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Abstract Episyrphus balteatus can provide dual ecosystem services including pest control and pollination, which the larvae are excellent predators of aphid pest whereas adults are efficient pollinator. In this study, we assembled a high-quality genome of E. balteatus from northern China geographical population at the chromosome level by using Illumina, PacBio long reads, and Hi-C technologies. The 467.42 Mb genome was obtained from 723 contigs, with a contig N50 of 9.16 Mb and Scaffold N50 of 118.85 Mb, and 90.25% (431.75 Mb) of the assembly was anchored to 4 pseudo-autosomes and one pseudo-heterosome. In total, 14,848 protein-coding genes were annotated, and 95.14% of genes were fully represented in NR, GO, KEGG databases. Besides, we also obtained the mitochondrial genome of E. balteatus of 16, 837 bp in length with 37 typical mitochondrial genes. Overall, this high-quality genome is valuable for evolutionary and genetic studies of E. balteatus and other Syrphidae hoverfly species.

IntroductionSymbiotic bacteria play key roles in a variety of important life processes of insects such as development, reproduction and environmental adaptation, and the elucidation of symbiont population structure and dynamics is crucial for revealing the underlying regulatory mechanisms. The marmalade hoverfly (Episyrphus balteatus) is not only a remarkable aphid predator, but also a worldwide pollinator second to honeybees. However, its symbiont composition and dynamics remain unclear.MethodsHerein, we investigate the symbiotic bacterial dynamics in marmalade hoverfly throughout whole life cycle, across two sexes, and in its prey Megoura crassicauda by 16S rRNA sequencing.ResultsIn general, the dominant phyla were Proteobacteria and Firmicutes, and the dominant genera were Serratia and Wolbachia. Serratia mainly existed in the larval stage of hoverfly with the highest relative abundance of 86.24% in the 1st instar larvae. Wolbachia was found in adults and eggs with the highest relative abundance of 62.80% in eggs. Significant difference in species diversity was observed between the adults feeding on pollen and larvae feeding on M. crassicauda, in which the dominant symbiotic bacteria were Asaia and Serratia, respectively. However, between two sexes, the symbionts exhibited high similarity in species composition. In addition, our results suggested that E. balteatus obtainded Serratia mainly through horizontal transmission by feeding on prey aphids, whereas it acquired Wolbachia mainly through intergeneration vertical transmission. Taken together, our study revealed the effects of development stages, diet types and genders of E. balteatus on symbionts, and explored transmission modes of dominant bacteria Serratia and Wolbachia.DiscussionOur findings lay a foundation for further studying the roles of symbiotic bacteria in E. balteatus life cycle, which will benefit for revealing the co-adaptation mechanisms of insects and symbiotic bacteria.

IntroductionExcessive application of nitrogen fertilizer in cotton field causes soil and water pollution as well as significant increase of aphid population. Reasonable fertilization is an important approach to improve agricultural production efficiency and reduce agriculture-derived pollutions. This study was aimed to explore the effects of nitrogen fertilizer on the Bt cotton physiological characteristics and the growth and development of A. gossypii, a sap-sucking cotton pest.MethodsFive different levels of Ca(NO3)2 (0.0 g/kg, 0.3 g/kg, 0.9 g/kg, 2.7 g/kg and 8.1 g/kg) were applied into vermiculite as nitrogen fertilizer in order to explore the effects of nitrogen fertilizer on the growth and development of Bt cotton and aphids.ResultsThe results showed that the medium level of nitrogen fertilizer (0.9 g/kg) effectively facilitated the growth of Bt cotton plant and suppressed the population expansion of aphids, whereas high and extremely high nitrogen application (2.7 and 8.1 g/kg) significantly increased the population size of aphids. Both high and low nitrogen application benefited aphid growth in multiple aspects such as prolonging nymph period and adult lifespan, enhancing fecundity, and improving adult survival rate by elevating soluble sugar content in host Bt cotton plants. Cotton leaf Bt toxin content in medium nitrogen group (0.9 g/kg) was significantly higher than that in high (2.7 and 8.1 g/kg) and low (0.3 g/kg) nitrogen groups, but Bt toxin content in aphids was very low in all the nitrogen treatment groups, suggesting that medium level (0.9 g/kg) might be the optimal nitrogen fertilizer treatment level for promoting cotton seedling growth and inhibiting aphids.DiscussionOverall, this study provides insight into trophic interaction among nitrogen fertilizer levels, Bt cotton, and cotton aphid, and reveals the multiple effects of nitrogen fertilizer levels on growth and development of cotton and aphids. Our findings will contribute to the optimization of the integrated management of Bt cotton and cotton aphids under nitrogen fertilization.

Abstract The oriental armyworm, Mythimna separata, is an extremely destructive polyphagous pest with a broad host range that seriously threatens the safety of agricultural production. Here, a high-quality chromosome-level genome was assembled using Illumina, PacBio HiFi long sequencing, and Hi-C scaffolding technologies. The genome size was 706.30 Mb with a contig N50 of 22.08 Mb, and 99.2% of the assembled sequences were anchored to 31 chromosomes. In addition, 20,375 protein-coding genes and 258.68 Mb transposable elements were identified. The chromosome-level genome assembly of M. separata provides a significant genetic resource for future studies of this insect and contributes to the development of management strategies.

The recent dominance of Adelphocoris suturalis Jakovlev as the primary cotton field pest in Bt-cotton-cultivated areas has generated significant interest in cotton pest control research. This study addresses the limited understanding of cotton defense mechanisms triggered by A. suturalis feeding. Utilizing LC-QTOF-MS, we analyzed cotton metabolomic changes induced by A. suturalis, and identified 496 differential positive ions (374 upregulated, 122 downregulated) across 11 categories, such as terpenoids, alkaloids, phenylpropanoids, flavonoids, isoflavones, etc. Subsequent iTRAQ-LC-MS/MS analysis of the cotton proteome revealed 1569 differential proteins enriched in 35 metabolic pathways. Integrated metabolome and proteome analysis highlighted significant upregulation of 17 (89%) proteases in the α-linolenic acid (ALA) metabolism pathway, concomitant with a significant increase in 14 (88%) associated metabolites. Conversely, 19 (73%) proteases in the fructose and mannose biosynthesis pathway were downregulated, with 7 (27%) upregulated proteases corresponding to the downregulation of 8 pathway-associated metabolites. Expression analysis of key regulators in the ALA pathway, including allene oxidase synthase (AOS), phospholipase A (PLA), allene oxidative cyclase (AOC), and 12-oxophytodienoate reductase3 (OPR3), demonstrated significant responses to A. suturalis feeding. Finally, this study pioneers the exploration of molecular mechanisms in the plant–insect relationship, thereby offering insights into potential novel control strategies against this cotton pest.

Abstract Background Apolygus lucorum is a worldwide omnivorous pest damaging a range of crops and causing great economic losses. Symbiotic bacteria living in insects play a key role in the nutrition, physiology, and behavior of hosts. Here, we present an experiment using Illumina HiSeq sequencing targeting the V3–V4 regions of bacteria’s 16S rRNA throughout the entire life cycle of A. lucorum. Results The first and second instar nymphs have the largest alpha diversity compared with other life stages of the insect. Bacterial phyla Proteobacteria (72.29%), Firmicutes (15.24%), Actinobacteria (7.76%) exhibit the largest relative abundance in all developmental stages. Erwinia (23.97%) and Lactococcus (10.62%) are the two genera with the highest relative abundance. The relative abundance of Erwinia in the nymph stage is significantly greater than the adult stage, and the relative abundance of Lactococcus in 6-day-old and 9-day-old adult females is higher compared with adult males. Conclusions These results reveal that microbial community composition and relative abundance shift dynamically at different life stages, implying that different bacterial phyla and genera may have specific roles in specific life stages such as metabolism, nutrition absorption, detoxification, and reproduction. This study reveals for the first time the community composition and ecological dynamics of symbiotic bacteria throughout the life stages of A. lucorum, and thus may provide insight to new strategies for pest control.

Insecticides play a crucial role in safeguarding crops from pest. However, prior to their release, it is essential to assess the efficacy and potential effects of these protectants. Flupyradifurone (FBF) and sulfoxaflor (SFX), two insecticides commonly used to control aphids, have been found to exert negative effects on the growth and development of Binodoxys communis Gahan, a beneficial parasitic wasp that targets aphids. However, it remains unknown if FBF and SFX influence the symbiotic bacteria residing within B. communis. In this study, 16SrRNA sequencing was utilized to assess the populations of B. communis symbionts exposed to sublethal concentrations (LC10 and LC25) of FBF and SFX. The control and treatment groups exhibited similar bacterial community structures, with Bacteroidota identified as the dominant phylum in each. After exposure to FBF and SFX, the diversity and richness of symbionts (Firmicutes, Bacteroidota, and Actinobacteriota) in B. communis changed notably. The two different insecticides and exposure durations exerted distinct effects on the microbial community. For instance, SFX stress resulted in a decrease in Akkermansia and an increase in Escherichia Shigella 1h after exposure, with both returning to baseline levels after 3d. In contrast, FBF exposure caused a reduction in Akkermansia presence after 1h, which persisted even after 3d. This study represents the first assessment of the effects of FBF and SFX exposure on symbiotic bacteria in B. communis, expanding our understanding of how insecticides influence natural enemies and their symbiotic bacterial relationships. This study provides theoretical guidance for field applications of FBF and SFX, as well as a reasonable basis for exploring chemical resistance conferred by symbiotic bacteria.

Cotton bollworm (Helicoverpa armigera) poses a global problem, causing substantial economic and ecological losses. Endosymbionts in insects play crucial roles in multiple insect biological processes. However, the interactions between H. armigera and its symbionts have not been well characterized to date. We investigated the symbionts of H. armigera in the whole life cycle from different geographical locations. In the whole life cycle of H. armigera, Proteobacteria, Firmicutes, Bacteroidetes, and Actinobacteria were the dominant bacteria at the phylum level, while Enterococcus, Enterobacter, Glutamicibacter, and Bacillus were the four dominant bacteria at the genus level. Furthermore, high similarity in symbiotic bacterial community was observed in different stages of H. armigera, which were dominated by Enterococcus and Enterobacter. In fields, the dominant bacteria were Proteobacteria and Bacteroidetes, whereas, in the laboratory, the dominant bacteria were Proteobacteria. At the genus level, the dominant bacteria in cotton bollworm eggs of wild populations were Enterobacter, Morganella, Lactococcus, Asaia, Apibacter, and Enterococcus, and the subdominant bacteria were Bartonella, Pseudomonas, and Orbus. Moreover, the symbionts varied with geographical locations, and the closer the geographical distance, the more similar the microbial composition. Taken together, our study identifies and compares the symbiont variation along with geographical gradients and host development dynamic and reveals the high flexibility of microbiome communities in H. armigera, which probably benefits for the successful survival in a complicated changing environment.

Integrated pest management is focused on combining biological and chemical controls. There is evidence of a negative impact of neonicotinoids on biological control, however, sulfoxaflor (SFX), a novel insecticide, its impact on parasitoid natural predator remain limited. Binodoxys communis is an important parasitic natural enemy of Aphis gossypii, which may have direct and indirect toxicity from the insecticides and aphids. Understanding the potential threat of SFX to B. communis is therefore essential to integrated pest management and the conservation of parasitoids. Here, the effects of sublethal doses of SFX on B. communis larvae and adults are presented for the first time. Sublethal SFX doses had a significant negative effect on the survival rate, adult life span, duration of development, and rate of parasitism. Moreover, exposure to sublethal SFX doses also had adverse effects on the biological performance of the next generation of B. communis. Based on the transcriptome analysis, the expression of genes involved in fatty acid metabolism, glycerolipid metabolism, glycerophospholipid metabolism, peroxidase, lysosomes, glutathione metabolism, drug metabolism, and CYP450 were significantly shifted by sublethal SFX exposure. These results indicate that sublethal SFX doses might adversely affect the biological performance of B. communis by altering gene expression related to the function of detoxification systems and energy metabolism. In conclusion, considering the beneficial ecological services of provided by parasitoids and the negative effects of sulfoxaflor across a greater usage scale, we emphasize the importance to optimize pesticide applications in IPM packages, in order to ensure the safety and survival of natural pest parasitoids.

Abstract Background Aphis gossypii Glover (Hemiptera: Aphididae), a worldwide polyphagous phloem-feeding agricultural pest, has three wing morphs (winged parthenogenetic female, gynopara, and male) in the life cycle. The exclusive males could fly from summer hosts to winter hosts, which are essential for gene exchanges of cotton aphid populations from different hosts or regions. However, the molecular mechanism of wing differentiation of male in A. gossypii remains unclear. Results Morphological observation of male A. gossypii showed that there is no distinct difference in the external morphologies of the 1st and 2nd instar nymphs. The obvious differentiation of wing buds started in the 3rd instar nymph and was visible via naked eyes in the 4th instar nymphal stage, then adult male emerged with full wings. According to morphological dynamic changes, the development of wings in males were divided into four stages: preliminary stage (the 1st instar to 2nd instar), prophase (the 3rd instar), metaphase (the 4th instar), anaphase (the 5th instar). Results of feeding behavior monitoring via EPG (electrical penetration graph) technology indicated that although the male cotton aphids had strong desire to feed (longer duration of C 55.24%, F 5.05% and Pd waves 2.56%), its feeding efficiency to summer host cotton was low (shorter E1 3.56% and E2 waves 2.63%). Dynamic transcriptome analysis of male aphid at 5 different developmental periods showed that in the 3rd instar nymph, the number of up-regulated DEGs was significant increased, and time-course gene transcriptional pattern analyses results also showed that numerous genes categorized in clusters 3, 5, and 8 had the highest expressed levels, which were consistent with morphological changes of wing buds. These results indicate that the 3rd instar nymph is the critical stage of wing bud differentiation in males. Furthermore, through pathway enrichment analysis of DEGs and WGCNA, it revealed that the neuroactive ligand-receptor interaction, Ras signaling pathway, dopaminergic synapse, circadian entrainment and the corresponding hub genes of PLK1, BUB1, SMC2, TUBG, ASPM, the kinesin family members (KIF23, KIF20, KIF18-19) and the novel subfamily of serine/threonine (Aurora kinase A and Aurora kinase B) probably played an important role in the critical stage of wing bud differentiation. Conclusion This study explored morphological changes and genes transcriptional dynamics males in cotton aphid, revealed the phenomenon of low feeding efficiency of winged males on summer host cotton, and identified key signaling pathways and potential hub genes potentially involved in wing bud differentiation of male in A. gossypii.

While genetically modified (GM) crops bring economic benefits to human beings, their impact on non-target organisms has become an important part of environmental safety assessments. Symbiotic bacteria play an important role in eukaryotic biological functions and can adjust host communities to adapt to new environments. Therefore, this study examined the effects of Cry1B protein on the growth and development of non-target natural enemies of Pardosa astrigera (L. Koch) from the perspective of symbiotic bacteria. Cry1B protein had no significant effect on the health indicators of P. astrigera (adults and 2nd instar spiderlings). 16S rRNA sequencing results revealed that Cry1B protein did not change the symbiotic bacteria species composition of P. astrigera, but did reduce the number of OTU and species diversity. In 2nd instar spiderlings, neither the dominant phylum (Proteobacteria) nor the dominant genus (Acinetobacter) changed, but the relative abundance of Corynebacterium-1 decreased significantly; in adult spiders, the dominant bacteria genera of females and males were different. The dominant bacterial genera were Brevibacterium in females and Corynebacterium-1 in males, but Corynebacterium-1 was the dominant bacteria in both females and males feeding on Cry1B. The relative abundance of Wolbachia also increased significantly. In addition, bacteria in other genera varied significantly by sex. KEGG results showed that Cry1B protein only altered the significant enrichment of metabolic pathways in female spiders. In conclusion, the effects of Cry1B protein on symbiotic bacteria vary by growth and development stage and sex.

Assessing the potential effects of insecticides on beneficial biological control agents is key to facilitating the success of integrated pest management (IPM) approaches. Flupyradifurone (FPF) is a novel neonicotinoid insecticide that is replacing traditional neonicotinoids over a large geographical range to control pests. Binodoxys communis, is the dominant parasitic natural enemy of aphids. To date, no reports have addressed sublethal effects of FPF on B. communis. In this study, the lethal and sublethal effects of FPF on B. communis were investigated by indirect exposure to larvae and direct exposure to adults. Results showed that the sublethal LC10 and LC25 of FPF had negative effects on the biological parameters of B. communis, including significantly reducing survival rate, adult longevity, parasitism rate, and emergence rate, and significantly prolonging the developmental stages from egg to cocoons. In addition, we observed a transgenerational effect of FPF on the next generation (F1). RNA-Seq transcriptomic analysis identified a total of 1429 differentially expressed genes (DEGs) that were significantly changed between FPF-treated and control groups. These DEGs are mainly enriched in metabolic pathways such as peroxisomes, glutamate metabolism, carbon metabolism, fatty acid metabolism, and amino acid metabolism. This report is the first comprehensive evaluation of how FPF effects B. communis, which adds to the methods of assessing pesticide exposure in parasitic natural enemies. We speculate that the significant changes in pathways, especially those related to lipid synthesis, may be the reason for weakened parasitoid biocontrol ability. The present study provides new evidence for the toxic effects and environmental residue risk of FPF.

Propylea japonica (Thunberg) (Coleoptera: Coccinellidae) is a dominant natural enemy of insect pests in farmland ecosystems. It also serves as an important non-target insect for environmental safety evaluations of transgenic crops. Widespread planting of transgenic crops may result in direct or indirect exposure of P. japonica to recombinant Bacillus thuringiensis (Bt) protein, which may in turn affect the biological performance of this natural enemy by affecting the P. japonica microflora. However, the effects of Bt proteins (such as Cry1B) on the P. japonica microbiota are currently unclear. Here, we used a high-throughput sequencing method to investigate differences in the P. japonica microbiota resulting from treatment with Cry1B compared to a sucrose control. The results demonstrated that the P. japonica microbiome was dominated by Firmicutes at the phylum level and by Staphylococcus at the genus level. Within-sample (α) diversity indices demonstrated a high degree of consistency between the microbial communities of P. japonica treated with the sucrose control and those treated with 0.25 or 0.5 mg/mL Cry1B. Furthermore, there were no significant differences in the abundance of any taxa after treatment with 0.25 mg/mL Cry1B for 24 or 48 h, and treatment with 0.5 mg/mL Cry1B for 24 or 48 h led to changes only in Staphylococcus, a member of the phylum Firmicutes. Treatment with a high Cry1B concentration (1.0 mg/mL) for 24 or 48 h caused significant changes in the abundance of specific taxa (e.g., Gemmatimonades, Patescibacteria, Thauera, and Microbacterium). However, compared with the control, most taxa remained unchanged. The statistically significant differences may have been due to the stimulatory effects of treatment with a high concentration of Cry1B. Overall, the results showed that Cry1B protein could alter endophytic bacterial community abundance, but not composition, in P. japonica. The effects of Bt proteins on endophytes and other parameters in non-target insects require further study. This study provides data support for the safety evaluation of transgenic plants.

The cotton aphid, Aphis gossypii Glover, is a species of polyphagous aphid with many biotypes, and its host transfer has always been the focus of research on the control of cotton aphid. An important factor affecting aphid specialization is the nutritional association with microbial symbionts that provide the host with nutrients lacking in the diet. We analyzed the microbial composition and biodiversity of reared on zucchini for 10 generations (T1–T10) and cotton as a control (CK), by high-throughput Illumina sequencing of 16S ribosomal RNA genes. The findings showed that the change in plant hosts decreased the richness and variety of microbial species. Regardless of whether the plant host is altered or not, Proteobacteria and Firmicutes are the predominate phyla in cotton-specialized aphid. Additionally, cotton-specialized aphids that live in zucchini had considerably lower relative abundances of non-dominant phyla (Bacteroidetes) than cotton hosts. At the genus level the dominant communities were Buchnera, Acinetobacter, and Arsenophonus. The relative abundance of Buchnera was significantly higher in aphids reared on zucchini than those on cotton, whereas the opposite was observed for Acinetobacter, as well as for some non-dominant communities (Stenotrophomonas, Pseudomons, Flavobacterium, Novosphingobium). Collectively, this study clarifies the dynamic changes of symbiotic bacteria in cotton-specialized aphids reared on zucchini for multiple generations. Among them, Buchnera is crucial for the cotton-specialized aphid to get nutrients during the transfer of the host and has a favorable impact on the colonization of cotton-specialized aphid populations on zucchini hosts. It not only enriches our understanding of the relationship between the bacterial microbiota of aphids and their adaptability to new hosts, zucchini, but also expands the current body of research on the mechanisms underlying the host shifting ability of cotton-specialized aphids.

IntroductionReproductive polymorphism and symbiotic bacteria are commonly observed in aphids, but their interaction remains largely unclear. In polymorphic aphid species (Aphis gossypii), offspring of parthenogenetic females (PFs) develops into sexuparae which produces gynoparae and males successively. Gynoparae further produces sexual females (SFs), and these sexual females mate with males to produce offspring.MethodsIn this study, we investigated the dynamic changes of symbiotic bacteria during the above-mentioned five reproductive morph switch in A. gossypii via 16S rRNA sequencing technology.ResultsThe results showed that species richness and community diversity of symbiotic bacteria in males were the highest. Proteobacteria was absolutely dominant bacterial phylum (with relative abundance of more than 90%) in the five reproductive morphs of A. gossypii, and Buchnera was absolutely dominant genus (with relative abundance of >90%), followed by Rhodococcus, Pseudomonas, and Pantoea. Male-killing symbiont Arsenophonus presented the highest relative abundance in gynoparae, a specific morph whose offsprings were exclusively sexual females. Both principal component analysis (PCA) and clustering analysis showed trans-generation similarity in microbial community structure between sexuparae and sexual females, between PFs and gynoparae. PICRUSt 2 analysis showed that symbiotic bacteria in the five reproductive morphs were mainly enriched in metabolic pathways.DiscussionReproductive morph switch induced by environmental changes might be associated with bacterial community variation and sexual polymorphism of aphids. This study provides a new perspective for further deciphering the interactions between microbes and reproductive polymorphism in host aphids.

Fipronil is widely used in the agricultural world as an efficient phenylpyrazole insecticide to control pests. Binodoxys communis is a key parasitic natural enemy of major homopteran pests and can successfully control the population of pests such as cotton aphids. It has not yet been studied what effects would sublethal doses of fipronil have on Binodoxys communis larvae. Here, this study evaluated the effect of fipronil on Binodoxys communis larvae and analyze the transcriptome results. The results showed that LC10 (1.19 mg/L) and LC25 (1.73 mg/L) had significant negative effects on the survival rate and parasitism rate of F0 generation. Moreover, exposure to high concentrations (LC25) of fipronil still had obvious passive effect on the F1 generation of Binodoxys communis. These results indicated that sublethal doses of fipronil have malignant effects on the biological functions of parasitoids and their offspring. The results of transcriptome analysis showed that differentially expressed genes (DEGs) of Binodoxys communis after LC10 treatment are mainly related to immunity and detoxification. LC25 treatment instead resulted in changes in the expression of genes related to nutrition, energy and metabolism reactions. Seven of the identified DEGs were selected for real-time fluorescence quantitative PCR analysis. To the best of our knowledge, this is the first report to evaluate the sublethal, intergenerational, and transcriptomic side effects of fipronil on larvae of parasitic natural pest enemies. Our findings provide data to accurately assess the risk of fipronil usage on Binodoxys communis larvae, and provide important theoretical support for the comprehensive prevention and control of natural enemies and pesticides.

IntroductionBacteria play critical roles in the reproduction, metabolism, physiology, and detoxification of their insect hosts. The ladybird beetle (Harmonia axyridis) harbors a myriad of endosymbiotic microbes. However, to date, little is known about how the microbial composition of H. axyridis varies throughout its life cycle.MethodsIn this study, 16S rRNA amplicon sequencing and quantitative PCR were employed to investigate the diversity and dynamics of bacterial symbionts across the egg, larval, pupae, and adults stages of H. axyridis.ResultsHigher bacterial community richness and diversity were observed in eggs, followed by those in adults and pupae. The community richness index differed significantly between second-instar larvae and other developmental stages. Proteobacteria, Firmicutes, and Actinobacteria were the dominant phyla. Staphylococcus, Enterobacter, Glutamicibacter, and Acinetobacter were the dominant bacteria genera; however, their relative abundances fluctuated across host developmental stages. Interestingly, the larval stage harbored high proportions of Firmicutes, whereas the adult microbial community largely consisted of Proteobacteria.DiscussionThis study is the first to determine the symbiotic bacterial composition across key life stages of H. axyridis. These outcomes can foster the development of environmental risk assessments and novel biological control strategies.

The lady beetle Propylea japonica is a dominant natural predator of insect pests in farmland ecosystems and an important non-target indicator insect for the environmental safety assessment of GM crops. The commercial cultivation of GM crops may cause P. japonica to frequently be exposed to the Bt protein environment. In this study, the biological characteristics, enzyme activity, and expression levels of detoxification and metabolism in P. japonica were studied after Cry1B protein treatment. No significant differences were observed in developmental duration, emergence rate, or body weight at different ages after feeding larvae 0.5 mg/mL of Cry1B protein compared with the control. Furthermore, there were no significant differences in the activities of glutathione S-transferase (GST), catalase (CAT), and peroxidase (POD) after feeding 0.25 mg/mL and 0.5 mg/mL Cry1B protein. However, when the concentration of Cry1B protein increased to 1.0 mg/mL, the activities of the GST, CAT, and POD increased significantly. Compared with the control group, there were no significant differences in the expression levels of most detoxification metabolism related genes; only a few genes had changed expression levels at the individual concentrations (CYP345B1, CYP4Q2, CYP9F2, GST, and microsomal GST). Overall, these results suggest that Cry1B protein has little or no effect on the biological characteristics of P. japonica. Genes related to enzyme activity and detoxification are differentially expressed at high concentration stimulation. Therefore, this research suggests that the potential risks of Cry1B for the predator P. japonica are negligible.

The vast majority of parasitoids are capable of precise and meticulous regulation of nutrition and metabolism within the host. An important building block of life, amino acids are critical to the development of parasitoids. To date, research on how parasitoids regulate host amino acid metabolism remains limited. In this study, Aphis gossypii and its dominant parasitoid Binodoxys communis were used as a study system to explore how parasitism may change the regulation of amino acids in A. gossypii with UHPLC-MS/MS and RT-qPCR techniques. Here, for the first 8 h of parasitism the abundance of almost all amino acids in cotton aphids increased, and after 16 h most of the amino acids decreased. An amino acid of parasitic syndrome, the content of Tyr increased gradually after being parasitized. The expression of genes related to amino acid metabolism increased significantly in early stages of parasitism and then significantly decreased gradually. At the same time, the abundance of Buchnera, a cotton aphid specific symbiont increased significantly. Our comprehensive analyses reveal impacts of B. communis on the amino acid regulatory network in cotton aphid from three aspects: amino acid metabolism, gene expression, and bacterial symbionts. Therefore, this research provides an important theoretical basis for parasitoid nutritional regulation in host, which is highly significant as it may inform the artificial reproduction of parasitoids and the biological control of insect pests.

Abstract Background The worldwide pest Aphis gossypii has three-winged morphs in its life cycle, namely, winged parthenogenetic female (WPF), winged gynopara (GP), and winged male, which are all produced by a wingless parthenogenetic female (WLPF). Most studies on A. gossypii have focused on WPF, while few have investigated GP and male. The shared molecular mechanism underlying the wing differentiation in the three wing morphs of A. gossypii remains unknown. The wing differentiation of WPF was explored in a previous study. Herein, GP and male were induced indoors. The characters of the body, internal genitals, wing veins, and fecundity of GP and male were compared with those of WPF or WLPF. Compared with WLPF, the shared and separate differentially expressed genes (DEGs) were identified in these three-wing morphs. Results Newly-born nymphs reared in short photoperiod condition (8 L:16D, 18 °C) exclusively produced gynoparae (GPe) and males in adulthood successively, in which the sex ratio was GP biased. A total of 14 GPe and 9 males were produced by one mother aphid. Compared with WLPF, the three-wing morphs exhibited similar morphology and wing vein patterns but were obviously discriminated in the length of fore- and underwings, reproductive system, and fecundity. A total of 37 090 annotated unigenes were obtained from libraries constructed using the four morphs via RNA sequencing (RNA-Seq). In addition, 10 867 and 19 334 DEGs were identified in the pairwise comparison of GP versus WLPF and male versus WLPF, respectively. Compared with WLPF, the winged morphs demonstrated 2 335 shared DEGs (1 658 upregulated and 677 downregulated). The 1 658 shared upregulated DEGs were enriched in multiple signaling pathways, including insulin, FoxO, MAPK, starch and sucrose metabolism, fatty acid biosynthesis, and degradation, suggesting their key roles in the regulation of wing plasticity in the cotton aphid. Forty-four genes that spanned the range of differential expression were chosen to validate statistical analysis based on RNA-Seq through the reverse transcription quantitative real time polymerase chain reaction (RT-qPCR). The comparison concurred with the expression pattern (either up- or downregulated) and supported the accuracy and reliability of RNA-Seq. Finally, the potential roles of DEGs related to the insulin signaling pathway in wing dimorphism were discussed in the cotton aphid. Conclusions The present study established an efficiently standardized protocol for GP and male induction in cotton aphid by transferring newly-born nymphs to short photoperiod conditions (8 L:16D, 18 °C). The external morphological characters, especially wing vein patterns, were similar among WPFs, GPe, and males. However, their reproductive organs were strikingly different. Compared with WLPF, shared (2 235) and exclusively (1 470 in WLPF, 2 419 in GP, 10 774 male) expressed genes were identified in the three-wing morphs through RNA-Seq, and several signaling pathways that are potentially involved in their wing differentiation were obtained, including insulin signaling, starch and sucrose metabolism.

Background: Aphis gossypii Glover is an important agricultural pest distributed worldwide. Binodoxys communis Gahan is the main parasitoid wasp of A. gossypii. Previous studies have shown that parasitization causes reduced egg production in A. gossypii, but the effects of parasitism on the symbiotic bacteria in the host ovaries are unknown. Results: In this study, we analyzed the microbial communities in the ovaries of A. gossypii without and after parasitization. Whether parasitized or not, Buchnera was the dominant genus of symbiotic bacteria in the ovaries, followed by facultative symbionts including Arsenophonus, Pseudomonas, and Acinetobacter. The relative abundance of Buchnera in the aphid ovary increased after parasitization for 1 d in both third-instar nymph and adult stages, but decreased after parasitization for 3 d. The shifts in the relative abundance of Arsenophonus in both stages were the same as those observed for Buchnera. In addition, the relative abundance of Serratia remarkably decreased after parasitization for 1 d and increased after parasitization for 3 d. A functional predictive analysis of the control and parasitized ovary microbiomes revealed that pathways primarily enriched in parasitization were “amino acid transport and metabolism” and “energy production and conversion.” Finally, RT-qPCR analysis was performed on Buchnera, Arsenophonus, and Serratia. The results of RT-qPCR were the same as the results of 16S rDNA sequencing. Conclusions: These results provide a framework for investigating shifts in the microbial communities in host ovaries, which may be responsible for reduced egg production in aphids. These findings also broaden our understanding of the interactions among aphids, parasitoid wasps, and endosymbionts.

The large-scale commercial cultivation of genetically modified (GM) cotton has brought significant economic and environmental benefits. However, GM crops must undergo strict environmental monitoring and long-term observation. An important natural enemy insect in cotton fields, Geocoris pallidipennis, can ingest the Bt protein expressed in GM cotton by feeding on herbivorous insects that feed on the cotton. However, the potential risk of GM cotton to G. pallidipennis is still unclear. We here evaluated the effects of Bt cotton expressing the Cry1Ac/1Ab protein on nymphs and adults G. pallidipennis. Cry1Ac protein was detected in the midgut of the cotton bollworm, Helicoverpa armigera, after it ingested Bt cotton, and in the midgut of G. pallidipennis nymphs and adults preying on Bt-fed H. armigera. However, the survival rate, growth, development, and fecundity of G. pallidipennis were not adversely affected. Furthermore, G. pallidipennis cadherins, and those genes related to detoxification, antioxidant activity, nutrient utilization, and immune function were not differentially expressed in response to Cry1Ac exposure. Finally, we showed that Cry1Ac could not bind to brush border membrane vesicles (BBMV) proteins in G. pallidipennis nymphs or adults. In summary, these results indicate that the potential negative effect of transgenic Cry1Ac/1Ab cotton on the insect redator G. pallidipennis is negligible.

Abstract Background Aphis gossypii is a worldwide sap-sucking pest with a variety of hosts and a vector of more than 50 plant viruses. The strategy of wing polyphenism, mostly resulting from population density increasing, contributes to the evolutionary success of this pest. However, the related molecular basis remains unclear. Here, we identified the effects of postnatal crowding on wing morph determination in cotton aphid, and examined the transcriptomic differences between wingless and wing morphs. Results Effect of postnatal crowding on wing determination in A. gossypii was evaluated firstly. Under the density of 5 nymphs·cm− 2, no wing aphids appeared. Proportion of wing morphs rised with the increase of density in a certain extent, and peaked to 56.1% at the density of 20 nymphs·cm− 2, and reduced afterwards. Then, transcriptomes of wingless and wing morphs were assembled and annotated separately to identify potentially exclusively or differentially expressed transcripts between these two morphs, in which 3 126 and 3 392 unigenes annotated in Nr (Non-redundant protein sequence) database were found in wingless or wing morphs exclusively. Moreover, 3 187 up- and 1 880 down-regulated genes were identified in wing versus wingless aphid. Pathways analysis suggested the involvement of differentially expressed genes in multiple cellular signaling pathways involved in wing morphs determination, including lipid catabolic and metabolism, insulin, ecdysone and juvenile hormone biosynthesis. The expression levels of related genes were validated by the reverse transcription quantitative real time polymerase chain reaction (RT-qPCR) soon afterwards. Conclusions The present study identified the effects of postnatal crowding on wing morphs induction and demonstrated that the critical population density for wing morphs formation in A. gossypii was 20 nymphs·cm− 2. Comparative transcriptome analysis provides transcripts potentially expressed exclusively in wingless or wing morph, respectively. Differentially expressed genes between wingless and wing morphs were identified and several signaling pathways potentially involved in cotton aphid wing differentiation were obtained.

The complete mitochondrial genome of Aphis gossypii Glover cucumber biotype was sequenced using traditional PCR amplification coupled with Sanger sequencing. The genome is 15,870 bp long, with 83.7% AT content (MW048625). The genome encodes 37 typical mitochondrial genes, including 13 protein-coding genes, 2 ribosomal RNA genes, 22 transfer RNAs, a repeat region of 784 bp, and a control region of 627 bp. The base composition of the genome is A (45.4%), T (38.3%), C (10.5%), and G (5.8%). An analysis of two biotypes A. gossypii mitogenomes identified 77 single nucleotide polymorphisms and 1 insertion and deletion.

Abstract Background Recently, due to the development of food security strategies, cotton has been planted in inland saline-alkali dry soils or in coastal some saline-alkali soils in China. Under the condition, to comprehensively prevent and control Helicoverpa armigera in cotton fields with saline-alkali soils, it is important to study the larval growth and development of H. armigera and to study adult oviposition selectivity in H. armigera adults that feed on NaCl-stressed cotton plants. Results In this study, Bt cotton GK19 was used for the experimental group and its nontransgenic parent Simian 3 was used for the control to study the effects of biochemical substances in cotton as well as larval growth and development and adult oviposition selectivity of H. armigera. The experiments were performed by growing cotton indoors under NaCl stress at concentrations of 0 mmol·L− 1, 75 mmol·L− 1 and 150 mmol·L− 1, respectively. The results showed that the expression of Bt protein was significantly inhibited for NaCl-stressed Bt cotton. The content of soluble protein and K+ in the leaves of cotton were decreased, while the content of gossypol and Na+ were increased. In addition, the 5th instar H. armigera larvae exhibited shorten the life span in a 13-day trial period. Under enclosure treatments and at different female densities, the adult oviposition of H. armigera decreased on high NaCl-stressed nontransgenic cotton, while the oviposition on Bt cotton tended to first increase but then decrease under low, moderate and high NaCl stress treatments. Conclusions Under certain content ranges of NaCl stress treatments, larval of H. armigera growth and development, and adult oviposition were no significant difference in the change for a certain period. However, under high NaCl stress, larval growth, development and adult oviposition were affected, which may provide insights for the prevention and control of H. armigera for Bt cotton in saline-alkali soils.

An amendment to this paper has been published and can be accessed via the original article.

Lysiphlebia japonica Ashmead (Hymenoptera, Braconidae) is an endophagous parasitoid and Aphis gossypii Glover (Hemiptera, Aphididae) is a major pest in cotton. The relationship between insect host-parasitoids and their hosts involves complex physiological, biochemical and genetic interactions. This study examines changes in the development and physiological metabolism of A. gossypii regulated by L. japonica. Our results demonstrated that both the body length and width increased compared to non-parasitized aphids. We detected significantly increases in the developmental period as well as severe reproductive castration following parasitization by L. japonica. We then used proteomics to characterize these biological changes, and when combined with transcriptomes, this analysis demonstrated that the differential expression of mRNA (up or downregulation) captured a maximum of 48.7% of the variations of protein expression. We assigned these proteins to functional categories that included immunity, energy metabolism and transport, lipid metabolism, and reproduction. We then verified the contents of glycogen and 6-phosphate glucose, which demonstrated that these important energy sources were significantly altered following parasitization. These results uncover the effects on A. gossypii following parasitization by L. japonica, additional insight into the mechanisms behind insect-insect parasitism, and a better understanding of host-parasite interactions.

The relationships between hosts and symbionts are complex and dynamic. Not only can symbionts significantly impact host phenotypes, but host interactions with symbionts may influence parasitoids. However, few studies to date have investigated the microbiomes of parasitized host offspring and considered the ecological implications of longitudinal microbiome dynamics. In this study, the parasitoid wasp Lysiphlebia japonica and its aphid host, Aphis gossypii, were used to examine the impact of parasitism on host offspring. Parental parasitism reduced the weight and reproductive capacity of the offspring; furthermore, the offspring showed reduced L. japonica parasitism rates. Moreover, the microbial composition of the parasitized and unparasitized aphid offspring were characterized with high-throughput 16S rRNA gene sequencing. This revealed that L. japonica parasitism strongly altered the microbial community in the offspring of parasitized aphids, particularly, the offspring of aphids parasitized for day 4. Overall, these results indicated that parasitoids can influence host--microbiome interactions by altering the microbiome of the host offspring. Our findings support the importance of further investigations into host--parasitoid--microbiome interactions. [ABSTRACT FROM AUTHOR]