Insect Vectors of Plant Pathogens

Huanglongbing (HLB) is a citrus disease of worldwide importance, associated with the presence of Candidatus Liberibacter asiaticus (Las) and vectored by the psyllid Diaphorina citri in Asia and the Americas. To properly manage HLB, removal of inoculum sources and control of the psyllid are undertaken. We evaluated the percentage of the psyllid population with Las, sampled from yellow sticky traps over a three-year period and its relationship with insect population, regions, season of the year, and HLB management in citrus areas in the southwestern, central, and northern regions of São Paulo (SP) and southwestern region of Minas Gerais states, Brazil. In each reading, up to 50 psyllids per region were collected and detection of Las in individual psyllids were made by quantitative polymerase chain reaction. The percentage of psyllids with Las—an average of 65.3%—was constant throughout the year in the southwestern region of SP state, while showing an increase from spring to autumn when sampled from central to northern regions. The proportion of psyllids carrying Las from each region and year period were compared by a proportion test and spectral density analysis. The proportion of psyllids carrying Las evaluated in the same region in different seasons presented statistical differences in central (Araraquara) and southwestern (Santa Cruz do Rio Pardo) regions in 2015, with higher values in the first semester (summer and autumn) than in the second semester (winter and spring). Orchards with poor HLB management had higher incidence of psyllids with Las. Spectral density analysis indicated that good management areas had 50% less relevant peaks of psyllids with Las than in areas with poor HLB management practices. The relationship between the percentage of psyllids carrying Las and the number of captured psyllids in the region in a given time denotes the most critical intake time for HLB spread in citrus orchards. The reduction in the population of psyllids carrying Las is a direct benefit from the use of good management practices. Full article

A combination of biological control and host plant resistance would be desirable for optimally controlling the greenhouse whitefly, Trialeurodes vaporariorum in tomato crops. Whitefly settlement preference, oviposition, and survivorship were evaluated on ABL 10-4 and ‘Moneymaker’, two nearly-isogenic tomato lines with, and without, whitefly-resistance traits based on type IV leaf glandular trichomes derived from the tomato wild species Solanum pimpinellifolium, respectively. Significantly reduced preference of T. vaporariorum adult whiteflies for ABL 10-4 leaves was observed. Moreover, T. vaporariorum altered its abaxial–adaxial settling performance on leaves of ABL 10-4 plants. A significantly lower tendency to settle on abaxial leaf surface was observed in ABL 10-4 compared to Moneymaker plants. Furthermore, T. vaporariorum deposited fewer eggs and exhibited a significantly reduced egg to adult survivorship in ABL 10-4 than in Moneymaker plants. Therefore, reduced fitness and distorted performance were observed for T. vaporariorum on ABL 10-4 tomato plants supporting that type IV leaf glandular trichomes might protect them from this pest and, indirectly, from the viruses it transmits. Full article

The meadow spittlebug, Philaenus spumarius (L.) (Hemiptera Aphrophoridae), the main vector of Xylella fastidiosa Wells et al. in Europe, has few known natural enemies. The endoparasitoid Verrallia aucta (Fallén) (Diptera, Pipunculidae) was first noticed a long time ago but very little is known about its biology and prevalence. In this study, the presence and prevalence of V. aucta were investigated in different regions of northern Italy, both in plain–foothill and montane zones. Parasitic larvae were identified by the dissection of spittlebug adults, P. spumarius and Neophilaenus campestris (Fallén), and by a new species-specific molecular tool targeting the ITS2 and COI genomic regions, developed in this work. A small-scale rearing was set up to gain information on the life cycle of V. aucta on its main host P. spumarius. During the four-year investigation (2016–2019) the pipunculid parasitoid displayed low prevalence, reaching a maximum parasitization rate of 17.5% (calculated over the adult spittlebug season) in vineyards of the Piemonte region. Over the whole period, no significant difference in the prevalence was found between male and female spittlebugs. Collected data and rearing observations suggest that V. aucta is monovoltine and synchronous with P. spumarius, laying eggs in newly emerged adults, developing as an endoparasitoid through two larval stages during the whole summer, and overwintering as a pupa in the soil. Full article

Hyalesthes obsoletus is the vector of “Candidatus Phytoplasma (Ca. P.) solani,” the causal agent of grapevine yellows Bois noir (BN). The relationships among the planthopper, its main herbaceous hosts as phytoplasma reservoirs (Convolvolus arvensis and Urtica dioica) and BN spreading were studied in northern Italy. In two areas the relationship between host plants and the phenology and survival of planthopper adults was investigated in potted plants and in field conditions. Moreover, H. obsoletus ecology, newly symptomatic grapevine occurrence and “Ca. P. solani” tuf-types’ presence were studied in two vineyards (2014–2019). An earlier occurrence of H. obsoletus adults on C. arvensis than U. dioica and better adult survival of the originating host were observed. When U. dioica was prevalent, the vector occurred almost exclusively along the ditch outside the vineyard. Hyalesthes obsoletus amount varied widely from year to year and nymphal mortality due to late frosts was supposed. In one vineyard, the amount of newly symptomatic grapevines was significantly correlated with vector abundance in the previous year. The “Ca. P. solani” tuf-type was influenced by vector population levels on the two hosts. Since the abundance of H. obsoletus populations on the two hosts influences BN epidemiology and dynamics and the “Ca. P. solani” tuf-type, this must be considered in BN control strategies. Full article

Phytoplasma transmission takes place by insect vectors through an Acquisition Access Period (AAP), Latency Period (LP) and Inoculation Access Period (IAP). Generally, phytoplasmas are believed to be transmitted more efficiently by nymphs because they need a long LP to reach the salivary glands before becoming infective. The transmission can start from adults as well, but in this case a long LP may exceed the insect’s lifespan. However, previous evidence has indicated that adults can undergo a shorter LP, even though little knowledge is available regarding the phytoplasma temporal dynamics during this period. Here, we investigate the minimum time required by the phytoplasma to colonize the vector midgut and salivary glands, and finally to be inoculated into a plant. We used the leafhopper Euscelidius variegatus to investigate the life cycle of flavescence dorée phytoplasma (FDP). Phytoplasma-free E. variegatus adults were left on broad beans (BBs) infected with FDP for an AAP of 7 days. Subsequently, they were individually transferred onto a healthy BB for seven different IAPs, each one lasting 24 h from day 8 to 14. Molecular analyses and fluorescence in situ hybridization were performed for FDP detection. FDP was found in the leafhopper midgut from IAP 1 with an infection rate reaching 50%, whereas in the salivary glands it was found from IAP 2 with an infection rate reaching 30%. FDP was also detected in BBs from IAP 4, with infection rates reaching 10%. Our results represent an important step to further deepen the knowledge of phytoplasma transmission and its epidemiology. Full article

Mixed infection of plant viruses is ubiquitous in nature and can affect virus–plant–vector interactions differently than single virus infection. While several studies have examined virus–virus interactions involving mixed virus infection, relatively few have examined effects of mixed virus infection on vector preference and fitness, especially when multiple vectors are involved. This study explored how single and mixed viral infection of a non-persistently transmitted cucumber mosaic virus (CMV) and propagative and persistently-transmitted tomato spotted wilt orthotospovirus (TSWV) in pepper, Capsicum annum L., influenced the preference and fitness of their vectors, the green peach aphid, Myzus persicae (Sulzer), and the tobacco thrips, Frankliniella fusca (Hinds), respectively. In general, mixed infected plants exhibited severe symptoms compared with individually infected plants. An antagonistic interaction between the two viruses was observed when CMV titer was reduced following mixed infection with TSWV in comparison with the single infection. TSWV titer did not differ between single and mixed infection. Myzus persicae settling preference and median developmental were not significantly different between CMV and/or TSWV-infected and non-infected plants. Moreover, M. persicae fecundity did not differ between CMV-infected and non-infected pepper plants. However, M. persicae fecundity was substantially greater on TSWV-infected plants than non-infected plants. Myzus persicae fecundity on mixed-infected plants was significantly lower than on singly-infected and non-infected plants. Frankliniella fusca fecundity was higher on CMV and/or TSWV-infected pepper plants than non-infected pepper plants. Furthermore, F. fusca-induced feeding damage was higher on TSWV-infected than on CMV-infected, mixed-infected, or non-infected pepper plants. Overall, our results indicate that the effects of mixed virus infection on vectors were not different from those observed following single virus infection. Virus-induced host phenotype-modulated effects were realized on both specific and non-specific vectors, suggesting crosstalk involving all vectors and viruses in this pathosystem. The driving forces of these interactions need to be further examined. The effects of interactions between two viruses and two vectors towards epidemics of one or both viruses also need to be examined. Full article

Plant pathogenic bacteria may influence vector behavior by inducing physiological changes in host plants, with implications for their spread. Here, we studied the effects of maize bushy stunt phytoplasma (MBSP) on the host selection behavior of the leafhopper vector, Dalbulus maidis (DeLong and Wolcott). Choice assays contrasting leaves of healthy (mock-inoculated) vs. infected maize (Zea mays L.) were conducted during the asymptomatic and symptomatic phases of plant infection, with leafhopper males or females previously exposed to infected plants (bacteriliferous insects) or not. In each assay, 40 adults were released in choice arenas where only the leaves of two plants from each treatment were offered and visible, and the insects landed on the leaves were counted 1, 2, 3, 5, 7, 9, 11 and 23 h after release. During the asymptomatic phase of plant infection, an effect was observed only on bacteriliferous females, who preferred leaves of healthy plants 5 h after release or later. The symptomatic phase triggered a pull–push effect on non-bacteriliferous females, who were first attracted to symptomatic leaves but hours later moved to healthy leaves. Non-bacteriliferous males initially preferred symptomatic leaves (up to 5 h after release) and later became equally distributed between treatments. Bacteriliferous males and females initially did not discriminate between healthy and symptomatic leaves, but only the females tended to move to healthy leaves 9 h after release. Oviposition was drastically reduced on symptomatic leaves. The changes in vector behavior induced by MBSP favor its primary spread, since bacteriliferous females prefer healthy leaves at early (asymptomatic) stages of the crop. At later stages, secondary spread may be favored because non-bacteriliferous females are initially attracted to infected (symptomatic) leaves, allowing pathogen acquisition and subsequent transmission as they move to healthy plants. Full article

The transmission of phytoplasmas is the result of an intricate interplay involving pathogens, insect vectors and host plants. Knowledge of the vector’s competence during its lifespan allows us to define more sustainable well-timed control strategies targeted towards the most worrisome life stages. We investigated the temporal dynamics of ‘Candidatus Phytoplasma mali’ load in Cacopsylla melanoneura in the different developmental stages in Northwest Italy. The phytoplasma load in the vector was evaluated in overwintering adults, nymphs and newly emerged adults after different acquisition access periods. Moreover, we followed the multiplication of the phytoplasma during the aestivation and the overwintering period on conifers. Our results confirmed the ability of remigrants to retain the phytoplasma until the end of winter. We also highlighted the high acquisition efficiency and vector competence, based on phytoplasma load, of nymphs and newly emerged adults. Therefore, particular attention should be paid to the management of overwintered C. melanoneura as soon as they return to the orchards, but also to newly emerged adults, particularly in orchards with a high infection rate and when the migration to conifers is delayed. Full article

Tomato chlorosis virus (ToCV) is a phloem-limited crinivirus transmitted by whiteflies and seriously affects tomato crops worldwide. As with most vector-borne viral diseases, no cure is available, and the virus is managed primarily by the control of the vector. This study determined the effects of the foliar spraying with the insecticides, acetamiprid, flupyradifurone and cyantraniliprole, on the feeding behavior, mortality, oviposition and transmission efficiency of ToCV by B. tabaci MEAM1 in tomato plants. To evaluate mortality, oviposition and ToCV transmission in greenhouse conditions, viruliferous whiteflies were released on insecticide-treated plants at different time points (3, 24 and 72 h; 7 and 14 days) after spraying. Insect mortality was higher on plants treated with insecticides; however, only cyantraniliprole and flupyradifurone differed from them in all time points. The electrical penetration graph (DC-EPG) technique was used to monitor stylet activities of viruliferous B. tabaci in tomato plants 72 h after insecticide application. Only flupyradifurone affected the stylet activities of B. tabaci, reducing the number and duration of intracellular punctures (pd) and ingestion of phloem sap (E2), a behavior that possibly resulted in the lower percentage of ToCV transmission in this treatment (0–60%) in relation to the control treatment (60–90%) over the periods evaluated. Our results indicate that flupyradifurone may contribute to management of this pest and ToCV in tomato crops. Full article

Bacteria belonging to ‘Candidatus Liberibacter spp.’ are associated with various severe diseases in the five continents. The African citrus psyllid Trioza erytreae (Hemiptera: Triozidae) is an efficient vector of citrus huanglongbing-HLB disease, absent in the Mediterranean basin. This psyllid is currently present in the islands and mainland Portugal and Spain, where the prevalence of ‘Ca. Liberibacter solanacearum’ (CaLsol) associated to a carrot disease is high. Trioza erytreae normally feeds on citrus plants but has also been observed on other crops. It would be a great concern to the Mediterranean citrus industry if T. erytreae could transmit this bacterium from carrots to citrus and cause disease; therefore, the transmission of CaLsol from carrot plants to citrus plants was experimentally assessed. Although CaLsol was initially detected on receptor citrus plants in transmission assays by dodder and budding, the infection was not established. The feeding behavior by electrical penetration graphs and oviposition of T. erytreae on carrot plants versus citrus plants was evaluated. Trioza erytreae only reached the phloem in citrus plants. However, it was able to acquire CaLsol from infected carrots but unable to transmit it to citrus plants. CaLsol was detected in some carrot plants immediately after 7 and 14 days (inoculation access period), but it was not detected after one month. Trioza erytreae was unable to complete its life cycle on carrot plants. In conclusion, the efficient vector of bacteria associated to huanglongbing was unable to transmit CaLsol from carrot to citrus plants, but it acquired and transmitted the bacterium from carrot to carrot plants with low efficiency. Full article

Xylella fastidiosa is a vector-borne bacterium that causes diseases in many plants of economic interest. The bacterium–vector initial interactions involve bacterial membrane-bound adhesins that mediate cell attachment to the foregut of insect vectors. We investigated the role of the afimbrial adhesin XadA2 in the binding and biofilm formation of X. fastidiosa subsp. pauca to vector surfaces in vitro, as well as its potential to disrupt pathogen transmission. We showed that XadA2 has binding affinity for polysaccharides on sharpshooter hindwings, used as a proxy for the interactions between X. fastidiosa and vectors. When in a medium without carbon sources, the bacterium used wing components, likely chitin, as a source of nutrients and formed a biofilm on the wing surface. There was a significant reduction in X. fastidiosa biofilm formation and cell aggregation on vector wings in competition assays with XadA2 or its specific antibody (anti-XadA2). Finally, pathogen acquisition and transmission to plant were significantly reduced when the vectors acquired X. fastidiosa from an artificial diet supplemented with anti-XadA2. These results show that XadA2 is important in mediating bacterial colonization in the insect and that it could be used as a target for blocking X. fastidiosa transmission. Full article

Climate change impacts crop production, pest and disease pressure, yield stability, and, therefore, food security. In order to understand how climate and atmospheric change factors affect trophic interactions in agriculture, we evaluated the combined effect of elevated carbon dioxide (CO₂) and temperature on the interactions among wheat (Triticum aestivum L.), Barley yellow dwarf virus species PAV (BYDV-PAV) and its vector, the bird cherry-oat aphid (Rhopalosiphum padi L.). Plant traits and aphid biological parameters were examined under two climate and atmospheric scenarios, current (ambient CO₂ and temperature = 400 ppm and 20 °C), and future predicted (elevated CO₂ and temperature = 800 ppm and 22 °C), on non-infected and BYDV-PAV-infected plants. Our results show that combined elevated CO₂ and temperature increased plant growth, biomass, and carbon to nitrogen (C:N) ratio, which in turn significantly decreased aphid fecundity and development time. However, virus infection reduced chlorophyll content, biomass, wheat growth and C:N ratio, significantly increased R. padi fecundity and development time. Regardless of virus infection, aphid growth rates remained unchanged under simulated future conditions. Therefore, as R. padi is currently a principal pest in temperate cereal crops worldwide, mainly due to its role as a plant virus vector, it will likely continue to have significant economic importance. Furthermore, an earlier and more distinct virus symptomatology was highlighted under the future predicted scenario, with consequences on virus transmission, disease epidemiology and, thus, wheat yield and quality. These research findings emphasize the complexity of plant–vector–virus interactions expected under future climate and their implications for plant disease and pest incidence in food crops. Full article

Candidatus Liberibacter asiaticus (CLas) is a phloem-limited bacterium that is associated with the Huanglongbing (HLB) disease of citrus and transmitted by the psyllid, Diaphorina citri. There are no curative methods to control HLB and the prevention of new infections is essential for HLB management. Therefore, the objective of our study was to determine the effects of systemic insecticides, such as the neonicotinoids imidacloprid, thiamethoxam, and a mixture of thiamethoxam and chlorantraniliprole (diamide) on the probing behavior of CLas-infected D. citri and their effect on CLas transmission. The electrical penetration graph (EPG-DC) technique was used to monitor the stylet penetration activities of CLas-infected D. citri on sweet orange [Citrus sinensis (L.) Osbeck] ‘Valencia’ treated with systemic insecticides. Systemic insecticides disrupted the probing behavior of CLas-infected D. citri, in a way that affected CLas transmission efficiency, particularly by negatively affecting the stylet activities related to the phloem phase. All insecticides reduced (by 57–73%) the proportion of psyllids that exhibited sustainable phloem ingestion (waveform E2 > 10 min), with significant differences observed on plants treated with thiamethoxam and thiamethoxam + chlorantraniliprole. The transmission rate of CLas with high inoculum pressure (five CLas-infected D. citri per plant and a seven-day inoculation access period) to untreated control plants was 93%. In contrast, CLas transmission was reduced to 38.8% when test plants were protected by systemic insecticides. Our results indicated that all insecticides tested presented a potential to reduce CLas inoculation by an average of 59%; therefore, these insecticides can be used to reduce the spread of HLB. Full article

Quantitative estimates of vector populations and their infectivity in the wild and in cultivated compartments of agroecosystems have been carried out to elucidate the role of the wild compartment in the epidemiology of Flavescence dorée (FD). Seven sites were selected for the investigations in the Piedmont Region of Italy. They were characterized by a high variety of agricultural and ecological landscape features, and included a vineyard surrounded by wild vegetation. In order to describe abundance and prevalence of FD-infected vectors in the cultivated and wild compartments of the vineyard agroecosystem, adults of Scaphoideus titanus were collected by yellow sticky traps inside and outside the vineyard over the period July 10th–September 9th, 2015. They were counted and singly analyzed for the presence of FD phytoplasmas by PCR. Multifactorial correlations among vector population level, prevalence of infected insects inside and outside the vineyards, disease prevalence in cultivated and wild Vitis plants, and location of wild Vitis plants with respect to the vineyard were analyzed. Abundance of S. titanus adults significantly decreased from the end of July onwards, particularly inside the vineyard (average range 22.7 ± 2.5 insects/trap). Percentage of FD-positive S. titanus was significantly higher outside the vineyard (up to 48% on average) compared to inside the vineyard (up to 34% on average), and increased during the season in both compartments. Full article

Tomato yellow leaf curl virus (TYLCV), which is transmitted by Bemisia tabaci in a persistent-circulative manner, threatens tomato production worldwide. Little is known about the complicated interaction during this process at the molecular level. In this study, viral AAPs at 0 h, 2 h, 6 h, 12 h and 48 h were investigated using a comparative transcriptome analysis to uncover the transcriptional responses of whiteflies to virus infection. Our results have shown that 755, 587, 1140 and 1347 differentially expressed genes (DEGs) were identified in the comparisons of the data of 0 h vs. 2 h, 0 h vs. 6 h, 0 h vs. 12 h and 0 h vs. 48 h, respectively. KEGG analysis showed that DEGs associated with metabolisms and signal transduction were down-regulated in virus-infected whiteflies. Additionally, 16 up-regulated putative transporter genes and 10 down-regulated genes associated with IL-17 signaling pathway were identified by time-associated gene cluster analysis. These data boost our comprehensions on whitefly-TYLCV interactions associated with different viral AAPs. Full article

Concanavalin A (ConA), a legume lectin, has been drawing increasing attention in recent years concerning its toxicity against insects and its potential application in pest management. In an attempt to evaluate the effect of ConA on potato psyllid (Bactericera cockerelli), an economically important pest of solanaceous crops, the effect of ConA on potato psyllid survival, psyllid gut nuclear morphology, and expression of psyllid caspase genes were evaluated. Our results determined that artificial diet-feeding assays using ConA had deleterious effects on potato psyllids, resulting in significant psyllid mortality following ingestion. We also found that an apoptotic response was induced by ConA in psyllid midgut cells, which was demonstrated by the DNA fragmentation and abnormal nuclear architecture in the midgut cells. Following ConA ingestion, there was also upregulation of caspase genes in the psyllid midguts. Therefore, a key mechanism behind ConA toxicity towards potato psyllid probably involves the induction of apoptosis in midgut cells. This study could provide a better understanding of the mechanisms underlying ConA toxicity in insects and be a stepping stone towards the development of new psyllid control strategies based on plant lectins. Full article

Potato psyllid (Bactericera cockerelli) is one of the most important pests in potatoes (Solanum tuberosum L.) due to its feeding behavior and the transmission of a bacterium (Candidatus Liberibacter solanacearum) that causes zebra chip disease, altering the quality of the potato tuber and the fried potato chip or french fry. This pest is thus a threat to the chip potato industry and often requires preventive measures including the use of costly insecticides. The objectives of this research were to monitor the variation in B. cockerelli adult abundance and to evaluate the risk of zebra chip disease in northwestern New Mexico, USA. Yellow sticky traps were used to collect the pest at the Agricultural Experiment Station at Farmington, NM and in nearby commercial fields at the Navajo Agricultural Products Industry (NAPI) and Navajo Mesa Farms during the 2017–2019 period. The collected adult pests were analyzed at Texas A & M University for the presence of Candidatus L. solanacearum (Lso). The results showed field infestation by B. cockerelli in early June and that the population peaked during the second half of July and decreased as the potato growing season progressed. However, a second less important peak of the pest was revealed around mid- to late-August, depending on the growing season and field. While the B. cockerelli population increased linearly with average air temperature, it showed strong third order polynomial relationships with the accumulated thermal units and the Julian days. The test of B. cockerelli for the Lso infection revealed a low incidence of the pathogen varying from 0.22% to 6.25% and the infected adult B. cockerelli were collected during the population peak period. The results of this study may be helpful to potato growers in pest management decision-making and control. However, more study is needed to evaluate zebra chip disease in terms of its prevention and economic impact, and to develop economic thresholds and pest management programs for northwestern New Mexico and neighboring regions. Full article

‘Candidatus Liberibacter asiaticus’ (CLas) is a major causal agent of citrus Huanglongbing (HLB), which is transmitted by Asian citrus psyllid (ACP), Diaphorina citri, causing severe losses in various regions of the world. Vector efficiency is higher when acquisition occurs by ACP immature stages and over longer feeding periods. In this context, our goal was to evaluate the progression of CLas population and infection rate over four ACP generations that continuously developed on infected citrus plants. We showed that the frequency of CLas-positive adult samples increased from 42% in the parental generation to 100% in the fourth generation developing on CLas-infected citrus. The bacterial population in the vector also increased over generations. This information reinforces the importance of HLB management strategies, such as vector control and eradication of diseased citrus trees, to avoid the development of CLas-infected ACP generations with higher bacterial loads and, likely, a higher probability of spreading the pathogen in citrus orchards. Full article