Doublesex is essential for masculinization but not feminization in Lygus hesperus.

In most holometabolous insects, sex differentiation occurs via a hierarchical cascade of transcription factors, with doublesex (dsx) regulating genes that control sex-specific traits. Although less is known in hemimetabolous insects, early evidence suggests that substantial differences exist from more evolutionarily advanced insects. Here, we identified and characterized dsx in Lygus hesperus (western tarnished plant bug), a hemipteran pest of many agricultural crops in western North America. The full-length transcript for L. hesperus dsx (Lhdsx) and several variants encode proteins with conserved DNA binding and oligomerization domains. Transcript profiling revealed that Lhdsx is ubiquitously expressed, likely undergoes alternative pre-mRNA splicing, and, unlike several model insects, is sex-biased rather than sex-specific. Embryonic RNA interference (RNAi) of Lhdsx only impacted sex development in adult males, which lacked both internal reproductive organs and external genitalia. No discernible impacts on adult female development or reproductivity were observed. RNAi knockdown of Lhdsx in nymphs likewise only affected adult males, which lacked the characteristic dimorphic coloration but had dramatically elevated vitellogenin transcripts. Gene knockout of Lhdsx by CRISPR/Cas9 editing yielded only females in G0 and strongly biased heterozygous G1 offspring to females with the few surviving males showing severely impaired genital development. These results indicate that L. hesperus male development requires Lhdsx, whereas female development proceeds via a basal pathway that functions independently of dsx. A fundamental understanding of sex differentiation in L. hesperus could be important for future gene-based management strategies of this important agricultural pest.

ß-tubulin functions in spermatogenesis in Lygus hesperus Knight.

Lygus hesperus Knight is an important insect pest of crops across western North America, with field management heavily reliant on the use of chemical insecticides. Because of the evolution of resistance to these insecticides, effective and environmentally benign pest management strategies are needed. Traditional sterile insect technique (SIT) has been successfully employed to manage or eradicate some insect pests but involves introducing irradiated insects with random mutations into field populations. New genetically-driven SIT techniques are a safer alternative, causing fixed mutations that manipulate individual genes in target pests to produce sterile individuals for release. Here, we identified seven ß-tubulin coding genes from L. hesperus and show that Lhßtub2 is critical in male sperm production and fertility. Lhßtub2 is expressed primarily in the male testes and targeting of this gene by RNA interference or gene editing leads to male sterility.

Impact of Bemisia tabaci MEAM1 (Hemiptera: Aleyrodidae) on Soybean Yield and Quality Under Field Conditions.

Bemisia tabaci MEAM1 (Hemiptera: Aleyrodidae) is a key insect pest in soybean fields in Brazil but data are lacking on the relationship between pest abundance and crop yield and quality. Controlled infestation studies were conducted on caged soybean plants in the field over a two year period at two sites in Brazil. Differences in temperature in the two years affected population growth of B. tabaci, reaching 413 nymphs per leaflet in the first year, and 179 the second year even when the average temperature was 3ºC higher. Higher temperatures promoted a shorter lifecycle and nearly one more generation. Yield was affected with losses up to 500 kg/ha in 2017/2018 and 1,147 kg/ha in 2018/2019. A decrease in the weight of a thousand grains of 18 and 33 g was observed in the first and second year, respectively. No significant differences were observed in grain germination, but estimated losses in protein content were up to 440 kg/ha at the highest infestation level. Pest density and yield data were used to estimate economic injury levels (EILs). EILs ranged from 2.5 to 25.67 nymphs per leaflet and 0.17-1.79 adults per leaflet over a range of control costs, soybean production values, and control efficacies. These results should provide data useful toward development of pest management decision making tools.

Correlation-Based Network Analysis of the Influence of Bemisia tabaci Feeding on Photosynthesis and Foliar Sugar and Starch Composition in Soybean.

Bemisia tabaci (MEAM1) represents a species of economic importance in soybean. One of the obstacles to the management of B. tabaci is the quantification of damage by the pest because damage is indirectly inferred through losses in productivity. The objective of this study was to characterize the influence of B. tabaci feeding on soybean by assessing effects on photosynthetic parameters and the sugar and starch content of soybean leaves. The goal was to identify the optimal parameter to directly quantify pest damage on crop yield. Correlation networks were created among data on sugar content (fructose, glucose, and sucrose), starch and photosynthetic parameters (initial fluorescence, performance index on absorption basis, and turn-over number), and the number of nymphs at each of three infestations level (low, medium, and high) during both the vegetative and reproductive stage of the crop. In general, nymphs were more abundant during the vegetative stage. Starch content was strongly correlated with nymph density. A strong positive correlation was observed between fructose and nymph density during the vegetative stage. Among the photosynthetic parameters, the turn-over number N was positively correlated with nymph density at a low-infestation level and negatively correlated with nymphs when they occurred at a high-infestation level. B. tabaci feeding affected the plant's physiology and its interaction is reflected in part by the relationships among photosynthetic parameters as well as the levels of sugars and starch. This understanding might be useful in developing better monitoring tools for pest management.

Monitoring Bemisia tabaci (Gennadius) (Hemiptera: Aleyrodidae) Infestation in Soybean by Proximal Sensing.

Although monitoring insect pest populations in the fields is essential in crop management, it is still a laborious and sometimes ineffective process. Imprecise decision-making in an integrated pest management program may lead to ineffective control in infested areas or the excessive use of insecticides. In addition, high infestation levels may diminish the photosynthetic activity of soybean, reducing their development and yield. Therefore, we proposed that levels of infested soybean areas could be identified and classified in a field using hyperspectral proximal sensing. Thus, the goals of this study were to investigate and discriminate the reflectance characteristics of soybean non-infested and infested with Bemisia tabaci using hyperspectral sensing data. Therefore, cages were placed over soybean plants in a commercial field and artificial whitefly infestations were created. Later, samples of infested and non-infested soybean leaves were collected and transported to the laboratory to obtain the hyperspectral curves. The results allowed us to discriminate the different levels of infestation and to separate healthy from whitefly infested soybean leaves based on their reflectance. In conclusion, these results show that hyperspectral sensing can potentially be used to monitor whitefly populations in soybean fields.

Toxicity of Baits and Their Effects on Population Suppression of Anastrepha fraterculus (Diptera: Tephritidae): Implications for Field Management.

Anastrepha fraterculus (Wiedemann, 1830) is the main pest of fruit in southern Brazil. The use of toxic baits is one of the alternatives for its management. In this study, the toxic baits Anamed + malathion (10,000 mg/liter), Flyral 1.25% + malathion (2,000 mg/liter), and Gelsura (alpha-cypermethrin, 2,000 and 4,000 mg/liter) were highly toxic to the adults of A. fraterculus (lethal time [LT50] < 7 h). In contrast, Success 0.02 CB had an LT50 of 48.4 h. In the absence of rain, all the formulations had residual effects (>90% mortality) on A. fraterculus adults up to 21 d after treatment (DAT). In the presence of 5, 25, and 50 mm of rainfall, there was a significant reduction in the residual effect over time. However, with up to 50 mm of rain, Anamed + malathion and Gelsura 2,000 and 4,000 mg/liter caused between 43.0 and 79.0% of mortality. In the field, during two consecutive seasons (2015/2016 and 2016/2017), applications of Gelsura 2,000 mg/liter (four applications/season) caused population suppression of the pest throughout the apple fruiting period. However, in the 2016/2017 season, in the area using Gelsura, a higher percentage (≈12%) of apple fruits damaged by A. fraterculus females was observed when compared with the area with insecticide application (damage <3%). The toxic bait Gelsura (2,000 and 4.000 mg/liter) was shown to be promising for use in the management of A. fraterculus, with results similar to those with the application of synthetic insecticides.