Identification of nectar sources foraged by female mosquitoes in Canada.

For many mosquito species, the females must obtain vertebrate blood to complete a gonotrophic cycle. These blood meals are frequently supplemented by feeding on sugary plant nectar, which sustains energy reserves needed for flight, mating, and overall fitness. Our understanding of mosquito nectar foraging behaviors is mostly limited to laboratory experiments and direct field observations, with little research into natural mosquito-host plant relationships done in North America. In this study, we collected nectar-fed female mosquitoes over a 2-year period in Manitoba, Canada, and amplified a fragment of the chloroplast rbcL gene to identify the plant species fed upon. We found that mosquitoes foraged from diverse plant families (e.g., grasses, trees, ornamentals, and legumes), but preferred certain species, most notably soybean and Kentucky blue grass. Moreover, there appeared to be some associations between plant feeding preferences and mosquito species, date of collection, landscape, and geographical region. Overall, this study implemented DNA barcoding to identify nectar sources forage by mosquitoes in the Canadian Prairies.

Molecular mechanisms underpinning quantitative resistance to Phytophthora sojae in Glycine max using a systems genomics approach.

Expression of quantitative disease resistance in many host-pathogen systems is controlled by genes at multiple loci, each contributing a small effect to the overall response. We used a systems genomics approach to study the molecular underpinnings of quantitative disease resistance in the soybean-Phytophthora sojae pathosystem, incorporating expression quantitative trait loci (eQTL) mapping and gene co-expression network analysis to identify the genes putatively regulating transcriptional changes in response to inoculation. These findings were compared to previously mapped phenotypic (phQTL) to identify the molecular mechanisms contributing to the expression of this resistance. A subset of 93 recombinant inbred lines (RILs) from a Conrad × Sloan population were inoculated with P. sojae isolate 1.S.1.1 using the tray-test method; RNA was extracted, sequenced, and the normalized read counts were genetically mapped from tissue collected at the inoculation site 24 h after inoculation from both mock and inoculated samples. In total, more than 100,000 eQTLs were mapped. There was a switch from predominantly cis-eQTLs in the mock treatment to an almost entirely nonoverlapping set of predominantly trans-eQTLs in the inoculated treatment, where greater than 100-fold more eQTLs were mapped relative to mock, indicating vast transcriptional reprogramming due to P. sojae infection occurred. The eQTLs were organized into 36 hotspots, with the four largest hotspots from the inoculated treatment corresponding to more than 70% of the eQTLs, each enriched for genes within plant-pathogen interaction pathways. Genetic regulation of trans-eQTLs in response to the pathogen was predicted to occur through transcription factors and signaling molecules involved in plant-pathogen interactions, plant hormone signal transduction, and MAPK pathways. Network analysis identified three co-expression modules that were correlated with susceptibility to P. sojae and associated with three eQTL hotspots. Among the eQTLs co-localized with phQTLs, two cis-eQTLs with putative functions in the regulation of root architecture or jasmonic acid, as well as the putative master regulators of an eQTL hotspot nearby a phQTL, represent candidates potentially underpinning the molecular control of these phQTLs for resistance.

The influence of weather on the population dynamics of common mosquito vector species in the Canadian Prairies.

BACKGROUND: Mosquito seasonal activity is largely driven by weather conditions, most notably temperature, precipitation, and relative humidity. The extent by which these weather variables influence activity is intertwined with the animal's biology and may differ by species. For mosquito vectors, changes in weather can also alter host-pathogen interactions thereby increasing or decreasing the burden of disease. METHODS: In this study, we performed weekly mosquito surveillance throughout the active season over a 2-year period in Manitoba, Canada. We then used Generalized Linear Mixed Models (GLMMs) to explore the relationships between weather variables over the preceding 2 weeks and mosquito trap counts for four of the most prevalent vector species in this region: Oc. dorsalis, Ae. vexans, Cx. tarsalis, and Cq. perturbans. RESULTS: More than 265,000 mosquitoes were collected from 17 sampling sites throughout Manitoba in 2020 and 2021, with Ae. vexans the most commonly collected species followed by Cx. tarsalis. Aedes vexans favored high humidity, intermediate degree days, and low precipitation. Coquillettidia perturbans and Oc. dorsalis activity increased with high humidity and high rainfall, respectively. Culex tarsalis favored high degree days, with the relationship between number of mosquitoes captured and precipitation showing contrasting patterns between years. Minimum trapping temperature only impacted Ae. vexans and Cq. perturbans trap counts. CONCLUSIONS: The activity of all four mosquito vectors was affected by weather conditions recorded in the 2 weeks prior to trapping, with each species favoring different conditions. Although some research has been done to explore the relationships between temperature/precipitation and Cx. tarsalis in the Canadian Prairies, to our knowledge this is the first study to investigate other commonly found vector species in this region. Overall, this study highlights how varying weather conditions can impact mosquito activity and in turn species-specific vector potential.

Black queen cell virus detected in Canadian mosquitoes.

Black queen cell virus (BQCV) is a ubiquitous honeybee virus and a significant pathogen to queen bee (Apis mellifera) larvae. However, many aspects of the virus remain poorly understood, including the transmission dynamics. In this study, we used next-generation sequencing to identify BQCV in Aedes vexans (n = 4,000) collected in 2019 and 2020 from Manitoba, Canada. We assembled de novo the nearly complete (>96%) genome sequence of the virus, which is the first available from North America and the first report of BQCV being harbored by mosquitoes. Phylogenetic tree reconstructions indicated that the genome had 95.5% sequence similarity to a BQCV isolate from Sweden. Sequences of a potential vector (Varroa destructor) and a microsporidian associated with BQCV (Nosema apis) were not identified in the mosquito samples, however, we did detect sequences of plant origin. We, therefore, hypothesize that the virus was indirectly acquired by mosquitoes foraging at the same nectar sources as honeybees.

Seasonal turnover and insights into the overwintering biology of wireworms (Coleoptera: Elateridae) in the Canadian Prairies.

BACKGROUND: The long-lived terricolous larvae of click beetles, colloquially called wireworms, pose a significant threat to agriculture worldwide. Several economically important pest species have been documented in the Canadian Prairies, including Hypnoidus bicolor, Limonius californicus and Hypnoidus abbreviatus. However, most monitoring activities are performed in the early spring and there is evidence from other geographical regions of seasonal shifts in wireworm species composition and prevalence. Further, little is known about the overwintering physiology or behaviors of wireworms, which undoubtedly contribute to their population dynamics. RESULTS: We surveyed wireworm populations from four Manitoban fields six times throughout the 2020 and 2021 growing seasons. Both Hypnoidus species were active throughout the spring and summer; however, L. californicus did not become active until later in the spring. Chill-coma recovery assays indicated Hypnoidus species recovered quicker than L. californicus from cold acclimation. Vertical migration assays simulating progressively lower ambient temperatures experienced by overwintering larvae identified H. bicolor throughout the soil profile, with L. californicus preferentially found at cooler, shallower depths. We speculate that these differences in species distribution within the soil column are due to the higher levels of putative cryoprotectants (for example, trehalose, sorbitol, glucose, glycerol) in L. californicus, as identified by targeted liquid chromatography tandem mass spectrometry. CONCLUSION: Our findings of a stark seasonal turnover in wireworm species prevalence and composition in the Canadian Prairies should be incorporated into future integrated pest management and surveillance activities. This study also advances our understanding of wireworm overwintering biology, which should be factored into current management approaches. © 2022 His Majesty the King in Right of Canada. Pest Management Science © 2022 Society of Chemical Industry. Reproduced with the permission of the Minister of Agriculture and Agri-Food Canada.

Diet and ontogeny drastically alter the larval microbiome of the invertebrate model Galleria mellonella.

Larvae of the greater wax moth (Galleria mellonella) are an emerging animal model to study the innate immune response and biodegradation of plastic polymers. Both of these complex biological processes are likely impacted by the plasticity of host-microbe interactions, which remains understudied in lepidopterans. Consequently, we carried out 16S rRNA sequencing to explore the effect diet (natural, artificial) has on the bacterial assemblages of G. mellonella in different tissues (gut, fat bodies, silk glands) throughout development (eggs, six instar stages, adults). The microbiome was rich in diversity, with Proteobacteria and Firmicutes being the most represented phyla. Contrary to other lepidopterans, G. mellonella appears to possess a resident microbiome dominated by Ralstonia. As larvae progress through development, the bacterial assemblages become increasingly shaped by the caterpillar's diet. In particular, a number of bacteria genera widely associated with the G. mellonella microbiome (e.g., Enterococcus and Enterbacter) were significantly enriched on an artificial diet. Overall, these results indicate that the G. mellonella microbiome is not as simplistic and homogenous as previously described. Rather, its bacterial communities are drastically affected by both diet and ontogeny, which should be taken into consideration in future studies planning to use G. mellonella as model species.

Evaluation of Standardized Bait Trapping Approaches and Climatic Factors That Influence Wireworm Catch in the Canadian Prairies.

Wireworms are significant pests of a variety of economically important crops grown in the Canadian Prairies. These soil-dwelling larvae of click beetles feed on and burrow into the accessible underground plant tissues, which can result in cosmetic injury, stunting, wilting, and plant death. Successful management of wireworms relies on accurate estimations of their abundance and activity in infested fields. Bait trapping is the most commonly used method for sampling wireworms and standardized approaches have been developed; however, little work has been done to optimize trapping efficacy in different geographical regions. In this study, we evaluated the effect of bait trapping duration, seed formulation, and the causal relationship with CO2 production and soil temperature on the wireworm catch in three fields located in Manitoba, Canada. As expected, wireworm catch increased with trapping duration and placing traps in ground for 8 d is adequate in most cases. Both barley and wheat were more effective baits than soybean; however, barley released more CO2 (i.e., an attractant for wireworms) and performed better at elevated soil temperatures. Overall, the results of this study will serve as valuable guidelines to improve current wireworm sampling methods, and can be integrated into strategies aimed at managing these important pests to crop production.

Changes in the phyllosphere and rhizosphere microbial communities of soybean in the presence of pathogens.

Soybean (Glycine max L.) is host to an array of foliar- and root-infecting pathogens that can cause significant yield losses. To provide insights into the roles of microorganisms in disease development, we evaluated the bacterial and fungal communities associated with the soybean rhizosphere and phyllosphere. For this, leaf and soil samples of healthy, Phytophthora sojae-infected and Septoria glycines-infected plants were sampled at three stages during the production cycle, and then subjected to 16S and Internal Transcribed Spacer (ITS) amplicon sequencing. The results indicated that biotic stresses did not have a significant impact on species richness and evenness regardless of growth stage. However, the structure and composition of soybean microbial communities were dramatically altered by biotic stresses, particularly for the fungal phyllosphere. Additionally, we cataloged a variety of microbial genera that were altered by biotic stresses and their associations with other genera, which could serve as biological indicators for disease development. In terms of soybean development, the rhizosphere and phyllosphere had distinct microbial communities, with the fungal phyllosphere most influenced by growth stage. Overall, this study characterized the phyllosphere and rhizosphere microbial communities of soybean, and described the impact of pathogen infection and plant development in shaping these bacterial and fungal communities.

Fat on plastic: Metabolic consequences of an LDPE diet in the fat body of the greater wax moth larvae (Galleria mellonella).

The caterpillar larvae of the greater wax moth (Galleria mellonella) are avid plastivores, as when provided a diet of low-density polyethylene (LDPE) they actively feed on it. Recent work has highlighted the importance of their microbiome in the putative biodegradation of this plastic polymer, though the impact of plastic metabolism on the insect host is less clear. In the present study, we undertook an integrative approach spanning all levels of biological organization to explore the effects of a plastic diet on the metabolic physiology of this animal model of plastic biodegradation. We demonstrate that an LDPE diet is not sufficient to maintain optimal larval growth and survival. In addition, we confirm that plastic fed waxworms retain their fat body lipid stores in a manner proportional to their individual polyethylene consumption suggesting a direct effect of LDPE biodegradation. At the functional level, the oxidative capacity of the fat body of LDPE-fed larvae is maintained reflecting unaltered metabolic function of the tissue. Finally, metabolomic analyses confirmed fat body lipid stores maintenance in LDPE-fed worms, but uncovered various other nutritional deficiencies. Overall, this work unveils novel insights in the complex interplay between LDPE biodegradation and the metabolic physiology of this model plastivore.

Transcriptional profiling of Dermacentor variabilis (Acari: Ixodidae) provides insights into the role of the Haller's organ in spatial DEET recognition.

DEET is the most common active ingredient in commercial repellents, providing effective protection against blood-sucking insects and ticks. However, its mode of action is not fully understood, with several theories put forward to explain its repellency effect. Unique to ticks, the Haller's organ recognizes a variety of external stimuli through non-contact mechanisms, yet the extent to which the organ plays a role in tick chemoreception is not fully known. We previously found that DEET inhibited the expression/activity of cytochrome P450s and cholinesterases in Dermacentor variabilis, however, our experimental design could not distinguish between sexes (males/females), method of exposure (volatile/tactile) or the roles of chemosensory tissues (Haller's organ). In this study, we used RNA sequencing to assess changes in transcript expression induced by volatile DEET in D. variabilis males/females with/without intact Haller's organs. Male ticks showed much greater transcriptional responses to DEET than females, which may be at least partially attributed to the sexual dimorphism of the Haller's organ. Female transcript expression profiles were most influenced by condition (i.e., intact/excised Haller's organs) with minimal changes due to repellent exposure. On the other hand, removal of the Haller's organs caused DEET treated male ticks to exhibit similar expression profiles as control (ethanol) ticks with intact Haller's organs. Consequently, the transcript-level responses to spatial DEET exposure appears largely based on males possessing their Haller's organs. The molecular signature of this response included the suppression of a large number of transcripts involved in detoxification, lipid metabolism and immunity. Taken collectively, this study furthers our understanding of the Haller's organ role in volatile DEET recognition.

Three years of surveillance associates agro-environmental factors with wireworm infestations in Manitoba, Canada.

BACKGROUND: Wireworms, the soil-dwelling larvae of click beetles, are a major threat to global agricultural production. This is largely due to their generalist polyphagous feeding capabilities, extended and cryptic life cycles, and limited management options available. Although wireworms are well-documented as economically important pests in the Canadian Prairies, including Manitoba, there are gaps in knowledge on species distributions, subterranean behaviour and life cycles, feeding ecology and damage capacity, and economic thresholds for crop yield loss. RESULTS: We carried out 3 years (2018-2020) of intensive surveillance of larval populations across Manitoba. A total of 31 fields (24 in ≥ 2 consecutive years) were surveyed in early spring using standardized bait trapping approaches. Wireworms were present in 94% of surveyed sites, but the catch within fields varied year to year. While Hypnoidus bicolor predominated (94% of larvae), several other pest species were identified. We then explored the relationships between wireworm trap numbers and agro-environmental factors. The larval catch tended to decrease under conditions of low soil temperatures and increased clay content, coupled with high soil moisture and precipitation during the trapping period. Treatment and cultural methods appeared less influential; however, wheat production in either of the previous two growing seasons was associated with increased wireworm catch. Our models failed to predict a relationship between wireworm catch and crop yields, although infestations were rare in our region. CONCLUSION: Our findings better infer the risks posed by wireworms to crop production in the Canadian Prairies, and the agro-environmental factors that represent the greatest contributors to these risks. This information should be incorporated into future integrated pest management (IPM) strategies for wireworms. © 2021 Her Majesty the Queen in Right of Canada Pest Management Science © 2021 Society of Chemical Industry Reproduced with the permission of the Minister of Agriculture and Agri-Food Canada.

Occurrence of the Mosquito Aedes triseriatus (Diptera: Culicidae) Beyond Its Most Northwestern Range Limits in Manitoba, Canada.

Native to the Eastern United States and Eastern Canada, Aedes triseriatus (eastern tree hole mosquito) is an important vector of La Crosse virus and dog heartworm. Although its range has been well characterized in the United States, few studies have surveyed its distribution within Canada. In this study, mosquitoes were collected from a variety of urban and rural communities throughout Manitoba, Canada between the years of 2018 and 2020. Aedes triseriatus was identified and confirmed molecularly to be present in 13 communities. This includes localities that expand the species known distribution to new northern and western areas, and suggests that past surveillance efforts have not been comprehensive or environmental factors have caused this mosquito species to be present in areas in which it was not found previously. As Canada is showing signs of a changing climate, this may be driving the broader occurrence of Ae. triseriatus.

Genetic structure and population demographics of Hypnoidus bicolor (Coleoptera: Elateridae) in the Canadian Prairies.

BACKGROUND: Following banning of the pesticide lindane in most counties, wireworms (i.e., the soil-living larval stages of click beetles) have become major pests of a variety of economically important field crops. Hypnoidus bicolor is a common pest species in the Canadian Prairies. However, little is known about its life history, which impedes the development of effective integrated pest management (IPM) strategies. Population genetic approaches have the potential to assist in the development of IPM. RESULTS: We sequenced a 622-bp fragment of the COX1 gene from 326 H. bicolor wireworm and click beetles collected from 13 localities on the Canadian Prairies. Two genetically distinct (>4.66% sequence divergence) clades were identified, suggesting that they may be part of a species complex. Clade A predominated and increased in prevalence the further east samples were collected, whereas the opposite was true for clade B. Clade B appears to be comprised of two mitochondrial DNA groups, however, one group was represented by only one haplotype. Both clades were characterized by uneven gene flow among populations with low levels of regional genetic structuring. Clade A appeared to have undergone population and range expansions, which may coincide with the advent of intensive agriculture practices in the prairies. CONCLUSION: Knowledge of species composition and population structure is important for the development of effective IPM strategies but is often lacking for wireworms. Our study fills these knowledge gaps for a predominant pest species in the prairies, H. bicolor, by providing robust evidence for cryptic forms and characterizing its dispersal patterns and population dynamics. © 2021 Society of Chemical Industry.

Amplicon Sequencing Reveals Extensive Coinfections of Foliar Pathogens in Soybean.

Soybean (Glycine max) is one of the most economically important crops grown in North America and in other regions worldwide. However, the plant is susceptible to a variety of foliar pathogenic microorganisms, some of which are a significant threat to production. Several molecular and serological approaches are currently available to diagnose plant pathogens, but all have limitations including their capability to accurately detect coinfections of individual plants. We therefore used 16S and internal transcribed spacer amplicon sequencing to identify the suite of bacterial and fungal organisms infecting 96 soybean leaf samples collected throughout southern Manitoba, Canada, at two growth stages (V2/3 and R6). We were able to confirm the presence of pathogens previously known to our sampling regions, such as Septoria glycines, Alternaria alternata, and Pseudomonas spp. Importantly, we found that most of plants were infected by more than one putative pathogen, with 64% of V2/3 and 26% of R6 plants infected by three or more pathogens. Amplicon sequencing also indicated the presence of residual pathogens that infect crops other than soybean, as well as nonfoliar pathogens and nonpathogenic microorganisms. We discuss some of the benefits and drawbacks of using amplicon sequencing to detect foliar pathogens of soybean.

A Very Hungry Caterpillar: Polyethylene Metabolism and Lipid Homeostasis in Larvae of the Greater Wax Moth (Galleria mellonella).

Larvae of the greater wax moth (Galleria mellonella) possess the remarkable ability to consume and rapidly degrade low-density polyethylene. Previous studies have investigated the involvement of the animal's microbiome, but little is known about the host's actual role and if it benefits from biodegradation of this synthetic polymer. We used a combination of RNA sequencing and biochemical approaches to assess caterpillars fed honeycomb, fed polyethylene (PE), or starved for up to 72 h. Sequencing of gut transcripts revealed PE-fed larvae retain an expression profile consistent with normal intestinal function but also show distinct molecular signatures indicative of enhanced fatty acid metabolism (FAM). Further, quantification of total lipid content validated the impact of a PE diet on FAM; in contrast to lipid-depleted starved animals, PE-fed caterpillars maintain lipid reserves similar to honeycomb-fed larvae. Additionally, we found the activity of putative enzymes involved in lipid oxidation (e.g., alcohol dehydrogenase) are considerably higher in PE-fed larvae, indicating that on a functional level, these caterpillars are inducing pathways to effectively metabolize PE. Overall, we put forward a hypothesized model where the similarity in chemical structure between PE and its natural honeycomb diet has endowed larvae of G. mellonella with the extraordinary capability to derive energy from PE as an exclusive food source through pre-existing metabolic pathways.

Natural insensitivity and the effects of concentration on the repellency and survival of American dog ticks (Dermacentor variabilis) by DEET.

N,N-Diethyl-m-toluamide (DEET) is by far the most used repellent worldwide. When applied topically to the skin, the active ingredient has been shown to provide protection from a variety of hematophagous insects, including mosquitoes and flies. DEET's effectiveness against ticks is influenced by a variety of factors (e.g., duration and concentration of application, drying time, route of exposure, tick species and developmental stage), and may differ from insects due to their unique chemosensory system that primarily involves the Haller's organ. We therefore used several approaches to investigate DEET's efficacy to repel Dermacentor variabilis at different concentrations (5, 30 or 75%), as well as explore its toxicological properties and natural variability in DEET insensitivity across populations from Manitoba, Canada. Climbing bioassays indicated that higher concentrations of DEET were more effective at repelling D. variabilis, and that ticks from some sampling localities were more sensitive to lower concentrations than others. Petri dish arena assays revealed ticks exposed to high concentrations of the repellent lose their ability to discriminate lower concentrations, perhaps due to overstimulation or habituation. Finally, our tactile assays demonstrated reduced tick survival after contact with high DEET concentrations, with mortality occurring more rapidly with increased concentration. Dermacentor variabilis from these tactile assays displayed a multitude of physiological and neurological symptoms, such as 'hot foot' and various bodily secretions. Overall, our study shows a strong association between repellency, concentration and the acaricidal effects of DEET on D. variabilis.

Effects of MP Polyethylene Microparticles on Microbiome and Inflammatory Response of Larval Zebrafish.

Plastic polymers have quickly become one of the most abundant materials on Earth due to their low production cost and high versatility. Unfortunately, some of the discarded plastic can make its way into the environment and become fragmented into smaller microscopic particles, termed secondary microplastics (MP). In addition, primary MP, purposely manufactured microscopic plastic particles, can also make their way into our environment via various routes. Owing to their size and resilience, these MP can then be easily ingested by living organisms. The effect of MP particles on living organisms is suspected to have negative implications, especially during early development. In this study, we examined the effects of polyethylene MP ingestion for four and ten days of exposure starting at 5 days post-fertilization (dpf). In particular, we examined the effects of polyethylene MP exposure on resting metabolic rate, on gene expression of several inflammatory and oxidative stress linked genes, and on microbiome composition between treatments. Overall, we found no evidence of broad metabolic disturbances or inflammatory markers in MP-exposed fish for either period of time. However, there was a significant increase in the oxidative stress mediator L-FABP that occurred at 15 dpf. Furthermore, the microbiome was disrupted by MP exposure, with evidence of an increased abundance of Bacteroidetes in MP fish, a combination frequently found in intestinal pathologies. Thus, it appears that acute polyethylene MP exposure can increase oxidative stress and dysbiosis, which may render the animal more susceptible to diseases.

Role of the intestinal microbiome in low-density polyethylene degradation by caterpillar larvae of the greater wax moth, Galleria mellonella.

Recently, a few insects, including the caterpillar larva of the greater wax moth Galleria mellonella, have been identified as avid 'plastivores'. These caterpillars are able to ingest and metabolize polyethylene at unprecedented rates. While it appears that G. mellonella plays an important role in the biodegradation process, the contribution of its intestinal microbiome remains poorly understood and contested. In a series of experiments, we present strong evidence of an intricate relationship between an intact microbiome, low-density polyethylene (LDPE) biodegradation and the production of glycol as a metabolic by-product. First, we biochemically confirmed that G. mellonella larvae consume and metabolize LDPE, as individual caterpillars fed on polyethylene excreted glycol, but those excretions were reduced by antibiotic treatment. Further, while the gut bacterial communities remained relatively stable regardless of diet, we showed that during the early phases of feeding on LDPE (24-72 h), caterpillars exhibited increased microbial abundance relative to those starved or fed on their natural honeycomb diet. Finally, by isolating and growing gut bacteria with polyethylene as their exclusive carbon source for over 1 year, we identified microorganisms in the genus Acinetobacter that appeared to be involved in this biodegradation process. Taken collectively, our study indicates that during short-term exposure, the intestinal microbiome of G. mellonella is intricately associated with polyethylene biodegradation in vivo.

Whole genome sequence of the soybean aphid, Aphis glycines.

Aphids are emerging as model organisms for both basic and applied research. Of the 5,000 estimated species, only three aphids have published whole genome sequences: the pea aphid Acyrthosiphon pisum, the Russian wheat aphid, Diuraphis noxia, and the green peach aphid, Myzus persicae. We present the whole genome sequence of a fourth aphid, the soybean aphid (Aphis glycines), which is an extreme specialist and an important invasive pest of soybean (Glycine max). The availability of genomic resources is important to establish effective and sustainable pest control, as well as to expand our understanding of aphid evolution. We generated a 302.9 Mbp draft genome assembly for Ap. glycines using a hybrid sequencing approach. This assembly shows high completeness with 19,182 predicted genes, 92% of known Ap. glycines transcripts mapping to contigs, and substantial continuity with a scaffold N50 of 174,505 bp. The assembly represents 95.5% of the predicted genome size of 317.1 Mbp based on flow cytometry. Ap. glycines contains the smallest known aphid genome to date, based on updated genome sizes for 19 aphid species. The repetitive DNA content of the Ap. glycines genome assembly (81.6 Mbp or 26.94% of the 302.9 Mbp assembly) shows a reduction in the number of classified transposable elements compared to Ac. pisum, and likely contributes to the small estimated genome size. We include comparative analyses of gene families related to host-specificity (cytochrome P450's and effectors), which may be important in Ap. glycines evolution. This Ap. glycines draft genome sequence will provide a resource for the study of aphid genome evolution, their interaction with host plants, and candidate genes for novel insect control methods.

Hybrid Genome Assembly of a Major Quantitative Disease Resistance Locus in Soybean Toward Fusarium graminearum.

Schwabe [teleomorph: Gibberella zeae (Schweintiz) Petch] has been identified as a pathogen of soybean [ (L.) Merr.] causing seed, seedling damping-off and root rot in North America. A major quantitative disease resistance locus (QDRL) that contributed 38.5% of the phenotypic variance toward in soybean was previously identified through mapping of a recombinant inbred line (RIL) population derived from a cross between 'Wyandot' and PI 567301B. This major QDRL mapped to chromosome 8 to a predicted 305 kb region harboring 36 genes. This locus maps near the locus for soybean cyst nematode (SCN) and the locus contributing to seed coat color. Long-read sequencing of the region was completed and variations in gene sequence and gene order compared with the 'Williams 82' reference were identified. Molecular markers were developed for genes within this region and mapped in the original population, slightly narrowing the region of interest. Analyses of the hybrid genome reassembly using three previously published bacterial artificial chromosome (BAC) sequences (BAC56G2, BAC104J7, and BAC77G7-a) combined with RNA-sequencing narrowed the region making candidate gene identification possible. The markers within this region may be used for marker-assisted selection (MAS). There were 10 differentially expressed genes between resistant and susceptible lines, with four of these candidates also located within the genomic interval defined by the flanking markers. These genes included an actin-related protein 2/3 complex subunit, an unknown protein, a hypothetical protein, and a chalcone synthase 3.