Sublethal effects of a commercial Bt product and Bt cotton flowers on the bollworm (Helicoverpa zea) with impacts to predation from a lady beetle (Hippodamia convergens).

Insecticidal Bacillus thuringiensis Berliner (Bt) toxins produced by transgenic cotton (Gossypium hirsutum L.) plants have become an essential component of cotton pest management. Bt toxins are the primary management tool in transgenic cotton for lepidopteran pests, the most important of which is the bollworm (Helicoverpa zea Boddie) (Lepidoptera: Noctuidae) in the United States (U.S.). However, bollworm larvae that survive after consuming Bt toxins may experience sublethal effects, which could alter interactions with other organisms, such as natural enemies. Experiments were conducted to evaluate how sublethal effects of a commercial Bt product (Dipel) incorporated into artificial diet and from Bt cotton flowers impact predation from the convergent lady beetle (Hippodamia convergens Guérin-Méneville) (Coleoptera: Coccinellidae), common in cotton fields of the mid-southern U.S. Sublethal effects were detected through reduced weight and slower development in bollworm larvae which fed on Dipel incorporated into artificial diet, Bollgard II, and Bollgard 3 cotton flowers. Sublethal effects from proteins incorporated into artificial diet were found to significantly alter predation from third instar lady beetle larvae. Predation of bollworm larvae also increased significantly after feeding for three days on a diet incorporated with Bt proteins. These results suggest that the changes in larval weight and development induced by Bt can be used to help predict consumption of bollworm larvae by the convergent lady beetle. These findings are essential to understanding the potential level of biological control in Bt cotton where lepidopteran larvae experience sublethal effects.

Cotton plants overexpressing the Bacillus thuringiensis Cry23Aa and Cry37Aa binary-like toxins exhibit high resistance to the cotton boll weevil (Anthonomus grandis).

The cotton boll weevil (CBW, Anthonomus grandis) stands as one of the most significant threats to cotton crops (Gossypium hirsutum). Despite substantial efforts, the development of a commercially viable transgenic cotton event for effective open-field control of CBW has remained elusive. This study describes a detailed characterization of the insecticidal toxins Cry23Aa and Cry37Aa against CBW. Our findings reveal that CBW larvae fed on artificial diets supplemented exclusively with Cry23Aa decreased larval survival by roughly by 69%, while supplementation with Cry37Aa alone displayed no statistical difference compared to the control. However, the combined provision of both toxins in the artificial diet led to mortality rates approaching 100% among CBW larvae (LC50 equal to 0.26 PPM). Additionally, we engineered transgenic cotton plants by introducing cry23Aa and cry37Aa genes under control of the flower bud-specific pGhFS4 and pGhFS1 promoters, respectively. Seven transgenic cotton events expressing high levels of Cry23Aa and Cry37Aa toxins in flower buds were selected for greenhouse bioassays, and the mortality rate of CBW larvae feeding on their T0 and T1 generations ranged from 75% to 100%. Our in silico analyses unveiled that Cry23Aa displays all the hallmark characteristics of ß-pore-forming toxins (ß-PFTs) that bind to sugar moieties in glycoproteins. Intriguingly, we also discovered a distinctive zinc-binding site within Cry23Aa, which appears to be involved in protein-protein interactions. Finally, we discuss the major structural features of Cry23Aa that likely play a role in the toxin's mechanism of action. In view of the low LC50 for CBW larvae and the significant accumulation of these toxins in the flower buds of both T0 and T1 plants, we anticipate that through successive generations of these transgenic lines, cotton plants engineered to overexpress cry23Aa and cry37Aa hold promise for effectively managing CBW infestations in cotton crops.

Identification of Crucial Modules and Genes Associated with Bt Gene Expression in Cotton.

The expression of Bacillus thuringiensis (Bt) toxins in transgenic cotton confers resistance to insect pests. However, it has been demonstrated that its effectiveness varies among cotton cultivars and different tissues. In this study, we evaluated the expression of Bt protein in 28 cotton cultivars and selected 7 cultivars that differed in Bt protein expression for transcriptome analysis. Based on their Bt protein expression levels, the selected cultivars were categorized into three groups: H (high Bt protein expression), M (moderate expression), and L (low expression). In total, 342, 318, and 965 differentially expressed genes were detected in the H vs. L, M vs. L, and H vs. M comparison groups, respectively. And three modules significantly associated with Bt protein expression were identified by weighted gene co-expression network analysis. Three hub genes were selected to verify their relationships with Bt protein expression using virus-induced gene silencing (VIGS). Silencing GhM_D11G1176, encoding an MYC transcription factor, was confirmed to significantly decrease the expression of Bt protein. The present findings contribute to an improved understanding of the mechanisms that influence Bt protein expression in transgenic cotton.

iJAZ-based approach to engineer lepidopteran pest resistance in multiple crop species.

The fall armyworm (FAW) poses a significant threat to global crop production. Here we showed that overexpression of jasmonate ZIM-domain (JAZ) protein GhJAZ24 confers resistance to cotton bollworm and FAW, while also causing sterility in transgenic cotton by recruiting TOPLESS and histone deacetylase 6. We identified the NGR motif of GhJAZ24 that recognizes and binds the aminopeptidase N receptor, enabling GhJAZ24 to enter cells and disrupt histone deacetylase 3, leading to cell death. To overcome plant sterility associated with GhJAZ24 overexpression, we developed iJAZ (i, induced), an approach involving damage-induced expression and a switch from intracellular to extracellular localization of GhJAZ24. iJAZ transgenic cotton maintained fertility and showed insecticidal activity against cotton bollworm and FAW. In addition, iJAZ transgenic rice, maize and tobacco plants showed insecticidal activity against their lepidopteran pests, resulting in an iJAZ-based approach for generating alternative insecticidal proteins with distinctive mechanisms of action, thus holding immense potential for future crop engineering.

Rapid detection of multi-scale cotton pests based on lightweight GBW-YOLOv5 model.

BACKGROUND: Pest infestation is one of the primary causes of decreased cotton yield and quality. Rapid and accurate identification of cotton pest categories is essential for producers to implement effective and expeditious control measures. Existing multi-scale cotton pest detection technology still suffers from poor accuracy and rapidity of detection. This study proposed the pruned GBW-YOLOv5 (Ghost-BiFPN-WIoU You Only Look Once version 5), a novel model for the rapid detection of cotton pests. RESULTS: The detection performance of the pruned GBW-YOLOv5 model for cotton pests was evaluated based on the self-built cotton pest dataset. In comparison with the original YOLOv5 model, the pruned GBW-YOLOv5 model demonstrated significant reductions in complexity, size, and parameters by 68.4%, 66.7%, and 68.2%, respectively. Remarkably, the mean average precision (mAP) decreased by a mere 3.8%. The pruned GBW-YOLOv5 model outperformed other classic object detection models, achieving an outstanding detection speed of 114.9 FPS. CONCLUSION: The methodology proposed by our research enabled rapid and accurate identification of cotton pests, laying a solid foundation for the implementation of precise pest control measures. The pruned GBW-YOLOv5 model provided theoretical research and technical support for detecting cotton pests under field conditions. © 2024 Society of Chemical Industry.

Interaction of Fusarium Wilt Race 4 with Root-Knot Nematode Increases Disease Severity in Cotton.

Fusarium wilt, caused by Fusarium oxysporum f. sp. vasinfectum, is a severe disease of cotton (Gossypium spp.). Strains of the wilt pathogen in the United States, such as race 1, require the presence of nematodes such as southern root-knot nematode (Meloidogyne incognita) to cause appreciable disease. The exception is the race 4 strain of the wilt pathogen, which can attack cotton without concomitant infection by plant-parasitic nematodes and was first identified in California in 2001 and in Texas and New Mexico since 2017. The effects of the interaction between M. incognita and race 1 or race 4 on wilt severity and nematode reproduction on two Gossypium hirsutum cultivars, Acala 44 and FM 966, and a G. barbadense cultivar, Pima S-4, were directly compared in growth chamber assays. All three cultivars were susceptible to M. incognita. Suppression of nematode reproduction by the wilt pathogen was detected only for race 4 on all three cultivars on a per plant basis but not on a per gram root tissue basis. The control, M. incognita alone, and race 1 alone treatments caused no symptoms. Inoculation with race 1 and M. incognita caused moderate wilt symptoms in 'Acala 44' and 'FM 966' and mild symptoms in 'Pima S-4'. However, race 4 treatment caused severe wilt in 'Pima S-4' and moderate wilt severity in 'Acala 44' and 'FM 966'. The symptom severity of 'Acala 44' and 'FM 966' further increased in the presence of M. incognita. Thus, race 4 is not only capable of causing wilt in the absence of M. incognita but can also interact with the nematode to further increase disease severity. Though control of wilt caused by race 1 can be achieved mainly through breeding for nematode resistance, it will be imperative to incorporate both southern root-knot nematode and race 4 resistance to effectively control the disease should race 4 expand into southern root-knot nematode-infested fields.

Mutation in the Cadherin Gene Is a Key Factor for Pink Bollworm Resistance to Bt Cotton in China.

Transgenic crops producing Bacillus thuringiensis (Bt) toxins are widely planted for insect control, but their efficacy may decrease as insects evolve resistance. Understanding the genetic basis of insect resistance is essential for developing an integrated strategy of resistance management. To understand the genetic basis of resistance in pink bollworm (Pectinophora gossypiella) to Bt cotton in the Yangtze River Valley of China, we conducted an F2 screening for alleles associated with resistance to the Bt (Cry1Ac) protein for the first time. A total of 145 valid single-paired lines were screened, among which seven lines were found to carry resistance alleles. All field parents in those seven lines carried recessive resistance alleles at the cadherin locus, including three known alleles, r1, r13 and r15, and two novel alleles, r19 and r20. The overall frequency of resistance alleles in 145 lines was 0.0241 (95% CI: 0.0106-0.0512). These results demonstrated that resistance was rare and that recessive mutation in the cadherin gene was the primary mechanism of pink bollworm resistance to Bt cotton in the Yangtze River Valley of China, which will provide a scientific basis for implementing targeted resistance management statics of pink bollworm in this region.

Plant age, crop stage and surrounding habitats: their impact on sucking pests and predators complex in cotton (Gossypium hirsutum L) field plots in arid climate at district Layyah, Punjab, Pakistan / Idade da planta, estágio da cultura e habitats circundantes: seu impacto no complexo de pragas sugadoras e predadores em parcelas de algodão (Gossypium hirsutum L) em clima árido no distrito de Layyah, Punjab, Paquistão

Sucking pests are major threat to cotton field crop which cause unbearable losses to the crop yield. Aim of the current study was to record seasonal dynamics of major sucking insect pests including whitefly, jassid, thrips and their natural arthropod predators i.e. green lacewings and spiders in cotton field plots. The effects of surrounding field crops on pests' density and predatory efficiency of predators were also recorded. For sampling and survey of insects, the visual counting was found to be the most efficient method for recording the abundance of insects, trailed by net sweeping and tapping. Whitefly was the most dominant sucking pest found on the vegetative stage of cotton, followed by jassid and thrips. Fluctuated populations of predatory arthropods, spiders and green lacewings were also recorded during whole cropping season however, the densities of pests and predators varied with crop phenology. Spiders' population was encouraging at both vegetative and flowering stage and also the same trend of jassid and whitefly were observed at both stages of the crop. Surrounding habitats showed non-significant effect on population densities of insect pests and predators. For abiotic factors, the spiders showed strong positive correlation with humidity and temperature. However, green lacewing was only positively correlated with humidity. On the other hand, the populations of whitefly, jassid and thrips showed non-significant correlation with both temperature and humidity. Overall densities of sucking insect pests were found above economic threshold level. The plant age, crop stage and surrounding habitats effect on the population fluctuation of pests as well as the predators' abundance. The future studies are also warranted to investigate the altered habitats and multiple trap cropping to find out their impact on unattended insect predators and parasitoids in cotton crop.

As pragas sugadoras são uma grande ameaça para a cultura do algodão, causando perdas insuportáveis no rendimento da cultura. O objetivo do estudo atual foi registrar a dinâmica sazonal das principais pragas de insetos sugadores, incluindo mosca-branca, jassid, tripes e seus artrópodes predadores naturais, ou seja, crisopídeos e aranhas verdes em parcelas de algodão. Os efeitos das plantações circundantes na densidade de pragas e na eficiência predatória de predadores também foram registrados. Para amostragem e pesquisa de insetos, a contagem visual foi considerada o método mais eficiente para registrar a abundância de insetos, seguido por varredura e batida de rede. A mosca-branca foi a praga sugadora mais dominante encontrada na fase vegetativa do algodoeiro, seguida pelo jassid e tripes. Populações flutuantes de artrópodes predadores, aranhas e crisálidas também foram registradas durante toda a safra, no entanto as densidades de pragas e predadores variaram com a fenologia da cultura. A população de aranhas foi encorajadora tanto na fase vegetativa como na floração e também a mesma tendência de jassid e mosca-branca foi observada em ambas as fases da cultura. Os habitats circundantes mostraram efeito não significativo nas densidades populacionais de insetos-praga e predadores. Para os fatores abióticos, as aranhas apresentaram forte correlação positiva com umidade e temperatura. No entanto, lacewing verde foi apenas positivamente correlacionado com a umidade. Por outro lado, as populações de mosca-branca, jassid e tripes apresentaram correlação não significativa com temperatura e umidade. As densidades gerais de pragas sugadoras de insetos foram encontradas acima do nível do limiar econômico. A idade da planta, o estágio da cultura e os habitats circundantes afetam a flutuação populacional de pragas, bem como a abundância de predadores. Os estudos futuros também são necessários para investigar os habitats alterados e cultivo com armadilhas múltiplas [...].

Path analysis based on genetic association of yield components and insects pest in upland cotton varieties.

Gossypium hirsutum L. is also called upland cotton or Mexican cotton. It is the most widely cultivated species of cotton in the whole world. Globally, about 90% of all cotton production comes from the cultivars derived from this species. Some genetic parameters like monopodial branches per plant, sympodial branches per plant, sympodial branch length, bolls per plant, boll weight, sympo-boll distance, Ginning Out Turn%, staple length (rg = 0.9199**), and fiber strength along with seed cotton yield were evaluated for their potential utilization via selection in seed cotton yield improvement. Significant positive genetic correlations were estimated for monopodial branches per plant (rg = 0.9722**), sympodial branches per plant (rg = 0.7098**), sympodial branch length (rg = 0.617**), bolls per plant (rg = 0.8271**), boll weight (rg = 0.8065**), sympo-boll distance (rg = 0.6507**), Ginning Out Turn (GOT)% (rg = 0.7541**), staple length (rg = 0.9199**), and fiber strength (rg = 0.7534**) with seed cotton yield. A path analysis of all the yield traits under study revealed strong positive direct effects of monopodial branch length (1.1556), sympo-boll distance (0.8173) and staple length (0.7633), while plant height exerted a highly strong direct negative effect (-1.2096) on yield. It is concluded that a direct selection based on monopodial branch length and sympo-boll distance, and staple length is effective, whereas, monopodial branch length, and sympodial branch length are good selection indicators via bolls per plant for yield improvement in cotton.

Foliar herbivory increases sucrose concentration in bracteal extrafloral nectar of cotton.

Cultivated cotton, such as Gossypium hirsutum L., produces extrafloral (EF) nectar on leaves (foliar) and reproductive structures (bracteal) as an indirect anti-herbivore defense. In exchange for this carbohydrate-rich substance, predatory insects such as ants protect the plant against herbivorous insects. Some EF nectar-bearing plants respond to herbivory by increasing EF nectar production. For instance, herbivore-free G. hirsutum produces more bracteal than foliar EF nectar, but increases its foliar EF nectar production in response to herbivory. This study is the first to test for systemically induced changes to the carbohydrate composition of bracteal EF nectar in response to foliar herbivory on G. hirsutum. We found that foliar herbivory significantly increased the sucrose content of bracteal EF nectar while glucose and fructose remained unchanged. Sucrose content is known to influence ant foraging behavior and previous studies of an herbivore-induced increase to EF nectar caloric content found that it led to increased ant activity on the plant. As a follow-up to our finding, ant recruitment to mock EF nectar solutions that varied in sucrose content was tested in the field. The ants did not exhibit any preference for either solution, potentially because sucrose is a minor carbohydrate component in G. hirsutum EF nectar: total sugar content was not significantly affected by the increase in sucrose. Nonetheless, our findings raise new questions about cotton's inducible EF nectar responses to herbivory. Further research is needed to determine whether an herbivore-induced increase in sucrose content is typical of Gossypium spp., and whether it constitutes a corollary of systemic sucrose induction, or a potentially adaptive mechanism which enhances ant attraction to the plant.

Engineering tolerance to CLCuD in transgenic Gossypium hirsutum cv. HS6 expressing Cotton leaf curl Multan virus-C4 intron hairpin.

Cotton leaf curl disease (CLCuD), caused by begomoviruses in combination with betasatellite molecule, has adversely affected cotton industry of Indian subcontinent. To devise a CLCuD-control strategy, RNAi-mediated approach was followed in this study. Gossypium hirsutum cv. HS6 plants were transformed with intron-hairpin RNAi (ihpRNAi-C4) construct carrying silencing suppressor C4 gene of Cotton leaf curl Multan virus (CLCuMuV). Efficacy of the construct in imparting CLCuD resistance was evaluated in transgenic (T0, T1) cotton lines. Accumulation of CLCuMuV/betasatellite and attenuation of CLCuD symptoms in the transgenic lines were monitored at different times interval after virus inoculation. Northern hybridization revealed the expression of C4-gene derived siRNA. Expression of the ihpRNAi transcript was recorded higher in transgenic lines expressing siRNA which supposedly targeted the C4 gene. A significant delay in detection of virus as well as betasatellite was observed in the transgenic lines. At 30 days post inoculation (dpi), none of the lines tested positive. At 45 dpi, however, it could be detected in few lines having much lower titre as compared to non-transformed control plants. Notably, till 60 dpi, no significant progression of the virus/betasatellite DNA was observed and the plants did not exhibit any characteristic CLCuD symptoms. A tolerance phenomenon leading to escape of CLCuD symptoms in the transformed cotton was described.

Field efficacy of Bt cotton containing events DAS-21023-5 × DAS-24236-5 × SYN-IR102-7 against lepidopteran pests and impact on the non-target arthropod community in Brazil.

The efficacy and non-target arthropod effects of transgenic DAS-21023-5 × DAS-24236-5 × SYN-IR102-7 Bt cotton, expressing proteins Cry1Ac, Cry1F and Vip3Aa19, was examined through field trials in Brazil. Fifteen field efficacy experiments were conducted from 2014 through the 2020 growing season across six different states in Brazil to evaluate performance against key lepidopteran pests through artificial infestations of Chrysodeixis includens (Walker), Spodoptera frugiperda (J.E. Smith,1797), Spodoptera cosmioides (Walker, 1858) and Chloridea virescens (F., 1781), and natural infestations of Alabama argillacea (Hübner) and S. frugiperda. The impact of this Bt cotton technology on the non-target arthropod community in Brazilian cotton production systems was also assessed in a multi-site experiment. DAS-21023-5 × DAS-24236-5 × SYN-IR102-7 cotton significantly reduced the feeding damage caused by S. frugiperda, S. cosmioides, C. includens, C. virescens and A. argillacea, causing high levels of mortality (greater than 99%) to all target lepidopteran pests evaluated during vegetative and/or reproductive stages of crop development. Non-target arthropod community-level analyses confirmed no unintended effects on the arthropod groups monitored. These results demonstrate the value of transgenic Bt cotton containing event DAS-21023-5 × DAS-24236-5 × SYN-IR102-7 for consideration as part of an integrated approach for managing key lepidopteran pests in Brazilian cotton production systems.

CRISPR-mediated mutations in the ABC transporter gene ABCA2 confer pink bollworm resistance to Bt toxin Cry2Ab.

Crops genetically engineered to produce insecticidal proteins from Bacillus thuringiensis (Bt) have many benefits and are important globally for managing insect pests. However, the evolution of pest resistance to Bt crops reduces their benefits. Understanding the genetic basis of such resistance is needed to better monitor, manage, and counter pest resistance to Bt crops. Previous work shows that resistance to Bt toxin Cry2Ab is associated with mutations in the gene encoding the ATP-binding cassette protein ABCA2 in lab- and field-selected populations of the pink bollworm (Pectinophora gossypiella), one of the world's most destructive pests of cotton. Here we used CRISPR/Cas9 gene editing to test the hypothesis that mutations in the pink bollworm gene encoding ABCA2 (PgABCA2) can cause resistance to Cry2Ab. Consistent with this hypothesis, introduction of disruptive mutations in PgABCA2 in a susceptible strain of pink bollworm increased the frequency of resistance to Cry2Ab and facilitated creation of a Cry2Ab-resistant strain. All Cry2Ab-resistant individuals tested in this study had disruptive mutations in PgABCA2. Overall, we found 17 different disruptive mutations in PgABCA2 gDNA and 26 in PgABCA2 cDNA, including novel mutations corresponding precisely to single-guide (sgRNA) sites used for CRISPR/Cas9. Together with previous results, these findings provide the first case of practical resistance to Cry2Ab where evidence identifies a specific gene in which disruptive mutations can cause resistance and are associated with resistance in field-selected populations.

Functional Analysis of Alkaline Phosphatase in Whitefly Bemisia tabaci (Middle East Asia Minor 1 and Mediterranean) on Different Host Plants.

Alkaline phosphatases (ALPs: EC 3.1.3.1) are ubiquitous enzymes and play crucial roles in the fundamental phosphate uptake and secretory processes. Although insects are regarded as the most diverse group of organisms, the current understanding of ALP roles in insects is limited. As one type of destructive agricultural pest, whitefly Bemisia tabaci, a phloem feeder and invasive species, can cause extensive crop damage through feeding and transmitting plant diseases. In this study, we retrieved five ALP genes in MEAM1 whitefly, nine ALP genes in MED whitefly via comparative genomics approaches. Compared with nine other insects, whiteflies' ALP gene family members did not undergo significant expansion during insect evolution, and whiteflies' ALP genes were dispersed. Moreover, whiteflies' ALP gene family was conserved among insects and emerged before speciation via phylogenetic analysis. Whiteflies' ALP gene expression profiles presented that most ALP genes have different expression patterns after feeding on cotton or tobacco plants. Female/male MED whiteflies possessed higher ALP activities on both cotton and tobacco plants irrespective of sex, relative to MEAM1 whiteflies. Meanwhile, adult MED whiteflies possessed higher ALP activity in both whole insect and salivary samples, relative to MEAM1 whiteflies. We also found that both MED and MEAM1 whiteflies could upregulate ALP activities after feeding on cotton compared with feeding on tobacco plants. These findings demonstrated the functions of whiteflies ALPs and will assist the further study of the genomic evolution of insect ALPs.

Electrical signalling on Bt and non-Bt cotton plants under stress by Aphis gossypii.

Plants have developed various mechanisms to respond specifically to each biotrophic attack. It has been shown that the electrical signals emitted by plants are associated with herbivory stress responses and can lead to the activation of multiple defences. Bt cotton is a genetically modified pest-resistant plant that produces an insecticide from Bacillus thuringiensis (Bt) to control Lepidopteran species. Surprisingly, there is no study-yet, that characterizes the signalling mechanisms in transgenic cotton plants attacked by non-target insects, such as aphids. In this study, we characterized the production of electrical signals on Bt and non-Bt cotton plants infested with Aphis gossypii and, in addition, we characterized the dispersal behaviour of aphids to correlate this behaviour to plant signalling responses. Electrical signalling of the plants was recorded with an extracellular measurement technique. Impressively, our results showed that both Bt and non-Bt cotton varieties, when attacked by A. gossypii, emitted potential variation-type electrical signals and clearly showed the presence of distinct responses regarding their perception and the behaviour of aphids, with evidence of delay, in terms of signal amount, and almost twice the amount of Cry1F protein was observed on Bt cotton plants at the highest density of insects/plant. We present in our article some hypotheses that are based on plant physiology and insect behaviour to explain the responses found on Bt cotton plants under aphid stress.

Prey removal in cotton crops next to woodland reveals periodic diurnal and nocturnal invertebrate predation gradients from the crop edge by birds and bats.

Factors influencing the efficacy of insectivorous vertebrates in providing natural pest control services inside crops at increasing distances from the crop edge are poorly understood. We investigated the identity of vertebrate predators (birds and bats) and removal of sentinel prey (mealworms and beetles) from experimental feeding trays in cotton crops using prey removal trials, camera traps and observations. More prey was removed during the day than at night, but prey removal was variable at the crop edge and dependent on the month (reflecting crop growth and cover) and time of day. Overall, the predation of mealworms and beetles was 1-times and 13-times greater during the day than night, respectively, with predation on mealworms 3-5 times greater during the day than night at the crop edge compared to 95 m inside the crop. Camera traps identified many insectivorous birds and bats over crops near the feeding trays, but there was no evidence of bats or small passerines removing experimental prey. A predation gradient from the crop edge was evident, but only in some months. This corresponded to the foraging preferences of open-space generalist predators (magpies) in low crop cover versus the shrubby habitat preferred by small passerines, likely facilitating foraging away from the crop edge later in the season. Our results are in line with Optimal Foraging Theory and suggest that predators trade-off foraging behaviour with predation risk at different distances from the crop edge and levels of crop cover. Understanding the optimal farm configuration to support insectivorous bird and bat populations can assist farmers to make informed decisions regarding in-crop natural pest control and maximise the predation services provided by farm biodiversity.

Insecticidal activity of some synthesized 1,3,4-oxadiazole derivatives grafted on chitosan and polymethylmethacrylate against the cotton leafworm Spodoptera littoralis.

To combat insect pests and vectors that are responsible for high losses in food and lives, insecticide discovery is of top priority. This study aimed to synthesize, characterize and investigate the insecticidal activity of 1,3,4-oxadiazole derivatives grafted on chitosan (CS) and modified polymethyl methacrylate (PMMA). 5-(pyridin-3-yl)-1,3,4-oxadiazole-2-thiol and 5-(pyridin-4-yl)-1,3,4-oxadiazole-2-thiol were respectively reacted with ethylchloroacetate and methyl-2-choloroacetoacetate. The resulted esters were grafted with CS and modified-PMMA. The products were characterized using FT-IR, 1H NMR, TGA, and XRD techniques. Four CS grafted ones were able to show good insecticidal activity against the cotton leafworm Spodoptera littoralis. Furthermore, the safety of these compounds was tested using MTT assay on a human cell line (WI-38). The results indicated that compounds 2a, 2b, 6a, and 6d are considered insecticide candidates to S. littoralis fourth-instar larvae. Cytotoxicity of 2b and 6d indicated that they are the least toxic to humans. It is concluded that both compounds may represent promising insecticide candidates.

Degree day-based model predicts pink bollworm phenology across geographical locations of subtropics and semi-arid tropics of India.

There is a global concern about the effects of climate change driven shifts in species phenology on crop pests. Using geographically and temporally extensive data set of moth trap catches and temperatures across the cotton growing states of India, we predicted the phenology of cotton pink bollworm Pectinophora gossypiella (Saunders). Our approach was centered on growing degree days (GDD), a measure of thermal accumulation that provides a mechanistic link between climate change and species' phenology. The phenology change was predicted by calculating absolute error associated with DD and ordinal date, an alternative predictor of phenology, for peak moth abundance. Our results show that GDD outperformed the ordinal dates in predicting peak moth abundance in 6 out of 10 selected locations. Using established thresholds of 13.0/34.0 °C, mean DD accumulated between the consecutive moth peaks across different years were estimated at 504.05 ± 4.84. Seven generations were determined for pink bollworm in a cropping season, the length of which varied between 35 and 73 days in response to temperature. Pink bollworm population reached its peak during third generation which can be the target for management actions. The study provides essential information for developing pink bollworm management strategies under climate change.

Efficacy of biorational insecticides against Bemisia tabaci (Genn.) and their selectivity for its parasitoid Encarsia formosa Gahan on Bt cotton.

The toxicity of seven biorational insecticides [five insect growth regulators (Buprofezin, Fenoxycarb, Pyriproxyfen, Methoxyfenozide, and Tebufenozide) and two oil-extracts of neem and bitter gourd seeds] against Bemisia tabaci and their selectivity for its parasitoid, Encarsia formosa were evaluated in laboratory and field conditions for 2 years (2018-2019) in Pakistan. Toxicity results demonstrate that Pyriproxyfen, Buprofezin, and Fenoxycarb proved to be effective (80-91% mortality and 66.3-84.2% population-reduction) against B. tabaci followed by Methoxyfenozide, Tebufenozide (50-75% mortality and 47.8-52.4% population-reduction), and then oil-extracts of neem and bitter gourd (25-50% mortality and 36.5-39.8% population-reduction) in the laboratory [72 h post-application exposure interval (PAEI)] and field trails (168 h PAEI), respectively. All tested biorationals, except Methoxyfenozide [(slightly-harmful/Class-II), i.e., causing mortality of parasitoids between a range of 25-50%] and Tebufenozide [(moderately-harmful/Class-III), i.e., causing mortality of parasitoids between the ranges of 51-75%], proved harmless/Class-I biorationals at PAEI of 7-days in the field (parasitism-reduction < 25%) and 3-days in the lab (effect < 30%). In laboratory bioassays, exposure of parasitized-pseudopupae and adult-parasitoids to neem and bitter gourd oils demonstrated that these compounds proved harmless/Class-I biorationals (< 30% mortality). Alternatively, Pyriproxyfen, Buprofezin, Fenoxycarb, Methoxyfenozide, and Tebufenozide were slightly-harmful biorationals (30-79% mortality) against the respective stages of E. formosa. We conclude that most of the tested biorationals proved harmless or slightly harmful to E. formosa, except tebufenozide after PAEI of 7-days (168 h) in the field and, therefore, may be used strategically in Integrated Pest Management (IPM) of B. tabaci.

Silencing of a LIM gene in cotton exhibits enhanced resistance against Apolygus lucorum.

Plant bugs (Miridae species) have become major agricultural pests that cause increasing and severe economic damage. Plant-mediated RNA interference (RNAi) is emerging as an eco-friendly, efficient, and reliable strategy for pest management. In this study, we isolated and characterized a lethal gene of Apolygus lucorum and named it Apolygus lucorum LIM (AlLIM), which produced A. lucorum mortality rates ranging from 38% to 81%. Downregulation of the AlLIM gene expression in A. lucorum by injection of a double-stranded RNA (dsRNA) led to muscle structural disorganization that resulted in metamorphosis deficiency and increased mortality. Then we constructed a plant expression vector that enabled transgenic cotton to highly and stably express dsRNA of AlLIM (dsAlLIM) by Agrobacterium-mediated genetic transformation. In the field bioassay, dsAlLIM transgenic cotton was protected from A. lucorum damage with high efficiency, with almost no detectable yield loss. Therefore, our study successfully provides a promising genetically modified strategy to overpower A. lucorum attack.