Leafhopper Scaphoideus Titanus Research Articles

Control of Scaphoideus titanus with Natural Products in Organic Vineyards.

The leafhopper Scaphoideus titanus is the vector of ‘Candidatus Phytoplasma vitis’, the causal agent of Flavescence dorée (FD) a key disease for European viticulture. In organic vineyards, the control of S. titanus relies mostly on the use of pyrethrins that have suboptimal efficacy. During 2016, three field trials were conducted to evaluate the efficacy of kaolin, orange oil, insecticidal soap and spinosad against S. titanus nymphs, in comparison with pyrethrins. The activity of kaolin was evaluated also in the laboratory. In all field trials, kaolin had an efficacy against nymphs comparable to pyrethrins, while the other products were not effective. Laboratory results confirmed that kaolin increased nymph mortality. In organic vineyards, kaolin and pyrethrins are valuable tools in the management of FD. Nevertheless, their efficacy is lower compared to that of the synthetic insecticides used in conventional viticulture. Therefore, further research should be conducted in order to identify alternatives to synthetic insecticides for S. titanus control in the context of organic viticulture.

Distribution of the American grapevine leafhopper (Scaphoideus titanus Ball 1932) in surroundings of Nagyvárad (Oradea, West Romania) and Debrecen (East Hungary)

The first European appearance of american grapevine leafhopper (Scaphoideus titanus), which is the vector of Grapevine Flavescence Dorée phytoplasma (‘Ca. Phytoplasma vitis’), were detected in France in 1924. The leafhopper has been distributed since 2006 in Hungary and now it occurs in the whole country, while the first record of the phytoplasma was published in 2013.We studied the distribution of the leafhopper in surroundings of Nagyvárad (Oradea, West Romania) in 2015 and surroundings of Debrecen (East Hungary) in 2016. During the studies 17 sites were sampled. The leafhoppers were found in six Romanian places with relatively low abundances while it occurs in all studied Hungarian sites and two of them showed extremely high frequenncy.

The reproductive strategy and the vibrational duet of the leafhopper Empoasca vitis.

The recent description of a new vibrational mating disruption method to control the leafhopper Scaphoideus titanus Ball opened questions about its possible application to other leafhopper pests. Since the prerequisite for the method's successful application is a deep knowledge of the species mating behavior and the exact role of associated signals, we conducted behavioral assays on the green leafhopper Empoasca vitis Göthe, a pest of grapevine and other crops in Europe and Asia. Laser vibrometer recordings of single and paired individuals (male and female) during a 24-h period enabled us to detect and describe 2 male and 1 female signal. The pair formation starts when the female replies to a male call and a duet is established, then it continues through 2 different behavioral stages: Location and Courtship. The proper courtship begins only when the male locates the female. The latter is characterized by a significant change in temporal parameters that regards both the signals and the duet structure. Although the male calling activity and the female replying rate were the same during the 24 h, a lower number of matings was recorded during the night. We discuss the possible role of vision and of the species ecology as factors of reproductive success and mating strategy. Our conclusion is that the mechanical mating disruption technique seems feasible for future application to this species.

A Phloem-Feeding Insect Transfers Bacterial Endophytic Communities between Grapevine Plants.

Bacterial endophytes colonize the inner tissues of host plants through the roots or through discontinuities on the plant surface, including wounds and stomata. Little is known regarding a possible role of insects in acquiring and transmitting non-phytopathogenic microorganisms from plant to plant, especially those endophytes that are beneficial symbionts providing plant protection properties and homeostatic stability to the host. To understand the ecological role of insects in the transmission of endophytic bacteria, we used freshly hatched nymphs of the American sap-feeding leafhopper Scaphoideus titanus (vector) to transfer microorganisms across grapevine plants. After contact with the vector, sink plants were colonized by a complex endophytic community dominated by Proteobacteria, highly similar to that present in source plants. A similar bacterial community, but with a higher ratio of Firmicutes, was found on S. titanus. Insects feeding only on sink plants transferred an entirely different bacterial community dominated by Actinobacteria, where Mycobacterium sp., played a major role. Despite the fact that insects dwelled mostly on plant stems, the bacterial communities in plant roots resembled more closely those inside and on insects, when compared to those of above-ground plant organs. We prove here the potential of insect vectors to transfer entire endophytic bacterial communities between plants. We also describe the role of plants and bacterial endophytes in establishing microbial communities in plant-feeding insects.

New insights on Flavescence dorée phytoplasma ecology in the vineyard agro‐ecosystem in southern Switzerland

AbstractPhytoplasmas associated with Flavescence dorée (FDp) grapevine disease are quarantine pathogens controlled through mandatory measures including the prompt eradication and destruction of diseased plants, and the insecticide treatments against the insect vector, the ampelophagous leafhopper Scaphoideus titanus. In the present study, a multidisciplinary approach has been applied to investigate the FDp ecological cycle in a test vineyard agro‐ecosystem in Canton Ticino, south Switzerland. Despite the scarce population density of S. titanus, a regular trend of new infections (3.4% of the total vines) through the years was observed. The leafhopper Orientus ishidae was found as the most abundant among the captured insect species known as phytoplasma vectors (245 out of 315 specimens). The population of O. ishidae was evidenced prevalently (167 specimens) in the south‐western side of the vineyard and within the neighbouring forest constituted mainly by hazel (Corylus avellana) and willow (Salix spp.). These plant species were found infected by FDp related strains (30% of analysed trees) for the first time in this study. Interestingly, O. ishidae was found to harbour FDp related strains in high percentage (26% of the analysed pools). In addition, 16SrV phytoplasma group was detected for the first time in the insect Hyalesthes obsoletus and a FDp related strain in Thamnotettix dilutior, present in low populations within the test vineyard. Molecular characterisation and phylogenetic analyses of methionine aminopeptidase (map) gene sequences of FDp and related strains, here identified, revealed the great prevalence of the map‐type FD2 in grapevines (97%) and in O. ishidae pools (72%). Such a map‐type was found also in hazel and in T. dilutior, but not in S. titanus. Moreover, map‐types FD1 and FD3 were identified for the first time in Switzerland in several host plants and phytoplasma vectors, including grapevine (FD1), S. titanus (FD1) and O. ishidae (FD1 and FD3). Based on the data obtained in this study, it is reasonable to hypothesise that the ecological cycle of FDp could be related not exclusively to the grapevine‐specific feeding diet of S. titanus, but it could include other insect vector(s) and/or plant host(s). Further studies will be needed to prove the role of O. ishidae as vector able to transmit FDp from wild plants (e.g. hazel) to grapevine.

First finding of a dual-meaning X wave for phloem and xylem fluid ingestion: Characterization of Scaphoideus titanus (Hemiptera: Cicadellidae) EPG waveforms

The leafhopper Scaphoideus titanus (Hemiptera: Cicadomorpha: Cicadellidae), an invasive deltocephaline species introduced into Europe from North America, is the vector of the most important phytoplasma disease in European viticulture, flavescence dorée. In this first electropenetrography (EPG) study of S. titanus, we characterized its feeding waveforms and defined their biological meanings. Four typical waveform phases (pathway, X wave, sustained ingestion, and interruption) and four families within those phases (A, B, C, and N) were characterized using DC EPG technology. We proposed biological meanings for these waveforms based on excreta pH-ingestion correlations, presence of X waves, and comparison with previous AC, DC, and AC-DC EPG waveforms conducted on Cicadomorpha. We observed that sustained (i.e., >10min) ingestion by a deltocephaline leafhopper can occur from both xylem and phloem vascular cells. Waveform C2x represented ingestion of xylem fluid, and two waveforms represented behaviors when stylets were inserted into phloem sieve elements: C2p variant 1 (C2p-1), which may represent salivation (perhaps simultaneous with ingestion), and C2p variant 2 (C2p-2), which represented active ingestion. Furthermore, we found that the EPG-recorded X wave has a dual meaning by occurring prior to sustained ingestion from either phloem or xylem. This X wave was very similar in appearance to the model X wave of sharpshooters, an entirely different leafhopper subfamily, Cicadellinae. All cicadellines are obligate xylem-ingesters. Such a “dual-meaning X wave” will provide insights into how the feeding tactics of S. titanus relate to other sheath-feeding hemipterans, and will provide support for future research to clarify the role of this leafhopper as a vector of plant pathogens.

Mixed xylem and phloem sap ingestion in sheath-feeders as normal dietary behavior: Evidence from the leafhopper Scaphoideus titanus

In phytophagous piercing-sucking insects, salivary sheath-feeding species are often described as xylem- or phloem-sap feeding specialists. Because these two food sources have very different characteristics, two feeding tactics are often associated with this supposed specialization. Studying the feeding behavior of insects provides substantial information on their biology, ecology, and evolution. Furthermore, study of feeding behavior is of primary importance to elucidate the transmission ability of insects that act as vectors of plant pathogens. In this study, we compared the durations of ingestion performed in xylem versus phloem by a leafhopper species, Scaphoideus titanus Ball, 1932. This was done by characterizing and statistically analyzing electrical signals recorded using the electropenetrography technique, derived from the feeding behaviors of males and females. We identified three groups of S. titanus based on their feeding behavior: 1) a group that reached the phloem quickly and probed for a longer time in phloem tissue than the other groups, 2) a group that reached the xylem quickly and probed for a longer time in xylem tissue than the other groups, and 3) a group where individuals did not ingest much sap. In addition, the numbers and durations of waveforms representing ingestion of xylem and phloem saps differed significantly depending on the sex of the leafhopper, indicating that the two sexes exhibit different feeding behaviors. Males had longer phloem ingestion events than did females, which indicates that males are greater phloem feeders than females. These differences are discussed, specifically in relation to hypotheses about evolution of sap feeding and phytoplasma transmission from plant to plant.

Host preferences of the leafhopperScaphoideus titanus, vector of “flavescence dorée” phytoplasma

The spread of “flavescence doree” (FD) phytoplasma among grapevines is made by the leafhopper Scaphoideus titanus (Hemiptera: Cicadellidae). Laboratory choice tests were performed to better understand insect vector preferences. The host preferences of nymphs and adults were compared between an experimental host plant, broadbean, and its natural host plant, grapevine, and between healthy and FD-infected plants. Grapevine was preferred to broadbean by nymphs and adults, but this preference was influenced by feeding experience. The presence of FD phytoplasma in plants made FD-infected grapevines more attractive than the healthy ones to both instar stages tested, while the preference for infected broadbean over healthy ones was only observed in adults. Plant yellowing due to the phytoplasma may increase the attractiveness of FD-infected plants. The experimental results showed that S. titanus was more attracted to yellow cues than to green, blue, or red ones. The attractiveness of FD-infected grapevines to the vector could be an efficient way to enhance disease spreading by increasing the proportion of infective vectors in the population.

Spatial Distribution and Sampling Plans for Grapevine Plant Canopy-Inhabiting Scaphoideus titanus (Hemiptera: Cicadellidae) Nymphs.

The paper deals with the study of the spatial distribution and the design of sampling plans for estimating nymph densities of the grape leafhopper Scaphoideus titanus Ball in vine plant canopies. In a reference vineyard sampled for model parameterization, leaf samples were repeatedly taken according to a multistage, stratified, random sampling procedure, and data were subjected to an ANOVA. There were no significant differences in density neither among the strata within the vineyard nor between the two strata with basal and apical leaves. The significant differences between densities on trunk and productive shoots led to the adoption of two-stage (leaves and plants) and three-stage (leaves, shoots, and plants) sampling plans for trunk shoots- and productive shoots-inhabiting individuals, respectively. The mean crowding to mean relationship used to analyze the nymphs spatial distribution revealed aggregated distributions. In both the enumerative and the sequential enumerative sampling plans, the number of leaves of trunk shoots, and of leaves and shoots of productive shoots, was kept constant while the number of plants varied. In additional vineyards data were collected and used to test the applicability of the distribution model and the sampling plans. The tests confirmed the applicability 1) of the mean crowding to mean regression model on the plant and leaf stages for representing trunk shoot-inhabiting distributions, and on the plant, shoot, and leaf stages for productive shoot-inhabiting nymphs, 2) of the enumerative sampling plan, and 3) of the sequential enumerative sampling plan. In general, sequential enumerative sampling was more cost efficient than enumerative sampling.

Plant-mediated interspecific horizontal transmission of an intracellular symbiont in insects.

Intracellular reproductive manipulators, such as Candidatus Cardinium and Wolbachia are vertically transmitted to progeny but rarely show co-speciation with the host. In sap-feeding insects, plant tissues have been proposed as alternative horizontal routes of interspecific transmission, but experimental evidence is limited. Here we report results from experiments that show that Cardinium is horizontally transmitted between different phloem sap-feeding insect species through plants. Quantitative PCR and in situ hybridization experiments indicated that the leafhopper Scaphoideus titanus releases Cardinium from its salivary glands during feeding on both artificial media and grapevine leaves. Successional time-course feeding experiments with S. titanus initially fed sugar solutions or small areas of grapevine leaves followed by feeding by the phytoplasma vector Macrosteles quadripunctulatus or the grapevine feeder Empoasca vitis revealed that the symbionts were transmitted to both species. Explaining interspecific horizontal transmission through plants improves our understanding of how symbionts spread, their lifestyle and the symbiont-host intermixed evolutionary pattern.

The first detection of leafhopper Scaphoideus titanus Ball (Hemiptera, Cicadellidae) in Slovakia

Tothova M., Bokor P., Cagaň Ľ. (2015): The first detection of leafhopper Scaphoideus titanus Ball (Hemiptera, Cicadellidae) in Slovakia. Plant Protect. Sci., 51: 88–93. Scaphoideus titanus Ball, the main vector of Flavescence doree phytoplasma, was monitored in Slovakia during 2013 and 2014. The species was present in the vineyards of eastern Slovakia but it was rarely found in the south of central part of the country. It was also found at many locations of western Slovakia but not in the southeast of western Slo vakia. Higher abundance of S. titanus was detected in the vineyards with biological control. Varietal preference of the leafhopper was not confirmed. The first nymphs were found on June 5 and the last nymphs on July 17, adults occurred from the end of June to the end of September. There is one generation of S. titanus per year in Slovakia.

Heterologous expression and processing of the flavescence dorée phytoplasma variable membrane protein VmpA in Spiroplasma citri.

BackgroundFlavescence dorée (FD) of grapevine is a phloem bacterial disease that threatens European vineyards. The disease is associated with a non-cultivable mollicute, a phytoplasma that is transmitted by the grapevine leafhopper Scaphoideus titanus in a persistent, propagative manner. The specificity of insect transmission is presumably mediated through interactions between the host tissues and phytoplasma surface proteins comprising the so-called variable membrane proteins (Vmps). Plant spiroplasmas and phytoplasmas share the same ecological niches, the phloem sieve elements of host plants and the hemocoel of insect vectors. Unlike phytoplasmas, however, spiroplasmas, and Spiroplasma citri in particular, can be grown in cell-free media and genetically engineered. As a new approach for studying phytoplasmas-insect cell interactions, we sought to mimic phytoplasmas through the construction of recombinant spiroplasmas exhibiting FD phytoplasma Vmps at the cell surface.ResultsHere, we report the expression of the FD phytoplasma VmpA in S. citri. Transformation of S. citri with plasmid vectors in which the vmpA coding sequence was under the control of the S. citri tuf gene promoter resulted in higher accumulation of VmpA than with the native promoter. Expression of VmpA at the spiroplasma surface was achieved by fusing the vmpA coding sequence to the signal peptide sequence of the S. citri adhesin ScARP3d, as revealed by direct colony immunoblotting and immunogold labelling electron microscopy. Anchoring of VmpA to the spiroplasma membrane was further demonstrated by Triton X-114 protein partitioning and Western immunoblotting. Using the same strategy, the secretion of free, functionally active β-lactamase (used as a model protein) into the culture medium by recombinant spiroplasmas was achieved.ConclusionsConstruction of recombinant spiroplasmas harbouring the FD phytoplasma variable membrane protein VmpA at their surface was achieved, which provides a new biological approach for studying interactions of phytoplasma surface proteins with host cells. Likewise, the secretion of functional β-lactamase by recombinant spiroplasmas established the considerable promise of the S. citri expression system for delivering phytoplasma effector proteins into host cells.Electronic supplementary materialThe online version of this article (doi:10.1186/s12866-015-0417-5) contains supplementary material, which is available to authorized users.

Examination of the American grapevine leafhopper (Scaphoideus titanus Ball) in Debrecen and Micske (Misca, West Romania)

Grapevine flavescence dorée (FD) was detected first in Hungary in 2013 in Zala County (South-West-Hungary). The disease is a serious danger for grapevine growing and grapevine propagating production. In 2014, the pathogen has been found in several new places in Hungary, viz. in Vas and Fejér Counties, and it was also detected in the former location in Zala County. The american grapevine leafhopper (Scaphoideus titanus) is the main vector of the disease. This pest was detected first in Hungary in 2006 and then it has spread all over the country. Since we have not detailed distribution data of this pest in surroundings of Debrecen, therefore we made observations in this region in 2014. The presence of the pest was confirmed by yellow sticky cards in two locations in Debrecen and another site in West Romania near to Hungarian border. We found that S. titanus is present in each sampled sites that cause serious potential danger for the appearance and spread of Grapevine flavescence dorée (FD) in this region.

The process of pair formation mediated by substrate-borne vibrations in a small insect

The ability to identify and locate conspecifics depends on reliable transfer of information between emitter and receiver. For a majority of plant-dwelling insects communicating with substrate-borne vibrations, localization of a potential partner may be a difficult task due to their small body size and complex transmission properties of plants. In the present study, we used the leafhopper Scaphoideus titanus as a model to investigate duetting and mate searching associated with pair formation. Studying these insects on a natural substrate, we showed that the spatio-temporal structure of a vibrational duet and the perceived intensity of partner's signals influence the mating behaviour. Identification, localization and courtship stages were each characterized by a specific duet structure. In particular, the duet structure differed in synchronization between male and female pulses, which enables identification of the partner, while the switch between behavioural stages was associated with the male-perceived intensity of vibrational signals. This suggests that males obtain the information about their distance from the female and optimize their strategy accordingly. More broadly, our results show that even in insects smaller than 1cm, vibrational signals provide reliable information needed to find a mating partner.

Dynamics of acquisition and transmission of “flavescence dorée” phytoplasma in grapevine

A dynamic model of “flavescence dorée” phytoplasma (FD) in grapevine transmitted by the leafhopper Scaphoideus titanus was developed and tested. The model accounted for the biological life cycle of S. titanus on Vitis vinifera, epidemics progression in V. vinifera, vineyard setting, and agronomic practices aimed to control the epidemics. It was comprehensively tested against data sets of FD presence gathered in various vineyards in Piemonte, Italy, over nine years. The model achieved a correlation coefficient (R) and residual (NRMSE) against observations in the range R ≥ 91.3% and NRMSE ≤ 12.59%, respectively. A stochastic sensitivity analysis showed that FD progression was slightly more susceptible to uncertainty in S. titanus life cycle parameters than equivalent uncertainty in FD pathogen acquisition and transmission between plant and insect and initial conditions. Examples of analysis of outbreak scenarios and assessment of effectiveness of mitigation measures showed that combinations of insecticide applications and plant roguing were optimal to reduce FD progression within time scales of about 10 to 15 years. On a wider perspective, evidence validated by comparison with empirical data is presented for the potential use of epidemiological models in decision making when control strategies should be implemented in agronomic practices.

The Johnston's organ of three homopteran species: A comparative ultrastructural study

A transmission electron-microscopy study has been carried out on the pedicel of three homopteran species, with particular focus on the leafhopper Scaphoideus titanus Ball. The two other species, the planthoppers Hyalesthes obsoletus Signoret and Metcalfa pruinosa Say, were investigated in order to compare the ultrastructure of the Johnston's organ (JO) among representatives of the Auchenorrhyncha group. The results showed the presence of a well developed JO located within the pedicel. Depending on the species the JO is made of 25 up to 72 scolopidia arranged in a coronal array. Each scolopidium is connective, heterodynal, amphinematic and hosts three structurally dissimilar sensory neurons. Two of them have a type 1 ciliary segment while the third bears a type 2 cilium. The type 2 dendrite tip is associated with a tubular cap and is longer than the others, ending into the cuticle at the base of the flagellum. Other scolopidia with one or two neurons were found in S. titanus, forming an accessory organ. The presence of such a well developed mechanosensory apparatus is discussed in relation with the lifestyle of the three species.

Testing the efficacy of single applications of five insecticides against Scaphoideus titanus on common grapevines

The appearance of the Flavescence Dorée phytoplasma and the possibility of its rapid spread by its vector, the American Grapevine Leafhopper (Scaphoideus titanus Ball) (AGVL), represent a serious threat to viticulture production in Slovenia and elsewhere in Europe. Insecticide treatment represents one of major means of reducing the abundance of AGVL in wine-growing areas. In the period from 2010 to 2011, five insecticides against AGVL were tested, and the efficacy of the substances was calculated according to the number of nymphs (at 2, 7, 14 and 21 d after application), which were counted on the grapevine leaves. In both years, the highest cumulative efficacy (96–97% or 0.01 nymphs/leaf) was confirmed for thiamethoxam, whereas chlorpyrifos-methyl showed a very high cumulative efficacy (87–89% or 0.01–0.02 nymphs/leaf). In 2010, indoxacarb exhibited only a 43% (0.09 nymphs/leaf) cumulative efficacy; however, its substitute in 2011, chlorpyrifos, was more effective (73% or 0.05 nymphs/leaf). A single application of pyrethrin demonstrated only 45–61% (0.05–0.09 nymphs/leaf) efficacy. Based on the results of our research, we recommend the use of thiamethoxam and chlorpyrifos-methyl against AGVL.

Spread of grapevine phytoplasma diseases in Europe

“Flavescence dorée” is a quarantine disease in Europe as it is epidemically transmitted by the grapevine leafhopper Scaphoideus titanus, an insect of North American origin now widely distributed in the vineyards of Southern Europe. Strains of FD phytoplasma mainly spread from grapevine to grapevine but surrounding wild Vitis regrowth, infected alders and Clematis may constitute potential epidemic reservoir. “Bois noir” disease is endemic in the Euro-Mediterranean area and is mainly transmitted by Hyalesthes obsoletus, a planthopper residing in weeds such as bindweeds and stinging nettles which also act as plant reservoirs for the BN phytoplasma. “Bois noir” is caused by the accidental transmission of different “stolbur” phytoplasma strains that are maintained by epidemiological systems based on different endemic weed species as reservoir plant. The geographic distribution of the two diseases and some of their epidemiological features are described.

Migration of Scaphoideus titanus Ball from the surrounding vineyards into the nursery

A one-year study of the mobility of leafhopper Scaphoideus titanus Ball (ST), vector of Flavescence dorée phytoplasma, was conducted in 2009 and included three Slovenian nurseries in the Primorska winegrowing region, as well as two Italian and one Swiss nursery, and was aiming at evaluating the risk of spreading ST with vine propagating material. The presence of ST was monitored by the means of yellow sticky traps that were placed at three distances in the nurseries and the nearby vineyards on three parallel lines (replicates). The traps were equidistantly placed from the nurseries’ and vineyards’ borders in order to determine the possible distance gradient. The monitoring started in the latest nymphal stages (middle of June) and terminated in the first decade of October. Insecticide treatments with deltamethrin (Decis 2.5 EC) were applied in nurseries, according to the instructions for use and recommendations for suppression of ST. On all Slovenian plots in the vineyards, the population level of ST was high (1796 adults in average / vineyard or 18 adults per trap / week), which indicated a great migration potential of the vector towards the nurseries. Catches were very low (54 adults in average / nursery or 0.4 adults / trap / week) on two nurseries’ plots (Lože and Slap), although high numbers of ST adults were caught in the adjacent untreated vineyards. On the Poreče nursery plot, insecticide treatments were not sufficient to avoid high numbers of ST catches (in total 481 adults or 3 adults / trap / week). This could be related to the sub-optimal timing of the first insecticide treatment and/or migration of adults from two nearby untreated vineyards located at a distance of 70 and 150 m. The distance between the nurseries and vineyards did not affect ST catches in the nurseries.

Microsatellite and mitochondrial data provide evidence for a single major introduction for the Neartic leafhopper Scaphoideus titanus in Europe.

Scaphoideus titanus, a leafhopper native to North America and invasive in Europe, is the vector of the Flavescence dorée phytoplasma, the causal agent of the most important form of grapevine yellows in European vineyards. We studied 10 polymorphic microsatellite loci and a 623 bp fragment of the mitochondrial cytochrome oxidase II gene in native S. titanus from north-eastern America and introduced European populations, to elucidate the colonization scenario. Consistent with their recent history, invasive European populations were less genetically diverse than American populations for both types of markers, suggesting a recent bottleneck. Significant isolation by distance was detected between American populations but not between European populations. None of the European mitochondrial haplotypes was found in the American vineyards, from which they are assumed to have originated. The precise source of the invasive S. titanus populations therefore remains unclear. Nevertheless, the high heterozygosity of North-East American populations (which contained 92% of the observed alleles) suggests that this region is part of the native range of S. titanus. Clustering population genetics analyses with microsatellite and mitochondrial data suggested that European populations originated from a single introduction event. Most of the introduced populations clustered with populations from Long Island, the Atlantic Coast winegrowing region in which Vitis aestivalis occurs.