Age-stage, two-sex life table, and population projection of Ostrinia furnacalis (Lepidoptera: Crambidae) at different temperatures.

Ostrinia furnacalis (Guenée) (Lepidoptera: Crambidae), a highly destructive pest in Asia, poses a significant threat to maize production by causing substantial yield losses. However, there is a lack of information regarding the impact of temperature variations on its population dynamics and the age-stage and two-sex life table. This study aimed to investigate the impact of 4 temperatures (20 °C, 24 °C, 28 °C, 32 °C) on the development, reproduction, and survival of O. furnacalis under controlled laboratory conditions. Our results revealed that O. furnacalis successfully developed, survived, and laid eggs across the tested temperatures (20-32 °C). The shortest developmental duration for all immature stages was observed at 32 °C. Conversely, increasing temperatures led to decreased longevity. Among the temperatures tested, 28 °C proved to be optimal for O. furnacalis, exhibiting the highest intrinsic rate of increase, finite rate of increase, and net reproductive rate. Our findings indicate that O. furnacalis thrives within a wide temperature range of 20-32 °C, with 28 °C being the most favorable for reproduction. These insights are crucial for predicting population dynamics under diverse climatic conditions and developing effective control strategies against O. furnacalis. This study enhances our understanding of O. furnacalis' life-history traits and provides valuable information for targeted pest management approaches.

Competitive interactions between invasive fall armyworm and Asian corn borer at intraspecific and interspecific level on the same feeding guild.

Interspecific competition is an important factor in the population dynamics and geographical distribution of insect populations. Fall armyworm (FAW), Spodoptera frugiperda, an invasive species, and the Asian corn borer (ACB), Ostrinia furnacalis, a native pest species are major pests on maize in China, posing a threat to maize yield and grain quality. A series of laboratory, greenhouse, and field experiments were conducted to elucidate the competitive interactions between FAW and ACB. In the laboratory experiments, FAW exhibited aggressive behaviors more frequently when compared with ACB, while the latter species exhibited defense behaviors more frequently. Higher intraspecific competition was recorded in the FAW conspecific rather than in ACB, particularly in the 6th larval stage. FAW had a higher interspecific competitive advantage through intraguild predation over ACB, resulting in partial or complete displacement ACB when initially the ratio of the 2 species was 1 : 1. The interspecific competition also had significantly influenced on the population parameters, defensive enzymes, and nutrient of these 2 species. Competitive interaction proved that the response of superoxide, catalase, and soluble protein in FAW were significantly increased, whereas the total sugar content in both species was substantially decreased. Survival rate, and the plant damage that co-infested by both species varied significantly among the sequential combinations under greenhouse and field conditions. FAW consistently exhibited stronger intraspecific aggression than ACB under laboratory and field conditions when co-existing on the same feeding guild. These findings contribute to efforts toward the improvement of integrated pest management programs for FAW, in decision making for invasive and native pests' management strategies to reduce the high risks of FAW and ACB outbreaks.

Genome-wide identification of TaCIPK gene family members in wheat and their roles in host response to Blumeria graminis f. sp. tritici infection.

Powdery mildew, caused by Blumeria graminis f. sp. tritici (Bgt), is a destructive disease affecting wheat crops worldwide. Functional genes can be activated in response to Bgt inoculations. Calcineurin B-like protein (CBL) together with CBL-interacting protein kinase (CIPK) forms the CBL-CIPK protein complex that participates in Ca2+ sensor kinase-related signaling pathways responding to abiotic and biotic stresses. In this study, we performed a genome-wide screening and identified 27 CIPK subfamilies (123 CIPK transcripts, TaCIPKs) including 55 new and 47 updated TaCIPKs in wheat. Phylogenetic analysis revealed that 123 TaCIPKs could be divided into four groups. Segmental duplications and tandem repeats promoted the expansion of the TaCIPK family. Gene function was further evidenced by differences in gene structure, cis-elements, and protein domains. TaCIPK15-4A was cloned in this study. TaCIPK15-4A contained 17 serine, seven tyrosine, and 15 threonine phosphorylation sites and localized in the plasma membrane and cytoplasm. TaCIPK15-4A expression was induced after Bgt inoculation. Virus-induced gene silencing and overexpression experiments indicated that TaCIPK15-4A could play a positive role in wheat disease resistance to Bgt. Overall, these results provide insights into the role of the TaCIPK gene family in wheat resistance and could be beneficial for further research to prevent Bgt infection.

Roles of Ear Injury, Infestation, and Vector Activity by Ostrinia furnacalis (Asian Corn Borer) and Conogethes punctiferalis (Yellow Peach Moth) in Fusarium verticillioides Infection and Kernel Fumonisin Level.

Field experiments were conducted to evaluate the roles of two corn borers, Asian corn borer (ACB), Ostrinia furnacalis and yellow peach moth (YPM), Conogethes punctiferalis, in Fusarium verticillioides infection using green fluorescent protein (GFP) as a marker. Effects of insect injury, manual injury, and insecticide application on fumonisin production also were assessed. In this study, third instars of ACB and YPM significantly increased GFP-tagged F. verticillioides infection compared with the control, regardless of the fungal inoculation method. Besides acquiring F. verticillioides spores from leaf surfaces and transmitting them to ears, larvae of the ACB and YPM also injure maize ears, which allows F. verticillioides from leaves or silk to infect ears more easily. This suggests that ACB and YPM larvae are vectors of F. verticillioides, which can increase the occurrence of ear rot. Manual injuries significantly increased GFP-tagged F. verticillioides infection of ears, while effective insect control significantly reduced F. verticillioides infection of ears. Insecticide control of borers also significantly reduced fumonisin content in kernels. Larval infestations significantly increased fumonisins in kernels to levels higher than or very close to the European Union threshold (4,000 µg kg-1). Significant and high correlations among corn borer attack, F. verticillioides severity, and kernel fumonisin levels were discovered, confirming the important role of ACB and YPM activity in F. verticillioides infection and kernel fumonisin production.

4-octyl itaconate improves the viability of D66H cells by regulating the KEAP1-NRF2-GCLC/HO-1 pathway.

As a novel nuclear factor E2-related factor 2 (NRF2) activator, the itaconate has shown significant therapeutic potential for oxidative stress diseases. However, its role in Vohwinkel syndrome in relation to the gap junction protein beta 2 (GJB2) mutation is still unclear. This study aimed at investigating the effect of 4-octyl itaconate (OI) on HaCaT and D66H cells and clarify its potential mechanism in vitro. The optimal concentration and treatment time of OI on HaCaT cells and D66H cells were determined by CCK-8 and LDH experiments. The effect of OI on cell proliferation was detected by EdU staining and FACS analysis of PI, while the apoptosis was evaluated by TUNEL staining and FACS analysis of Annexin V. The ROS staining was performed, and the levels of SOD, MDA, GSH and GSH/GSSG were detected to evaluate the effect of OI on oxidative damage induced by D66H-type mutation. CO-IP, Western blot, immunofluorescence and qPCR analyses were employed to detect the activation of KEAP1-NRF2-GCLC/HO-1 pathway by OI. Finally, sh-NRF2 was used to confirm the activation of this pathway by OI. Results showed that OI could improve the cell viability decreased by GJB2 gene mutation by regulating the balance between cell growth and apoptosis induced by oxidative damage. Furthermore, this alleviation process was regulated by the KEAP1-NRF2-HO-1/GCLC pathway. In conclusion, OI could improve the viability of HaCaT and D66H cells via regulating the KEAP1-NRF2-GCLC/HO-1 pathway, which provided a wide spectrum of potential targets for effective therapeutic treatments of Vohwinkel syndrome in the clinic.

Secondary symbionts affect aphid fitness and the titer of primary symbiont.

Bacterial symbionts associated with aphids are important for their ecological fitness. The corn leaf aphid, Rhopalosiphum maidis (Fitch), is one of the most damaging aphid pests on maize and has been reported to harbor Hamiltonella defensa and Regiella insecticola while the effects of the secondary symbionts (S-symbionts) on host ecology and primary symbiont Buchnera aphidicola remain unclear. Here, four aphid strains were established, two of which were collected from Langfang - Hebei Province, China, with similar symbiont pattern except for the presence of H. defensa. Two other aphid strains were collected from Nanning - Guangxi Province, China, with the same symbiont infection except for the presence of R. insecticola. Phylogenetic analysis and aphid genotyping indicated that the S-symbiont-infected and free aphid strains from the same location had identical genetic backgrounds. Aphid fitness measurement showed that aphid strain infected with H. defensa performed shortened developmental duration for 1st instar and total nymph stages, reduced aphid survival rate, offspring, and longevity. While the developmental duration of H-infected strains was accelerated, and the adult weight was significantly higher compared to the H-free strain. Infection with R. insecticola did not affect the aphid's entire nymph stage duration and survival rate. As the H-strain does, aphids infected with R. insecticola also underwent a drop in offspring, along with marginally lower longevity. Unlike the H-infected strain, the R-infected strain performed delayed developmental duration and lower adult weight. The B. aphidicola titers of the H-infected strains showed a steep drop during the aphid 1st to 3rd instar stages, while the augmentation of B. aphidicola titers was found in the R-infected strain during the aphid 1st to 3rd instar. Our study investigated for the first time the effect of the S-symbionts on the ecology fitness and primary symbiont in R. maidis, indicating that infection with secondary symbionts leads to the modulation of aphid primary symbiont abundance, together inducing significant fitness costs on aphids with further impact on environmental adaptation and trophic interactions.

Seven Years of Monitoring Susceptibility to Cry1Ab and Cry1F in Asian Corn Borer.

Resistance monitoring in the Asian corn borer, Ostrinia furnacalis, is necessary to accommodate the commercial introduction and stewardship of Bt maize in China. The susceptibility of 56 O. furnacalis field populations, collected between 2015 and 2021 from the corn belt regions of China, to Cry1Ab and Cry1F toxins was determined. Neonate larvae (within 12 h after hatching) were placed on the surface of semi-artificial agar-free diet incorporating a series of concentrations of purified toxins, and mortality was evaluated after 7d. The median lethal concentration (LC50) values of Cry1Ab and Cry1F were 0.05 to 0.37 µg/g (protein/diet) and 0.10 to 1.22 µg/g, respectively. Although interpopulation variation in susceptibility to the toxins was observed, the magnitude of the differences was 5.8-fold and 8.3-fold for Cry1Ab and Cry1F, respectively. These results suggested that the observed susceptibility differences reflect natural geographical variation in response and not variation caused by prior exposure to selection pressures. Therefore, the O. furnacalis populations were apparently still susceptible to Cry1Ab and Cry1F across their range within China. The monitoring data established here will serve as a comparative reference for early warning signs of field-evolved resistance after the cultivation of Bt maize in China.

Intra-Population Alteration on Voltinism of Asian Corn Borer in Response to Climate Warming.

The Asian corn borer (ACB) Ostrinia furnacalis (Guenée) can occur in one to seven generations annually from cool (48°00' N) to warm (18°10' N) region of corn cultivation in China. Although ACB is commonly known as a facultative larval diapause insect, the co-existence of various voltinism suggests that intra-population variation may have evolved for the nature of diapause, i.e., voltinism plasticity. Here, we conducted recurrent selection efforts to establish three strains of, respectively, univoltine (with obligate diapause), multivoltine (with facultative diapause), and non-diapausing ACB under various temperature and photoperiod environments. The univoltine (Lu) strain has evolved a stable univoltinism under a diapause suppressing condition (16 h daylength at 28 °C), with the diapause incidence constantly over 80% after three generations of selection. The multivoltine strain (Lm) under the high temperature (28 °C) was shown to have a typical facultative diapause induced by a range of short-day lengths (11-13.5 h). Diapause incidence was constantly <2.6% under the long day length (16 h) when the temperature was from 18 to 28 °C, i.e., low temperature could not enhance the diapause response in the Lm strain. However, the development was prolonged from 14.2 ± 0.3 d to 46.0 ± 0.8 d when the temperature was reduced from 28 °C to 18 °C. The majority (94.4%) of the developed Ln strain still maintained the non-diapausing nature under a diapause enhancing condition, i.e., a short (13 h) daylength at a low temperature (22 °C). Lm and Ln were able to complete their second generation in Heihe (50°14' N) if the first-generation moth oviposits before 18 June. The study suggests that ACB has evolutionary intra-population variation in voltinism. Under the climate change scenario warmer spring and summer might affect the proportion of sympatric voltine biotype populations that evolve toward being multivoltine.

Analysis of the Genetic Diversity of Two Rhopalosiphum Species from China and Europe Based on Nuclear and Mitochondrial Genes.

Population genetic studies can reveal clues about the evolution of adaptive strategies of aphid species in agroecosystems and demonstrate the influence of environmental factors on the genetic diversity and gene flow among aphid populations. To investigate the genetic diversity of two Rhopalosiphum aphid species from different geographical regions, 32 populations (n = 535) of the bird cherry-oat aphid (Rhopalosiphum padi Linnaeus) and 38 populations (n = 808) of the corn leaf aphid (Rhopalosiphum maidis Fitch) from China and Europe were analyzed using one nuclear (elongation factor-1 alpha) and two mitochondrial (cytochrome oxidase I and II) genes. Based on the COI-COII sequencing, two obvious clades between Chinese and European populations and a low level of gene flow (Nm = 0.15) were detected in R. padi, while no geographical-associated genetic variation was found for EF-1α in this species. All genes in R. maidis had low genetic variation, indicating a high level of gene flow (Nm = 5.31 of COI-COII and Nm = 2.89 of EF-1α). Based on the mitochondrial result of R. padi, we concluded that the long distance between China and Europe may be interrupting the gene flow. The discordant results of nuclear gene analyses in R. padi may be due to the slower evolution of nuclear genes compared to mitochondrial genes. The gene exchange may occur gradually with the potential for continuous migration of the aphid. This study facilitates the design of control strategies for these pests.

Rotation of Multiple Single-Gene Transgenic Crops Did Not Slow the Evolution of Resistance to Cry1F or Cry1Ie in Ostrinia furnacalis.

A common strategy for delaying the evolution of resistance to transgenic crops that produce insecticidal proteins from Bacillus thuringiensis is to ensure that insect pests are exposed to multiple toxins with different mechanisms of action (MoAs). This can take the form of planting crops in a rotation pattern when different crops expressing single toxins are available on the market. The efficacy of a rotation strategy is reliant on mathematical models based on biological assumptions. Here, we designed laboratory evolution experiments to test whether Bt-based insecticidal proteins with different MoAs used in rotation could delay resistance from developing in Asian corn borer (ACB), Ostrinia furnacalis. We investigated the proteins Cry1Ab, Cry1F, and Cry1Ie, which are widely utilized for commercial insect control. We found that rotation of multiple toxins did not slow the evolution of resistance to Cry1F or Cry1Ie. Furthermore, the evolution of ACB to the Cry1Ab toxin develops faster when Cry1F or Cry1Ie is present, as compared to Cry1Ab exposure only. Our results suggest that toxins used in a rotation fashion do not work as an effective strategy in delaying ACB resistance evolution to Cry toxins over one-toxin exposure. Our result highlights the need to better understand the biological factors leading to insecticidal protein resistance and to develop IRM strategies against target insects.

Dynamics of gut microflora across the life cycle of Spodoptera frugiperda and its effects on the feeding and growth of larvae.

BACKGROUND: Spodoptera frugiperda is an important invasive agricultural pest that causes huge economic losses worldwide. Gut microorganisms play a vital role in host feeding, digestion, nutrition, immunity, growth and insecticide resistance. Illumina high-throughput sequencing was used to study the gut microbial community dynamics across the life cycle (egg, 1st to 6th instar larvae, pupae, and male and female adults) of S. frugiperda fed on maize leaves. Furthermore, the gut microbial community and food intake of the 5th instar S. frugiperda larvae were studied after feeding them antibiotics. RESULTS: Enterobacteriaceae and Enterococcaceae dominated the gut during growth and feeding of the larvae. The relative abundance of Enterobacteriaceae was higher in the 4th and 6th instar larvae. With the increase in larval feeding, the relative abundance of Enterococcaceae gradually increased. In addition, principal coordinate analysis and linear discriminant effect size analysis confirmed differences in the structure of gut microbiota at different developmental stages. After antibiotic treatment, the relative abundance of Firmicutes, Proteobacteria and Fusobacteriota decreased. The relative abundance of Enterococcus and Klebsiella decreased significantly. Antibiotic treatment inhibited the gut flora of S. frugiperda, which decreased larval food intake and body weight gain, and prolonged the larval stage. CONCLUSION: The composition of the gut bacterial community plays an important role in the growth, development, and feeding of S. frugiperda. The results have a certain theoretical value for the development of bio-pesticides targeting intestinal flora. © 2022 Society of Chemical Industry.

Genotypic variation in field-grown maize eliminates trade-offs between resistance, tolerance and growth in response to high pressure from the Asian corn borer.

Insect herbivory challenges plant survival, and coordination of the interactions between growth, herbivore resistance/tolerance is a key problem faced by plants. Based on field experiments into resistance to the Asian corn borer (ACB, Ostrinia furnacalis), we selected 10 inbred maize lines, of which five were resistant and five were susceptible to ACB. We conducted ACB larval bioassays, analysed defensive chemicals, phytohormones, and relative gene expression using RNA-seq and qPCR as well as agronomic traits, and found resistant lines had weaker inducibility, but were more resistant after ACB attack than susceptible lines. Resistance was related to high levels of major benzoxazinoids, but was not related to induced levels of JA or JA-Ile. Following combination analyses of transcriptome, metabolome and larval performance data, we discovered three benzoxazinoids biosynthesis-related transcription factors, NAC60, WRKY1 and WRKY46. Protoplast transformation analysis suggested that these may regulate maize defence-growth trade-offs by increasing levels of benzoxazinoids, JA and SA but decreasing IAA. Moreover, the resistance/tolerance-growth trade-offs were not observed in the 10 lines, and genotype-specific metabolic and genetic features probably eliminated the trade-offs. This study highlights the possibility of breeding maize varieties simultaneously with improved defences and higher yield under complex field conditions.

Asian Corn Borer (Ostrinia furnacalis) Infestation Increases Fusarium verticillioides Infection and Fumonisin Contamination in Maize and Reduces the Yield.

Field trials based on manual infestation of the Asian corn borer (ACB) (Ostrinia furnacalis [Guenée]) and Fusarium verticillioides (Nirenberg) atomization were conducted on four maize hybrids to investigate the relationship between ACB infestation and F. verticillioides infection, yield loss, and fumonisin contamination in maize. Analysis of fumonisins B1 and B2 was carried out using an LC-MS/MS system. In this study, manual ACB infestation significantly promoted F. verticillioides infection (both symptomatic and symptomless) and grain fumonisin levels. Ear rot incidence and severity, symptomless kernel infection, and fumonisin contamination were significantly correlated to each other and to ACB damage severity. Manual ACB infestation increased fumonisin levels from 580 to 4,418 µg/kg in 2018; 6,059 to 10,681 µg/kg in 2019 spring-sown maize (2019A); and 2,042 to 5,060 µg/kg in 2019 summer-sown maize (2019B), with the threshold of the European Union (EU) being 4,000 µg/kg. The threshold was exceeded in spring of 2019 in untreated controls. Regarding yield, significant negative correlation between ACB damage and ear weight was observed in three seasons. These results indicated that ACB infestation can lead to severe quality degradation and yield loss of maize. Kernel fumonisin levels may exceed the concentration threshold of the EU in certain conditions, threatening the health of livestock and humans. Measures should be taken to reduce ACB infestation to ensure food and feed security.

Seed Myco-priming improves crop yield and herbivory induced defenses in maize by coordinating antioxidants and Jasmonic acid pathway.

BACKGROUND: Seed Myco-priming based on consortium of entomopathogenic fungi is very effective seed treatment against Ostrinia furnacalis herbivory. Maize regulates defense responses against herbivory by the production of defense-related enzymatic and non-enzymatic antioxidants, phytohormones, and their corresponding genes. Jasmonic acid (JA) plays a key role in plant-entomopathogenic fungi-herbivore interaction. RESULTS: To understand how a consortium of the entomopathogenic fungi Beauveria bassiana and Trichoderma asperellum induce changes in the response of maize to herbivory and increase the crop yield, 2-year field experiment, antioxidant enzymes, leaf transcriptome, and phytohormone were performed. Fungal inoculation enhanced the production of antioxidant enzymes and JA signaling pathway more than the normal herbivory. The comparison between single inoculated, consortium inoculated, and non-inoculated plants resulted in distinct transcriptome profiles representing a considerable difference in expression of antioxidant- and JA- responsive genes identified through Weighted gene co-expression network analysis (WGCNA) and expression analysis, respectively. Seed priming with a consortium of B. bassiana and T. asperellum significantly enhanced the expression of genes involved in antioxidants production and JA biosynthesis cascade, with the highest expression recorded at 24-h post O. furnacalis larval infestation. They reduced the larval nutritional indices and survival up to 87% and enhancing crop yield and gross return up to 82-96% over the year 2018 and 2019. CONCLUSION: From our results we suggest that a consortium of B. bassiana and T. asperellum can be used synergistically against O. furnacalis in maize under field condition and can mediate antioxidants- and JA- associated maize defense response by boosting up the expression of their responsive genes, thereby enhancing crop yield.

Hemocyte response to treatment of susceptible and resistant Asian corn borer (Ostrinia furnacalis) larvae with Cry1F toxin from Bacillus thuringiensis.

Midgut receptors have been recognized as the major mechanism of resistance to Cry proteins in lepidopteran larvae, while there is a dearth of data on the role of hemocyte's response to Cry intoxication and resistance development. We aimed at investigating the role of circulating hemocytes in the intoxication of Cry1F toxin in larvae from susceptible (ACB-BtS) and resistant (ACB-FR) strains of the Asian corn borer (ACB), Ostrinia furnacalis. Transcriptome and proteome profiling identified genes and proteins involved in immune-related (tetraspanin and C-type lectins) and detoxification pathways as significantly up-regulated in the hemocytes of Cry1F treated ACB-FR. High-throughput in vitro assays revealed the binding affinity of Cry1F with the tetraspanin and C-type lectin family proteins. We found significant activation of MAPKinase (ERK 1/2, p38α, and JNK 1/2) in the hemocytes of Cry1F treated ACB-FR. In testing plausible crosstalk between a tetraspanin (CD63) and downstream MAPK signaling, we knocked down CD63 expression by RNAi and detected an alteration in JNK 1/2 level but a significant increase in susceptibility of ACB-FR larvae to Cry1F toxin. Information from this study advances a change in knowledge on the cellular immune response to Cry intoxication and its potential role in resistance in a lepidopteran pest.

Resistance of Spodoptera frugiperda to Cry1, Cry2, and Vip3Aa Proteins in Bt Corn and Cotton in the Americas: Implications for the Rest of the World.

The fall armyworm, Spodoptera frugiperda, is an economically important pest of corn, cotton, and soybean, and a major target of transgenic crops expressing Bacillus thuringiensis (Bt) proteins. In recent years, this insect has invaded most countries in Africa, Southeastern Asia, and Oceania, posing a great threat to food security. Successful use of Bt crops in the U.S. indicates that Bt technology can be an effective tool for management of S. frugiperda in other countries. Evolution of insect resistance is the primary threat to the long-term efficacy of Bt technology. There are many factors that may affect the rate of evolution of insect resistance to Bt crops, which include initial resistance allele frequency, the dose of Bt protein in Bt crops, cross-resistance, complete/incomplete resistance, and fitness costs associated with resistance. Currently, the high dose/refuge and gene-pyramiding approaches are the two main IRM strategies used in the U.S. to combat evolution of insect resistance. In this paper, we review research on resistance of S. frugiperda to Cry1, Cry2, and Vip3Aa proteins. Specifically, we discuss the resistance allele frequencies of S. frugiperda to these three proteins in the field, the genetic basis of resistance, the patterns of cross-resistance, and the fitness costs associated with resistance. Experience and knowledge gained from these studies provide valuable information for the successful use of Bt crop technology for control of S. frugiperda worldwide.

Impact of Temperature Change on the Fall Armyworm, Spodoptera frugiperda under Global Climate Change.

The fall armyworm (FAW), Spodoptera frugiperda (J. E. Smith, 1797), known as an important agricultural pest around the world, is indigenous to the tropical-subtropical regions in the Western Hemisphere, although its distribution has expanded over large parts of America, Africa, Asia and Oceania in the last few years. The pest causes considerable costs annually coupled with its strong invasion propensity. Temperature is identified as the dominant abiotic factor affecting herbivorous insects. Several efforts have reported that temperature directly or indirectly influences the geographic distribution, phenology and natural enemies of the poikilothermal FAW, and thus may affect the damage to crops, e.g., the increased developmental rate accelerates the intake of crops at higher temperatures. Under some extreme temperatures, the FAW is likely to regulate various genes expression in response to environmental changes, which causes a wider viability and possibility of invasion threat. Therefore, this paper seeks to review and critically consider the variations of developmental indicators, the relationships between the FAW and its natural enemies and the temperature tolerance throughout its developmental stage at varying levels of heat/cold stress. Based on this, we discuss more environmentally friendly and economical control measures, we put forward future challenges facing climate change, we further offer statistical basics and instrumental guidance significance for informing FAW pest forecasting, risk analyses and a comprehensive management program for effective control globally.

RNA Interference of Phenoloxidases of the Fall Armyworm, Spodoptera frugiperda, Enhance Susceptibility to Bacillus thuringiensis Protein Vip3Aa19.

Phenoloxidase (PO) is an important enzyme in the cellular immune system and is involved in defense against a wide range of pathogens, including Bacillus thuringiensis. Vip3Aa19 is secreted and expressed by Bacillus thuringiensis (Bt) at the middle exponential growth phase and is a kind of protein with efficient insecticidal activity against Spodoptera frugiperda. However, immune responses of the target insects have been regarded as a hindrance to Bt pathogenicity. This paper reports two phenoloxidase (PO) genes (SfPAE and SfPO2) identified from the hemocyte transcriptome data of the fall armyworm, Spodoptera frugiperda. qRT-PCR validation results showed that the expression levels of two PO genes were significantly upregulated after Vip3Aa19 (LC50 = 4.98 µg/g) toxin treatment compared with those of S. frugiperda fed an insecticide-free artificial diet. Meanwhile, two PO genes were expressed from the egg to adult stages even without an immune challenge. We noticed that at all developmental stages investigated in the S. frugiperda, SfPAE was generally expressed at a higher level than SfPO2. However, after Vip3Aa19 treatment, the SfPO2 gene mRNA expression level was significantly elevated in response to the toxin challenge. An injection of a specific double-stranded RNA (dsRNA) against POs could suppress its expression. The third instar larvae of S. frugiperda treated with dsRNA were much more susceptible to Vip3Aa19 toxin than the control larvae were. Notably, the mortality rate was nearly 90% after a dsPO2 injection. These results proved that SfPO2 was more important for the survival of S. frugiperda. Finally, RNA interference and then PO activity detection revealed that PO genes mainly existed in the hemolymph and played an important role in immune defense against Bt toxin.

Synergistic Effects of Graphene Oxide and Pesticides on Fall Armyworm, Spodoptera frugiperda.

Fall armyworm Spodoptera frugiperda, a native insect pest in tropical and subtropical America, has rapidly spread to most parts of China and become a major pest of corn and other crops since invading in early January 2019. As an emergency and important control measure, chemical control of S. frugiperda has the advantages of quick effect and low cost. However, long-term and large-scale use of pesticides might pollute the environment and increase pest resistance. By improving the control effect and reducing the dosage of chemical pesticides, graphene oxide (GO) is used synergistically with insecticides to increase control efficacy to achieve low-cost and sustainable management of insect pests as a new type of synergist. In this study, graphene oxide was compounded with insecticides to form nanocomposites. To clarify pest physiological responses, the laboratory toxicity of graphene oxide-insecticide nanocomposites was measured on the larvae of S. frugiperda. The results demonstrated that GO could enhance the activity of four selected pesticides: chlorantraniliprole (Chl), beta cypermethrin (Bet), methoxyhydrazide (Met) and spinetoram (Spi). Compared with pesticides alone, the toxicity of Chl-GO, Bet-GO, Met-GO and Spi-GO mixtures to the third instar larvae of S. frugiperda increased by 1.56, 1.54, 2.53 and 1.74 times, respectively. The easy preparation and higher bioactivity of GO-pesticide nanocomposites indicated their promising application potential in pest control.

Transcription dynamics of heat shock proteins in response to thermal acclimation in Ostrinia furnacalis.

Acclimation to abiotic stress plays a critical role in insect adaption and evolution, particularly during extreme climate events. Heat shock proteins (HSPs) are evolutionarily conserved molecular chaperones caused by abiotic and biotic stressors. Understanding the relationship between thermal acclimation and the expression of specific HSPs is essential for addressing the functions of HSP families. This study investigated this issue using the Asian corn borer Ostrinia furnacalis, one of the most important corn pests in China. The transcription of HSP genes was induced in larvae exposed to 33°C. Thereafter, the larvae were exposed to 43°C, for 2 h, and then allowed to recover at 27 C for 0, 0.5, 1, 2, 4, 6, and 8 h. At the recovery times 0.5-4 h, most population tolerates less around 1-3 h than without recovery (at 0 h) suffering continuous heat stress (43 C). There is no difference in the heat tolerance at 6 h recovery, with similar transcriptional levels of HSPs as the control. However, a significant thermal tolerance was observed after 8 h of the recovery time, with a higher level of HSP70. In addition, the transcription of HSP60 and HSC70 (heat shock cognate protein 70) genes did not show a significant effect. HSP70 or HSP90 significantly upregulated within 1-2 h sustained heat stress (43 C) but declined at 6 h. Our findings revealed extreme thermal stress induced quick onset of HSP70 or HSP90 transcription. It could be interpreted as an adaptation to the drastic and rapid temperature variation. The thermal tolerance of larvae is significantly enhanced after 6 h of recovery and possibly regulated by HSP70.