Beet Armyworm Spodoptera Exigua Research Articles

Functional characterization of fatty acyl-coenzyme A reductase, RcFAR1, specially expressed by Rosa chinensis consequent to infection by rose powdery mildew, Podosphaera pannosa

Rose (Rosa chinensis), as the most popular cut flowers in the world, is perennially attacked by rose powdery mildew (Podosphaera pannosa) and beet armyworm (Spodoptera exigua). The indirect plant-mediated interactions between these two pests always exist in the ternary system consisting of these three organisms, but little is known about the molecular mechanisms for such interactions. In this paper, we investigated the function of an insect-resistance-related gene in rose, RcFAR1, encoding a fatty acyl coenzyme A reductase. The gene was expressed in the roses when the plants were infected by P. pannosa, and as a consequence, the resistance of the roses against S. exigua was significantly induced. The results from heterologous expression of RcFAR1 in Saccharomyces cerevisiae showed that RcFAR1 primarily accumulated primary alcohols, including C12:0, C16:0, and C18:0 by using GC-MS analyses. Overexpression in Arabidopsis thaliana resulted in 4.0-, 3.6-, 8.2-, and 3.1-fold increases in the contents of fatty alcohols in T1 generation plants of A. thaliana for C12:0, C14:0, C16:0, and C18:0, respectively, suggesting that RcFAR1 catalyzes the biosynthesis of primary alcohols of C12:0, C14:0, C16:0, and C18:0 in the plants. Transient expression in the tobacco showed that RcFAR1 localized to the cytoplasm or the cell membrane. Our study has elucidated the molecular mechanism for the synthesis of dodecanol, tetradecanol, hexadecanol and other primary alcohols in P. pannosa-infected roses, and thus revealed one of the important molecular regulatory mechanisms by which the resistances of the roses against S. exigua are induced by the infection of the roses by P. pannosa.

Population dynamic and pheromone use for early monitoring of Spodoptera exigua Hübner (Lepidoptera: Noctuidae) in Indonesia

Population dynamic and pheromone use for early monitoring of Spodoptera exigua Hübner (Lepidoptera: Noctuidae) in Indonesia

Toxicity of Ocimum basilicum L. leaf extract against Spodoptera exigua Hübner (Lepidoptera: Noctuidae)

The beet armyworm Spodoptera exigua Hübner (Lepidoptera: Noctuidae) is one of the most significant shallot pests worldwide, which affects agricultural production by approximately 100% in Indonesia. The growing awareness of sustainable agriculture encourages the search for natural alternatives to synthetic pesticides that promote resistance and contaminate the environment. Therefore, this study aims to evaluate the efficacy of basil (Ocimum basilicum L) leaf extract against the 2nd and 3rd instar larvae of S. exigua. The extracts were prepared using maceration and hydrodistillation and the pathogenicity was carried out against the 2nd and 3rd instar of S. exigua using three replicates with several doses of extract ranging from 0- 2.0%. The extracts were applied to the larvae using contact and topical methods. The results showed that maceration extraction yields a greater extract with different types compared to those of the hydrodistillation method. Maceration extract of basil leaves with the contact application method to 2nd instar showed better results than the topical application with the highest mortality rate of 75% (F=24.464; P< 0.001) and LC50 of 0.007%. This indicated that basil leaf extract with the contact application method has great potential to be developed as a botanical insecticide to control S. exigua in the field as part of Integrated Pest Management (IPM).

Insectivorous bats provide more pest suppression services than disservices - a case study in China

Many studies have investigated the role of insectivorous bats as natural enemies of agricultural pests. However, insectivorous bats have a broad diet, and whether their prey species are beneficial or harmful is unknown; thus, accurately assessing the services provided by bats is challenging. In China, most people, including farmers and plant protection workers, have an insufficient understanding of the ecological value of bats in biological control. Therefore, we collected the fecal samples of 10 bat species from three different sampling sites located in Guizhou Province, China, and conducted DNA metabarcoding to determine the diet composition of bats. We also categorized the prey species from the diet of bats as harmful or beneficial, based on previous studies, and recorded the industries they affect. Our results showed that the 10 bat species consumed prey from 19 orders and 96 families; most of them being lepidopterans. Among 564 total prey species recorded, 390 (69.15%) were pests and 28 (4.96%) were beneficial insects. Among them, 326 (57.80%) were agriculture or forestry pests, such as globally present Cotton bollworm (Helicoverpa armigera) and Beet armyworm (Spodoptera exigua). These results indicated that insectivorous bats have more beneficial effects than harmful effects, especially in the context of agriculture and forestry, as they suppressed very few beneficial arthropod mesopredators. By comparing the differences in the diet of I. io in different regions, we found that bats present in different regions can provide different predation services. Our study showed that insectivorous bats provide excellent pest suppression services in various industries and regions. It might be helpful for increasing awareness among people and bat conservation in China and other regions.

Controlling Armyworm (Spodoptera exigua) (Lepidoptera: Noctuidae) on Shallot using Nano Husk Ash

The Beet Armyworm (Spodoptera exigua) is a polyphagous defoliator that poses a significant threat to cultivated plants, particularly shallot crops. Its attacks during the vegetative phase cause substantial damage. To control this pest, farmers often resort to synthetic pesticides, leading to negative consequences such as resistance development. To address consequences, internal control measures, like enhancing plant resistance through beneficial nutrients like silica, are crucial. Silica has been known to improve plant resistance against pest attacks. This study aims to investigate the effects of various methods of applying nano-sized rice husk ash fertilizer on the growth and resistance of Biru Lancor cultivar of shallots against S. exigua. Nano-rice husk ash fertilizer with an average particle size of 65,326 nm was produced and applied using three methods: placement, foliar, and seed coating. The results demonstrated that all methods of nano-rice husk ash application increased plant resistance, reducing leaf damage and pest mortality. The foliar application yielded the best results, enhancing plant height and epidermal cell wall thickness. Furthermore, nano-rice husk ash application positively influenced shallot productivity. This study highlights the potential of nano-sized rice husk ash as a sustainable and effective internal control measure against S. exigua, complementing synthetic pesticides for shallot pest management.

Anthranilic diamide compounds: Discovery of tetrachlorantraniliprole

Given the development of insects’ resistance and deteriorative ecological environment, the discovery of insecticides with a new mode of action has been proven to be an important tool for comprehensive pest management. Herein, a novel anthranilic diamides containing 3,5-dichloropyridylpyrazole group were designed and synthesized. Their insecticidal potency was assayed against beet armyworm (Spodoptera exigua). Moreover, the structure-activity relationship of these new compounds was also discussed. Of them, compound 15g exhibited potential insecticidal activity. Because of the excellent efficacy against a series of Lepidoptera and low toxicity to mammals, compound 15g, namely tetrachlorantraniliprole, has been developed and launched into the market as an insecticide.

Rational Design of Insecticidal Isoxazolines Containing Sulfonamide or Sulfinamide Structure as Antagonists of GABA Receptors with Reduced Toxicities to Honeybee and Zebrafish.

To develop highly effective, nontarget organism-friendly insecticides based on the isoxazoline scaffold, we rationally designed and synthesized 25 isoxazoline derivatives containing sulfonamides and sulfinamides. Their insecticidal activities against the diamondback moth (Plutella xylostella), fall armyworm (Spodoptera frugiperda), beet armyworm (Spodoptera exigua), and Spodoptera litura Fabricius (S. litura) were evaluated. The trifluoromethyl sulfinamide-containing compound 7w displayed excellent activities with LC50 values being 0.09, 0.84, 0.87, and 0.68 mg/L against P. xylostella, S. frugiperda, S. exigua, and S. litura, respectively, which were superior to fluxametamide (LC50 = 0.09, 1.24, 1.10, and 0.65 mg/L, respectively) and maintained at the same order of magnitude LC50 values as fluralaner (LC50 = 0.02, 0.17, 0.12, and 0.19 mg/L, respectively). Importantly, compound 7w showed a medium toxicity level of acute toxicity to honeybee (LD50 = 2.22 μg/adult), which is significantly lower than the fluralaner (high toxicity level, LD50 = 0.09 μg/adult). Acute toxicity experiments with zebrafish (Danio rerio) indicated that compound 7w was safe with the LC50 value being 42.4 mg/L (low toxicity level). Furthermore, electrophysiological experiments and molecular docking studies preliminarily verified that compound 7w acts on the insect GABA receptor, and the theoretical calculations explained that the sulfinamide structure may play an important role in exhibiting biological activities. The above results suggest that compound 7w could be employed as a potentially highly effective, environmentally friendly insecticide to control multiple agricultural pests.

SeBLOS2 knockout via CRISPR/Cas9 leads to the loss of larval integument coloration in Spodoptera exigua (Lepidoptera: Noctuidae).

CRISPR/Cas9 technology is a precise and powerful tool for functionally exploring insect genes. The present study tested CRISPR/Cas9 as a way of undertaking effective gene mutagenesis in an important agricultural pest, the beet armyworm Spodoptera exigua. Based on a S. exigua transcriptome database, the entire complementary DNA sequence of SeBLOS2 encoding 140 amino acid residues was cloned. The gene was highly expressed in late larval stages (L3-L5). Using the CRISPR/Cas9 method, SeBLOS2 was knocked out by altering two sites in the coding region. This resulted in 70%-74% of the G0 generation (L4-L5) larvae displaying mosaic translucent integument. Four different mutations occurred at SeBLOS2-specific target sites, as demonstrated by further polymerase chain reaction-based genotypic analysis. Homozygote mutant L3 larvae were obtained in the G1 generation, with complete loss of white stripes and spots on their larval integument. These results demonstrate a crucial role of SeBLOS2 in integument pigmentation and suggest that the gene can act as a suitable nonlethal marker for functional research on genes in S. exigua and other Lepidopteran pests.

Structural optimization based on 4,5-dihydropyrazolo[1,5-a]quinazoline scaffold for improved insecticidal activities

The long-term and irrational application of insecticides has increased the rate of development of pest resistance and caused numerous environmental issues. To address these problems, our previous work reported that 4,5-dihydropyrazolo[1,5-a]quinazoline (DPQ) is a class of gelled heterocyclic compounds that act on insect γ-aminobutyric acid receptors (GABAR). DPQ scaffold has no cross-resistance to existing insecticides, so the development of this scaffold is an interesting task for integrated pest management. In the present study, a novel series of 4,5-dihydropyrazolo[1,5-a]quinazolines (DPQs) were designed and synthesized based on pyraquinil, a highly insecticidal compound discovered in our previous work. Insecticidal activities of the target compounds against diamondback moth (Plutella xylostella), beet armyworm (Spodoptera exigua), fall armyworm (Spodoptera frugiperda), and red imported fire ant (Solenopsis invicta Buren) were evaluated. Compounds 6 and 12 showed the best insecticidal activity against Plutella xylostella (P. xylostella) (LC50 = 1.49 and 0.97 mg/L), better than pyraquinil (LC50 = 1.76 mg/L), indoxacarb and fipronil (LC50 = 1.80 mg/L). Meanwhile, compound 12 showed slow toxicity to Solenopsis invicta Buren (S. invicta), with a 5 d mortality rate of 98.89% at 0.5 mg/L that is similar to fipronil. Moreover, Electrophysiological studies against the PxRDL1 GABAR heterologously expressed in Xenopus oocytes indicated that compound 12 could act as a potent GABA receptor antagonist (2 μΜ, inhibition rate, 68.25%). Molecular docking results showed that Ser285 (chain A) and Thr289 (chain D) of P. xylostella GABAR participated in hydrogen bonding interactions with compound 12, and density functional theory (DFT) calculations suggested the importance of pyrazolo[1,5-a]quinazoline core in potency. This systematic study provides valuable clues for the development of DPQ scaffold in the field of agrochemicals, and compound 12 can be further developed as an insecticide and bait candidate.

Group V Chitin Deacetylases Influence the Structure and Composition of the Midgut of Beet Armyworm, Spodoptera exigua.

Chitin deacetylase (CDA) can accelerate the conversion of chitin to chitosan, influencing the mechanical properties and permeability of the cuticle structures and the peritrophic membrane (PM) in insects. Putative Group V CDAs SeCDA6/7/8/9 (SeCDAs) were identified and characterized from beet armyworm Spodoptera exigua larvae. The cDNAs of SeCDAs contained open reading frames of 1164 bp, 1137 bp, 1158 bp and 1152 bp, respectively. The deduced protein sequences showed that SeCDAs are synthesized as preproteins of 387, 378, 385 and 383 amino acid residues, respectively. It was revealed via spatiotemporal expression analysis that SeCDAs were more abundant in the anterior region of the midgut. The SeCDAs were down-regulated after treatment with 20-hydroxyecdysone (20E). After treatment with a juvenile hormone analog (JHA), the expression of SeCDA6 and SeCDA8 was down-regulated; in contrast, the expression of SeCDA7 and SeCDA9 was up-regulated. After silencing SeCDAV (the conserved sequences of Group V CDAs) via RNA interference (RNAi), the layer of intestinal wall cells in the midgut became more compact and more evenly distributed. The vesicles in the midgut were small and more fragmented or disappeared after SeCDAs were silenced. Additionally, the PM structure was scarce, and the chitin microfilament structure was loose and chaotic. It was indicated in all of the above results that Group V CDAs are essential for the growth and structuring of the intestinal wall cell layer in the midgut of S. exigua. Additionally, the midgut tissue and the PM structure and composition were affected by Group V CDAs.

A chromosome-level genome assembly of the beet armyworm Spodoptera exigua

BackgroundThe beet armyworm Spodoptera exigua is a polyphagous caterpillar that causes serious damage to many species of crops and vegetables. To gain insight into how this polyphagous insect differs from less harmful oligophagous species, we generated a chromosome-level assembly and compared it to closely related species with the same or different feeding habits. ResultsBased on Illumina and Pacific Biosciences data and Hi-C technology, 425.6 Mb of genome sequences were anchored and oriented into 31 linkage groups, with an N50 length of 14.8 Mb. A total of 24,649 gene models were predicted, of which 97.4% were identified in the genome assembly. Chemosensory genes are vital for locating food: of the four main families, odorant-binding proteins, chemosensory proteins and olfactory receptors showed little difference, whereas gustatory receptors are greatly expanded in S. exigua. Examination of other polyphagous insects confirmed this difference from oligophagous congeners and further identified the bitter receptor subfamily as being particularly affected. ConclusionOur high-quality genome sequence for beet armyworm identified a key expansion of the bitter gustatory receptor subfamily in this and other pests that differs crucially from more benign relatives and offers insight into the biology and possible future means of control for these economically important insects.

Host-Specific larval lepidopteran mortality to pathogenic Serratia mediated by poor diet

Insect guts often harbor an abundance of bacteria. Many of these members are commensal, but some may emerge as opportunistic pathogens when the host is under stress. In this study, we evaluated how dietary nutritional concentration mediates a shift from commensal to pathogenic, and if host species influences those interactions. We used the lepidopterans (Noctuidae) fall armyworm (Spodoptera frugiperda), beet armyworm (Spodoptera exigua), and corn earworm (Helicoverpa zea) as hosts and a Serratia strain initially isolated from healthy fall armyworm. Diet concentration was altered by bulk reduction in nutritional content with dilution using cellulose. Our experiments revealed that low nutrient diet increased mortality from Serratia for beet armyworm and corn earworm. However, for fall armyworm, little mortality was observed in any of the diet combinations. Dietary nutrition and oral inoculation with Serratia did not change the expression of two antimicrobial peptides in fall and beet armyworm, suggesting that other mechanisms that mediate mortality were involved. Our results have implications for how pathogens may persist as commensals in the digestive tract of insects. These findings also suggest that diet plays a very important role in the switch from commensal to pathogen. Finally, our data indicate that the host response to changing conditions is critical in determining if a pathogen may overtake its host and that these three lepidopteran species have different responses to opportunistic enteric pathogens.

Signatures of plant defense response specificity mediated by herbivore-associated molecular patterns in legumes.

Chewing herbivores activate plant defense responses through a combination of mechanical wounding and elicitation by herbivore-associated molecular patterns (HAMPs). HAMPs are wound response amplifiers; however, specific defense outputs may also exist that strictly require HAMP-mediated defense signaling. To investigate HAMP-mediated signaling and defense responses, we characterized cowpea (Vigna unguiculata) transcriptome changes following elicitation by inceptin, a peptide HAMP common in Lepidoptera larvae oral secretions. Following inceptin treatment, we observed large-scale reprogramming of the transcriptome consistent with three different response categories: (i) amplification of mechanical wound responses, (ii) temporal extension through accelerated or prolonged responses, and (iii) examples of inceptin-specific elicitation and suppression. At both early and late timepoints, namely 1 and 6 h, large sets of transcripts specifically accumulated following inceptin elicitation. Further early inceptin-regulated transcripts were classified as reversing changes induced by wounding alone. Within key signaling- and defense-related gene families, inceptin-elicited responses included target subsets of wound-induced transcripts. Transcripts displaying the largest inceptin-elicited fold changes included transcripts encoding terpene synthases (TPSs) and peroxidases (POXs) that correspond with induced volatile production and increased POX activity in cowpea. Characterization of inceptin-elicited cowpea defenses via heterologous expression in Nicotiana benthamiana demonstrated that specific cowpea TPSs and POXs were able to confer terpene emission and the reduced growth of beet armyworm (Spodoptera exigua) herbivores, respectively. Collectively, our present findings in cowpea support a model where HAMP elicitation both amplifies concurrent wound responses and specifically contributes to the activation of selective outputs associated with direct and indirect antiherbivore defenses.

Bt GS57 Interaction With Gut Microbiota Accelerates Spodoptera exigua Mortality.

The Beet armyworm Spodoptera exigua (Lepidoptera: Noctuidae, Spodoptera) is an important global polyphagous pest. Pathogen infection could destroy the intestinal microbial homeostasis of insects, leading to the death of the host. However, the effect of the host intestinal microbial community on the insecticidal effect of Bacillus thuringiensis is rarely studied. In this study, the genome characteristics of Bt GS57 and the diversity and functions of the gut bacteria in S. exigua are investigated using crystal morphology, biological activity, and Illumina HiSeq high-throughput sequencing. The total size of the Bt GS57 genome is 6.17 Mbp with an average G + C content of 35.66%. Furthermore, the Bt GS57 genome contains six cry genes: cry1Ca, cry1Da, cry2Ab, cry9Ea, cry1Ia, and cry1Aa, and a vegetative insecticidal protein gene vip3Aa. The Bt GS57 strain can produce biconical crystals, mainly expressing 70 kDa and 130 kDa crystal proteins. The LC50 value of the Bt GS57 strain against the S. exigua larvae was 0.339 mg mL–1. Physiological and biochemical reactions showed that Bt GS57 belongs to B.t. var. thuringiensis. In addition, we found that B. thuringiensis can cause a dynamic change in the gut microbiota of S. exigua, with a significant reduction in bacterial diversity and a substantial increase in bacterial load. In turn, loss of gut microbiota significantly decreased the B. thuringiensis susceptibility of S. exigua larvae. Our findings reveal the vital contribution of the gut microbiota in B. thuringiensis-killing activity, providing new insights into the mechanisms of B. thuringiensis pathogenesis in insects.

Characterization of the gut microbiome in the beet armyworm Spodoptera exigua in response to the short-term thermal stress

• Enterococcus was the most abundant in the gut of S. exigua larvae. • The temperature stress had certain effects on the microbial composition of S. exigua. • The symbiotic bacteria in S. exigua were more sensitive to low temperatures. The gut microbiota is critical for energy and nutrient utilization and plays a role in host immunity in response to environmental changes. The beet armyworm Spodoptera exigua is a worldwide polyphagous agricultural pest and has frequently experienced potentially stressful temperature fluctuations under natural environmental conditions. However, little is known about the effects of thermal stress on the gut microbiome of this moth pest. Therefore, we investigated the gut microbiome variations, composition and community structure of S. exigua among low-temperature (10 °C), control (26 °C) and high temperature (35 °C) treatments using 16S amplicon sequencing. Overall, 1,192,707 high-quality reads and 762 operational taxonomic units (OTUs) were detected from 15 samples. A total of 289 genera belonging to 19 bacterial phyla were captured, with Firmicutes and Proteobacteria being the most prominent phyla. Alpha diversity metrics indicated no significant differences in the gut bacterial diversity of S. exigua among the three temperature treatments. Principal coordinates and hierarchical cluster analysis revealed significant differences in the structure of gut microbiota between the low-temperature treatment and the other two temperature treatments. In addition, PICRUSt2 analysis demonstrated that the predicted metagenomes associated with the gut microbiome were amino carbohydrate transport and metabolism, acid transport and metabolism, inorganic ion transport and metabolism and cellular processes. Our study showed that thermal stress induced changes in the gut microbiome of the beet armyworm, which may contribute to better understanding the ecological adaptation of S. exigua under changing temperature trends and to evaluating the use of gut microorganisms as biocontrol agents for this pest.

Metabolic-based insecticide resistance mechanism and ecofriendly approaches for controlling of beet armyworm Spodoptera exigua: a review.

The beet army worm, Spodoptera exigua, is a widely distributed polyphagous pest of economically important crops worldwide. The management of this pest insect continues to face many challenges. Despite synthetic chemicals posing a serious threat to the environment, these remain the conventional approach for controlling S. exigua in the field. An over-reliance on chemical control has not only led to selection for resistance to insecticides and to a reduction of natural enemies, but has also polluted various components of ecosystem. Given these increasing pressures on the ecosystem, there is a need to implement integrated pest management (IPM) approaches exploiting a wider range of tools (biotechnological approaches, microbial control, biological control, cultural control, and use of host plant resistance) for an alternative to chemical control. The IPM approach can not only reduce the hazard of chemical residues in the environment and associated health problems, but may also provide best strategies to control insect pests. This review synthesizes published information on insecticide resistance of S. exigua and explores alternative IPM approaches to control S. exigua.

Residual toxicity and sublethal effects of fenvalerate on the development and physiology of Spodoptera exigua reared on different hosts

Beet armyworm Spodoptera exigua (Hübner) (Lepidoptera: Noctuidae) is one of the most devastating pests of different key crops. The present study deals with the toxic effect of fenvalerate insecticide on the development, reproduction and the detoxification enzyme’s activity of S. exigua feeding on tomato, cabbage, and artificial diet. The developmental period of the second instar to adult emergence was prolonged (16.74 days) reared on tomato followed by cabbage (15.97 days) and artificial diet (15.62 days). The fecundity of S. exigua was lowest on the tomato plant (279 eggs), followed by an artificial diet (347 eggs), while the highest number of eggs was observed on cabbage after the control diet (421eggs). Moreover, the lowest survival rate of the second to the third instar, fourth to the fifth instar, and hatchability were recorded in insects reared on tomato (70.89%) compared to that reared on cabbage (77.08%) and artificial diet (79.66%). S. exigua reared on tomato had the lowest intrinsic rate of increase (r: 0.133) and the highest mean generation time (T: 31.36). Furthermore, detoxifying enzyme activity in S. exigua was much lower on tomato than on cabbage and diet. Moreover, the fenvalerate toxicity was synergized by either piperonyl butoxide (PBO) or S,S,S tributylphosphorotrithioate (DEF) due to the likely participation of monooxygenases or esterase; this suggests a general metabolic resistance. This research provides a solid base for devising an applicable and successful strategy for the controlling the resistance development in beet armyworms.

UJI BEBERAPA KONSENTRASI EKSTRAK KULIT JENGKOL (Pithecellobium lobatum Benth) UNTUK MENGENDALIKAN ULAT DAUN BAWANG (Spodoptera exigua Hubner)

Beet armyworm (Spodoptera exigua Hubner) is one of the main problems in shallot cultivation which can cause production losses of up to 100% if not controlled. The use of botanical pesticides from jengkol rind extract (Pithecellobium lobatum Benth) is a solution to control beet armyworms (Spodoptera exigua Hubner). This study aims to obtain a concentration of jengkol rind extract which is effective to controlling beet armyworm. The study was conducted at the Laboratory of Plant Pest and Experimental Garden at the Faculty of Agriculture, University of Riau from January until March 2020. This research was conducted experimentally using a completely randomized design (CRD) with five treatments and four replications. The treatments given were jengkol rind extract concentration 0 g.l-1 water, 25 g.l-1 water, 50 g.l-1 water, 75 g.l-1 water and 100 g.l-1 water. The results indicate that the application of jengkol rind extract 100 gl-1 water is an effective concentration to control beet armyworm with an initial time of death 12.75 hours, a lethal time 50 at 37.50 hours, the highest daily mortality on the second day at 37.5% and total mortality is 92.5% in the 5 days after application.

Accelerated delivery of dsRNA in lepidopteran midgut cells by a Galanthus nivalis lectin (GNA)-dsRNA-binding domain fusion protein

Lepidopteran insects are highly refractory to oral RNA interference (RNAi). Degradation, impaired cellular uptake and intracellular transport of double-stranded RNA (dsRNA) are considered the major factors responsible for the reduced RNAi efficiency in these insects. In this study, the potential of lectins to improve dsRNA delivery and RNAi efficacy was evaluated. First, a fusion protein consisting of the Galanthus nivalis agglutinin (GNA) and a dsRNA binding domain was developed, further referred to as GNA:dsRBD (GNAF). Then, its ability to increase dsRNA uptake and transfection efficiency in lepidopteran midgut cells was evaluated, as well as its ability to protect and promote the RNAi response in the beet armyworm Spodoptera exigua. Confocal microscopy analysis showed that GNAF-complexed dsRNA was internalized faster in Choristoneura fumiferana midgut CF1 cells (1 min) compared to naked dsRNA (>1 h). The faster uptake was also correlated with an increased RNAi efficiency in these CF1 cells. In vivo feeding bioassays with GNAF-complexed dsRNA led to an increased mortality in S. exigua compared to the controls. By targeting the essential gene V-ATPase A, we observed that the mortality increased to 48% in the GNAF-dsRNA treatment compared to only 8.3% and 6.6% in the control treatments with the naked dsRNA and the GNAF, respectively.

Effects of cadmium stress at different concentrations on the reproductive behaviors of beet armyworm Spodoptera exigua (Hübner)

Insects are exposed to cadmium stress since cadmium pollution has increasingly become a serious global environmental issue. However, until now few studies have paid attention to the effect of heavy metals on insect reproductive behaviors. In our study, the courtship behaviors, mating behaviors and fecundity of beet armyworm Spodoptera exigua (Lepidoptera: Noctuidae) exposed to different concentrations of cadmium in artificial diets at larval stage were studied. The results showed that cadmium stress changed the courtship rhythm by significantly advancing or delaying the courtship starting time. Low dose of cadmium (0.2 mg/kg) increased the courtship frequency in the first two scotophases, but in the fourth phase, the two cadmium treatments reduced the frequency. The total courtship duration was significantly shortened in the first six scotophases except high dose of cadmium treatment (51.2 mg/kg) in the sixth dark phase. Paired adults did not mate after the seventh scotophase under low cadmium exposure, while high cadmium stress made the paired adults just copulate in the first four scotophases. The daily mating rate and total mating rate decreased with the increase in cadmium concentration. The number of eggs of low cadmium treatment was higher than that of control, but the difference was not significant; the number of eggs in high cadmium treatment was lower than that of control and low cadmium treatment. Our results indicate that cadmium exposure can disrupt the courtship rhythm for females and has negative influences on copulation behavior and high cadmium stress can reduce fecundity. Hence, the insect population increase will be affected by heavy metal pollution. Our study will provide scientific reference for environmental risk assessment of heavy metal pollution.