Correction: Nickel-catalyzed γ-alkylation of cyclopropyl ketones with unactivated primary alkyl chlorides: balancing reactivity and selectivity via halide exchange.

[This corrects the article DOI: 10.1039/D4RA02616K.].

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.

Revealing the difference of α-amylase and CYP6AE76 gene between polyphagous Conogethes punctiferalis and oligophagous C. pinicolalis by multiple-omics and molecular biological technique.

BACKGROUND: Conogethes pinicolalis has been thought as a Pinaceae-feeding variant of the yellow peach moth, Conogethes punctiferalis. The divergence of C. pinicolalis from the fruit-feeding moth C. punctiferalis has been reported in terms of morphology, ecology, and genetics, however there is a lack of detailed molecular data. Therefore, in this study, we investigated the divergence of C. pinicolalis from C. punctiferalis from the aspects of transcriptomics, proteomics, metabolomics and bioinformatics. RESULTS: The expression of 74,611 mRNA in transcriptome, 142 proteins in proteome and 218 metabolites in metabolome presented significantly differences between the two species, while the KEGG results showed the data were mainly closely related to metabolism and redox. Moreover, based on integrating system-omics data, we found that the α-amylase and CYP6AE76 genes were mutated between the two species. Mutations in the α-amylase and CYP6AE76 genes may influence the efficiency of enzyme preference for a certain substrate, resulting in differences in metabolic or detoxifying ability in both species. The qPCR and enzyme activity test also confirmed the relevant gene expression. CONCLUSIONS: These findings of two related species and integrated networks provide beneficial information for further exploring the divergence in specific genes, metabolism, and redox mechanism. Most importantly, it will give novel insight on species adaptation to various diets, such as from monophagous to polyphagous.

Toxicity of chemical pesticides commonly used in maize to Trichogramma ostriniae (Hymenoptera: Trichogrammatidae), an egg parasitoid of Asian corn borer.

Chemical pesticides are largely applied worldwide for the control of agricultural pests, and often cause lethal and sublethal side effects on natural enemies. To determine the compatibility of chemical pesticides, commonly used by maize growers, with Trichogramma ostriniae, the toxicities of four herbicides (pendimethalin, topramezone, glyphosate, and atrazine), three insecticides (imidacloprid, emamectin benzoate, and ethiofencarb), and three fungicides (propiconazole, benzoyl cyclazole, and difenoconazole) were tested under laboratory conditions, measuring residual toxicity to T. ostriniae adults, their lethal effects on the parasitoid's eggs inside the host egg, and on the reproduction of the parasitoid's female offspring. Three tested pesticides including ethiofencarb, glyphosate, and benzoyl cyclazole were slightly harmful to T. ostriniae adults, whereas the other seven pesticides were moderately harmful. The pesticide residues of ethiofencarb, and emamectin benzoate, showed slightly persistence of toxicity to T. ostriniae adults, in contrast to the remaining pesticides. Glyphosate, emamectin benzoate, imidacloprid, ethiofencarb, and propiconazole were classified as harmless, causing < 30% reduction in emergence rate of wasps from host eggs, while atrazine, pendimethalin, topramezone, benzoyl cyclazole, and difenoconazole were classified as slightly harmful, causing 31-52% reduction in parasitoid emergence rates. Furthermore, the number of eggs subsequently laid by T. ostriniae females that had emerged from parasitized host eggs treated with atrazine was significantly reduced compared to which of untreated parasitized host eggs. No negative effects were observed with atrazine treatment on the emergence rates of F1 wasps, their wing deformity rates and sex ratios. The data presented here indicate that the safety interval between pesticide applications and releases of T. ostriniae wasps is 2-3 weeks, which provide valuable information in designing a sound maize integrated pest management program that combines chemical pesticide applications with augmentative releases of T. ostriniae wasps.

Synergistic interaction of Cry1Ah and Vip3Aa19 proteins combination with midgut ATP-binding cassette subfamily C receptors of Conogethes punctiferalis (Guenée) (Lepidoptera: Crambidae).

Bacillus thuringiensis Cry and Vip proteins are highly effective at controlling agricultural pests and could be used in pyramided transgenic crops. However, the molecular mechanism underlying the Cry1Ah and Vip3Aa19 synergistic interaction has never been investigated at the molecular level in Yellow peach moth (YPM) Conogethes punctiferalis. Binding affinity and synergism of Cry1Ah and Vip3Aa19 proteins with ABC transporter subfamily C receptors ABCC1, ABCC2 and ABCC3 proteins from the midgut of YPM larva by using surface plasmon resonance (SPR) and pull-down assays. Both assays revealed that Cry1Ah could interact with ABCC1, ABCC2, and ABCC3, whereas Vip3Aa19 only interacts with ABCC1 and ABCC3, but not with ABCC2. Hence, when compared to the Vip3Aa19 protein, Cry1Ah had a higher binding affinity for ABCC1, ABCC2, and ABCC3. Furthermore, competitive binding assay between Cry1Ah and Vip3Aa19 protein with ABC transporter subfamily C receptors resulted in the final eluted protein samples displaying vibrant blue bands of Cry1Ah and very faint bands of Vip3Aa19. Suggesting that Cry and Vip proteins could deliver a synergistic effect after cleaving the midgut proteases. Therefore, this finding indicated that the Cry1Ah and Vip3Aa19 do not compete for interacting with midgut receptors and thus provide strong synergism against YPM.

Myco-Synergism Boosts Herbivory-Induced Maize Defense by Triggering Antioxidants and Phytohormone Signaling.

BACKGROUND: Biocontrol strategies are the best possible and eco-friendly solution to develop resistance against O furnacalis and improve the maize yield. However, the knowledge about underlying molecular mechanisms, metabolic shifts, and hormonal signaling is limited. METHODS: Here, we used an axenic and a consortium of entomopathogenic Beauveria bassiana OFDH1-5 and a pathogen-antagonistic Trichoderma asperellum GDFS1009 in maize and observed that consortium applications resulted in higher chlorophyll contents and antioxidants activities [superoxide dismutase (SOD), peroxidase (POD), proline, protease, and polyphenol oxidase (PPO)] with a decrease in O. furnacalis survival. We performed a comprehensive transcriptome and an untargeted metabolome profiling for the first time at a vegetative stage in fungal inoculated maize leaves at 0-, 12-, 24-, 48-, and 72-h post insect infestation. RESULTS: The consortium of B. bassiana and T. asperellum leads to 80-95% of O. furnacalis mortality. A total of 13,156 differentially expressed genes were used for weighted gene coexpression network analysis. We identified the six significant modules containing thirteen candidate genes [protein kinase (GRMZM2G025459), acyl-CoA dehydrogenase (GRMZM5G864319), thioredoxin gene (GRMZM2G091481), glutathione S-transferase (GRMZM2G116273), patatin-like phospholipase gene (GRMZM2G154523), cytochrome P450 (GRMZM2G139874), protease inhibitor (GRMZM2G004466), (AC233926.1_FG002), chitinase (GRMZM2G453805), defensin (GRMZM2G392863), peroxidase (GRMZM2G144153), GDSL- like lipase (AC212068.4_FG005), and Beta-glucosidase (GRMZM2G031660)], which are not previously reported that are highly correlated with Jasmonic acid - Ethylene (JA-ET) signaling pathway and antioxidants. We detected a total of 130 negative and 491 positive metabolomic features using a ultrahigh-performance liquid chromatography ion trap time-of-flight mass spectrometry (UHPLC-QTOF-MS). Intramodular significance and real time-quantitative polymerase chain reaction (RT-qPCR) expressions showed that these genes are the true candidate genes. Consortium treated maize had higher jasmonic acid (JA), salicylic acid (SA), and ethylene (ET) levels. CONCLUSION: Our results provide insights into the genetics, biochemicals, and metabolic diversity and are useful for future biocontrol strategies against ACB attacks.

Managing the Invasive Fall Armyworm through Biotech Crops: A Chinese Perspective.

In late 2018, the highly destructive and polyphagous fall armyworm was first detected in China. It is now a major economic threat to corn production. In this article, the main control strategies that are available are reviewed and prospects to manage this pest with Bacillus thuringiensis (Bt) corn in China are discussed.

Comparison of larval performance and oviposition preference of Spodoptera frugiperda among three host plants: Potential risks to potato and tobacco crops.

Fall armyworm (FAW), Spodoptera frugiperda (J.E. Smith), is a major polyphagous pest with the potential to seriously damage various crops. A better understanding of FAW's performance on different hosts may help to predict which plants will be attacked when the preferred host is absent, and facilitate the development of effective pest management practices. We compared the larval performance of FAW fed on maize with that of FAW fed on potato and tobacco, which are important crops in China, using an age-stage two-sex life table and adult female oviposition preference experiments. In cage experiments with potato, tobacco, or maize as the host, FAW reared on maize exhibited the strongest performance with shorter developmental time in the larval stage, longer longevity, and a higher reproductive rate in adults. Females oviposited on maize in preference to potato or tobacco. Compared with larvae fed on maize, those fed on potato and tobacco exhibited significantly lower survival, with only 31.61% and 8.13% developing to the adult stage, respectively. Several life table parameters, including the mean generation time (T), net reproductive rate (R0 ), finite rate of increase (λ), and intrinsic rate of natural increase (r) were negatively affected in FAW fed on potato and tobacco. Our results support the preference-performance hypothesis, that is, that herbivore females maximize fitness by choosing host plants associated with strong larval performance. Although larvae and adults performed poorly on potato and tobacco, their offspring will be capable of establishing populations on them, posing a potential threat to these crops in China.

Synergistic Effect of Beauveria bassiana and Trichoderma asperellum to Induce Maize (Zea mays L.) Defense against the Asian Corn Borer, Ostrinia furnacalis (Lepidoptera, Crambidae) and Larval Immune Response.

Ostrinia furnacalis, is the major pest of maize causing significant yield losses. So far, many approaches have been used to increase the virulence of entomopathogenic fungal isolates. The current study is an attempt to estimate synergistic effect of Beauveria bassiana and Trichoderma asperellum in order to explore larval immune response through RNA sequencing and differentially expression analysis. In vivo synergism was examined in seven proportions (B. bassiana: T. asperellum = 1:1, 1:2, 1:3, 1:4, 4:1, 3:1, 2:1) and in the in vitro case, two inoculation methods were applied: seed coating and soil drenching. Results revealed significant decrease in plant damage and high larval mortality in fungal treatments. Fungal isolates mediated the plant defense by increasing proline, superoxide dismutase (SOD), peroxidase (POD), polyphenol oxidase (PPO) and protease activities. Seed coating method was proved to be the most effective in case of maize endophytic colonization. In total, 59 immune-related differentially expressed genes DEGs were identified including, cytochrome P450, heat shock protein, ABC transporter, cadherin, peptidoglycan recognition protein (PGRP), cuticlular protein, etc. Further, transcriptomic response was confirmed by qRT-PCR. Our results concluded that, coculture of B. bassiana and T. asperellum has the synergistic potential to suppress the immune response of O. furnacalis and can be used as sustainable approach to induce plant resistance through activation of defense-related enzymes.

Contribution of phenoloxidase activation mechanism to Bt insecticidal protein resistance in Asian corn borer.

Phenoloxidase (PO) is a crucial enzyme in the Arthropods melanization process, in which synthesized melanin rapidly acts at the site of injury and infection. In this study, we observed significant changes in humoral and cellular responses after exposing susceptible and resistant strains to a sub-lethal concentration of Cry1Ah toxin. Based on STRING v 11.0 computational protein-protein interaction analysis, we selected seven immune genes namely Prophenoloxidase PPO1b, PP03, Serpin-3, Serpin-5, Beta-1,3-glucan recognition protein, Immulectin-3 and Serine protease SP105 reported in Asian corn borer. Quantitative real-time PCR gene expression studies showed Cry1Ah resistant strain had higher expression of PPO1b, PP03, Serpin-3, Beta-1,3-glucan recognition protein, Immulectin-3 and Serine protease SP105 genes in midgut and hemocyte samples. This study also investigated and found that the level of prophenoloxidation (proPO) activity in Cry1Ah resistant strains was significantly higher than susceptible strains. Cry1Ah toxin significantly increased the resistant strain's immune responses, the difference was observed through assays of bacterial agglutination and phagocytosis. Additionally, immune response induced by Cry1Ah toxin influences the microbiome composition associated with the host system. These parameters seem to explain the contribution of PO/PO regulating proteins render the host to resist the Cry1Ah toxin.

Differential wing polyphenism adaptation across life stages under extreme high temperatures in corn leaf aphid.

Polyphenism, a common phenomenon in nature, is an important form of adaptation in a diverse environment. Corn leaf aphid (CLA), Rhopalosiphum maidis, (Hemiptera: Aphididae), exhibit wing polyphenism in response to poor habitat quality. In this study, we focused on the effects of crowding and thermal cues on morph determination of CLA. Five developmental stages of aphids (1st to 4th nymphs and maternal adults) with increased population densities, were tested under two kinds of temperature patterns, i.e., A) a constant temperature of 22 °C with 2 h exposure to high temperature in the range of 35 to 39 °C during mid-photophase and B) different constant temperatures in the range of 22-30 °C with 2 h exposure to high temperature of 39 °C during mid-photophase. Crowding was found to directly impact winged induction. The 1st and 2nd nymphs were more sensitive for alate morphs induction under high density. In addition, temperature played a significant role in wing production, with the temperature setting of 26/39 °C in pattern B inducing higher alate morphs and survival than other temperature settings. Therefore, we hypothesize that warmer climate with brief high temperature is more favourable for survival and alate morphs production, but cool weather and transient extreme high temperature (>39 °C) is detrimental for CLA. Our results provide a new perspective on understanding the interactions between changes in extreme high temperatures and insect densities that differentially affect wing polymorphism for further demographic and distribution rates of species across temporal and spatial scales.

Mechanistic effect of the human GJB6 gene and its mutations in HaCaT cell proliferation and apoptosis.

We constructed lentiviral vectors containing the human wild-type GJB6 gene and the mutant variants A88V and G11R. The three proteins were stably expressed by the Tet-on system in the HaCaT cell line and used to study the functional effect of the variants. The CCK-8 assay and flow cytometric analyses were used to determine the levels of cell proliferation and apoptosis. Western blot analyses were performed to analyze the relevant clinical indicators of hidrotic ectodermal dysplasia and markers of apoptosis in transfected HaCaT cells. The CCK8 assay and the flow cytometry results showed a significant increase (P<0.05) in the apoptosis of HaCaT cells expressing the A88V and G11R mutants. In addition, we demonstrated that the A88V and G11R mutants induced the apoptosis of transfected HaCaT cells via the activation of caspase-3, -8, -9, and PARA. No change was observed in the activity of BAX compared with the control. This study provides further clarification on the mechanisms underlying the effect of the mutant variants A88V and G11R of the GJB6 gene on the induction of HaCaT cell apoptosis.

Clip domain prophenoloxidase activating protease is required for Ostrinia furnacalis Guenée to defend against bacterial infection.

The prophenoloxidase (PPO) activating system in insects plays an important role in defense against microbial invasion. In this paper, we identified a PPO activating protease (designated OfPAP) containing a 1203 bp open reading frame encoding a 400-residue protein composed of two clip domains and a C-terminal serine protease domain from Ostrinia furnacalis. SignalP analysis revealed a putative signal peptide of 18 residues. The mature OfPAP was predicted to be 382 residues long with a calculated Mr of 44.8 kDa and pI of 6.66. Multiple sequence alignment and phylogenetic analysis indicated that OfPAP was orthologous to the PAPs in the other lepidopterans. A large increase of the transcript levels was observed in hemocytes at 4 h post injection (hpi) of killed Bacillus subtilis, whereas its level in integument increased continuously from 4 to 12 hpi in the challenged larvae and began to decline at 24 hpi. After OfPAP expression had been silenced, the median lethal time (LT50) of Escherichia coli-infected larvae (1.0 day) became significantly lower than that of E. coli-infected wild-type (3.0 days, p < 0.01). A 3.5-fold increase in E. coli colony forming units occurred in larval hemolymph of the OfPAP knockdown larvae, as compared with that of the control larvae not injected with dsRNA. There were notable decreases in PO and IEARase activities in hemolymph of the OfPAP knockdown larvae. In summary, we have demonstrated that OfPAP is a component of the PPO activation system, likely by functioning as a PPO activating protease in O. furnacalis larvae.

Mechanistic effect of the human GJB6 gene and its mutations in HaCaT cell proliferation and apoptosis

We constructed lentiviral vectors containing the human wild-type GJB6 gene and the mutant variants A88V and G11R. The three proteins were stably expressed by the Tet-on system in the HaCaT cell line and used to study the functional effect of the variants. The CCK-8 assay and flow cytometric analyses were used to determine the levels of cell proliferation and apoptosis. Western blot analyses were performed to analyze the relevant clinical indicators of hidrotic ectodermal dysplasia and markers of apoptosis in transfected HaCaT cells. The CCK8 assay and the flow cytometry results showed a significant increase (P<0.05) in the apoptosis of HaCaT cells expressing the A88V and G11R mutants. In addition, we demonstrated that the A88V and G11R mutants induced the apoptosis of transfected HaCaT cells via the activation of caspase-3, -8, -9, and PARA. No change was observed in the activity of BAX compared with the control. This study provides further clarification on the mechanisms underlying the effect of the mutant variants A88V and G11R of the GJB6 gene on the induction of HaCaT cell apoptosis.

Physiological Responses Induced by Ostrinia furnacalis (Lepidoptera: Crambidae) Feeding in Maize and Their Effects on O. furnacalis Performance.

λ: Plants damaged by herbivorous insects often respond by mounting a series of defense responses that can inhibit the insect's fitness. Ostrinia furnacalis (Guenée) (Lepidoptera: Crambidae) is a major insect pest in maize throughout much of Asia, Australia, and the western Pacific islands. We examined the effects of O. furnacalis -induced maize defenses on O. furnacalis fitness, and explained the effects from biochemical changes that occur in maize leaves in response to O. furnacalis feeding. The results of the age-stage, two-sex life table showed that significantly longer larval and pupal life spans, and total preoviposition period (TPOP) occurred. A decrease in the longevity and fecundity of female adults was observed in O. furnacalis fed on O. furnacalis -damaged leaves. The mean generation time ( T ), finite rate of increase ( ), net reproductive rate ( R 0 ), and intrinsic rate of increase ( r ) were also correspondingly affected. Biochemical assays indicated that 24 h of O. furnacalis herbivory resulted in decreased levels of the benzoxazinoids, 2,4-dihydroxy-7-methoxy-1,4-benzoxazin-3-one (DIMBOA), and 2-(2,4-dihydroxy-7-methoxy-1,4-benzoxazin-3-one)-ß-D-glucopyranose (DIMBOA-Glc), and a corresponding increase in 2-(2-hydroxy-4,7-dimethoxy-1,4-benzoxazin-3-one)-ß-D-glucopyranose (HDMBOA-Glc). Maize also exhibited higher activities of the defensive enzymes-peroxidase (POD), superoxide dismutase (SOD), catalase (CAT), and polyphenol oxidase (PPO)-after 24 h of herbivory. We concluded that exposure to O. furnacalis -damaged leaves had an inhibitory impact on the fitness of the neonate to pupa stages of O. furnacalis . The observed higher level of HDMBOA-Glc and higher enzymatic activities of POD, SOD, CAT, and PPO may account, in part, for the observed inhibitory effects on O. furnacalis fitness.

Identification of putative chemosensory receptor genes from yellow peach moth Conogethes punctiferalis (Guenée) antennae transcriptome.

The yellow peach moth, Conogethes punctiferalis, is an extremely important polyphagous insect in Asia. The chemosensory systems of moth play an important role in detecting food, oviposition sites and mate attraction. Several antennal chemosensory receptors are involved in odor detection. Our study aims to identify chemosensory receptor genes for potential applications in behavioral responses of yellow peach moth. By transcriptomic analysis of male and female antennae, 83 candidate chemosensory receptors, including 62 odorant receptors, 11 ionotropic receptors and 10 gustatory receptors were identified. Through Blast and sequence alignment, the highly conserved co-receptor Orco was annotated, eight unigenes clustered into pheromone receptors, and two clustered as sugar receptor. Among the IRs, one unigenes was similar with co-receptors IR25a. Expression levels of 50 odorant receptors were further evaluated by quantitative real-time PCR in antennae. All the ORs tested were detected in antennae and some of which were associated with sex-biased expression. The chemosensory receptors identified in C. punctiferalis provide a foundational resource for further analysis on olfaction for behavior. The expression profiles of ORs in antennae indicated variant functions in olfactory recognition, and our results provided the possibility for the potential application of semiochemical to control this pest moth.

Molecular cloning, characterization and expression profiling of a ryanodine receptor gene in Asian corn borer, Ostrinia furnacalis (Guenée).

Ryanodine receptor (RyR) Ca(2+) release channel is the target of diamide insecticides, which show selective insecticidal activity against lepidopterous insects. To study the molecular mechanisms underlying the species-specific action of diamide insecticides, we have cloned and characterized the entire cDNA sequence of RyR from Ostrinia furnacalis (named as OfRyR). The OfRyR mRNA has an Open Reading Frame of 15324 bp nucleotides and encodes a 5108 amino acid polypeptide that displays 79-97% identity with other insects RyR proteins and shows the greatest identity with Cnaphalocrocis medinalis RyR (97%). Quantitative real-time PCR showed that the OfRyR was expressed at the lowest level in egg and the highest level in adult. The relative expression level of OfRyR in first, third and fifth-instar larva were 1.28, 1.19 and 1.99 times of that in egg. Moreover, two alternative splicing sites were identified in the OfRyR gene. One pair of mutually exclusive exons (a/b) were present in the central part of the predicted SPRY domain, and an optional exon (c) was located between the third and fourth RyR domains. Diagnostic PCR demonstrated that exons a and b existed in all developmental stages of OfRyR cDNA, but exon c was not detected in the egg cDNA. And the usage frequencies of these exons showed a significant difference between different developmental stages. These results provided the crucial basis for the functional expression of OfRyR and for the discovery of compound with potentially selective insect activtity.