grape bunch composition, which enhanced unfavourable outcomes in wines. Differences in fruit composition and wine quality between healthy and BN affected grapevines were most pronounced in years with optimal weather conditions and the negative effect was covered up in poor years. In wine regions where mechanical harvesting is applied, harvesting of phytoplasma diseased vineyards can therefore not be selective and qualitative. According to the three years study presented BN induce severe loss, and BN-affected plants compromise the yield and wine quality, and therefore the overall profitability of a vineyard.

International audience; Little is known about the mechanisms of insects - phytoplasma interactions which are driving the ecological diversification of phytoplasmas. Phytoplasma surface proteins play an important role in phytoplasma life cycle and polymorphism of this protein might determine the transmission ability of different insect species. Strains of Candidatus Phytoplasma solani' from several European countries, based on variable membrane protein gene (stamp), were classified into four distinct phylogenetic clusters which clusters are present in different geographical areas. This may reflect the distribution of the different known insect vectors and the adaptation of the various 'Ca. P. solani' strain to a specific vector. To detect STAMP protein, monoclonal antibody 2A10 is available. This antibody was produced against StolburC, which is a representative strain of stamp clusters I. In this work we performed heterologous expression antigenic membrane proteins STAMP from different 'Ca. P. solani' genetic clusters and tested their recognition by the 2A10 monoclonal antibody.

International audience; In order to evaluate the risk represented by the wild reservoir as a possible source of 'flavescence doree' outbreaks in Hungary, diverse wild perennial plants growing in vineyard areas were tested for the presence of 16SrV-C and D subgroup phytoplasmas. 16SrV phytoplasmas were detected by nested PCR-RFLP on the 16SrDNA in alders (86% infected) and in clematis (71% infected). Further characterisation by sequencing of the map gene revealed in both plants strains having the same map gene sequence as 'flavescence doree' strains.