Optimizing a rapid LAMP assay for discrimination of Drosophila suzukii (Diptera: Drosophilidae) from common drosophilids captured in monitoring traps from the Midwest, United States.

Spotted-wing Drosophila, Drosophila suzukii, is an economically important pest of small fruits worldwide. Currently, the timing of management strategies relies on detection of adult flies captured in baited monitoring traps; however, identifying D. suzukii in trap catch based on morphology can be challenging for growers. DNA-based diagnostic methods such as loop-mediated isothermal amplification (LAMP) have the potential to improve D. suzukii detection. This study evaluated a LAMP assay as a diagnostic tool to discriminate between D. suzukii and closely related drosophilid species found commonly in monitoring traps in the Midwestern United States. Targeting the mitochondrial cytochrome oxidase I (COI) gene, we found the LAMP assay accurately detected D. suzukii with as little as 0.1 ng/µl of DNA at 63 °C for 50 min. Under these optimal incubation conditions, D. suzukii could be discriminated from D. affinis and D. simulans consistently, when specimens collected from liquid monitoring traps were tested independently. Compared to other DNA-based diagnostic tools for D. suzukii, LAMP offers unique benefits: DNA extraction is not required, testing occurs at one temperature in less than 1 h, and positive results are visible as a colorimetric change from pink to yellow. The LAMP assay for D. suzukii can reduce reliance on morphological identification, enhance the adoption of monitoring tools, and improve accuracy of detection. Further optimization can be conducted to evaluate the accuracy and sensitivity of results when a mixture of DNA from both D. suzukii and congener flies are tested in a single LAMP reaction.

Genetic mutations linked to field-evolved Cry1Fa-resistance in the European corn borer, Ostrinia nubilalis.

Transgenic corn, Zea mays (L.), expressing insecticidal toxins such as Cry1Fa, from Bacillus thuringiensis (Bt corn) targeting Ostrinia nubilalis (Hübner) (Lepidoptera: Crambidae) resulted in over 20 years of management success. The first case of practical field-evolved resistance by O. nubilalis to a Bt corn toxin, Cry1Fa, was discovered in Nova Scotia, Canada, in 2018. Laboratory-derived Cry1Fa-resistance by O. nubilalis was linked to a genome region encoding the ATP Binding Cassette subfamily C2 (ABCC2) gene; however, the involvement of ABCC2 and specific mutations in the gene leading to resistance remain unknown. Using a classical candidate gene approach, we report on O. nubilalis ABCC2 gene mutations linked to laboratory-derived and field-evolved Cry1Fa-resistance. Using these mutations, a DNA-based genotyping assay was developed to test for the presence of the Cry1Fa-resistance alleles in O. nubilalis strains collected in Canada. Screening data provide strong evidence that field-evolved Cry1Fa-resistance in O. nubilalis maps to the ABCC2 gene and demonstrates the utility of this assay for detecting the Cry1Fa resistance allele in O. nubilalis. This study is the first to describe mutations linked to Bt resistance in O. nubilalis and provides a DNA-based detection method that can be used for monitoring.

Presence-Absence Sampling Plans for Stink Bugs (Hemiptera: Pentatomidae) in the Midwest Region of the United States.

Stink bugs represent an increasing risk to soybean production in the Midwest region of the United States. The current sampling protocol for stink bugs in this region is tailored for population density estimation and thus is more relevant to research purposes. A practical decision-making framework with more efficient sampling effort for management of herbivorous stink bugs is needed. Therefore, a binomial sequential sampling plan was developed for herbivorous stink bugs in the Midwest region. A total of 146 soybean fields were sampled across 11 states using sweep nets in 2016, 2017, and 2018. The binomial sequential sampling plans were developed using combinations of five tally thresholds at two proportion infested action thresholds to identify those that provided the best sampling outcomes. Final assessment of the operating characteristic curves for each plan indicated that a tally threshold of 3 stink bugs per 25 sweeps, and proportion infested action thresholds of 0.75 and 0.95 corresponding to the action thresholds of 5 and 10 stink bugs per 25 sweeps, provided the optimal balance between highest probability of correct decisions (≥ 99%) and lowest probability of incorrect decisions (≤ 1%). In addition, the average sample size for both plans (18 and 12 sets of 25 sweeps, respectively) was lower than that for the other proposed plans. The binomial sequential sampling plan can reduce the number of sample units required to achieve a management decision, which is important because it can potentially reduce risk/cost of management for stink bugs in soybean in this region.

Transcriptome Analysis Reveals Functional Diversity in Salivary Glands of Plant Virus Vector, Graminella nigrifrons.

Insect salivary glands play an important role for host feeding, specifically by secreting salivary proteins for digestion and potentially modulating host defenses. Compared to other hemipterans, the significance of salivary glands is less studied in the black-faced leafhopper, Graminella nigrifrons, a crop pest that vectors several agronomically important plant viruses. To identify functionally important genes in the salivary glands of the black-faced leafhopper, we compared transcriptomes between adult salivary glands (SG) and the remaining carcasses. We identified 14,297 salivary gland-enriched transcripts and 195 predicted secretory peptides (i.e., with a signal peptide and extracellular localization characteristics). Overall, the SG transcriptome included functions such as 'oxidoreduction', 'membrane transport', and 'ATP-binding', which might be important for the fundamental physiology of this tissue. We further evaluated transcripts with potential contributions in host feeding using RT-qPCR. Two SG-enriched transcripts (log2 fold change > 5), GnP19 and GnE63 (a putative calcium binding protein), were significantly upregulated in maize-fed adults relative to starved adults, validating their importance in feeding. The SG-enriched transcripts of the black-faced leafhopper could play a potential role for interacting with maize and could be targets of interest for further functional studies and improve pest control and disease transmission.

Thiamethoxam Differentially Impacts the Survival of the Generalist Predators, Orius insidiosus (Hemiptera: Anthocoridae) and Hippodamia convergens (Coleoptera: Coccinellidae), When Exposed via the Food Chain.

Insect predators are seldom considered during toxicological trophic assessments for insecticide product development. As a result, the ecological impact of novel insecticides on predators is not well understood, especially via the food chain, i.e., when their prey is exposed to insecticides. Neonicotinoids are systemic insecticides widely used in agriculture to control herbivorous insects, but their effects on predatory insects via the food chain have not been well characterized. In this study, we documented the time-course effects of the neonicotinoid thiamethoxam on the survival of two predators, the insidiosus flower bug Orius insidiosus (Say) and the convergent lady beetle Hippodamia convergens Guérin-Méneville, when preying upon the aphids Aphis glycines Matsumura (Hemiptera: Aphididae), Aphis gossypii Glover (Hemiptera: Aphididae), and Myzus persicae (Sulzer) (Hemiptera: Aphididae). Aphids were exposed to thiamethoxam-treated or untreated plants every week over the course of 5 wk. After transferring aphids to Petri dishes, predators were allowed to feed on aphids. We found that the survival of the insidiosus flower bug, but not the convergent lady beetle, was reduced after consuming aphids reared on thiamethoxam-treated plants compared to untreated plants. Survival reduction of the insidiosus flower bug was observed only during the first weeks after thiamethoxam application; no reduction occurred 28 d after treatment or beyond. These results demonstrate that a systemic application of thiamethoxam could be compatible with convergent lady beetles and insidiosus flower bugs, if the time of predator release does not coincide with thiamethoxam activity. These findings are critical for the development of future pest control programs that integrate biological and chemical control.

Genome scan detection of selective sweeps among biotypes of the soybean aphid, Aphis glycines, with differing virulence to resistance to A. glycines (Rag) traits in soybean, Glycine max.

Multiple biotypes of soybean aphid, Aphis glycines, occur in North America adapted for survival (virulence) on soybean, Glycine max, with one or more different resistance to A. glycines (Rag) traits. The degree of genome-wide variance between biotypes and the basis of virulence remains unknown, but the latter is hypothesized to involve secreted effector proteins. Between 167,249 and 217,750 single nucleotide polymorphisms (SNPs) were predicted from whole genome re-sequencing of A. glycines avirulent biotype 1 (B1) and virulent B2, B3 and B4 colony-derived iso-female lines when compared to the draft B1 genome assembly, Ag_bt1_v6.0. Differences in nucleotide diversity indices (π) estimated within 1000 bp sliding windows demonstrated that 226 of 353 (64.0%) regions most differentiated between B1 and ≥ 2 virulent biotypes, representing < 0.1% of the 308 Mb assembled genome size, are located on 15 unordered scaffolds. Furthermore, these 226 intervals were coincident and show a significant association with 326 of 508 SNPs with significant locus-by-locus FST estimates between biotype populations (r = 0.6271; F1,70 = 45.36, P < 0.001) and genes showing evidence of directions selection (πN/πS > 2.0; r = 0.6233; F1,70 = 50.20, P < 0.001). A putative secreted effector glycoprotein is encoded in proximity to genome intervals of low estimated π (putative selective sweep) within avirulent B1 compared to all three virulent biotypes. Additionally, SNPs are clustered in or in proximity to genes putatively involved in intracellular protein cargo transport and the regulation of secretion. Results of this study indicate that factors on a small number of scaffolds of the A. glycines genome may contribute to variance in virulence towards Rag traits in G. max.

Hybridization and introgression between Helicoverpa armigera and H. zea: an adaptational bridge.

BACKGROUND: Invasion of organisms into new ecosystems is increasingly common, due to the global trade in commodities. One of the most complex post-invasion scenarios occurs when an invasive species is related to a native pest, and even more so when they can hybridize and produce fertile progeny. The global pest Helicoverpa armigera was first detected in Brazil in 2013 and generated a wave of speculations about the possibility of hybridization with the native sister taxon Helicoverpa zea. In the present study, we used genome-wide single nucleotide polymorphisms from field-collected individuals to estimate hybridization between H. armigera and H. zea in different Brazilian agricultural landscapes. RESULTS: The frequency of hybridization varied from 15 to 30% depending on the statistical analyses. These methods showed more congruence in estimating that hybrids contained approximately 10% mixed ancestry (i.e. introgression) from either species. Hybridization also varied considerably depending on the geographic locations where the sample was collected, forming a 'mosaic' hybrid zone where introgression may be facilitated by environmental and landscape variables. Both landscape composition and bioclimatic variables indicated that maize and soybean cropland are the main factors responsible for high levels of introgression in agricultural landscapes. The impact of multiple H. armigera incursions is reflected in the structured and inbred pattern of genetic diversity. CONCLUSIONS: Our data showed that the landscape composition and bioclimatic variables influence the introgression rate between H. armigera and H. zea in agricultural areas. Continuous monitoring of the hybridization process in the field is necessary, since agricultural expansion, climatic fluctuations, changing composition of crop species and varieties, and dynamic planting seasons are some factors in South America that could cause a sudden alteration in the introgression rate between Helicoverpa species. Introgression between invasive and native pests can dramatically impact the evolution of host ranges and resistance management.

Performance of Field-Collected Spodoptera frugiperda (Lepidoptera: Noctuidae) Strains Exposed to Different Transgenic and Refuge Maize Hybrids in Argentina.

Spodoptera frugiperda (J. E. Smith) is one of the major pests of maize in Argentina. The main tool for its control is the use of genetically modified maize hybrids expressing Bacillus thuringiensis (Bt) insecticidal proteins. Maize growers in Argentina initially controlled this pest with Bt maize that expressed a single Bt protein (Cry1Ab or Cry1F). Currently it is necessary to plant maize cultivars that produce two Bt proteins to achieve the satisfactory control. Recently, Cry1F field-evolved resistant populations of this species were confirmed in Argentina. The objective of this study was to evaluate the performance of S. frugiperda field-collected strains on different Bt and non-Bt maize hybrids. Strains were collected from non-Bt maize (T1), Agrisure TDMax (T2), Agrisure Viptera (T3), Agrisure Viptera 3110 (T4), Genuity VT Triple Pro (T5), and Power Core (T6). Three experiments were performed to 1) determine the survivorship and reproduction of field-collected larvae (F0) from Bt maize hybrids, 2) evaluate Cry1F resistance using an F1 screen, and 3) assess the performance of F1 strains on different maize hybrids. In the F0, the survivorship from larva to adult ranged from 0 to 63%. We obtained adults from only the T1, T2, T5, and T6 strains with no significant differences in the reproductive parameters. Continuously rearing F1 larvae on their collected hosts affected larval duration, which was significantly shorter for a known-laboratory Bt-susceptible strain than the field-collected strains. Our results support the existence of Cry1F-resistance alleles in S. frugiperda field populations in Argentina.

Modalidades de Expressão dos Jovens Gamers nas Redes Sociais Digitais / Modes of Expression of the Young Gamers in the Digital Social Networks

O presente estudo, de caráter netnográfico, visou analisar as modalidades de expressão da subjetividade dos jovens gamers nas redes sociais e suas relações com a saúde. Foram entrevistados oito jovens na rede social facebook, em comunidades relacionadas ao console Playstation 4. Para sistematização dos resultados foi utilizada a análise de conteúdo. A análise evidenciou as seguintes categorias: comunicação escrita, interesse grupal, agressividade, valorização da imagem e do semblante, sinalizando modos de habitar o espaço cibernético. Tais dimensões devem ser consideradas ao se planejar políticas públicas e ações de promoção da saúde voltadas à realidade dos jovens gamers, tendo-se como apoio os princípios da cultura de paz e de respeito às diferenças, em busca do desenvolvimento de espaços saudáveis à expressão das singularidades. Palavras-chave: Jovens; jogos de computador; promoção da saúde.

This study aimed to analyze the subjectivity expression modes of young gamers in social networks and their relationships with health, using the netnographic method. Eight young people were interviewed on the facebook social network, in communities related to the Playstation 4. The analysis showed the following categories: written communication, group interest, aggressiveness, image and semblance valorization, signaling ways of inhabiting the cyber space. Such dimensions should be considered when planning public policies and health promotion actions focused on the reality of young gamers, having as support the principles of peace culture and respect for differences, in search of the development of healthy spaces for the expression of singularities.

Community Composition, Abundance, and Phenology of Stink Bugs (Hemiptera: Pentatomidae) in Soybean in the North Central Region of the United States.

Stink bugs (Hemiptera: Pentatomidae) are an increasing threat to soybean (Fabales: Fabaceae) production in the North Central Region of the United States, which accounts for 80% of the country's total soybean production. Characterization of the stink bug community is essential for development of management programs for these pests. However, the composition of the stink bug community in the region is not well defined. This study aimed to address this gap with a 2-yr, 9-state survey. Specifically, we characterized the relative abundance, richness, and diversity of taxa in this community, and assessed phenological differences in abundance of herbivorous and predatory stink bugs. Overall, the stink bug community was dominated by Euschistus spp. (Hemiptera: Pentatomidae) and Chinavia hilaris (Say) (Hemiptera: Pentatomidae). Euschistus variolarius (Palisot de Beauvois) (Hemiptera: Pentatomidae), C. hilaris and Halyomorpha halys (Stål) (Hemiptera: Pentatomidae) were more abundant in the northwestern, southeastern and eastern parts, respectively, of the North Central Region of the United States. Economically significant infestations of herbivorous species occurred in fields in southern parts of the region. Species richness differed across states, while diversity was the same across the region. Herbivorous and predatory species were more abundant during later soybean growth stages. Our results represent the first regional characterization of the stink bug community in soybean fields and will be fundamental for the development of state- and region-specific management programs for these pests in the North Central Region of the United States.

Spatial Patterns and Sequential Sampling Plans for Estimating Densities of Stink Bugs (Hemiptera: Pentatomidae) in Soybean in the North Central Region of the United States.

Stink bugs are an emerging threat to soybean (Fabales: Fabaceae) in the North Central Region of the United States. Consequently, region-specific scouting recommendations for stink bugs are needed. The aim of this study was to characterize the spatial pattern and to develop sampling plans to estimate stink bug population density in soybean fields. In 2016 and 2017, 125 fields distributed across nine states were sampled using sweep nets. Regression analyses were used to determine the effects of stink bug species [Chinavia hilaris (Say) (Hemiptera: Pentatomidae) and Euschistus spp. (Hemiptera: Pentatomidae)], life stages (nymphs and adults), and field locations (edge and interior) on spatial pattern as represented by variance-mean relationships. Results showed that stink bugs were aggregated. Sequential sampling plans were developed for each combination of species, life stage, and location and for all the data combined. Results for required sample size showed that an average of 40-42 sample units (sets of 25 sweeps) would be necessary to achieve a precision of 0.25 for stink bug densities commonly encountered across the region. However, based on the observed geographic gradient of stink bug densities, more practical sample sizes (5-10 sample units) may be sufficient in states in the southeastern part of the region, whereas impractical sample sizes (>100 sample units) may be required in the northwestern part of the region. Our findings provide research-based sampling recommendations for estimating densities of these emerging pests in soybean.

Weekly Survivorship Curves of Soybean Aphid Biotypes 1 and 4 on Insecticidal Seed-Treated Soybean.

Thiamethoxam, an insecticide used in soybean seed treatments, effectively suppresses soybean aphids (Aphis glycines) Matsumura (Hemiptera: Aphididae) for a short time after planting. However, exactly when and how quickly soybean aphid populations could increase is unknown. Likewise, we lack data on virulent soybean aphid biotypes (that can overcome soybean resistance) when fed on seed-treated soybean. Determining the survival of soybean aphids over time on insecticidal seed-treated soybean is critical for improving soybean aphid management and may provide insights to manage aphid virulence to aphid resistant-soybean. In greenhouse and field experiments, aphid-susceptible soybean plants (with and without an insecticidal seed treatment) were infested at 7, 14, 21, 28, 35, and 42 days after planting (DAP). We compared aphid survival among biotypes 1 (avirulent) and 4 (virulent) and insecticide treatment 72 h after infestation. We also measured thiamethoxam concentrations in plant tissue using liquid chromatography-tandem mass spectrometry. As expected, soybean aphid survival was significantly lower on seed-treated soybean up to 35 DAP for both biotypes, which correlates with the decrease of thiamethoxam in the plant over time. Moreover, we found no significant difference between avirulent and virulent biotype survivorship on insecticidal seed-treated soybean plants, although we did find significantly greater survival for the virulent biotype compared with the avirulent biotype on untreated soybean in the field. In conclusion, our study further characterized the relative short duration of seed treatment effectiveness on soybean aphid and showed that survivorship of virulent aphids on seed-treated soybean is similar to avirulent aphids.

Genome-wide genotyping of a novel Mexican Chile Pepper collection illuminates the history of landrace differentiation after Capsicum annuum L. domestication.

Studies of genetic diversity among phenotypically distinct crop landraces improve our understanding of fruit evolution and genome structure under domestication. Chile peppers (Capsicum spp. L.) are economically valuable and culturally important species, and extensive phenotypic variation among landraces exists in southern Mexico, a center of C. annuum diversity. We collected 103 chile pepper seed accessions from 22 named landraces across 27 locations in southern Mexico. We genotyped these accessions with genotyping by sequencing (GBS), yielding 32,623 filtered single-nucleotide polymorphisms. Afterward, we genotyped 32 additional C. annuum accessions from a global collection for comparison to the Mexican collection. Within the Mexican collection, genetic assignment analyses showed clear genetic differentiation between landraces and clarified the unique nature of the Tusta landrace. Further clustering analyses indicated that the largest fresh-use Chile de Agua and dry-use Costeño landraces were part of separate clades, indicating that these two landraces likely represent distinct populations. The global accessions showed considerable admixture and limited clustering, which may be due to the collapse of use-type divisions outside of Central America. The separation of the Mexican landraces in part by fruit morphology related to use highlights the relevance of this use-type morphological diversity for plant breeders and the utility of fruit development variation for evolutionary biologists.

Inward rectifier potassium (Kir) channels in the soybean aphid Aphis glycines: Functional characterization, pharmacology, and toxicology.

Inward rectifier K+ (Kir) channels contribute to a variety of physiological processes in insects and are emerging targets for insecticide development. Previous studies on insect Kir channels have primarily focused on dipteran species (e.g., mosquitoes, fruit flies). Here we identify and functionally characterize Kir channel subunits in a hemipteran insect, the soybean aphid Aphis glycines, which is an economically important insect pest and vector of soybeans. From the transcriptome and genome of Ap. glycines we identified two cDNAs, ApKir1 and ApKir2, encoding Kir subunits that were orthologs of insect Kir1 and Kir2, respectively. Notably, a gene encoding a Kir3 subunit was absent from the transcriptome and genome of Ap. glycines, similar to the pea aphid Acyrthosiphon pisum. Heterologous expression of ApKir1 and ApKir2 in Xenopus laevis oocytes enhanced K+-currents in the plasma membrane; these currents were inhibited by barium and the small molecule VU041. Compared to ApKir2, ApKir1 mediated currents that were larger in magnitude, more sensitive to barium, and less inhibited by small molecule VU041. Moreover, ApKir1 exhibited stronger inward rectification compared to ApKir2. Topical application of VU041 in adult aphids resulted in dose-dependent mortality within 24 h that was more efficacious than flonicamid, an established insecticide also known to inhibit Kir channels. We conclude that despite the apparent loss of Kir3 genes in aphid evolution, Kir channels are important to aphid survival and represent a promising target for the development of new insecticides.

Defoliation of Soybean Expressing Cry1Ac by Lepidopteran Pests.

Lepidoptera, stink bugs, and weevils are important pests in soybean. For lepidopteran control, insecticides and seed treatments are used. As an alternative, Bt soybean was developed to control primary pests of Lepidoptera such as Rachiplusia nu (Guenée) (Noctuidae), Chrysodeixisincludens (Walker) (Noctuidae), Anticarsia gemmatalis Hübner (Erebidae), Helicoverpa gelotopoeon (Dyar) (Noctuidae), and Crocidosema aporema (Walsingham) (Tortricidae). However, the use of transgenic plants, and the resulting reduction of insecticide against target pests, may allow other pest species to become more prevalent in agricultural environments. Soybean expressing Cry1Ac against different lepidopteran nontarget and target insect pests was evaluated, and its performance was compared with non-Bt soybean with seed treatment. The treatments were Bt soybean, non-Bt soybean with seed treatment (Fortenza® diamide insecticide, Syngenta, Buenos Aires, Argentina), and non-Bt soybean without seed treatment. Larvae of H. gelotopoeon, Spodoptera albula (Walker) (Noctuidae), Spodoptera cosmiodes (Walker) (Noctuidae), Spodoptera eridania (Stoll) (Noctuidae), and Spodoptera frugiperda (J. E. Smith) (Lep.: Noctuidae) were used. The plants of each treatment were infested with larvae of each species, and the percentage of leaf damage produced by each species was recorded. The results showed that Bt soybean provided control of H. gelotopoeon and had a suppressive effect on S. frugiperda and S. albula. However, S. eridania and S. cosmiodes were not susceptible to the Cry1Ac protein in MON 87701 × MON 89788 soybean when evaluated by greenhouse infestation. Considering the performance of each species using non-Bt soybean without seed treatment, S. eridania would represent a potential risk in soybean crops.

Influence of historical land use and modern agricultural expansion on the spatial and ecological divergence of sugarcane borer, Diatraea saccharalis (Lepidoptera: Crambidae) in Brazil.

Human-mediated changes in landscapes can facilitate niche expansion and accelerate the adaptation of insect species. The interaction between the evolutionary history of the sugarcane borer, Diatraea saccharalis Fabricius, and historical and modern agricultural activity in Brazil shaped its spatial genetic structure, facilitating ecological divergence and incipient host shifting. Based on microsatellite data, STRUCTURE analyses identified two (K = 2) and three (K = 3) significant genetic clusters that corresponded to: (a) a strong signal of spatial genetic structure and, (b) a cryptic signal of host differentiation. We inferred that K = 2 reflects the footprint of agricultural activity, such as expansion of crop production (sugarcane and maize), unintentional dispersion of pests, and management practices. In contrast, K = 3 indicated incipient host differentiation between larvae collected from sugarcane or maize. Our estimates of population size changes indicated that a historical bottleneck was associated with a reduction of sugarcane production ≈200 years ago. However, a more recent population expansion was detected (>1950s), associated with agricultural expansion of large crop production into previously unfarmed land. Partial Mantel tests supported our hypothesis of incipient host adaptation, and identified isolation-by-environment (e.g., host plant) in São Paulo and Minas Gerais states, where sugarcane has been traditionally produced in Brazil. The impact of agricultural production on D. saccharalis may continue, as the current population structure may hinder the efficacy of refuge plants in delaying insect resistance evolution to Bt toxin.

Transcriptomic dynamics in soybean near-isogenic lines differing in alleles for an aphid resistance gene, following infestation by soybean aphid biotype 2.

BACKGROUND: Genetic resistance of soybean [Glycine max (L.) Merr] against Aphis glycines provides effective management of this invasive pest, though the underlying molecular mechanisms are largely unknown. This study aimed to investigate genome-wide changes in gene expressions of soybean near-isogenic lines (NILs) either with the Rag5 allele for resistance or the rag5 allele for susceptibility to the aphid following infestation with soybean aphid biotype 2. RESULTS: The resistant (R)-NIL responded more rapidly to aphid infestation than the susceptible (S)-NIL, with differential expressions of 2496 genes during first 12 h of infestation (hai), compared to the aphid-free control. Although the majority of the differentially expressed genes (DEGs) in the R-NIL also responded to aphid infestation in S-NIL, overall the response time was longer and/or the magnitude of change was smaller in the S-NIL. In addition, 915 DEGs in R-NIL continued to be regulated at all time points (0, 6, 12, and 48 hai), while only 20 DEGs did so in S-NIL. Enriched gene ontology of the 2496 DEGs involved in plant defense responses including primary metabolite catalysis, oxidative stress reduction, and phytohormone-related signaling. By comparing R- vs. S-NIL, a total of 556 DEGs were identified. Of the 13 genes annotated in a 120-kb window of the Rag5 locus, two genes (Glyma.13 g190200 and Glyma.13 g190600) were differentially expressed (upregulated in S- or R-NIL), and another gene (Glyma.13 g190500) was induced up to 4-fold in the R-NIL at 6 and 12 h following aphid infestation. CONCLUSIONS: This study strengthens our understanding of the defense dynamics in compatible and incompatible interactions of soybean and soybean aphid biotype 2. Several DEGs (e.g., Glyma.13 g190200, Glyma.13 g190500, and Glyma.13 g190600) near the Rag5 locus are strong candidate genes for further investigations.

Whole Genome Sequence of the Soybean Aphid Endosymbiont Buchnera aphidicola and Genetic Differentiation among Biotype-Specific Strains.

Endosymbiosis with microorganisms is common in insects, with more than 10% of species requiring the metabolic capabilities of intracellular bacteria for their nutrient acquisition. Aphids harbor an obligate mutualism with the vertically transferred endosymbiont, Buchnera aphidicola, which produces key nutrients lacking in the aphid's phloem-based diet that are necessary for normal development and reproduction. It is thought that, in some groups of insects, bacterial symbionts may play key roles in biotype evolution against host-plant resistance. The genome of Buchnera has been sequenced in several aphid strains but little genomic data is currently available for the soybean aphid (Aphis glycines), one of the most important pests of soybean in North America. In this study, DNA sequencing was used to assemble and annotate the genome sequence of the Buchnera A. glycines strain and to reconstruct phylogenetic relationships among different strains. In addition, we identified several fixed Buchnera SNPs between Aphis glycines biotypes that were avirulent or virulent to a soybean aphid resistance gene (Rag1). The results of this study describe the genetic and evolutionary relationships of the Buchnera A. glycines strain, and begin to define the roles of an aphid symbiont in host-plant resistance.

Nanoparticles Containing High Loads of Paclitaxel-Silicate Prodrugs: Formulation, Drug Release, and Anticancer Efficacy.

We have investigated particle size, interior structure, drug release kinetics, and anticancer efficacy of PEG-b-PLGA-based nanoparticles loaded with a series of paclitaxel (PTX)-silicate prodrugs [PTX-Si(OR)3]. Silicate derivatization enabled us to adjust the hydrophobicity and hydrolytic lability of the prodrugs by the choice of the alkyl group (R) in the silicate derivatives. The greater hydrophobicity of these prodrugs allows for the preparation of nanoparticles that are stable in aqueous dispersion even when loaded with up to ca. 75 wt % of the prodrug. The hydrolytic lability of silicates allows for facile conversion of prodrugs back to the parent drug, PTX. A suite of eight PTX-silicate prodrugs was investigated; nanoparticles were made by flash nanoprecipitation (FNP) using a confined impingement jet mixer with a dilution step (CIJ-D). The resulting nanoparticles were 80-150 nm in size with a loading level of 47-74 wt % (wt %) of a PTX-silicate, which corresponds to 36-59 effective wt % of free PTX. Cryogenic transmission electron microscopy images show that particles are typically spherical with a core-shell structure. Prodrug/drug release profiles were measured. Release tended to be slower for prodrugs having greater hydrophobicity and slower hydrolysis rate. Nanoparticles loaded with PTX-silicate prodrugs that hydrolyze most rapidly showed in vitro cytotoxicity similar to that of the parent PTX. Nanoparticles loaded with more labile silicates also tended to show greater in vivo efficacy.

Shifts in Buchnera aphidicola density in soybean aphids (Aphis glycines) feeding on virus-infected soybean.

Vertically transmitted bacterial symbionts are common in arthropods. Aphids undergo an obligate symbiosis with Buchnera aphidicola, which provides essential amino acids to its host and contributes directly to nymph growth and reproduction. We previously found that newly adult Aphis glycines feeding on soybean infected with the beetle-transmitted Bean pod mottle virus (BPMV) had significantly reduced fecundity. We hypothesized that the reduced fecundity was attributable to detrimental impacts of the virus on the aphid microbiome, namely Buchnera. To test this, mRNA sequencing and quantitative real-time PCR were used to assay Buchnera transcript abundance and titre in A. glycines feeding on Soybean mosaic virus-infected, BPMV-infected, and healthy soybean for up to 14 days. Our results indicated that Buchnera density was lower and ultimately suppressed in aphids feeding on virus-infected soybean. While the decreased Buchnera titre may be associated with reduced aphid fecundity, additional mechanisms are probably involved. The present report begins to describe how interactions among insects, plants, and plant pathogens influence endosymbiont population dynamics.