Aphid gene expression following polerovirus acquisition is host species dependent.

Upon acquisition of persistent circulative viruses such as poleroviruses, the virus particles transcytose through membrane barriers of aphids at the midgut and salivary glands via hemolymph. Such intricate interactions can influence aphid behavior and fitness and induce associated gene expression in viruliferous aphids. Differential gene expression can be evaluated by omics approaches such as transcriptomics. Previously conducted aphid transcriptome studies used only one host species as the source of virus inoculum. Viruses typically have alternate hosts. Hence, it is not clear how alternate hosts infected with the same virus isolate alter gene expression in viruliferous vectors. To address the question, this study conducted a transcriptome analysis of viruliferous aphids that acquired the virus from different host species. A polerovirus, cotton leafroll dwarf virus (CLRDV), which induced gene expression in the cotton aphid, Aphis gossypii Glover, was assessed using four alternate hosts, viz., cotton, hibiscus, okra, and prickly sida. Among a total of 2,942 differentially expressed genes (DEGs), 750, 310, 1,193, and 689 genes were identified in A. gossypii that acquired CLRDV from infected cotton, hibiscus, okra, and prickly sida, respectively, compared with non-viruliferous aphids that developed on non-infected hosts. A higher proportion of aphid genes were overexpressed than underexpressed following CLRDV acquisition from cotton, hibiscus, and prickly sida. In contrast, more aphid genes were underexpressed than overexpressed following CLRDV acquisition from okra plants. Only four common DEGs (heat shock protein, juvenile hormone acid O-methyltransferase, and two unannotated genes) were identified among viruliferous aphids from four alternate hosts. Gene ontology (GO) enrichment analysis and Kyoto Encyclopedia of Genes and Genomes (KEGG) annotations indicated that the acquisition of CLRDV induced DEGs in aphids associated with virus infection, signal transduction, immune systems, and fitness. However, these induced changes were not consistent across four alternate hosts. These data indicate that alternate hosts could differentially influence gene expression in aphids and presumably aphid behavior and fitness despite being infected with the same virus isolate.

Correction: Mahas et al. Seasonal Dynamics of Aphid Flights and Cotton Leafroll Dwarf Virus Spread in Alabama. Insects 2023, 14, 604.

In the original publication [...].

Effects of Food Source Availability, Host Egg:Parasitoid Ratios, and Host Exposure Times on the Developmental Biology of Megacopta cribraria Egg Parasitoids.

Parasitoids forage for hosts in dynamic ecosystems and generally have a short period of time to access hosts. The current study examined the optimal reproductive attributes of two egg parasitoids, Paratelenomus saccharalis Dodd (Hymenoptera: Platygastridae) and Ooencyrtus nezarae Ishii (Hymenoptera: Encyrtidae), of the kudzu bug, Megacopta cribraria Fabricius (Hemiptera: Plataspidae). The proportion of O. nezarae and P. saccharalis adult offspring that emerged from M. cribraria eggs and the sex ratio of the parasitoid offspring were compared among treatments for the effects of different adult parasitoid food sources, host egg-to-adult parasitoid ratios, and host exposure times. Our results suggest that honey solution as a food source, a 21:7 host-to-parasitoid ratio, and three-to-five days of exposure time optimized the production of female O. nezarae offspring. For P. saccharalis, honey solution as a food source, a 21:7 host-to-parasitoid ratio, and one day were optimal for maximizing female offspring production. These findings provide new information about the biology of these egg parasitoids.

Seasonal Dynamics of Aphid Flights and Cotton Leafroll Dwarf Virus Spread in Alabama.

Cotton leafroll dwarf virus (CLRDV) is an introduced Polerovirus (Family: Solemoviridae) of cotton, Gossypium hirsutum L., in the U.S. The only vector known to transmit this virus to cotton is the cotton aphid, Aphis gossypii Glover; however, there are seven other species of aphids (Hemiptera: Aphididae) reported to colonize cotton in the southeastern U.S.: Protaphis middletonii (Thomas), Rhopalosiphum rufiabdominale (Sasaki), Aphis craccivora Koch, Macrosiphum euphorbiae Thomas, Myzus persicae (Sulzer), Smythurodes betae Westwood, and Aphis fabae Scopoli. Little to no information is available on annual population dynamics of these species in the southeastern U.S. The timing of CLRDV spread to cotton plantings is also unknown. The objective of this study was to monitor the population dynamics of eight cotton-feeding aphid species concurrent with the spread of CLRDV at three different locations in Alabama. Aphids were monitored weekly for two years with yellow pan traps, and sentinel plants were deployed weekly to monitor CLRDV spread throughout the cotton-growing season. During the two years, most CLRDV spread at all locations occurred when A. gossypii was actively dispersing in the field. Early season spread at sites in south and central Alabama, when A. gossypii was not abundant, suggests additional aphid vectors are possible.

The Spatiotemporal Distribution, Abundance, and Seasonal Dynamics of Cotton-Infesting Aphids in the Southern U.S.

Cotton leafroll dwarf virus (CLRDV) is an emerging aphid-borne pathogen infecting cotton, Gossypium hirsutum L., in the southern United States (U.S.). The cotton aphid, Aphis gossypii Glover, infests cotton annually and is the only known vector to transmit CLRDV to cotton. Seven other species have been reported to feed on, but not often infest, cotton: Protaphis middletonii Thomas, Aphis craccivora Koch, Aphis fabae Scopoli, Macrosiphum euphorbiae Thomas, Myzus persicae Sulzer, Rhopalosiphum rufiabdominale Sasaki, and Smynthurodes betae Westwood. These seven have not been studied in cotton, but due to their potential epidemiological importance, an understanding of the intra- and inter-annual variations of these species is needed. In 2020 and 2021, aphids were monitored from North Carolina to Texas using pan traps around cotton fields. All of the species known to infest cotton, excluding A. fabae, were detected in this study. Protaphis middletonii and A. gossypii were the most abundant species identified. The five other species of aphids captured were consistently low throughout the study and, with the exception of R. rufiabdominale, were not detected at all locations. The abundance, distribution, and seasonal dynamics of cotton-infesting aphids across the southern U.S. are discussed.

Genome segment ratios change during whitefly transmission of two bipartite cassava mosaic begomoviruses.

Cassava mosaic disease is caused by a complex of whitefly-transmitted begomoviruses, which often occur in co-infections. These viruses have bipartite genomes consisting of DNA-A and DNA-B that are encapsidated into separate virions. Individual viruses exist in plants and whitefly vectors as populations comprising both genome segments, which can occur at different frequencies. Both segments are required for infection, and must be transmitted for virus spread to occur. Cassava plants infected with African cassava mosaic virus (ACMV) and/or East African cassava mosaic Cameroon virus (EACMCV), in which the ratios of DNA-A:DNA-B titers differed between plants, were used to examine how titers of the segments in a plant relate to their respective probabilities of acquisition by whiteflies and to the titers of each segment acquired and subsequently transmitted by whiteflies. The probabilities of acquiring each segment of ACMV did not reflect their relative titers in the source plant but they did for EACMCV. However, for both viruses, DNA-A:DNA-B ratios acquired by whiteflies differed from those in the source plant and the ratios transmitted by the whitefly did not differ from one - the ratio at which the highest probability of transmitting both segments is expected.

Pest status, molecular evolution, and epigenetic factors derived from the genome assembly of Frankliniella fusca, a thysanopteran phytovirus vector.

BACKGROUND: The tobacco thrips (Frankliniella fusca Hinds; family Thripidae; order Thysanoptera) is an important pest that can transmit viruses such as the tomato spotted wilt orthotospovirus to numerous economically important agricultural row crops and vegetables. The structural and functional genomics within the order Thysanoptera has only begun to be explored. Within the > 7000 known thysanopteran species, the melon thrips (Thrips palmi Karny) and the western flower thrips (Frankliniella occidentalis Pergrande) are the only two thysanopteran species with assembled genomes. RESULTS: A genome of F. fusca was assembled by long-read sequencing of DNA from an inbred line. The final assembly size was 370 Mb with a single copy ortholog completeness of ~ 99% with respect to Insecta. The annotated genome of F. fusca was compared with the genome of its congener, F. occidentalis. Results revealed many instances of lineage-specific differences in gene content. Analyses of sequence divergence between the two Frankliniella species' genomes revealed substitution patterns consistent with positive selection in ~ 5% of the protein-coding genes with 1:1 orthologs. Further, gene content related to its pest status, such as xenobiotic detoxification and response to an ambisense-tripartite RNA virus (orthotospovirus) infection was compared with F. occidentalis. Several F. fusca genes related to virus infection possessed signatures of positive selection. Estimation of CpG depletion, a mutational consequence of DNA methylation, revealed that F. fusca genes that were downregulated and alternatively spliced in response to virus infection were preferentially targeted by DNA methylation. As in many other insects, DNA methylation was enriched in exons in Frankliniella, but gene copies with homology to DNA methyltransferase 3 were numerous and fragmented. This phenomenon seems to be relatively unique to thrips among other insect groups. CONCLUSIONS: The F. fusca genome assembly provides an important resource for comparative genomic analyses of thysanopterans. This genomic foundation allows for insights into molecular evolution, gene regulation, and loci important to agricultural pest status.

Characterizing the vector competence of Aphis gossypii, Myzus persicae and Aphis craccivora (Hemiptera: Aphididae) to transmit cotton leafroll dwarf virus to cotton in the United States.

Cotton leafroll dwarf virus (CLRDV) is a yield-limiting, aphid-transmitted virus that was identified in cotton, Gossypium hirsutum L., in the United States of America in 2017. CLRDV is currently classified in the genus Polerovirus, family Solemoviridae. Although 8 species of aphids (Hemiptera: Aphididae) are reported to infest cotton, Aphis gossypii Glover is the only known vector of CLRDV to this crop. Aphis gossypii transmits CLRDV in a persistent and nonpropagative manner, but acquisition and retention times have only been partially characterized in Brazil. The main objectives of this study were to characterize the acquisition access period, the inoculation access period, and retention times for a U.S. strain of CLRDV and A. gossypii population. A sub-objective was to test the vector competence of Myzus persicae Sulzer and Aphis craccivora Koch. In our study, A. gossypii apterous and alate morphs were able to acquire CLRDV in 30 min and 24 h, inoculate CLRDV in 45 and 15 min, and retain CLRDV for 15 and 23 days, respectively. Neither M. persicae nor A. craccivora acquired or transmitted CLRDV to cotton.

Imidacloprid-resistant Aphis gossypii populations are more common in cotton-dominated landscapes.

BACKGROUND: Widespread reports of reduced efficacy of imidacloprid for managing cotton aphids (Aphis gossypii Glover) in cotton (Gossypium hirsutum L.) prompted an investigation to characterize the susceptibility of 43 populations over a 2-year period. The susceptibility of A. gossypii populations to imidacloprid was examined by calculating LC50 values. Further analyses related resistance assay results to a gradient of cotton production intensity. RESULTS: Concentration-mortality bioassays documencted populations that were 4.26-607.16 times more resistant than the susceptible laboratory population. There was a significant positive relationship between LC50 values and percentage of cotton within 2.5- and 5-km buffers surrounding collection sites. No significant relationship was detected between LC50 values and the percentage of alternative crop and noncrop hosts. CONCLUSION: Variable and high levels of resistance were detected in A. gossypii populations, and this variation was positively associated with cotton production intensity. Cotton is a host that may receive multiple applications of neonicotinoids (via seed treatment and foliar sprays) annually for seedling and mid-season pests. Rotating modes of action and limiting insecticide use should be implemented to delay the evolution of insecticide resistance in A. gossypii populations. © 2022 Society of Chemical Industry.

Vector acquisition and co-inoculation of two plant viruses influences transmission, infection, and replication in new hosts.

This study investigated the role of vector acquisition and transmission on the propagation of single and co-infections of tomato yellow leaf curl virus (TYLCV,) and tomato mottle virus (ToMoV) (Family: Geminiviridae, Genus: Begomovirus) by the whitefly vector Bemisia tabaci MEAM1 (Gennadius) in tomato. The aim of this research was to determine if the manner in which viruses are co-acquired and co-transmitted changes the probability of acquisition, transmission and new host infections. Whiteflies acquired virus by feeding on singly infected plants, co-infected plants, or by sequential feeding on singly infected plants. Viral titers were also quantified by qPCR in vector cohorts, in artificial diet, and plants after exposure to viruliferous vectors. Differences in transmission, infection status of plants, and titers of TYLCV and ToMoV were observed among treatments. All vector cohorts acquired both viruses, but co-acquisition/co-inoculation generally reduced transmission of both viruses as single and mixed infections. Co-inoculation of viruses by the vector also altered virus accumulation in plants regardless of whether one or both viruses were propagated in new hosts. These findings highlight the complex nature of vector-virus-plant interactions that influence the spread and replication of viruses as single and co-infections.

Prospective Alternate Hosts of an Emerging Polerovirus in Cotton Landscapes in the Southeastern United States.

The identification of alternate hosts that can act as virus inoculum sources and vector reservoirs in the landscape is critical to understanding virus epidemics. Cotton leafroll dwarf virus (CLRDV) is a serious pathogen in cotton production and is transmitted by the cotton/melon aphid, Aphis gossypii, in a persistent, circulative, and non-propagative manner. CLRDV was first reported in the United States in Alabama in 2017, and thereafter in several cotton-producing states. CLRDV has since established itself in the southeastern United States. The role of alternate hosts in CLRDV establishment is not clear. Fourteen common plant species in the landscape, including crops, weeds, and ornamentals (cotton, hollyhock, marshmallow, country mallow, abutilon, arrowleaf sida, okra, hibiscus, squash, chickpea, evening primrose, henbit, Palmer amaranth, and prickly sida) were tested as potential alternate hosts of CLRDV along with an experimental host (Nicotiana benthamiana) via aphid-mediated transmission assays. CLRDV was detected following inoculation in hibiscus, okra, N. benthamiana, Palmer amaranth, and prickly sida by RT-PCR, but not in the others. CLRDV accumulation determined by RT-qPCR was the highest in N. benthamiana compared with cotton and other hosts. However, aphids feeding on CLRDV-infected prickly sida, hibiscus, and okra alone were able to acquire CLRDV and back-transmit it to non-infected cotton seedlings. Additionally, some of the alternate CLRDV hosts supported aphid development on par with cotton. However, in a few instances, aphid fitness was reduced when compared with cotton. Overall, this study demonstrated that plant hosts in the agricultural landscape can serve as CLRDV inoculum sources and as aphid reservoirs and could possibly play a role in the reoccurring epidemics of CLRDV in the southeastern United States.

Sampling Optimization and Crop Interface Effects on Lygus lineolaris Populations in Southeastern USA Cotton.

Tarnished plant bug, Lygus lineolaris (Hemiptera: Miridae), is an economically damaging pest in cotton production systems across the southern United States. We systematically scouted 120 commercial cotton fields across five southeastern states during susceptible growth stages in 2019 and 2020 to investigate sampling optimization and the effect of interface crop and landscape composition on L. lineolaris abundance. Variance component analysis determined field and within-field spatial scales, compared with agricultural district and state, accounted for more variation in L. lineolaris density using sweep net and drop cloth sampling. This result highlights the importance of field-level scouting efforts. Using within-field samples, a fixed-precision sampling plan determined 8 and 23 sampling units were needed to determine L. lineolaris population estimates with 0.25 precision for sweep net (100 sweeps per unit) and drop cloth (1.5 row-m per unit) sampling, respectively. A spatial Bayesian hierarchical model was developed to determine local landscape (<0.5 km from field edges) effects on L. lineolaris in cotton. The proportion of agricultural area and double-crop wheat and soybeans were positively associated with L. lineolaris density, and fields with more contiguous cotton areas negatively predicted L. lineolaris populations. These results will improve L. lineolaris monitoring programs and treatment management decisions in southeastern USA cotton.

Direct and Indirect Competitive Interactions between Ooencyrtus nezarae and Paratelenomus saccharalis Parasitizing Megacopta cribraria Egg Patches.

The present study investigated egg parasitoid interspecific interactions between a generalist, Ooencyrtus nezarae Ishii (Hymenoptera: Encyrtidae) and a specialist, Paratelenomus saccharalis Dodd (Hymenoptera: Platygastridae) in a laboratory setting using kudzu bug (Megacopta cribraria Fabricius, (Hemiptera: Plataspidae)) eggs as their shared host. Three experiments were conducted to evaluate the emergence of wasps from parasitized hosts after the simultaneous and sequential release of wasps, monitor aggressive behavior of P. saccharalis, and quantify intraguild predation of O. nezarae larvae on heterospecific P. saccharalis larvae. Results showed that total host egg parasitism was higher when both wasps were released simultaneously than if wasps were released sequentially. Ooencyrtus nezarae produced more total offspring than P. saccharalis in all sequential/simultaneous treatments but produced male offspring in most cases. In the aggressive behavioral experiment, specialist, P. saccharalis used head butting to fight O. nezarae, but no other aggressions were observed. In an experiment examining intraguild predation, O. nezarae was able to develop in host eggs parasitized by P. saccharalis four days earlier, acting as a superior larval competitor. These findings shed light on the potential interspecific interactions between O. nezarae and P. saccharalis, which may determine their relative abundance and influence their compatibility in kudzu bug biological control programs.

Natural Enemies, Mediated by Landscape and Weather Conditions, Shape Response of the Sorghum Agroecosystem of North America to the Invasive Aphid Melanaphis sorghi.

The sorghum (Sorghum bicolor [L.]) agroecosystem of North America provided an opportunity to evaluate agroecosystem response to an invading insect herbivore, Melanaphis sorghi (Theobald) (sorghum aphid) (previously published as Melanaphis sacchari Zehntner) (Hemiptera: Aphididae) onto a widely planted crop that experiences a range of agro-landscape and weather conditions. Initial sorghum risk assessments after M. sorghi's invasion in the mid-2010s provided forecasts of range expansion and annual migration, which were based on aphid life history, extent of sorghum cultivation and susceptibility to M. sorghi, and weather (aphid-plant-weather [APW] risk scenario). A more comprehensive risk assessment proposed here brings top-down forces of M. sorghi-natural enemy interactions to the forefront as mediated by agro-landscape and weather conditions (aphid-enemy/landscape-weather mediated [AE/LW] risk scenario). A hypothesis of regional differences in aphids and natural enemies and sensitivity to agro-landscape and weather was tested using empirical data of insect, landscape, and weather data across 5 years and four regions (two in the U.S. Great Plains [South GP and North GP], one farther south (South), and one in the southeast U.S. [South E]). Natural enemies were widespread with two parasitoids and four coccinellid species common across regions, but regional variation in M. sorghi and natural enemy abundance was detected. The AE/LW risk scenario accounted for natural enemy abundance and activity that was highest in the South region, functioned well across agro-landscape and weather conditions, and was accompanied by average low M. sorghi abundance (~23 M. sorghi per leaf). Positive correlations of natural enemy-M. sorghi abundance also occurred in the South GP region where M. sorghi abundance was low (~20 M. sorghi per leaf), and selected natural enemy activity appeared to be mediated by landscape composition. Melanaphis sorghi abundance was highest in the South E region (~136 aphids/leaf) where natural enemy activity was low and influenced by weather. The AE/LW risk scenario appeared suited, and essential in the South region, in assessing risk on a regional scale, and sets the stage for further modeling to generate estimates of the degree of influence of natural enemies under varying agro-landscape and weather conditions considered in the AE/LW risk scenario. Broadly, these findings are relevant in understanding agroecosystem resilience and recommending supportive management inputs in response to insect invasions in context of natural enemy activity and varied environmental conditions.

Evaluating Sugarcane Aphid, Melanaphis sacchari (Hemiptera: Aphididae), Population Dynamics, Feeding Injury, and Grain Yield Among Commercial Sorghum Varieties in Alabama.

The sugarcane aphid, Melanaphis sacchari (Zehntner), emerged as a severe pest of sorghum, Sorghum bicolor (L.), in Texas and Louisiana in 2013 and currently threatens nearly all sorghum production in the United States. Proper management of populations is critical as sugarcane aphid has a high reproductive potential and can rapidly damage plants, resulting in extensive yield losses. The overall objective of this work was to investigate sugarcane aphid population dynamics, and subsequent sorghum injury and grain yield on commercially available grain sorghum varieties in Alabama. This research includes three-site years of data that show variation in plant injury, physiological maturity, and yields among varieties tested. Although performance of each variety was variable among locations, potentially due to abiotic factors, four varieties including DKS 37-07, 1G588, 1G855, and 83P17 exhibited characteristics consistent with resistance and corroborates reports of resistance from other states.

Relationships of virus titers and transmission rates among sympatric and allopatric virus isolates and thrips vectors support local adaptation.

Plant viruses rely on insect vectors for transmission among plant hosts, but many of the specifics of virus-vector interactions are not fully understood. Thrips tabaci, which transmits Tomato spotted wilt virus (TSWV) in a persistent and propagative manner, varies greatly in its ability to transmit different isolates of TSWV. Similarly, TSWV isolates are transmitted at different efficiencies by different populations of T. tabaci. This study characterizes differences in virus titers in the vector among TSWV isolate-T. tabaci isoline pairings in relation to differences in transmission rates, and demonstrates that although transmission rates were higher for sympatric than allopatric TSWV isolate-T. tabaci isoline pairings, virus titers in the thrips vector were significantly lower in the sympatric pairings. Results further demonstrate that TSWV titers in the vector were unrelated to virus titers in the leaf tissue from which they acquired the virus and provide evidence for the importance of specific vector-virus interactions and local adaptation in determining transmission efficiency of TSWV by T. tabaci.

Behavioral Responses of Thrips (Thysanoptera: Thripidae) and Tarnished Plant Bug (Hemiptera: Miridae) to a New Bt Toxin, Cry51Aa2.834_16 in Cotton.

Thrips (Thysanoptera: Thripidae) and tarnished plant bug, Lygus lineolaris (Hemiptera: Miridae), are among the most important insect pests of cotton, Gosssypium hirsutum, in the mid-southern United States. These pests are currently managed primarily by insecticides; however, a new Bt toxin, Cry51Aa2.834_16 is under evaluation for control of thrips and tarnished plant bug. Experiments were conducted to evaluate the behavioral response of thrips and tarnished plant bug to Bt Cry51Aa2.834_16. Adult thrips avoided Bt Cry51Aa2.834_16 cotton in field choice tests and in separate field tests of Bt and non-Bt cotton not treated with insecticides. In a greenhouse choice test, approximately twice as many adult thrips and eggs were found on non-Bt compared with Bt Cry51Aa2.834_16 cotton. Similarly, in a field test of nontreated Bt Cry51Aa2.834_16 and non-Bt cotton, 68% of adult thrips collected were found on non-Bt cotton. In cotton that was not sprayed with insecticides, Bt Cry51Aa2.834_16 did not affect the distribution of tarnished plant bug within the canopy, although more square and flower injury was caused by tarnished plant bug in non-Bt cotton. Adult tarnished plant bug exhibited a nonpreference for diet containing lyophilized Bt Cry51Aa2.834_16 leaves and for excised Bt Cry51Aa2.834_16 squares in choice tests with non-Bt squares. The behavioral responses of these pests when exposed to this new Bt toxin will play a key role in the efficacy and potential resistance management strategies if this new technology is incorporated in an overall cotton insect pest management system.

Monitoring Seasonal Distribution of Thrips Vectors of Soybean Vein Necrosis Virus in Alabama Soybeans.

Soybean vein necrosis virus (SVNV), a new virus in the genus Orthotospovirus, has been found in all soybean-growing regions in the United States and Ontario, Canada. Soybean thrips, Neohydatothrips variabilis (Beach) (Thysanoptera: Thripidae), tobacco thrips, Frankliniella fusca (Hinds) (Thysanoptera: Thripidae), and eastern flower thrips, Frankliniella tritici (Fitch) (Thysanoptera: Thripidae) are reported vectors of this virus, but there are no reports on their distribution in Alabama. A monitoring study was conducted in 2015 and 2016 to determine thrips species composition and abundance in Alabama soybean agroecosystems. Thrips were monitored weekly by collecting them on yellow sticky traps and soybean plant parts including foliage and reproductive structures. All three reported vectors of SVNV were identified in Alabama, with N. variabilis and F. tritici as the predominant species, while F. fusca was not consistently collected from soybean plants. Four additional thrips species were collected, of which Echinothrips americanus (Morgan) (Thysanoptera: Thripidae) was commonly found on soybean at all three locations. Results presented in this study provide new information about seasonal thrips species abundance in soybean agroecosystems in Alabama, and is an important first step to understanding thrips vector species of epidemiological importance in the Southern United States.

Molecular Identification of Thrips Species Infesting Cotton in the Southeastern United States.

Traditional identification of thrips species based on morphology is difficult, laborious, and especially challenging for immature thrips. To support monitoring and management efforts of thrips as consistent and widespread pests of cotton (Gossypium hirsutum L.), a probe-based quantitative PCR (qPCR) assay with crude DNA extraction was developed to allow efficient and specific identification of the primary species of thrips infesting cotton. The assay was applied to identify over 5,000 specimens of thrips (including 3,366 immatures) collected on cotton seedlings from Alabama, Georgia, North Carolina, South Carolina, and Virginia in 2016. One half of all adult samples were examined by morphological identification, which provided a statistically equivalent species composition as the qPCR method. Frankliniella fusca (Hinds) (Thysanoptera: Thripidae) was the dominant species across all the locations (76.8-94.3% of adults and 81.6-98.0% of immatures), followed by Frankliniella occidentalis (Pergande) (Thysanoptera: Thripidae) in Georgia, North Carolina, and Virginia (4.6-19% of adults and 1.7-17.3% of immatures) or Frankliniella tritici (Fitch) (Thysanoptera: Thripidae) in South Carolina (10.8% of adults and 7.8% of immatures). Thrips tabaci (Lindeman) (Thysanoptera: Thripidae) and Neohydatothrips variabilis (Beach) (Thysanoptera: Thripidae) were occasionally found among adults but were rarely present among immature thrips. These five species of thrips represented 98.2-100% of samples collected across the Southeast. The qPCR assay was demonstrated to be a valuable tool for large-scale monitoring of species composition of thrips at different life stages in cotton. The tool will contribute to a better understanding of thrips population structure in cotton and could assist with development and application of improved management strategies.

Insecticide Resistance Signals Negative Consequences of Widespread Neonicotinoid Use on Multiple Field Crops in the U.S. Cotton Belt.

The intensification of industrial agriculture has been enabled by improved crop varieties, genetically engineered crops, fertilizers, and pesticides. Over the past 15 years, neonicotinoid seed treatments have been adopted worldwide and are used on a large proportion of U.S. field crops. Although neonicotinoids are used widely, little is known about how large-scale deployment affects pest populations over long periods. Here, we report a positive relationship between the deployment of neonicotinoid seed-dressings on multiple crops and the emergence of insecticide resistance in tobacco thrips (Frankliniella fusca), a polyphagous insect herbivore that is an important pest of seedling cotton but not soybean or maize. Using a geospatial approach, we studied the relationship between neonicotinoid resistance measured in 301 F. fusca populations to landscape-scale crop production patterns across nine states in the southeastern U.S. cotton production region, in which soybean, maize and cotton are the dominant crops. Our research linked the spatiotemporal abundance of cotton and soybean production to neonicotinoid resistance in F. fusca that is leading to a dramatic increase in insecticide use in cotton. Results demonstrate that cross-crop resistance selection has important effects on pests and, in turn, drives pesticide use and increases environmental impacts associated with their use.