Bioecological Traits of Spittlebugs and Their Implications for the Epidemiology and Control of the Xylella fastidiosa Epidemic in Apulia (Southern Italy).

Spatial-temporal dynamics of spittlebug populations, together with transmission biology, are of major importance to outline the disease epidemiology of Xylella fastidiosa subsp. pauca in Apulian olive groves. The spread rate of X. fastidiosa is mainly influenced by (i) the pathogen colonization of the host plant; (ii) the acquisition of the pathogen by the vector from an infected plant, and its inoculation to healthy plants; (iii) the vector population dynamics and abundance at different spatial scales; and (iv) the dispersal of the vector. In this contribution we summarize the recent advances in research on insect vectors' traits-points ii, iii, and iv-focusing on those most relevant to X. fastidiosa epidemic in Apulia. Among the vectors' bioecological traits influencing the X. fastidiosa epidemic in olive trees, we emphasize the following: natural infectivity and transmission efficiency, phenological timing of both nymphal and adult stage, the role of seminatural vegetation as a vector reservoir in the agroecosystem and landscape, and preferential and directional dispersal capabilities. Despite the research on X. fastidiosa vectors carried out in Europe in the last decade, key uncertainties on insect vectors remain, hampering a thorough understanding of pathogen epidemiology and the development of effective and targeted management strategies. Our goal is to provide a structured and contextualized review of knowledge on X. fastidiosa vectors' key traits in the Apulian epidemic, highlighting information gaps and stimulating novel research pathways on X. fastidiosa pathosystems in Europe. [Formula: see text] Copyright © 2023 The Author(s). This is an open access article distributed under the CC BY-NC-ND 4.0 International license.

Morpho-functional Analysis of the Head Glands in Three Auchenorrhyncha Species and Their Possible Biological Significance.

The Cicadomorpha Philaenus spumarius, Neophilaenus campestris, and Cicadella viridis are known transmitters of the bacterium Xylella fastidiosa. Here, we studied the ultrastructural organization of their cephalic glands. Our investigations with scanning, transmission, focused ion beam-scanning electron microscopes and light microscope revealed for the first time in Auchenorrhyncha the presence of two types of cephalic glands. Both belonged to the Class III epidermal glands, according to the Noirot and Quennedey classification. Type A glands were the most common, being mainly located around antennae, lorum, and gena. Moreover, these glands were observed also on the abdomen and thorax, always in association with sensilla trichoidea. The second type of glands (type B) were located exclusively at the apical part of the postclypeus in P. spumarius and N. campestris. The ultrastructural organization was similar in both types, being composed of a secretory cell and a conducting canal. Differences were observed in the width of the cuticular opening, being smaller in the type II glands. In addition, we have recorded the presence of a maxillary sensory pit in all species and described sensilla trichoidea ultrastructural organization. Finally, we discussed the ultrastructural organization of the glands and their potential biological role.

Diversity of the Bacterial Community Associated with Hindgut, Malpighian Tubules, and Foam of Nymphs of Two Spittlebug Species (Hemiptera: Aphrophoridae).

Spittlebugs are xylem-sap feeding insects that can exploit a nutrient-poor diet, thanks to mutualistic endosymbionts residing in various organs of their body. Although obligate symbioses in some spittlebug species have been quite well studied, little is known about their facultative endosymbionts, especially those inhabiting the gut. Recently, the role played by spittlebugs as vectors of the phytopathogenetic bacterium Xylella fastidiosa aroused attention to this insect group, boosting investigations aimed at developing effective yet sustainable control strategies. Since spittlebug nymphs are currently the main target of applied control, the composition of gut bacterial community of the juveniles of Philaenus spumarius and Lepyronia coleoptrata was investigated using molecular techniques. Moreover, bacteria associated with their froth, sampled from different host plants, were studied. Results revealed that Sodalis and Rickettsia bacteria are the predominant taxa in the gut of P. spumarius and L. coleoptrata nymphs, respectively, while Rhodococcus was found in both species. Our investigations also highlighted the presence of recurring bacteria in the froth. Furthermore, the foam hosted several bacterial species depending on the host plant, the insect species, or on soil contaminant. Overall, first findings showed that nymphs harbor a large and diverse bacterial community in their gut and froth, providing new accounts to the knowledge on facultative symbionts of spittlebugs.

Cicadomorpha Community (Hemiptera: Auchenorrhyncha) in Portuguese Vineyards with Notes of Potential Vectors of Xylella fastidiosa.

Cicadomorpha (Hemiptera) insects are currently responsible for a growing negative impact on the agricultural economy due to their ability to directly damage crops or through the capacity to act as vectors for plant pathogens. The phytopathogenic bacterium Xylella fastidiosa, the causal agent of Pierce's disease in vineyards, is exclusively transmitted by insects of this infraorder. Therefore, knowledge of the Cicadomorpha species and understanding their biology and ecology is crucial. In this work, in 2018 and 2019, the canopy and inter-row vegetation of 35 vineyards distributed in mainland Portugal were sampled to investigate species composition, richness, and diversity of the Cicadomorpha community, with a special focus given to vectors and potential vectors of X. fastidiosa. A total of 11,834 individuals were collected, 3003 in 2018 and 8831 in 2019. Of the 81 species/morphospecies identified, only five are considered vectors or potential vectors of this pathogen, namely, Cicadella viridis (Linnaeus, 1758), Philaenus spumarius (Linnaeus, 1758), Neophilaenus campestris (Fallén, 1805), Lepyronia coleoptrata (Linnaeus, 1758), and N. lineatus (Linnaeus, 1758). Cicadella viridis was the most abundant xylem sap feeder, followed by P. spumarius. In addition, Cicadomorpha that cause direct damage to vines and vectors of grapevine yellows' phytoplasmas were also collected and identified in the sampled vineyards. The results suggested that vectors and potential vectors of X. fastidiosa and a large proportion of the population of Cicadomorpha have a positive correction with inter-row vegetation.

Assessing the driving role of the anthropogenic landscape on the distribution of the Xylella fastidiosa-driven "olive quick decline syndrome" in Apulia (Italy).

The plant pathogen Xylella fastidiosa (Xf) is a significant threat to various economically important tree cash crops. Although previously found only in the Americas, the bacterium responsible for olive quick decline syndrome was detected in Apulia, Italy, in 2013. Since then, it has spread to approximately 54,000 ha of olive trees in the region, causing dramatic concern throughout the Mediterranean basin. As a result, it is crucial to comprehend its distribution and forecast its potential diffusion. The effect of the anthropogenic component of the landscape on the distribution of Xf remains little explored. The present study used an ecological niche model to identify how different land uses, used as proxies of different levels of human pressure across the Apulia territory, impacted the distribution of the Xf-infected olive trees in 2015-2021. Results demonstrated that the anthropogenic component significantly contributed to the epidemic, with the road system representing the main driver of diffusion and natural/seminatural areas hampering Xf spread at the landscape scale. This evidence highlighted the importance of explicitly considering the effects of the anthropogenic landscape when modelling Xf distribution and support the design of landscape-informed monitoring strategies to prevent Xf spread in Apulia and other Mediterranean countries.

Vectors as Sentinels: Rising Temperatures Increase the Risk of Xylella fastidiosa Outbreaks.

Global change is expected to modify the threat posed by pathogens to plants. However, little is known regarding how a changing climate will influence the epidemiology of generalist vector-borne diseases. We developed a high-throughput screening method to test for the presence of a deadly plant pathogen, Xylella fastidiosa, in its insect vectors. Then, using data from a four-year survey in climatically distinct areas of Corsica (France), we demonstrated a positive correlation between the proportion of vectors positive to X. fastidiosa and temperature. Notably, a higher prevalence corresponded with milder winters. Our projections up to 2100 indicate an increased risk of outbreaks. While the proportion of vectors that carry the pathogen should increase, the climate conditions will remain suitable for the bacterium and its main vector, with possible range shifts towards a higher elevation. Besides calling for research efforts to limit the incidence of plant diseases in the temperate zone, this work reveals that recent molecular technologies could and should be used for massive screening of pathogens in vectors to scale-up surveillance and management efforts.

Evaluation of Control Strategies for Xylella fastidiosa in the Balearic Islands.

The emergence of Xylella fastidiosa (Xf) in the Balearic Islands in October 2016 was a major phytosanitary challenge with international implications. Immediately after its detection, eradication and containment measures included in Decision 2015/789 were implemented. Surveys intensified during 2017, which soon revealed that the pathogen was widely distributed on the islands and eradication measures were no longer feasible. In this review, we analyzed the control measures carried out by the Balearic Government in compliance with European legislation, as well as the implementation of its control action plan. At the same time, we contrasted them with the results of scientific research accumulated since 2017 on the epidemiological situation. The case of Xf in the Balearic Islands is paradigmatic since it concentrates on a small territory with one of the widest genetic diversities of Xf affecting crops and forest ecosystems. We also outline the difficulties of anticipating unexpected epidemiological situations in the legislation on harmful exotic organisms on which little biological information is available. Because Xf has become naturalized in the islands, coexistence alternatives based on scientific knowledge are proposed to reorient control strategies towards the main goal of minimizing damage to crops and the landscape.

Transmission of Xylella fastidiosa subsp. pauca ST53 by the Sharpshooter Cicadella viridis From Different Source Plants and Artificial Diets.

The sharpshooter Cicadella viridis L. (Hemiptera: Cicadellidae) is the most common sharpshooter in Europe and, given its xylem feeding behavior, is considered a potential vector of the plant pathogenic bacterium Xylella fastidiosa Wells et al. (Xanthomonadales: Xanthomonadaceae). We tested X. fastidiosa subsp. pauca ST53 (Xfp) transmission capabilities of C. viridis adults, namely 1) acquisition efficiency from four host plant species-periwinkle, milkwort, lavender, alfalfa-and from two artificial diets (PD3 and Xfm), 2) inoculation efficiency to periwinkle at different times post acquisition from different plant and artificial diet sources. The main European vector species-Philaenus spumarius L. (Hemiptera: Aphrophoridae)-was used as a control. C. viridis was able to acquire Xfp from periwinkle, milkwort, and lavender, although with low efficiency (3-16%) and from artificial diets (23-25%). Successful inoculation on periwinkle was extremely rare, being observed only three times, following feeding on milkwort plant and PD3 artificial diet sources. Our study shows that C. viridis is not a relevant vector of Xfp, given the very low transmission rate in controlled conditions, and the inability to feed on olive. The low efficiency reported here correlates with ecological constraints of the vector (mainly monocots host plants, humid environments) that make it difficult to forecast a relevant role in dispersing X. fastidiosa, at least within the present distribution of the exotic bacterium in Europe. However, a possible role of this species in spreading Xf in other agroecosystems, e.g., vineyard and stone fruits grown in humid areas, cannot be excluded.

Preliminary Molecular Survey of the Possible Presence of Xylella fastidiosa in the Upper Ionian Coasts of Calabria, Italy, through the Capture and Analysis of Its Main Vector Insects.

Xylella fastidiosa subsp. pauca, strain CoDiRO is the bacterium responsible for the onset of the disease known as the olive quick decline syndrome, which has been causing a phytosanitary and economic emergency in the Apulia region since 2013. To date, three insect species have been identified as pathogenic carriers of X. fastidiosa. With the advancement of the infection front, and the possibility of pathogenic insects being "hitchhiked" over long distances, the monitoring of the vectors of X. fastidiosa in the Italian regions bordering Apulia is an increasingly contingent issue for the rapid containment of the bacterium and the protection of the olive-growing heritage. Accordingly, the present research concerned the capture and recognition of the vector insects of X. fastidiosa in the upper Ionian coasts of Calabria (Italy) to evaluate the possible presence of the bacterium through molecular diagnostic techniques. The sampling allowed us to ascertain the presence of Philaenus spumarius and Neophilaenus campestris and their preferential distribution in olive groves and meadows, whereas all the 563 individuals tested negative for the pathogen.

Xylella fastidiosa in Olive: A Review of Control Attempts and Current Management.

Since 2013, Xylella fastidiosa Wells et al. has been reported to infect several hosts and to be present in different areas of Europe. The main damage has been inflicted on the olive orchards of southern Apulia (Italy), where a severe disease associated with X. fastidiosa subspecies pauca strain De Donno has led to the death of millions of trees. This dramatic and continuously evolving situation has led to European and national (Italian and Spanish) measures being implemented to reduce the spread of the pathogen and the associated olive quick decline syndrome (OQDS). Research has been also carried out to find solutions to better and directly fight the bacterium and its main insect vector, Philaenus spumarius L. In the course of this frantic effort, several treatments based on chemical or biological substances have been tested, in addition to plant breeding techniques and integrated pest management approaches. This review aims to summarize the attempts made so far and describe the prospects for better management of this serious threat, which poses alarming questions for the future of olive cultivation in the Mediterranean basin and beyond.

Population structure, adaptation and divergence of the meadow spittlebug, Philaenus spumarius (Hemiptera, Aphrophoridae), revealed by genomic and morphological data.

Understanding patterns of population differentiation and gene flow in insect vectors of plant diseases is crucial for the implementation of management programs of disease. We investigated morphological and genome-wide variation across the distribution range of the spittlebug Philaenus spumarius (Linnaeus, 1758) (Hemiptera, Auchenorrhyncha, Aphrophoridae), presently the most important vector of the plant pathogenic bacterium Xylella fastidiosa Wells et al., 1987 in Europe. We found genome-wide divergence between P. spumarius and a very closely related species, P. tesselatus Melichar, 1899, at RAD sequencing markers. The two species may be identified by the morphology of male genitalia but are not differentiated at mitochondrial COI, making DNA barcoding with this gene ineffective. This highlights the importance of using integrative approaches in taxonomy. We detected admixture between P. tesselatus from Morocco and P. spumarius from the Iberian Peninsula, suggesting gene-flow between them. Within P. spumarius, we found a pattern of isolation-by-distance in European populations, likely acting alongside other factors restricting gene flow. Varying levels of co-occurrence of different lineages, showing heterogeneous levels of admixture, suggest other isolation mechanisms. The transatlantic populations of North America and Azores were genetically closer to the British population analyzed here, suggesting an origin from North-Western Europe, as already detected with mitochondrial DNA. Nevertheless, these may have been produced through different colonization events. We detected SNPs with signatures of positive selection associated with environmental variables, especially related to extremes and range variation in temperature and precipitation. The population genomics approach provided new insights into the patterns of divergence, gene flow and adaptation in these spittlebugs and led to several hypotheses that require further local investigation.

Phenology, Seasonal Abundance, and Host-Plant Association of Spittlebugs (Hemiptera: Aphrophoridae) in Vineyards of Northwestern Italy.

Spittlebugs (Hemiptera: Aphrophoridae) are the vectors of the bacterium Xylella fastidiosa (Xf) in Europe. Xf may cause severe epidemics in cultivated plants, including grapevines. To assess the threat represented by the bacterium to grapevines, detailed information on the vectors' phenology, density, and ecology in vineyards is needed. The aim of the present work was to describe spittlebug diversity, phenology, and host-plant association in the vineyard agroecosystem. Two separate field surveys of nymphal and adult spittlebug populations, i.e., a two-year survey of a single site and a one-year survey of three sites, were performed in vineyards of northwestern Italy in three consecutive years. Philaenus spumarius was the most common species, reaching average nymph densities on herbaceous cover up to 60-130 nymphs/m2. Adults were sampled on grapevines from May to September, with a peak in June (up to 0.43 insects/sweep). Herbaceous cover was colonized after egg hatching and in late summer for oviposition, while wild woody hosts represented a refuge during summer. The results show that spittlebugs can reach high population levels in vineyards, at least in the areas where the ground is covered by herbaceous plants for the whole season and the use of insecticides is moderate. The extended presence of P. spumarius adults on grapevines represents a serious risk factor for the spread of Xf. The scenarios of Xf establishment in vineyards in northwestern Italy and Europe are discussed in relation to the abundance, phenology, and plant association of spittlebugs.

Electrophysiological responses of Philaenus spumarius and Neophilaenus campestris females to plant volatiles.

The spittlebugs Philaenus spumarius and Neophilaenus campestris (Hemiptera: Aphrophoridae) are xylem-feeder insects that have been identified as vectors of Xylella fastidiosa in Europe. In the present study, we aim to identify volatile organic compounds (VOCs) that may act as semiochemicals for these species. Using the dynamic headspace technique, we collected VOCs from Olea europaea L. and Polygala myrtifolia L., highly susceptible plant species to X. fastidiosa, Pinus halepensis Mill., a common plant where N. campestris is found during summer, and from host plant species that are used as cover crops or exist as natural vegetation in olive orchards, such as Cistus creticus L., Medicago sativa L., Cynodon dactylon (L.) Pers., Lolium arundinaceum (Schreb.) Darbysh., Apium graveolens L. and Petroselinum crispum (Mill.) Fuss. We tested the response of female antennae on those blends with Gas Chromatography-Electroantennographic Detection (GC-EAD). The chemical profile of C. creticus and P. halepensis was rich in terpenes, alcohols, aldehydes, and esters. In the O. europaea profile, the main compounds were terpenes. As for P. myrtifolia, the volatile profile consisted mostly of alkanes and their substitutes. In the volatile profile of M. sativa, C. dactylon and L. arundinaceum, common volatile compounds were detected. Petroselinum crispum and Apium graveolens chemical profiles were dominated by terpenes. Several compounds elicited a consistent response to the female antennae of both species. In total, 65 compounds elicited consistent EAD responses for P. spumarius and 16 compounds for N. campestris. α-pinene was found in all tested plants and elicited consistent EAD responses of P. spumarius in five plants. In addition, antennae of P. spumarius females responded to camphor, limonene, 4-methyl octane and sabinene. These compounds were found in the volatile profile of at least 5 out of 8 examined plant species. Behavioral bioassays using Y-tube olfactometry were performed on volatiles that elicited antennal responses during electrophysiological studies. Among the compounds tested in behavioral studies, namely (-)-α-pinene, (+)-α-pinene, sabinene, (-)-S-limonene and (1R)-(+)-camphor, only the last one elicited a significant attraction response by P. spumarius females. The results achieved shed light on the VOCs from selected host plant species of X. fastidiosa that are perceived by two important insect vectors and a non-host plant, P. crispum. The identification of semiochemicals for manipulating spittlebugs' behavior contribute to the development of efficient monitoring tools for X. fastidiosa vectors.

Populations and Host/Non-Host Plants of Spittlebugs Nymphs in Olive Orchards from Northeastern Portugal.

The Aphrophoridae family contains important vectors of Xylella fastidiosa, a serious bacterial plant disease. In olive orchards, nymphs usually feed on the ground-cover vegetation. However, detailed information about their populations and host/non-host plants in some regions threatened by Xylella, such as the northeast of Portugal, is very limited. The goal of our work was to identify the vector species, nymphal development period, and their host and non-host herbaceous plants in olive orchards from northeastern Portugal. Ground-cover plant species hosting or not hosting nymphs were identified during the spring of 2017 to 2019 in olive orchards. Nymphal development period, nymph aggregation, and nymph's preferred feeding height of the ground-cover plants were recorded. The most abundant Aphrophoridae species was Philaenus spumarius followed by Neophilaenus sp. Nymphs developed from April to early May and showed a low number of individuals per foam (generally between one and three). They preferred the middle part of the plants. Philaenus spumarius feeds preferentially on Asteraceae and Fabaceae, and Neophilaenus sp. on Poaceae. Some abundant plants, such as Bromus diandrus, Astragalus pelecinus, Chrysanthemum segetum, Trifolium spp., Caryophyllaceae, and Brassicaceae, were barely colonized by Aphrophoridae nymphs. This knowledge is essential for the selection of the species composition of ground-cover vegetation to minimize the presence of vectors of X. fastidiosa in olive groves.

Development of an FTP-LAMP assay based on TaqMan real-time PCR and LAMP for the specific detection of Xylella fastidiosa De Donno and mulberry strains in both plants and insect vectors.

We developed two real-time detection assays, TaqMan real-time PCR and LAMP, using primers and probe designed based on a sequence annotated to code for a Haemagglutinin-related protein (Hg) of Xylella fastidiosa (Xf), a gene uniquely present in the Italian olive (De Donno of olive) and American mulberry strains, for specific detection of the target Xf strains. These assays were validated with DNA samples extracted from Xf-infected plant samples and from two species of insect vectors (Philaenus spumarius, Ps; and Neophilaenus campestris, Nc). Both techniques were proven to be highly sensitive (100 fg of Xf-genomic DNA) and specific to the Italian De Donno and American mulberry strains of Xf. When our LAMP was utilized in a duplex manner by combining with previously published universal primers and probe for detection of all Xf-subspecies and strains, the duplex LAMP showed high versatility in the simultaneous detection and differentiation of the Italian De Donno and American mulberry stains form other subspecies/strains. Furthermore, the Hg gene-specific LAMP primers and TaqMan probe were exploited to develop a new approach; henceforth referred to as the Fluorescence of TaqMan Probe upon Dequenching - Loop mediated Isothermal Amplification (FTP-LAMP). In the FTP-LAMP, the Xf-Hg specific fluorophore-quenched probe was added to a standard LAMP reaction and fluoresces only when bound to its target, allowing for a sequence-specific detection of the Xf-Italian De Donno and American mulberry strains in a LAMP context. Our FTP-LAMP assay showed to be highly sensitive detecting down to 100 fg genomic DNA of Xf, when tested on Xf-genomic DNA extracted from infected plants, DAS-ELISA-crude saps and insect vectors. Furthermore, the assay showed high specificity (98.7% vs 89% for LAMP) when applied on DNA templates from insect vectors. With the addition of an extra target sequence-specific probe acting as a direct Xf-specific dye, the FTP-LAMP has gained more specificity and reduced one of the main problems of the LAMP assay (false positives) when used for detecting of Xf in insect vectors. To the best of our knowledge, this study reports the development of the first LAMP assay and the first novel FTP-LAMP method for specific detection of the Italian De Donno and the American mulberry strains of Xf. Together with the Xf universal LAMP primers in a duplex approach, the FTP-LAMP could represent a useful tool not only for the specific detection of the olive-associated strain in Italy, but also to differentiate the De Donno strain from other strains of Xf already reported in Italy and Europe (Germany, France, Spain and Portugal).

Spittlebugs of Mediterranean Olive Groves: Host-Plant Exploitation throughout the Year.

Spittlebugs are the vectors of the bacterium Xylella fastidiosa Wells in Europe, the causal agent of olive dieback epidemic in Apulia, Italy. Selection and distribution of different spittlebug species on host-plants were investigated during field surveys in 2016-2018 in four olive orchards of Apulia and Liguria Regions of Italy. The nymphal population in the herbaceous cover was estimated using quadrat samplings. Adults were collected by sweeping net on three different vegetational components: herbaceous cover, olive canopy, and wild woody plants. Three species of spittlebugs were collected: Philaenus spumarius L., Neophilaenus campestris (Fallén), and Aphrophora alni (L.) (Hemiptera: Aphrophoridae). Philaenus spumarius was the predominant species both in Apulia and Liguria olive groves. Nymphal stages are highly polyphagous, selecting preferentially Asteraceae Fabaceae plant families, in particular some genera, e.g., Picris, Crepis, Sonchus, Bellis, Cichorium, and Medicago. Host-plant preference of nymphs varies according to the Region and through time and nymphal instar. In the monitored sites, adults peak on olive trees earlier in Apulia (i.e., during inflorescence emergence) than in Liguria (i.e., during flowering and beginning of fruit development). Principal alternative woody hosts are Quercus spp. and Pistacia spp. Knowledge concerning plant selection and ecological traits of spittlebugs in different Mediterranean olive production areas is needed to design effective and precise control strategies against X. fastidiosa vectors in olive groves, such as ground cover modifications to reduce populations of spittlebug vectors.

Distribution and Relative Abundance of Insect Vectors of Xylella fastidiosa in Olive Groves of the Iberian Peninsula.

The phytosanitary emergency caused by the spread of Xylella fastidiosa in the Mediterranean has raised demands for a better understanding of the ecology of its presumed and candidate insect vectors. Here, we present the results of a two-year survey carried out in olive groves across southern, eastern and Central Spain and northeastern Portugal. Several sampling methods were tested and compared to select the most appropriate to estimate population levels of potential vectors of X. fastidiosa. The spittlebugs Philaenus spumarius and Neophilaenus campestris (Hemiptera: Aphrophoridae) were the main species associated with olive groves. Both species were widely present on herbaceous ground vegetation within the olive groves; P. spumarius mainly associated with Asteraceae and N. campestris with Poaceae. Due to the patchy distribution of spittlebugs within the olive groves, sweep nets were the most effective and least time-consuming sampling method for the estimation of population size both in the ground cover and tree canopies. Trends in population density showed that spittlebugs can be abundant on ground vegetation but very rare on olive canopies. Spittlebugs disperse in late spring to non-cultivated hosts that act as natural reservoirs. In late fall, adults return to the olive groves for oviposition. However, olive trees may act as transient hosts for spittlebugs and high population densities of these insect vectors should be avoided in areas where X. fastidiosa is present.

Seasonal Fluctuations of Sap-Feeding Insect Species Infected by Xylella fastidiosa in Apulian Olive Groves of Southern Italy.

A study on seasonal abundance of Auchenorrhyncha species and their infectivity by Xylella fastidiosa in the Apulia region of Italy was conducted to identify ideal periods for monitoring and adoption of potential control measures against insect vectors. Adult populations of Auchenorrhyncha species were monitored monthly over a 2-yr period from five olive groves. A total of 15 species were captured, identified, and tested for presence of X. fastidiosa by polymerase chain reaction (PCR). For three species, Philaenus spumarius L., Neophilaenus campestris (Fallèn), and Euscelis lineolatus Brullé, positive reactions to X. fastidiosa were obtained, on average, in 16.3, 15.9 and 18.4% of adult insects, respectively. Philaneous spumarius was the dominant species (39.8% of total Auchenorrhyncha captured) with the highest adult abundance in summer months. Adult P. spumarius and N. campestris were first detected between March and May in both years, and all insects tested during these periods (year 1: n = 42, year 2: n = 132) gave negative reactions to X. fastidiosa by PCR. Similarly, first adults of E. lineolatus that appeared from October to November (year 1: n = 20, year 2: n = 15) tested negative for presence of X. fastidiosa Given the lack of transstadial and transovarial transmission of X. fastidiosa and considering that P. spumarius is univoltine, control measures against nymphal stages of P. spumarius should be investigated as means of population suppression to reduce spread of X. fastidiosa in olive groves.

Top-down effects of insect herbivores during early succession: influence on biomass and plant dominance.

We tested the hypothesis that phytophagous insects would have a strong top-down effect on early successional plant communities and would thus alter the course of succession. To test this hypothesis, we suppressed above-ground insects at regular intervals with a broad-spectrum insecticide through the first 3 years of old-field succession at three widely scattered locations in central New York State. Insect herbivory substantially reduced total plant biomass to a similar degree at all three sites by reducing the abundance of meadow goldenrod, Solidago altissima. As a result, Euthamia graminifolia dominated control plots whereas S. altissima dominated insecticide-treated plots by the third year of succession. S. altissima is the dominant old-field herbaceous species in this region but typically requires at least 5 years to become dominant. Past explanations for this delay have implicated colonization limitation whereas our data demonstrate that insect herbivory is a likely alternative explanation. A widespread, highly polyphagous insect, the xylem-tapping spittlebug, Philaenus spumarius, appeared to be the herbivore responsible for the reduction in standing crop biomass at all three sites. Insect herbivory typically caused little direct leaf tissue loss for the ten plant species we examined, including S. altissima. Consequently, the amount of leaf area removed was not a reliable indicator of the influence of insect herbivory on standing crop biomass or on early succession. Overall, we found a strong top-down effect of insect herbivores on biomass at several sites, so our results may be broadly applicable. These findings run counter to generalizations that top-down effects of herbivores, particularly insects, are weak in terrestrial systems. These generalizations may not apply to insects, such as spittlebugs, that can potentially mount an effective defense (i.e., spittle) against predators and subsequently reach relatively high abundance on common plant species. Our results suggest that insect herbivory may play an important but often overlooked role during early old-field succession.

APHID DISTRIBUTION AND THE EVOLUTION OF GOLDENROD RESISTANCE.

Although there is considerable evidence indicating that herbivory is detrimental to plant fitness, some recent studies of the evolution of plant resistance have concluded that insects do not impose selection on their host plants. A previously untested assumption that underlies most studies of the evolution of plant resistance is that insect distribution patterns are controlled directly by the effects of plant genotype on insect preference and performance. The experiments described here explicitly tested this assumption using the specialist herbivore Uroleucon tissoti (Homoptera: Aphididae) and its host plant Solidago altissima (Asteraceae). Measures of aphid preference and performance were used to predict aphid distribution patterns, and then the predicted distribution patterns were compared with the natural distribution pattern. Although goldenrod genotype had a strong effect on aphid distribution, aphid distribution was not controlled directly by the effect of goldenrod genotype on aphid preference and performance. Instead, a second experiment demonstrated that aphid and spittlebug (Philaenus spumarius and Lepyronia quadrangularis Homoptera: Cercopidae) distribution is controlled largely by genetic variation for resistance to a suite of "branch-causing" herbivores. These herbivores induce branching and aphids and spittlebugs are more abundant on branched plants than unbranched plants. These results indicate that any natural selection imposed by aphids and spittlebugs on goldenrod will depend on the presence or absence of branch-causing herbivores. Thus, selection for plant resistance may depend as much on the assemblage of insect species present as on the identity of each individual species.