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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. [ABSTRACT FROM AUTHOR]

Background Understanding the genotype of pest species provides an important baseline for designing integrated pest management (IPM) strategies. Recently developed long-read sequence technologies make it possible to compare genomic features of nonmodel pest species to disclose the evolutionary path underlying the pest species profiles. Here we sequenced and assembled genomes for 3 agricultural pest gelechiid moths: Phthorimaea absoluta (tomato leafminer), Keiferia lycopersicella (tomato pinworm), and Scrobipalpa atriplicella (goosefoot groundling moth). We also compared genomes of tomato leafminer and tomato pinworm with published genomes of Phthorimaea operculella and Pectinophora gossypiella to investigate the gene family evolution related to the pest species profiles. Results We found that the 3 solanaceous feeding species, P. absoluta, K. lycopersicella , and P. operculella , are clustered together. Gene family evolution analyses with the 4 species show clear gene family expansions on host plant–associated genes for the 3 solanaceous feeding species. These genes are involved in host compound sensing (e.g. gustatory receptors), detoxification (e.g. ABC transporter C family, cytochrome P450, glucose-methanol-choline oxidoreductase, insect cuticle proteins, and UDP-glucuronosyl), and digestion (e.g. serine proteases and peptidase family S1). A gene ontology enrichment analysis of rapid evolving genes also suggests enriched functions in host sensing and immunity. Conclusions Our results of family evolution analyses indicate that host plant adaptation and pathogen defense could be important drivers in species diversification among gelechiid moths. [ABSTRACT FROM AUTHOR]

Phytoplasmas are insect-vectored, difficult-to-culture bacterial pathogens that infect a wide variety of crop and non-crop plants, and are associated with diseases that can lead to significant yield losses in agricultural production worldwide. Phytoplasmas are currently grouped in the provisional genus 'Candidatus Phytoplasma', which includes 49 'Candidatus' species. Further differentiation of phytoplasmas into ribosomal groups is based on the restriction fragment length polymorphism (RFLP) pattern of the 16S rRNA-encoding operon, with more than 36 ribosomal groups (16Sr) and over 100 subgroups reported. Since disease symptoms on plants are not associated with phytoplasma identity, accurate diagnostics is of critical importance to manage disease associated with these microorganisms. Phytoplasmas are typically detected from plant and insect tissue using PCR-based methods targeting universal taxonomic markers. Although these methods are relatively sensitive, specific and are widely used, they have limitations, since they provide limited resolution of phytoplasma strains, thus necessitating further assessment of biological properties and delaying implementation of mitigation measures. Moreover, the design of PCR primers that can target multiple loci from phytoplasmas that differ at the sequence level can be a significant challenge. To overcome these limitations, a PCR-independent, multilocus sequence typing (MLST) assay to characterize an array of phytoplasmas was developed. Hybridization probe s targeting cpn60, tuf, secA, secY, and nusA genes, as well as 16S and rp operons, were designed and used to enrich DNA extracts from phytoplasma-infected samples for DNA fragments corresponding to these markers prior to Illumina sequencing. This method was tested using different phytoplasmas including 'Ca. P. asteris' (16SrI-B), 'Ca. P. pruni' (16SrIII-A),'Ca. P. prunorum' (16SrX-B), 'Ca. P. pyri' (16SrX-C), 'Ca. P. mali' (16SrX-A), and 'Ca. P. solani' (16SrXII-A). Thousands of reads were obtained for each gene with multiple overlapping fragments, which were assembled to generate full-length (typically >2kb), high-quality sequences. Phytoplasma groups and subgroups were accurately determined based on 16S ribosomal RNA and cpn60 gene sequences. Hybridization-based MLST facilitates the enrichment of target genes of phytoplasmas and allows the simultaneous determination of sequences corresponding to seven different markers. In this proof-of-concept study, hybridization-based MLST was demonstrated to be an efficient way to generate data regarding 'Ca. Phytoplasma' species/strain differentiation. [ABSTRACT FROM AUTHOR]

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. [ABSTRACT FROM AUTHOR]

Over the past 20 years, ash trees (Oleaceae) in parts of the western United States of America and Canada have been subject to infestations with the psyllid Psyllopsis discrepans (Flor) (Hemiptera: Psyllidae). Infested trees show a series of symptoms, including pseudogalls, canopy loss, and in many cases, tree death. This is an expensive problem for urban forests, particularly in the context of emerald ash borer (Coleoptera: Buprestidae) and Dutch elm disease (Ophiostomataceae), which also impact the diversity of urban forests. This paper presents results from a study on the efficacy of two tree-injected insecticides, Orthene® (acephate) and TreeAzin® (azadirachtin). Trees were treated with these insecticides, and egg and adult psyllids were counted. In addition, canopy cover and severity of pseudogalling were visually assessed. Orthene reduced canopy loss and severity and amount of pseudogalling compared to what occurred on control trees; however, there were more eggs on Orthene-treated trees, indicating that any potential benefit was offset by higher egg counts after treatment. Due to the rapid decline of the ash trees, TreeAzin could not be successfully injected. [ABSTRACT FROM AUTHOR]

Orange wheat blossom midge, Sitodiplosis mosellana (Géhin) (Diptera: Cecidomyiidae), has been successfully reared in the laboratory for more than 20 years in Winnipeg, Manitoba, Canada. The rearing method has been developed to the point where it efficiently produces large numbers of wheat midge continuously under laboratory conditions for use in experiments on wheat midge biology and for screening wheat lines for crop resistance. Adult survival was extended by providing high humidity, and oviposition was increased by simulating natural dawn and dusk conditions and by supplying preflowering spring wheat to adults. Preventing desiccation of the wheat midge larvae in the wheat spikes before overwintering in soil and providing optimal cold conditions for a long enough period to break larval diapause enabled successful adult emergence. We provide data to facilitate the coordination of timing of wheat midge emergence from diapause with the wheat susceptible period. The method can be readily scaled up for screening many lines for resistance or scaled down for small experiments. Here, we report details of the rearing method so that others can implement it for research on the management of this internationally important pest. [ABSTRACT FROM AUTHOR]

The orange blossom wheat midge, Sitodiplosis mosellana (Géhin) (Diptera: Cecidomyiidae), is a significant pest of wheat (Triticum spp.) grown in the Northern Hemisphere. It was accidently introduced to North America over 200 years ago and has subsequently spread throughout the northern Great Plains. Since 2010, several Canadian spring wheat varieties containing the resistance gene Sm1 have been released. Due to the potential of wheat midge populations to evolve virulent biotypes to Sm1, cultivars containing Sm1 are grown with a susceptible cultivar in an interspersed refuge. An understanding of the genetic diversity of wheat midge populations could provide important information on the potential development of resistance to Sm1. In the current study, we used two mitochondrial genes (CO1 and ND4) from wheat midge collected across the northern Great Plains and Québec in North America to assess population structure and genetic diversity. We found limited genetic diversity and population structure across the sampled North American populations. We also assessed North American haplotype similarity to wheat midge collected from Europe and China and found high similarity between North American and European populations, although sampling in Europe was limited. This supports the hypothesis that North American populations originated from Europe. [ABSTRACT FROM AUTHOR]

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. [ABSTRACT FROM AUTHOR]

In agricultural areas, the climatic conditions at the northern limit of crop cultures could impact pest regulation. In this paper, we study the aphid-parasitoid-hyperparasitoid network in cereal fields around Saskatoon (SK, Canada), one of the most harsh climatic conditions for cereal crops in the world, with an extremely short growing season. We have investigated the aphid-parasitoid-hyperparasitoid species relative abundances over two consecutive years. In 2014, a cold spring year, aphids were only found late in the growing season, and their arrival probably corresponded to late Southern winds. In 2015, aphids, parasitoids and hyperparasitoids arrived in the cereal fields in July and August, whereas the cereal harvest is typically complete by the end of the month of August. Three species of primary parasitoids were found for a total parasitism rate of 7.62%, and four genera of hyperparasitoids, with a hyperparasitism rate of 20.1%. There were 87.4% apterous aphids, and the proportion of apterous and winged (alate) aphids did not differ among the 3 aphid species, or among use by parasitoid species and hyperparasitoid species. The sex ratio of parasitoids in the complex was biased towards females for most of the species. In areas with extreme weather conditions and short growing seasons, effective biocontrol through parasitism is limited, although the synchronization level of parasitoids to their aphid hosts is high. We discuss the consequences of the limited length of the growing season on the provision of ecosystem services and on the seasonal ecology of insects of agronomic interest. We also provide information about morphological criteria for identification of parasitoids from the genus Aphidius. [ABSTRACT FROM AUTHOR]

Native corn in Mexico can be affected by maize bushy stunt disease (MBS), which can negatively impact maize-farming operations. Maize bushy stunt symptoms in native corn were linked to the presence of phytoplasmas through molecular analysis of symptomatic native corn plants in Mexico. These findings motivated a socioeconomic characterization and identification of the management strategies developed by the “subsistence farmers” of those communities to answer the questions: What elements of farming affect the presence of MBS disease in “subsistence farmers” fields? What do “subsistence farmers” know about MBS disease and the pathogen associated with MBS disease? How best can MBS disease be managed in small farming communities in Mexico? Through face-to-face interviews with growers and the corresponding analyses of their responses, we identified a widespread lack of knowledge about MBS disease, its causal agent and its vector(s). This study highlighted factors useful to guide future management decisions and education programs, as well as further research to transmit information about MBS disease among “subsistence farmers” in rural communities in Mexico. [ABSTRACT FROM AUTHOR]

Volatile chemicals produced by plants mediate host location, mate-finding and oviposition behavior in insects. State-dependent response to plant cues allows for timing of foraging, mating and oviposition on ephemeral host plants or plant parts. Caloptilia fraxinella is a herbivorous specialist on the foliage of ash trees (Fraxinus). Adults are long-lived and undergo a nine-month reproductive diapause over the fall and winter. Mating and oviposition occur in the spring when volatile chemicals released by ash leaves mediate host location. This study tested the plasticity of olfactory response of C. fraxinella to host plant volatiles using both electroantennogram and behavioral bioassays. The effect of moth physiological state on olfactory response was tested on male and female moths in different nutritional, mating, and diapause states. Antennal responses to host plant volatiles were plastic and depended on moth physiological state, and were highest when moths were reproductively active and would be seeking oviposition hosts. Moth sex and nutritional status also impacted antennal response to host plant volatiles. Oriented flight of females to ash seedlings varied with physiological state and nutritional status, with fed, reproductively active females having the highest response. Physiological state impacted oriented flight of males to female-produced sex pheromone signals whether or not a host plant was present, and there was no increase in behavioral response to sex pheromone in the presence of an ash host. [ABSTRACT FROM AUTHOR]

Quinoa, Chenopodium quinoa Willdenow (Amaranthaceae), cultivation has expanded beyond its historical range in South America into Europe and North America due to its high nutritional properties for human consumption. With the introduction of a crop into a novel range comes the potential for insect pest issues. Here, using traditional morphology and DNA barcoding we identify larvae of Scrobipalpa atriplicella (von Röslerstamm) (Lepidoptera: Gelechiidae), an introduced Eurasian moth, feeding on quinoa throughout the Prairie region of western Canada. Larvae can feed within the stem, on foliage, and directly on seed within the panicles, which can result in up to a 100% yield loss. We summarise distinguishing adult characteristics, potential host plants, and give initial remarks on the life cycle of this moth and its known host range in Canada. The potential for high yield losses makes further investigation of S. atriplicella a priority to maintain and expand sustainable quinoa production in Canada. [ABSTRACT FROM AUTHOR]

The preference-performance or 'mother-knows-best' hypothesis states that female insects choose to oviposit on a host plant that increases the performance of their offspring. This positive link between host plant choice and larval performance is especially important for leaf miners with non-motile larvae that are entirely dependent upon the oviposition choice of the female for host plant location. Preference and performance of the ash leaf coneroller, Caloptilia fraxinella ( Ely) ( Lepidoptera: Gracillariidae), a specialist on ash trees, Fraxinus spp. ( Oleaceae), were tested in a series of laboratory and field experiments. Female C. fraxinella were exposed to two closely related hosts, black ash, Fraxinus nigra Marshall, and green ash, Fraxinus pennsylvanica Marshall var. subintegerrima ( Vahl), in oviposition choice and wind tunnel flight experiments to determine which host is most attractive for oviposition. Caloptilia fraxinella females were inconsistent in host choice, yet performance of larvae was greater on green than black ash. In preference studies, C. fraxinella preferred to oviposit on black ash when leaflets were removed from the tree, but preferred intact green ash over black ash seedlings for oviposition and host location in a wind tunnel. In the field, however, more C. fraxinella visited black ash var. 'Fallgold' at leaf flush than green ash at the same sites. Age of the ash leaflet also influences oviposition in this leaf miner and females preferred new over old leaflets for oviposition. Performance of C. fraxinella larvae was evaluated in field and laboratory experiments and was greater on green ash than on black ash in both experiments based on larval survival and development time parameters. The stronger oviposition and host location preference in the field for black ash were not linked to enhanced performance of offspring, as green ash was the superior host, supporting higher larval survival and faster development. A stronger host location preference in the wind tunnel for green ash over black ash, however, suggests that under certain circumstances with this moth species, 'mother (may) know best'. [ABSTRACT FROM AUTHOR]

Inflorescences (heads or capitula) of the putative self-incompatible species, purple coneflower (Echinacea angustifolia (DC) Cronq. (Asteraceae) ), were visited by insects representing the Coleoptera, Diptera, Hymenoptera, and Lepidoptera, in accordance with a generalist pollination syndrome. Measurement of the effectiveness of insect species as pollinators was accomplished by permitting solitary visits to receptive, central disc florets of virgin (previously bagged) heads. Four parameters were quantified: total stigmatic pollen load and proportion of pollen grains germinated, numbers of pollen tubes at style bases, and percentages of total receptive florets that had retracted (shrivelled) styles. Quantifying total and germinated pollen grains proved ineffective, partly owing to the tendency of self-pollen to initiate pollen tubes. The most effective pollinators were Apidae, especially bumble bees (Bombus spp.) and the European honeybee (Apismellifera L.) (mean: 39-61% of styles retracted). Other noteworthy pollinators were cloudless sulfur butterflies (Phoebis sennae L.--Pieridae; mean 47% of style bases with pollen tubes), golden blister beetles (Epicauta ferruginea Say-Meloidae; 44%), and grasshopper bee flies (Systoechus vulgaris Loew--Bombyliidae; 22%). Sunflower leafcutter bees (Megachile pugnata Say) were less effective (4% of styles retracted). Promisingly, analysis of the proportion of retracted styles provided similar results to the established technique of pollen-tube quantification, but had the significant advantages of being completed more rapidly, without a microscope, and in the field. The quantitative technique of retracted-style analysis appears well suited for prompt measurement of inflorescence-visiting insects as pollinators of many asteraceans. [ABSTRACT FROM AUTHOR]

‘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. [ABSTRACT FROM AUTHOR]

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., (© The Author(s) 2023. Published by Oxford University Press on behalf of Entomological Society of America. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.)

A variety of sensitive and specific molecular diagnostic assays has been described for detecting nucleic acids in biological samples that may harbor pathogens of interest. These methods include very rapid, isothermal nucleic acid amplification methods that can be deployed outside of the laboratory environment, such as loop-mediated isothermal DNA amplification (LAMP) and recombinase-polymerase amplification (RPA). However, all molecular diagnostic assays must be preceded by nucleic acid extraction from the biological samples of interest, which provides suitable template molecules for the assays. To exploit the features of the amplification assays and be utilized outside of the lab, these methods must be rapid and avoid the need for typical laboratory chemicals and equipment. We describe a protocol for the extraction of DNA from field-collected insects that can be implemented at the point of collection and used to detect the presence of DNA sequences from potential plant pathogens that may be vectored by the insects. This protocol provides template DNA that is suitable for PCR, LAMP, and RPA. The FTA PlantSaver card-based DNA extraction product was also confirmed to amplify the mitochondrial cytochrome oxidase 1 (CO1) universal barcode that could later be sequenced to identify any insect. Lastly, we provide an example using field-collected insects, Neokolla (Graphocephala) heiroglyphica, and demonstrate the detection of the plant pathogen Xylella fastidiosa in carrier insects using PCR, RPA, and LAMP., (© 2022. The Author(s), under exclusive license to Springer Science+Business Media, LLC, part of Springer Nature.)

A draft genome sequence is presented for a strain of "Candidatus Phytoplasma asteris" affecting canola plants in Saskatoon, Canada. This phytopathogenic bacterium was determined to be a 16SrI strain and features 16S rRNA-encoding gene sequence heterogeneity.

Aster yellows (AY) is an important disease of Brassica crops and is caused by Candidatus Phytoplasma asteris and transmitted by the insect vector, Aster leafhopper (Macrosteles quadrilineatus). Phytoplasma-infected Aster leafhoppers were incubated at various constant and fluctuating temperatures ranging from 0 to 35 °C with the reproductive host plant barley (Hordium vulgare). At 0 °C, leafhopper adults survived for 18 days, but failed to reproduce, whereas at 35 °C insects died within 18 days, but successfully reproduced before dying. Temperature fluctuation increased thermal tolerance in leafhoppers at 25 °C and increased fecundity of leafhoppers at 5 and 20 °C. Leafhopper adults successfully infected and produced AY-symptoms in canola plants after incubating for 18 days at 0-20 °C on barley, indicating that AY-phytoplasma maintains its virulence in this temperature range. The presence and number of AY-phytoplasma in insects and plants were confirmed by droplet digital PCR (ddPCR) quantification. The number of phytoplasma in leafhoppers increased over time, but did not differ among temperatures. The temperatures associated with a typical crop growing season on the Canadian Prairies will not limit the spread of AY disease by their predominant insect vector. Also, ddPCR quantification is a useful tool for early detection and accurate quantification of phytoplasma in plants and insects.