The Role of Plant Defense Signaling Pathways in Phytoplasma-Infected and Uninfected Aster Leafhoppers' Oviposition, Development, and Settling Behavior.

In plant-microbe-insect systems, plant-mediated responses involve the regulation and interactions of plant defense signaling pathways of phytohormones jasmonic acid (JA), ethylene (ET), and salicylic acid (SA). Phytoplasma subgroup 16SrI is the causal agent of Aster Yellows (AY) disease and is primarily transmitted by populations of aster leafhoppers (Macrosteles quadrilineatus Forbes). Aster Yellows infection in plants is associated with the downregulation of the JA pathway and increased leafhopper oviposition. The extent to which the presence of intact phytohormone-mediated defensive pathways regulates aster leafhopper behavioral responses, such as oviposition or settling preferences, remains unknown. We conducted no-choice and two-choice bioassays using a selection of Arabidopsis thaliana lines that vary in their defense pathways and repeated the experiments using AY-infected aster leafhoppers to evaluate possible differences associated with phytoplasma infection. While nymphal development was similar among the different lines and groups of AY-uninfected and AY-infected insects, the number of offspring and individual female egg load of AY-uninfected and AY-infected insects differed in lines with mutated components of the JA and SA signaling pathways. In most cases, AY-uninfected insects preferred to settle on wild-type (WT) plants over mutant lines; no clear pattern was observed in the settling preference of AY-infected insects. These findings support previous observations in other plant pathosystems and suggest that plant signaling pathways and infection with a plant pathogen can affect insect behavioral responses in more than one manner. Potential differences with previous work on AY could be related to the specific subgroup of phytoplasma involved in each case.

Characterization and correlation of the probing behaviors of Macrosteles quadrilineatus (Hemiptera: Cicadellidae) with electropenetrography (EPG) waveforms.

Aster leafhopper (Hemiptera: Cicadellidae: Macrosteles quadrilineatus Forbes) is a polyphagous insect species that migrates into the upper Midwest of the United States and the Western Canadian Prairies. Populations of this insect are associated with the transmission of a plant pathogen (Candidatus Phytoplasma asteris, 16SrI) to several annual crops and perennial plant species. Previous studies suggest that aster leafhoppers can sometimes prefer less suitable hosts for their development and survival, yet it is unclear if this lower performance on certain plant species is associated with reduced or impaired probing behaviors due to characteristics of the plants. To characterize the probing behaviors of aster leafhoppers, direct current electropenetrography recordings of male and female adults on barley (Polaes: Poaceae: Hordeum vulgare L.) were combined with plant histology, allowing the identification of nine waveforms and their proposed biological meanings. For each waveform, the number of waveform events per insect (NWEI), the waveform duration per insect (WDI), the waveform duration per event per insect (WDEI), and the percentage of recording time were calculated and statistically compared between sexes. Male and female aster leafhoppers exhibited similar behavioral responses for most of these variables, except for the NWEI for waveforms associated with nonprobing activities and the pathway phase. In these cases, male aster leafhoppers exhibited a higher number of events than females. Comparison of the proposed waveforms in this study with previous work on other hemipteran species provided additional support to the interpretation of the biological activities associated with each waveform.

Development of economic thresholds for pea aphid (Hemiptera: Aphididae) management in lentil (Fabaceae) based on in-field insecticide efficacy trials.

Pea aphid (Acyrthosiphom pisum Harris, Hemiptera: Aphididae) presents a significant economic challenge to lentil (Lens culinaris Medik.) production in the major growing region of Saskatchewan, Canada. During 2019-2020, field experiments were conducted to optimize the management tools for pea aphid control on lentils. A randomized split-plot design was used with main plots consisting of different pea aphid pressures and subplots consisting of different insecticide treatments. The main plot design was aimed to assess the impact of A. pisum feeding on lentil yields during the late vegetative to early reproductive stages. Subplots of the study evaluated the efficacy of 3 insecticides in suppressing pea aphid populations on lentils. Lentil is susceptible to A. pisum feeding and requires management at low pest densities. The economic threshold for pea aphids on lentil crops varied depending on environmental conditions, ranging from 20 to 66 aphids per sweep, calculated using a discrete daily growth rate of 1.116. The estimated economic thresholds provided a 7-day lead time before aphid populations achieved the economic injury level (EIL). The EIL was defined as 78 ± 14 aphids per sweep net sample or 743 ± 137 cumulative aphid days from the first aphid present in the field. In addition, the results of the study found that, on average, foliar applications of insecticides containing the pyrethroid active ingredient lambda-cyhalothrin (IRAC group: 3A) reduced pea aphid populations by 83% compared with untreated control.

Seed choice in ground beetles is driven by surface-derived hydrocarbons.

Ground beetles (Coleoptera: Carabidae) are among the most prevalent biological agents in temperate agroecosystems. Numerous species function as omnivorous predators, feeding on both pests and weed seeds, yet the sensory ecology of seed perception in omnivorous carabids remains poorly understood. Here, we explore the sensory mechanisms of seed detection and discrimination in four species of omnivorous carabids: Poecilus corvus, Pterostichus melanarius, Harpalus amputatus, and Amara littoralis. Sensory manipulations and multiple-choice seed feeding bioassays showed olfactory perception of seed volatiles as the primary mechanism used by omnivorous carabids to detect and distinguish among seeds of Brassica napus, Sinapis arvensis, and Thlaspi arvense (Brassicaceae). Seed preferences differed among carabid species tested, but the choice of desirable seed species was generally guided by the olfactory perception of long chain hydrocarbons derived from the seed coat surface. These olfactory seed cues were essential for seed detection and discrimination processes to unfold. Disabling the olfactory appendages (antennae and palps) of carabid beetles by ablation left them unable to make accurate seed choices compared to intact beetles.

A comprehensive review of zebra chip disease in potato and its management through breeding for resistance/tolerance to 'Candidatus Liberibacter solanacearum' and its insect vector.

Zebra chip disease (ZC), associated with the plant pathogenic bacterium 'Candidatus Liberibacter solanacearum' (psyllaurous) (CLso), is a major threat to global potato production. In addition to yield loss, CLso infection causes discoloration in the tubers, rendering them unmarketable. CLso is transmitted by the potato psyllid, Bactericera cockerelli (Sulc) (Hemiptera: Triozidae). ZC is managed by prophylactic insecticide applications to control the vector, which is costly and carries environmental and human health risks. Given the expense, difficulty, and unsustainability of managing vector-borne diseases with insecticides, identifying sources of resistance to CLso and developing varieties that are resistant or tolerant to CLso and/or potato psyllids has become a major goal of breeding efforts. These efforts include field and laboratory evaluations of noncultivated germplasm and cultivars, studies of tubers in cold storage, detailed quantifications of biochemical responses to infection with CLso, possible mechanisms underlying insect resistance, and traditional examination of potato quality following infections. This review provides a brief history of ZC and potato psyllid, a summary of currently available tools to manage ZC, and a comprehensive review of breeding efforts for ZC and potato psyllid management within the greater context of Integrated Pest Management (IPM) strategies. © 2022 Society of Chemical Industry. This article has been contributed to by U.S. Government employees and their work is in the public domain in the USA.

Do Options Matter? Settling Behavior, Stylet Sheath Counts, and Oviposition of Aster Leafhoppers (Hemiptera: Cicadellidae) in Two-Choice Bioassays.

Polyphagous insects are characterized by a broad diet comprising plant species from different taxonomic groups. Within these insects, migratory species are of particular interest, given that they encounter unpredictable environments, with abrupt spatial and temporal changes in plant availability and density. Aster leafhoppers (Hemiptera: Cicadellidae: Macrosteles quadrilineatus Forbes) arrive in the Canadian Prairies in spring and early summer and are the main vector of a prokaryotic plant pathogen known as Aster Yellows Phytoplasma (AYp) (Candidatus Phytoplasma asteris). Host choice selection behavior of Aster leafhoppers was evaluated through two-choice bioassays, using domesticated and wild plants species commonly found in the Canadian Prairies. Leaf tissues from these plants were collected and stained to quantify the number of stylet sheaths and eggs. To assess possible effects due to insect infection, two-choice bioassays were repeated using leafhoppers infected with AYp and a subset of plant species. When two domesticated or wild plant species were presented together, similar numbers of uninfected Aster leafhoppers were observed on both plant species in most combinations. In domesticated-wild plant bioassays, uninfected Aster leafhoppers preferred to settle on the domesticated species. There was little to no association between settling preferences and stylet sheath and egg counts. These findings provide a better understanding of AY epidemiology and suggest that after domesticated species germination, leafhoppers could move from nearby wild plants into the preferred cereals (Poales: Poaceae) to settle on them, influencing the risk of AYp infection in some of these species.

Detection and Identification of a 'Candidatus Liberibacter solanacearum' Species from Ash Tree Infesting Psyllids.

'Candidatus Liberibacter' species are associated with severe, economically important diseases. Nearly all known species are putatively insect transmitted, specifically by psyllids. Detection of 'Ca. Liberibacter' in plants is complicated by their uneven distribution in host plants and largely fastidius nature. The death of black (Fraxinus nigra) and mancana (Fraxinus mandshurica) ash trees in Saskatchewan, Canada has been associated with infestation by the cottony ash psyllid (Psyllopsis discrepans). A combination of conventional PCR amplification and Sanger sequencing of the 16S recombinant DNA was used to detect and identify 'Ca. Liberibacter' in psyllids collected from ash trees in Saskatchewan. BLAST analysis of two 16S sequences that were 1,058 and 1,085 bp long (NTHA 5, GenBank accession number MK942379 and NTHA 6, GenBank accession number MK937570, respectively) revealed they were 99 to 100% similar to a 'Ca. Liberibacter solanacearum' sequence (GenBank accession number KX197200) isolated from the Nearctic psyllid (Bactericera maculipennis) of U.S. provenance. Sequencing the psyllid genes CO1 and Cyt-b confirmed that the psyllids from which the bacterial DNA was isolated were P. discrepans, based on comparisons with sequences in GenBank and BOLD and a reference sample from the United Kingdom. These results provide the first evidence that 'Ca. Liberibacter solanacearum' species are associated with psyllids collected from ash trees and specifically P. discrepans. The recent episodes of dieback of ash in Saskatchewan associated with psyllid feeding are consistent with disease symptoms caused by 'Ca. Liberibacter' pathogens, and this possibility warrants further study.

Interspecific interactions within a vector-borne complex are influenced by a co-occurring pathosystem.

Potato virus Y (PVY) and zebra chip (ZC) disease are major threats to solanaceous crop production in North America. PVY can be spread by aphid vectors and through vegetative propagation in potatoes. ZC is associated with "Candidatus Liberibacter solanacearum" (Lso), which is transmitted by the tomato/potato psyllid, Bactericera cockerelli Sulc (Hemiptera: Triozidae). As these two pathosystems may co-occur, we studied whether the presence of one virus strain, PVY°, affected the host preference, oviposition, and egg hatch rate of Lso-free or Lso-carrying psyllids in tomato plants. We also examined whether PVY infection influenced Lso transmission success by psyllids, Lso titer and plant chemistry (amino acids, sugars, and phytohormones). Lso-carrying psyllids showed a preference toward healthy hosts, whereas the Lso-free psyllids preferentially settled on the PVY-infected tomatoes. Oviposition of the Lso-carrying psyllids was lower on PVY-infected than healthy tomatoes, but Lso transmission, titer, and psyllid egg hatch were not significantly affected by PVY. The induction of salicylic acid and its related responses, and not nutritional losses, may explain the reduced attractiveness of the PVY-infected host to the Lso-carrying psyllids. Although our study demonstrated that pre-existing PVY infection can reduce oviposition by the Lso-carrying vector, the preference of the Lso-carrying psyllids to settle on healthy hosts could contribute to Lso spread to healthy plants in the presence of PVY infection in a field.

Oviposition Behavior and Development of Aster Leafhoppers (Hemiptera: Cicadellidae) on Selected Host Plants From the Canadian Prairies.

Some plant pathogens are capable of manipulating their insect vectors and plant hosts in a way that disease transmission is enhanced. Aster leafhopper (Macrosteles quadrilineatus Forbes) (Hemiptera: Cicadellidae) is the main vector of Aster Yellows Phytoplasma (Candidatus Phytoplasma asteris) in the Canadian Prairies, which causes Aster Yellows (AY) disease in over 300 plant species including cereals and oilseeds. However, little is known about the host range of Aster leafhoppers or their host-choice selection behavior in this geographical region. Several crop and noncrop species commonly found in the Canadian Prairies were evaluated as food and reproductive hosts for Aster leafhoppers through no-choice bioassays. To study possible effects of pathogen infection, AY-uninfected and AY-infected insects were used. Cereals and some noncrops like fleabane were suitable reproductive hosts for Aster leafhoppers, with numbers of offspring observed in treatments using both AY-uninfected and AY-infected insects, suggesting an egg-laying preference on these plant species. Development was similar across the different plant species, except for canola and sowthistle, where growth indexes were lower. Sex-ratios of Aster leafhopper adults did not differ among the plant species or with respect to AY infection. Potential fecundity differed across plant species and was affected by the infection status of the insect. These findings have implications for AY epidemiology and suggest that while cereals can be suitable host plants for Aster leafhopper oviposition and development, some noncrop species could act as alternate hosts for leafhoppers that migrate into the Canadian Prairies before emergence of cereal and canola crops.

Functional Analysis of the Chemosensory Protein MsepCSP8 From the Oriental Armyworm Mythimna separata.

Chemosensory proteins (CSPs) play important roles in chemosensation in insects, but their exact physiological functions remain elusive. In order to investigate the functions of CSPs in the oriental armyworm Mythimna separata, in the present study we explored expression patterns and binding characteristics of the CSP, MsepCSP8. The distinctive functions of MsepCSP8 were also validated by RNAi. The results showed that MsepCSP8 shares high sequence similarity with CSPs of other insect family members, including the characteristic four-cysteine signature motif. MsepCSP8 mRNA was specifically expressed in antennae of females at levels well above those in other tissues. Competitive binding assays confirmed that 20 out of 56 ligands bound more strongly to MsepCSP8 at pH 7.4 than at pH 5.0. Protein structure modeling and molecular docking analyses identified amino acid residues involved in binding volatile compounds, and behavioral response experiments showed that M. separata elicited significant responses to five volatiles from compounds displaying high binding affinity to MsepCSP8. MsepCSP8 transcript abundance was decreased by dsMsepCSP8 injection, which affected the behavioral responses of M. separata to representative semiochemicals. Our findings demonstrate that MsepCSP8 likely contributes to mediating responses of M. separata adults to plant volatiles.

Culex quinquefasciatus larval microbiomes vary with instar and exposure to common wastewater contaminants.

Like many insects, mosquitoes, rely on endosymbionts to grow and develop. These can be acquired from the environment. We used next generation 454 pyrosequencing to discern the whole-body microbiome of the mosquito species Culex quinquefasciatus in various larval stadia and following exposure to common pharmaceutical and personal care products (PPCPs) found in wastewater. PPCP treatments included environmentally-relevant concentrations; 1) a combination of common antibiotics, 2) a combination of mammalian hormones, 3) a mixture of the antibiotic and hormone treatments plus acetaminophen and caffeine and, 4) an untreated control. Within control groups, the predominant families of bacterial symbionts change with each larval instar despite consistent diets and rearing conditions. This trend was also seen in hormone treatments but not in the antibiotic or the mixture treatments. Richness and evenness were reduced in both antibiotic and mixture treatments, suggesting that antibiotics remove certain bacteria or inhibit them from increasing to proportions seen in the control treatment. Interestingly, the mixture treatments had greater richness and evenness compared to antibiotic alone treatments, possibly due to the other contaminants facilitating growth of different bacteria. These findings illuminate the complexity of the microbiome of C. quinquefasciatus and may have implications for more effective control strategies.

Indirect Effects of One Plant Pathogen on the Transmission of a Second Pathogen and the Behavior of its Potato Psyllid Vector.

Plant pathogens can influence the behavior and performance of insect herbivores. Studies of these associations typically focus on tripartite interactions between a plant host, a plant pathogen, and its insect vector. An unrelated herbivore or pathogen might influence such interactions. This study used a model system consisting of Tobacco mosaic virus (TMV), the psyllid Bactericera cockerelli Sulc, and tomatoes to investigate multipartite interactions among a pathogen, a nonvector, and a plant host, and determine whether shifts in host physiology were behind potential interactions. Additionally, the ability of TMV to affect the success of another pathogen, 'Candidatus Liberibacter solanacearum,' which is transmitted by the psyllid, was studied. In choice trials, psyllids preferred nearly fourfold noninfected plants to TMV-infected plants. No-choice bioassays demonstrated that there was no difference in psyllid development between TMV-infected and control plants; oviposition was twice as high on control plants. Following inoculation by psyllids, 'Candidatus Liberibacter solanacearum' titers were lower in TMV-infected plants than control plants. TMV-infected plants had lower levels of amino acids and sugars but little differences in phenolics and terpenoids, relative to control plants. Possibly, these changes in sugars are associated with a reduction in psyllid attractiveness in TMV-infected tomatoes resulting in decreased infection of 'Candidatus Liberibacter solanacearum.'

Pharmaceuticals and personal care products alter the holobiome and development of a medically important mosquito.

The increasing demand for fresh water has forced many countries to use reclaimed wastewater for agricultural purposes. This water contains pharmaceuticals and personal care products (PPCPs) that remain biologically active following passage through wastewater treatment plants. Run-off from farms and contaminated water from treatment facilities exposes aquatic ecosystems to PPCPs. This study examined the effects of PPCPs on a lower trophic organism. Culex quinquefasciatus larvae were reared in water contaminated with environmentally relevant concentrations of common PPCPs. Acetaminophen alone and a mixture of contaminants were found to increase developmental time of larvae. Susceptibility to Bti increased in larvae exposed to antibiotics, acetaminophen, or a mixture of PPCPs. Antibiotics, hormones, and the mixture altered the mosquito bacterial microbiome. Overall, the results indicate that at environmentally relevant concentrations, PPCPs in reclaimed water can have biologically important effects on an ecologically and medically important lower trophic level insect.

Factors influencing host plant choice and larval performance in Bactericera cockerelli.

Among the many topics of interest to ecologists studying associations between phytophagous insects and their host plants are the influence of natal host plant on future oviposition decisions and the mechanisms of generalist versus specialist host selection behavior. In this study, we examined the oviposition preferences, behavior and larval development of the tomato/potato psyllid, Bactericera cockerelli. By rearing psyllids with two distinct geographically-linked haplotypes on different host plants, we were able to examine the role of natal host plant and potential local adaptation on host plant usage. Choice bioassays among three host species demonstrated that psyllids from California had clear preferences that were influenced by natal plant. We further found that patterns in choice bioassays corresponded to observed feeding and movement responses. No-choice bioassays demonstrated that there is little to no association between development and host-plant choice for oviposition, while also indicating that host choice varies between haplotypes. These findings support the concept that mothers do not always choose oviposition sites optimally and also add support for the controversial Hopkins' host selection principle.

A binomial sequential sampling plan for Bactericera cockerelli (Hemiptera: Triozidae) in Solanum lycopersicum (Solanales: Solanacea).

The tomato-potato psyllid Bactericera cockerelli (Sulc) (Hemiptera: Triozidae) is a pest of many solanaceous plants, including tomato (Solanum lycopersicum L.) and potato (Solanum tuberosum L.). In tomato, feeding by nymphs is associated with "psyllid yellows." B. cockerelli also vectors "Candidatus Liberibacter psyllaurous," an infectious bacterium that causes "vein greening" disease. Decisions about management action are much more effective when guided by robust sampling. However, there are few previous studies of potato psyllid spatial distribution in tomato fields, and no published sequential sampling plans for the pest in tomato. We studied B. cockerelli in various tomato fields in California and used these data to generate a sequential sampling plan. We found that juvenile B. cockerelli in tomato fields exhibit an edge effect, an aggregated distribution, and individuals are primarily located on the bottom of leaves. Psyllids were concentrated in the upper segments of plants, but this changed over time. Finally, we present three binominal sequential sampling plans for managing tomato psyllids in tomato fields. These plans differed from both those for bell pepper (Capsicum annum L.) and potato, indicating that B. cockerelli needs to be sampled using crop-specific sampling plans.

The influence of maturity and variety of potato plants on oviposition and probing of Bactericera cockerelli (Hemiptera: Triozidae).

The ecological theory on host plant choice by herbivores suggests that mothers should choose plants that will maximize their offspring's success. In annual host plants, physiology (and therefore host suitability) is sometimes influenced by maturity and growth stage, which may influence female choice. Potato plants were grown under greenhouse conditions and used in choice and no-choice bioassays to determine the effect of plant maturity and variety on oviposition and number of stylet sheaths (which approximate stylet insertions) by tomato/potato psyllids. No-choice bioassays suggested that maturity (time since planting) did not influence oviposition behavior, but oviposition varied significantly among potato plant varieties. There was a significant effect of both maturity and variety on the number of stylet sheaths, which peak toward the middle of the growing season. We also examined tomato/potato psyllid responses to plants grown in a commercial field and again found no effect on oviposition but differences in stylet sheaths. The results suggest that differential susceptibility to zebra chip disease may be associated with unequal feeding rates. Future studies should examine whether the maturity of plants influences larval fitness. Finally, potato variety has an influence on both oviposition and "probing," and has implications for management strategies and the development of resistant potato varieties.

A sequential binomial sampling plan for potato psyllid (Hemiptera: Triozidae) on bell pepper (Capsicum annum).

BACKGROUND: Potato psyllids (Bactericera cockerelli Sulc) are a pest on solanaceous crop plants, including bell peppers. Potato psyllids vector Candidatus Liberibacter psyllaurous, but bell peppers (Capsicum annum L.) do not exhibit symptoms from infection. Potato psyllids show variation in spatial patterns and host choice with cultivar and plant species. Consequently, a study of spatial distribution and sampling plan specific to bell peppers is necessary for management of this insect pest, as those developed for other crops are unlikely to transfer among crops. RESULTS: Potato psyllids were evenly distributed on both sides of leaves but prefer the top two-thirds of pepper plants. Within fields, psyllids demonstrated an aggregated spatial distribution, but the edge effect observed in other crop plants was absent. Eggs and nymphs had similar spatial distributions that differed from adults. A series of nymph-based sampling plans were examined. Sampling plans based on an infestation of less than 41% of plants infested (5 nymphs plant(-1)) were statistically unacceptable, while little difference was found between the 41% infestation plan and 56% (20 nymphs plant(-1)) infestation plan. At 41%, an average of 11 and maximum of 49 samples would be necessary to make a treatment decision. CONCLUSION: The binomial sequential sampling plan presented here offers an important yet simple tool for managing potato psyllids in bell pepper.