Insect Resistance Research Articles

CgSSR marker based trait associations for Sitotroga cerealella (Olivier) resistance in stored paddy rice

Rice is stored for a longer time and many primary and secondary insect pests attack grains, causing both quantitative and qualitative losses. Physicochemical characteristics of grain influence stored grain insect susceptibility and resistance. Identification of gene/Quantitative Trait Loci (QTLs) and introgression in the cultivars would provide the desired solution for varietal resistance against S. cerealella. A group of 80 rice varieties was used for genetic analysis for the identification of candidate genes responsible for Sitotroga cerealella resistance. Heat maps derived from values in the kinship matrix were used to group the varieties into three subpopulations. The largest subpopulation has 34 varieties, while the lowest had 19. The first three principal components (PCs) explained 16, 9, and 6 percent of total variation, respectively, as indicated by the Principal Component Analysis (PCA). The PCA results complemented the heat map results in defining population structure. Genome-wide association mapping can detect marker-trait associations/QTLs in a heterogeneous population of breeding lines, despite population structure and unequal family relatedness. As shown by the Genomic Association and Prediction Integrated Tool (GAPIT) heat map, there are three subpopulations in the association mapping panel. Two cgSSR markers, related to gene SDG725 and FLO2, were linked to the attributes and responsible for conferring resistance against S. cerealella infestation. This is the first report of association and that would help in the development of cultivar with S. cerealella resistance.

Temperature influences desiccation resistance of bumble bees

Ongoing climate change has increased temperatures and the frequency of droughts in many parts of the world, potentially intensifying the desiccation risk for insects. Because resisting desiccation becomes more difficult at higher temperatures and lower humidity, avoiding water loss is a key challenge facing terrestrial insects. However, few studies have examined the interactive effects of temperature and environmental humidity on desiccation resistance in insects. Such studies on bees (Hymenoptera: Apoidea: Anthophila) are especially rare, despite their ecological and economic importance. Here, we crossed temperature (20, 25, and 30 °C) with humidity (<5, 50, >95 % RH) manipulations and measured time to mortality, water loss rates, and the water content at mortality of bumble bees (Bombus impatiens). We found that both higher temperature and lower humidity increased water loss rates, while warmer temperatures reduced survival time and lower humidity decreased water content at mortality. Additionally, we observed large intraspecific variation in water balance traits between colonies, and larger individuals survived longer and could tolerate more water loss before mortality. This study raises important questions about the mechanisms underpinning water loss in bumble bees and suggests that frequent access to nectar may be especially important for bumble bees’ water balance and survival in a warming and drying climate.

Nicotinamide deficiency promotes imidacloprid resistance via activation of ROS/CncC signaling pathway-mediated UGT detoxification in Nilaparvata lugens

Metabolic alternation is a typical characteristic of insecticide resistance in insects. However, mechanisms underlying metabolic alternation and how altered metabolism in turn affects insecticide resistance are largely unknown. Here, we report that nicotinamide levels are decreased in the imidacloprid-resistant strain of Nilaparvata lugens, may due to reduced abundance of the symbiotic bacteria Arsenophonus. Importantly, the low levels of nicotinamide promote imidacloprid resistance via metabolic detoxification alternation, including elevations in UDP-glycosyltransferase enzymatic activity and enhancements in UGT386B2-mediated metabolism capability. Mechanistically, nicotinamide suppresses transcriptional regulatory activities of cap ‘n’ collar isoform C (CncC) and its partner small muscle aponeurosis fibromatosis isoform K (MafK) by scavenging the reactive oxygen species (ROS) and blocking the DNA binding domain of MafK. In imidacloprid-resistant N. lugens, nicotinamide deficiency re-activates the ROS/CncC signaling pathway to provoke UGT386B2 overexpression, thereby promoting imidacloprid detoxification. Thus, nicotinamide metabolism represents a promising target to counteract imidacloprid resistance in N. lugens.

Colonization of Piriformospora indica enhances rice resistance against the brown planthopper Nilaparvata lugens.

Piriformospora indica is an endophytic fungus that can promote the growth and confer the resistance against diverse stresses in host plants by root colonization. However, the effects of P. indica colonization on improving plant resistance to insect pests are still less explored. The brown planthopper (BPH) Nilaparvata lugens is a serious monophagous pest that causes extensive damage to rice plants. Here, we aimed to evaluate the effects of P. indica colonization on rice resistance against BPH. The colonization of P. indica in the rice roots resisted the damage from BPH. The age-stage, two-sex life table analyses showed that feeding on P. indica-colonized rice plants affected BPH's female adult longevity, oviposition period, fecundity, population parameters and population size. BPH female adults feeding on P. indica-colonized plants excreted less honeydew. P. indica colonization remarkably increased the duration of np, N2, and N3 waveform, as well as the occurrences of N1 and N2, and decreased the duration of N4-b for BPH on rice plants. Meanwhile, the weight of BPH on the colonized plants was significantly lower than the control. In addition, the feeding and oviposition preferences of BPH to P. indica-colonized plants were reduced. qRT-RCR analyses revealed that P. indica colonization induced the expressions of jasmonic acid (JA)- and salicylic acid (SA)-related genes in rice plants. P. indica colonization can reduce BPH performance on rice plants with potentially inhibitory effects on population growth. Collectively, these results support the potential for endophytically colonized P. indica as an effective strategy to improve insect resistance of crops. This article is protected by copyright. All rights reserved.

Genome-wide identification of G-protein coupled receptors (GPCRs) and their expression profile in response to β-cypermethrin stress in Zeugodacus cucurbitae

G-protein coupled receptors (GPCRs) are the largest and most diverse transmembrane receptor family in the cell. They are involved in regulating a wide range of biological processes, including behavior, reproduction, and development. However, GPCRs have not yet been identified in Zeugodacus cucurbitae. The current study focuses on the GPCRs identification, classification, distribution, and their expression analysis under β-cypermethrin stress to uncover novel targets for pest management and assist in the development of effective strategies for controlling the melon fly population. We identified 80 GPCRs genes including 50 GPCRs identified in family A, 17 GPCRs identified in family B, 8 identified in family C, and 5 identified in family F. Z. cucurbitae GPCRs showed significant differences in both the number of genes in families or subfamilies, as well as the sequencing of the genes. Interestingly, newly identified GPCRs genes are expressed differently at various developmental stages of Z. cucurbitae. Further, we evaluated these 80 GPCRs using Realtime quantitative PCR to confirm their expression between β-cypermethrin-resistant (RS) strain and susceptible strain (SS) of Z. cucurbitae. We identified 50 GPCR genes were highly overexpressed in a RS. Among these genes, eight genes were strongly induced by the 30% lethal concentration (LC) while two genes were significantly increased by the 50% LC of β-cypermethrin. This first genome-wide profiling and characterization of GPCRs could lay foundation for unraveling detoxification mechanism and target site modifications which may improve the insect resistance and could be effective insecticide targets for Z. cucurbitae management.

Continental-scale migration patterns and origin of Helicoverpa zea (Lepidoptera: Noctuidae) based on a biogeochemical marker.

Insect migrations have ecological and economic impacts, particularly in agriculture. However, there is limited knowledge about the migratory movements of pests at the continental scale, which is an important factor influencing the spread of resistance genes. Understanding the migratory patterns of economic pests, like Helicoverpa zea (Boddie), is essential for improving Integrated Pest Management (IPM) and Insect Resistance Management (IRM) strategies. In this study, we used stable hydrogen isotopic ratios in wing tissue as a biogeochemical marker to examine migratory patterns and estimate the native origins of H. zea individuals collected across a wide latitudinal gradient in North America. Samples collected at higher latitudes (Ontario, Canada and Minnesota, USA) exhibited a greater proportion (60%-96%) of nonlocal individuals, with an increased probability of origin from the southeastern United States. Populations from mid-latitudes (Florida, North Carolina, and South Carolina) showed a blend of local and nonlocal (40%-60%) individuals. Finally, 15% of the southernmost population individuals (Puerto Rico) were classified as migratory, with some having a probability of origin at higher latitudes (>30°). Overall, our results provide evidence of a northward spring/summer migration of H. zea in North America and underscore the significance of the southeastern United States as a hub for genetic flow. In addition, based on stable hydrogen isotopic ratios, there is strong evidence of reverse (southward) migration of H. zea from the continental United States to Puerto Rico. Our study highlights the implications for IPM and IRM programs and the need for management strategies that account for both northward and southward migration patterns.

The growth promotion in endophyte symbiotic plants does not penalise the resistance to herbivores and bacterial microbiota.

A trade-off between growth and defence against biotic stresses is common in plants. Fungal endophytes of the genus Epichloë may relieve this trade-off in their host grasses since they can simultaneously induce plant growth and produce antiherbivore alkaloids that circumvent the need for host defence. The Epichloë ability to decouple the growth-defence trade-off was evaluated by subjecting ryegrass with and without Epichloë endophytes to an exogenous treatment with gibberellin (GA) followed by a challenge with Rhopalosiphum padi aphids. In agreement with the endophyte-mediated trade-off decoupling hypothesis, the GA-derived promotion of plant growth increased the susceptibility to aphids in endophyte-free plants but did not affect the insect resistance in endophyte-symbiotic plants. In line with the unaltered insect resistance, the GA treatment did not reduce the concentration of Epichloë-derived alkaloids. The Epichloë mycelial biomass was transiently increased by the GA treatment but at the expense of hyphal integrity. The response of the phyllosphere bacterial microbiota to both GA treatment and Epichloë was also evaluated. Only Epichloë, and not the GA treatment, altered the composition of the phyllosphere microbiota and the abundance of certain bacterial taxa. Our findings clearly demonstrate that Epichloë does indeed relieve the plant growth-defence trade-off.

The Bombyx mori singed Gene Is Involved in the High-Temperature Resistance of Silkworms.

Temperature is an important factor in the growth, development, survival, and reproduction of organisms. The high-temperature resistance mechanism of insects may be significant for use in the prevention and control of insect pests. The silkworm, Bombyx mori, is an important Lepidoptera model species for studies on pest control in agriculture and forestry. We identified a gene in B. mori, the B. mori singed (Bmsn) gene, which is involved in the high-temperature resistance of silkworms. Sn proteins are highly conserved among species in many taxonomic groups. The overexpression of the Bmsn gene promoted the proliferation of silkworm cells, reduced oxidation, and reduced the accumulation of reactive oxygen species under stress. Interfering with the Bmsn gene had the opposite result. We constructed a transgenic B. mori strain that overexpressed the Bmsn gene. The physiological traits of the transgenic strain were significantly improved, and it had stronger high-temperature resistance. The Bmsn gene is involved in the process by which fat bodies respond to high-temperature stress. These findings provide insights into the mechanism of high-temperature resistance of insects and offer a new perspective on agricultural and forestry pest control.

Pecan secondary metabolites influenced the population of Zeuzera coffeae by affecting the structure and function of the larval gut microbiota.

The plant secondary metabolites (PSMs), as important plant resistance indicators, are important targets for screening plant insect resistance breeding. In this study, we aimed to investigate whether the population of Zeuzera coffeae (ZC) is affected by different varieties of Carya illinoinensis PSMs content. At the same time, the structure and function of the gut microbiome of ZC were also analyzed in relation to different pecan varieties. We counted the populations of ZC larvae in four pecan varieties and determined the content of four types of PSMs. The structure and function of the larval gut microbiota were studied in connection to the number of larvae and the content of PSMs. The relationships were investigated between larval number, larval gut microbiota, and PSM content. We found that the tannins, total phenolics, and total saponins of 4 various pecans PSMs stifled the development of the ZC larval population. The PSMs can significantly affect the diversity and abundance of the larval gut microbiota. Enrichment of ASV46 (Pararhizobium sp.), ASV994 (Olivibacter sp.), ASV743 (Rhizobium sp.), ASV709 (Rhizobium sp.), ASV671 (Luteolibacter sp.), ASV599 (Agrobacterium sp.), ASV575 (Microbacterium sp.), and ASV27 (Rhizobium sp.) in the gut of larvae fed on high-resistance cultivars was positively associated with their tannin, total saponin, and total phenolic content. The results of the gut microbiome functional prediction for larvae fed highly resistant pecan varieties showed that the enriched pathways in the gut were related to the breakdown of hazardous chemicals. Our findings provide further evidence that pecan PSMs influence the structure and function of the gut microbiota, which in turn affects the population stability of ZC. The study's findings can serve as a theoretical foundation for further work on selecting ZC-resistant cultivars and developing green management technology for ZC.

Assessment of Balkan Pepper (Capsicum annuum L.) Accessions for Agronomic, Fruit Quality, and Pest Resistance Traits

To maintain the continuous genetic variation and increase the genetic gain, appreciable germplasm diversity and its comprehensive characterization is necessary to further utilize gene sources for pre-breeding. The diversity of pepper forms, cultivation traditions and diverse fruit usages are typical for Balkan countries. Considering this rich diversity, 21 pepper accessions from the Balkan region were evaluated for morphological, biochemical, and insect resistance traits during 2018 and 2019 at Maritsa Vegetable Crops Research Institute, Plovdiv, Bulgaria. Among the studied accessions, the highest productivity was observed in pumpkin shape K1115 and kapia type K1081 accessions, with 0.74 kg and 0.70 kg per plant, respectively. Concerning fruit quality, the highest total polyphenols and ferric reducing antioxidant power (FRAP) were observed in pumpkin shape K712 (203.44 mg GAE/100 g FW) and K1103A (11.49 µmol Fe2+/g FW) accessions, respectively. Concerning insect resistance, 38% of studied accessions showed no infestation of green peach aphid. The kapia type K697 accession was seen as the most reliable resistance source, as it was not infested by aphids and had the least thrips (20% on plants) and cotton bollworm (6.67% on plants and 8.34% on fruit) damage. Based on examined traits, accessions were identified for enhanced fruit quality and promising insect resistance and have been included in further pre-breeding efforts.

The phyllosphere microbiome in Tsuga canadensis: Relationships with resistance to the insect Adelges tsugae and effects of the pesticide dinotefuran

Tsuga canadensis (Eastern hemlock) is an important tree species in North America, but it has been subject to threats from invasive insects such as Adelges tsugae (hemlock woolly adelgid-A. tsugae). A. tsugae is native to Asia and Tsuga species from Asia are typically resistant to this insect. In this study, we examined the phyllosphere microbiome of two hemlock species native to North America and susceptible to A. tsugae (T. canadensis and T. caroliniana), as well as two species native to Asia and resistant to A. tsugae (T. chinensis and T. diversifolia). Since pesticide application is often used to treat native Tsuga species impacted by A. tsugae, we also examined the response of the microbiome and overall plant physiology to the pesticide dinotefuran in T. canadensis. We found significant differences in the bacterial and fungal microbiome among the four species of Tsuga. Dinotefuran also significantly affected bacterial communities, but not fungal communities, of T. canadensis. The genus Methylobacterium, which uses methanol produced during plant cell elongation and growth, was a significant indicator of trees not subject to dinotefuran. This suggests that dinotefuran may have reduced cell growth in treated trees. Despite changes in the microbiome, dinotefuran application did not significantly affect plant physiology; however, the coefficient of variation (CV) in dinotefuran treated trees was larger than non-treated trees. Our results suggest that the microbiome could play a role in mediating insect resistance in Tsuga species, and that pesticide application could affect the phyllosphere microbiome with implications for plant growth and resistance.

Contribution of the transcription factor SfGATAe to Bt Cry toxin resistance in Spodoptera frugiperda through reduction of ABCC2 expression

Insect resistance evolution poses a significant threat to the advantages of biopesticides and transgenic crops utilizing insecticidal Cry-toxins from Bacillus thuringiensis (Bt). However, there is limited research on the relationship between transcriptional regulation of specific toxin receptors in lepidopteran insects and their resistance to Bt toxins. Here, we report the positive regulatory role of the SfGATAe transcription factor on the expression of the ABCC2 gene in Spodoptera frugiperda. DNA regions in the SfABCC2 promoter that are vital for regulation by SfGATAe, utilizing DAP-seq technology and promoter deletion mapping. Through yeast one-hybrid assays, DNA pull-down experiments, and site-directed mutagenesis, we confirmed that the transcription factor SfGATAe regulates the core control site PBS2 in the ABCC2 target gene. Tissue-specific expression analysis has revealed that SfGATAe is involved in the regulation and expression of midgut cells in the fall armyworm. Silencing SfGATAe in fall armyworm larvae resulted in reduced expression of SfABCC2 and decreased sensitivity to Cry1Ac toxin. Overall, this study elucidated the regulatory mechanism of the transcription factor SfGATAe on the expression of the toxin receptor gene SfABCC2 and this transcriptional control mechanism impacts the resistance of the fall armyworm to Bt toxins.

The Role of Organic Farming in Sustainable Agriculture

Organic farming, as a sustainable alternative to conventional high-input practices reliant on synthetic fertilizers, fungicides, and pesticides, prioritizes the vitality of soil as a living ecosystem. Embracing practices such as crop rotation, animal manures, crop residues, green manures, and biological pest and disease control, organic agriculture aims to uphold soil health and productivity. By meeting present food demands in an environmentally conscientious manner, organic farming also safeguards the needs of future generations, thereby preserving and nurturing our ecosystem. Modern agriculture has detrimentally impacted the environment, leading to issues such as decreased soil fertility, elevated water hardness, insect resistance development, reduced genetic diversity in plants, heightened levels of toxic residues in the food chain and animal feed, contributing to health problems and environmental degradation. In contrast, organic farming nurtures plants with essential macronutrients and micronutrients while simultaneously enhancing soil physical, chemical, and biological characteristics. Guided by the ethos of sustainability, organic farming prioritizes mitigating health risks linked to synthetic inputs and cultivating a healthier ecosystem. The minimal environmental footprint of organic farming positions it as a practical solution for tackling urgent concerns like soil degradation, water pollution, and global ecological well-being. Essentially, organic farming stands out as a beacon of sustainable agriculture, paving the way to reconcile food production with environmental conservation. Moreover, organic farming's low environmental impact presents an opportunity to rehabilitate and enhance degraded agricultural lands.

Utilization of Diverse Molecules as Receptors by Cry Toxin and the Promiscuous Nature of Receptor-Binding Sites Which Accounts for the Diversity.

By 2013, it had been shown that the genes cadherin-like receptor (Cad) and ATP-binding cassette transporter subfamily C2 (ABCC2) were responsible for insect resistance to several Cry1A toxins, acting as susceptibility-determining receptors, and many review articles have been published. Therefore, this review focuses on information about receptors and receptor-binding sites that have been revealed since 2014. Since 2014, studies have revealed that the receptors involved in determining susceptibility vary depending on the Cry toxin subfamily, and that binding affinity between Cry toxins and receptors plays a crucial role. Consequently, models have demonstrated that ABCC2, ABCC3, and Cad interact with Cry1Aa; ABCC2 and Cad with Cry1Ab and Cry1Ac; ABCC2 and ABCC3 with Cry1Fa; ABCB1 with Cry1Ba, Cry1Ia, Cry9Da, and Cry3Aa; and ABCA2 with Cry2Aa and Cry2Ba, primarily in the silkworm, Bombyx mori. Furthermore, since 2017, it has been suggested that the binding sites of BmCad and BmABCC2 on Cry1Aa toxin overlap in the loop region of domain II, indicating that Cry toxins use various molecules as receptors due to their ability to bind promiscuously in this region. Additionally, since 2017, several ABC transporters have been identified as low-efficiency receptors that poorly induce cell swelling in heterologously expressing cultured cells. In 2024, research suggested that multiple molecules from the ABC transporter subfamily, including ABCC1, ABCC2, ABCC3, ABCC4, ABCC10, and ABCC11, act as low-efficiency receptors for a single Cry toxin in the midgut of silkworm larvae. This observation led to the hypothesis that the presence of such low-efficiency receptors contributes to the evolution of Cry toxins towards the generation of highly functional receptors that determine the susceptibility of individual insects. Moreover, this evolutionary process is considered to offer valuable insights for the engineering of Cry toxins to overcome resistance and develop countermeasures against resistance.

Insecticidal bisindole alkaloids from leaves of Tabernaemontana divaricata 'Dwaft'

Six undescribed bisindole alkaloids, namely taberdisines A–F (1–6), were isolated from the leaves of Tabernaemontana divaricata 'Dwaft'. Among them, alkaloids 1 and 2 were the first examples of strychnos–iboga type alkaloid with both C–C linkage patterns. Alkaloid 3, a new type of aspidosperma–iboga with a furan-ring, as well as other three undescribed ones was disclosed. Their structures were elucidated by comprehensive spectroscopic analyses. Alkaloids 1 and 5 showed insecticide activity on Sf9 cell and eggs of Spodoptera frugiperda in vivo, which might explain the potential of the plants for insect resistance.

Gene Cloning, Heterologous Expression, and In Silico Analysis of Chitinase B from Serratia marcescens for Biocontrol of Spodoptera frugiperda Larvae Infesting Maize Crops.

Spodoptera frugiperda, the fall armyworm (FAW), is a highly invasive polyphagous insect pest that is considered a source of severe economic losses to agricultural production. Currently, the majority of chemical insecticides pose tremendous threats to humans and animals besides insect resistance. Thus, there is an urgent need to develop new pest management strategies with more specificity, efficiency, and sustainability. Chitin-degrading enzymes, including chitinases, are promising agents which may contribute to FAW control. Chitinase-producing microorganisms are reported normally in bacteria and fungi. In the present study, Serratia marcescens was successfully isolated and identified from the larvae of Spodoptera frugiperda. The bacterial strain NRC408 displayed the highest chitinase enzyme activity of 250 units per milligram of protein. Subsequently, the chitinase gene was cloned and heterologously expressed in E. coli BL21 (DE3). Recombinant chitinase B was overproduced to 2.5-fold, driven by the T7 expression system. Recombinant chitinase B was evaluated for its efficacy as an insecticidal bioagent against S. frugiperda larvae, which induced significant alteration in subsequent developmental stages and conspicuous malformations. Additionally, our study highlights that in silico analyses of the anticipated protein encoded by the chitinase gene (ChiB) offered improved predictions for enzyme binding and catalytic activity. The effectiveness of (ChiB) against S. frugiperda was evaluated in laboratory and controlled field conditions. The results indicated significant mortality, disturbed development, different induced malformations, and a reduction in larval populations. Thus, the current study consequently recommends chitinase B for the first time to control FAW.

Two cuticle-enriched chemosensory proteins confer multi-insecticide resistance in Spodoptera frugiperda

Recent studies revealed that insect chemosensory proteins (CSPs) both play essential roles in insect olfaction and insect resistance. However, functional evidence supporting the crosslink between CSP and insecticide resistance remains unexplored. In the present study, 22 SfruCSP transcripts were identified from the fall armyworm (FAW) and SfruCSP1 and SfruCSP2 are enriched in the larval cuticle and could be induced by multiple insecticides. Both SfruCSP1 and SfruCSP2 are highly expressed in the larval inner endocuticle and outer epicuticle, and these two proteins exhibited high binding affinities with three insecticides (chlorfenapyr, chlorpyrifos and indoxacarb). The knockdown of SfruCSP1 and SfruCSP2 increased the susceptibility of FAW larvae to the above three insecticides, and significantly increased the penetration ratios of these insecticides. Our in vitro and in vivo evidence suggests that SfruCSP1 and SfruCSP2 are insecticide binding proteins and confer FAW larval resistance to chlorfenapyr, chlorpyrifos and indoxacarb by an insecticide sequestration mechanism. The study should aid in the exploration of larval cuticle-enriched CSPs for insect resistance management.

NEW MULTIPLEX SNP GENOTYPING ASSAY TO SIMUTANEOUSLY SCREEN FOR EIGHT VOLTAGE-GATED SODIUM CHANNEL MUTATIONS IN AEDES AEGYPTI

Aedes aegypti has been implicated as the vector responsible for transmission of dengue, chikungunya, and Zika viruses during disease outbreaks in Florida within the past 15 years. Recently, locally acquired dengue cases have increased dramatically, with more than 450 cases documented in Florida since 2019. This mosquito is known to be resistant to pyrethroid-based insecticides in Florida. Resistance of insects to pyrethroids due to knockdown resistance (kdr) is the result of single nucleotide polymorphisms (SNPs) in the voltage-gated sodium channel gene (vgsc). Recently, two novel SNPs, F174I and E478K, and four known SNPs, V410L, S723T, D1763Y, and Q1853R were reported circulating in Floridian Ae. aegypti populations for the first time. The present study provides a more comprehensive estimate of these SNP frequencies through the screening of a larger number of Floridian Ae. aegypti samples using a new custom multiplex SNP assay we developed using the Agena Biosciences iPLEX Assay platform to facilitate the rapid screening of multiple SNPs at an affordable cost. Our assay was successful in screening 162 Ae. aegypti mosquitoes for 8 SNPs from 4 counties in Southern Florida (Broward, Collier, Palm Beach, and Monroe Counties). This new assay can be used for studies examining the association between genetic mutations and pyrethroid-resistant phenotypes in Ae. aegypti populations such as increased time of survival after insecticide exposure.

Laboratory evaluation of a bio-insecticide candidate from tangerine peel extracts against Trialeurodes vaporariorum (Homoptera: Aleyrodidae).

The excessive use of synthetic insecticides in modern agriculture has led to environmental contamination and the development of insect resistance. Also, the prolonged use of chemical insecticides in producing flowers and tomatoes in greenhouses has caused health problems for workers and their offspring. In this study, we analyzed the efficacy of mandarin peel (Citrus reticulata L.) essential oil (EO) as a natural insecticide against greenhouse whitefly (Trieurodes vaporariorum W., Homoptera: Aleyrodidae), a common pest in greenhouse production of different crops. Petroleum ether (PET) and n-hexane (HEX) were used as solvents to extract essential oil (EO) from tangerine peels. The yield of EO was 1.59% and 2.00% (m/m) for PET and HEX, respectively. Additionally, the insect-killing power of EO was tested by checking how many greenhouse whiteflies died at different times. The results showed that PET and HEX extracts of tangerine EO effectively controlled greenhouse whiteflies. Furthermore, with both solvents, a 12.5% (v/v) application was as practical as the commercial insecticide imidacloprid. Further characterization tests with the polarimeter, FTIR, HPLC-RP, and GC-MS showed that the essential oil (EO) contained about 41% (v/v) of d-limonene and that this compound may be responsible for the observed insecticidal properties. Therefore, tangerine peel essential oil is an excellent botanical insecticide candidate for controlling greenhouse whiteflies.

HairNet2: deep learning to quantify cotton leaf hairiness, a complex genetic and environmental trait

BackgroundCotton accounts for 80% of the global natural fibre production. Its leaf hairiness affects insect resistance, fibre yield, and economic value. However, this phenotype is still qualitatively assessed by visually attributing a Genotype Hairiness Score (GHS) to a leaf/plant, or by using the HairNet deep-learning model which also outputs a GHS. Here, we introduce HairNet2, a quantitative deep-learning model which detects leaf hairs (trichomes) from images and outputs a segmentation mask and a Leaf Trichome Score (LTS).ResultsTrichomes of 1250 images were annotated (AnnCoT) and a combination of six Feature Extractor modules and five Segmentation modules were tested alongside a range of loss functions and data augmentation techniques. HairNet2 was further validated on the dataset used to build HairNet (CotLeaf-1), a similar dataset collected in two subsequent seasons (CotLeaf-2), and a dataset collected on two genetically diverse populations (CotLeaf-X). The main findings of this study are that (1) leaf number, environment and image position did not significantly affect results, (2) although GHS and LTS mostly correlated for individual GHS classes, results at the genotype level revealed a strong LTS heterogeneity within a given GHS class, (3) LTS correlated strongly with expert scoring of individual images.ConclusionsHairNet2 is the first quantitative and scalable deep-learning model able to measure leaf hairiness. Results obtained with HairNet2 concur with the qualitative values used by breeders at both extremes of the scale (GHS 1-2, and 5-5+), but interestingly suggest a reordering of genotypes with intermediate values (GHS 3-4+). Finely ranking mild phenotypes is a difficult task for humans. In addition to providing assistance with this task, HairNet2 opens the door to selecting plants with specific leaf hairiness characteristics which may be associated with other beneficial traits to deliver better varieties.