Search Results (28)

Agasicles hygrophila is the most effective natural enemy for the control of the invasive weed Alternanthera philoxeroides (Mart.) Griseb. However, research on the gene function and potential genetic improvement of A. hygrophila is limited due to a lack of effective genetic tools. In this study, we employed the A. hygrophila white (AhW) gene as a target gene to develop a CRISPR/Cas9-based gene editing method applicable to A. hygrophila. We showed that injection of Cas9/sgRNA ribonucleoprotein complexes (RNPs) of the AhW gene into pre-blastoderm eggs induced genetic insertion and deletion mutations, leading to white eyes. Our results demonstrate that CRISPR/Cas9-mediated gene editing is possible in A. hygrophila, offering a valuable tool for studies of functional genomics and genetic improvement of A. hygrophila, which could potentially lead to more effective control of invasive weeds through the development of improved strains of this biocontrol agent. In addition, the white-eyed mutant strain we developed could potentially be useful for other transgenic research studies on this species. Full article

Glutathione S-transferase (GST) is a class of detoxifying enzymes in the second stage of insect metabolism and plays a key role in insecticide resistance. In this study, based on the transcriptome sequences of S. frugiperda, the full-length cDNA of SfGSTD1 was cloned and characterized. The temporal and spatial expression pattern showed that SfGSTD1 was highly expressed in Malpighian tubules, which are key excretion organs. Knocking down SfGSTD1 reduced S. frugiperd tolerance to abamectin and beta-cypermethrin. The overexpression of SfGSTD1 enhanced the viability of Sf9 cell under abamectin and beta-cypermethrin treatment. Furthermore, SfGSTD1 was overexpressed in Drosophila melanogaster using the GAL4/UAS binary expression system, and this overexpression strain was also less susceptible to abamectin and beta-cypermethrin. The enzyme activity of recombinant SfGSTD1 could also be significantly inhibited by abamectin and beta-cypermethrin. Taken together, our findings indicate that SfGSTD1 might be involved in the tolerance of abamectin and beta-cypermethrin in S. frugiperda. And these results provide theoretical foundations for understanding the resistance mechanism of S. frugiperda to abamectin and beta-cypermethrin. Full article

At the microscale, the three-dimensional morphological features of contact surfaces have a significant impact on the performance of electrical contacts. This paper aims to reconstruct the microscopic contact state of contact groups and to deeply study the effect of contact morphological features on electrical contact performance. To fully obtain multimodal data such as the three-dimensional micro-morphological features and chemical composition distribution of contact surfaces, this paper proposes a contact surface feature-matching method based on entropy rate superpixel seed point adaptive morphological reconstruction. This method can adaptively retain meaningful seed points while filtering out invalid seed points, effectively solving the problem of over-segmentation in traditional superpixel segmentation method. Experimental results show that the proposed method achieves a segmentation accuracy of 92% and reduces over-segmentation by 30% compared to traditional methods. Subsequently, on the basis of the moving and static contact group difference plane model and the W-M model, this paper constructs a three-dimensional surface fractal contact model with an irregular base. This model has the ability to layer simulate multi-parameter elastic and plastic and to extract fractal parameter point cloud height, which can more accurately reflect the actual contact state of the contact group. The model demonstrates a 15% improvement in contact area prediction accuracy and a 20% reduction in contact resistance estimation error compared to existing models. Finally, this paper compares and verifies the theoretical feasibility of the model, providing a new theoretical contact model for the study of the impact of three-dimensional micro-morphology on the electrical contact reliability. Full article

Cry2Ab is a significant alternative Bacillus thuringiensis (Bt) protein utilized for managing insect resistance to Cry1 toxins and broadening the insecticidal spectrum of crops containing two or more Bt genes. Unfortunately, the identified receptors fail to fully elucidate the mechanism of action underlying Cry2Ab. Previous studies have demonstrated the involvement of vacuolar H⁺-ATPase subunits A, B, and E (V-ATPase A, B, and E) in Bt insecticidal activities. The present study aims to investigate the contribution of V-ATPase C to the toxicities of Cry2Ab against Helicoverpa armigera. The feeding of Cry2Ab in H. armigera larvae resulted in a significant decrease in the expression of V-ATPase C. Further investigations confirmed the interaction between V-ATPase C and activated Cry2Ab protein according to Ligand blot and homologous and heterologous competition assays. Expressing endogenous HaV-ATPase C in Sf9 cells resulted in an increase in Cry2Ab cytotoxicity, while the knockdown of V-ATPase C by double-stranded RNAs (dsRNA) in midgut cells decreased Cry2Ab cytotoxicity. Importantly, a higher toxicity of the mixture containing Cry2Ab and V-ATPase C against insects was also observed. These findings demonstrate that V-ATPase C acts as a binding receptor for Cry2Ab and is involved in its toxicity to H. armigera. Furthermore, the synergy between V-ATPase C protein and Cry2Ab protoxins provides a potential strategy for enhancing Cry2Ab toxicity or managing insect resistance. Full article

Expressions of a wide range of cytoprotective counter-defense genes are mainly regulated by the Keap1-Nrf2-ARE signaling pathway in response to oxidative stress from xenobiotics. Gossypol is the major antiherbivore secondary metabolite of cotton, but how the polyphagous pest Helicoverpa armigera copes with this phytochemical to utilize its favorite host plant cotton remains largely elusive. In this study, we first suppressed the Keap1 gene in newly hatched larvae of cotton bollworm by feeding them the siRNA diet for 4 days. All of the larvae were subsequently fed the artificial diet supplied with gossypol or the control diet for 5 days. We identified that the knockdown of the Keap1 gene significantly decreased larval mortality and significantly increased the percentages of larval survival, reaching the fourth instar, compared with ncsiRNA when exposed to a diet containing gossypol. Three counter-defense genes CYP9A17, CYP4L11 and UGT41B3, which were related to the induction or metabolism of gossypol according to the report before, were all significantly up-regulated after the knockdown of the Keap1 gene. The Antioxidant Response Elements (AREs) were also detected in the promoter regions of the three counter-defense genes above. These data indicate that the suppression of the Keap1 gene activates the Keap1-Nrf2-ARE signaling pathway, up-regulates the expressions of counter-defense genes involved in the resistance of oxidative stress and finally contributes to reducing the susceptibility of gossypol. Our results provide more knowledge about the transcriptional regulation mechanisms of counter-defense genes that enable the cotton bollworm to adapt to the diversity of host plants including cotton. Full article

The polyphagous fall armyworm (FAW), Spodoptera frugiperda, has become an invasive pest worldwide in recent years. To develop maize germplasm with multiple pest resistance and understand genetic inheritance, 12 experimental hybrids (six pairs of reciprocal crosses) with diverse genetic backgrounds and four commercial checks were examined for FAW resistance in 2013 and 2014. The experiment utilized a randomized complete block design with four replications as the block factor. FAW injury on maize plants was assessed at 7 and 14 d after the artificial infestation at the V6 stage, and predatory arthropod taxa and abundance on maize seedlings were recorded 7 d after the infestation. Spodoptera frugiperda resistance varied significantly among the 16 hybrids. Two reciprocal crosses (‘FAW1430’ × ‘Oh43’ and ‘CML333’ × ‘NC358’) showed the least FAW injury. Eleven arthropod predators [i.e., six coleopterans, three hemipterans, earwigs (dermapterans), and spiders (or arachnids)] were also recorded; the two most common predators were the pink spotted ladybeetle, Coleomegilla maculata, and the insidious flower (or minute pirate) bug, Orius spp. Predator abundance was not correlated to FAW injury but varied greatly between 2013 and 2014. Principal component analysis demonstrated that, when compared with FAW resistant (or Bt-transgenic) checks (‘DKC69-71’, ‘DKC67-88’, and ‘P31P42’), five pairs of the reciprocal crosses had moderate FAW resistance, whereas a pair of reciprocal crosses (‘NC350’ × ‘NC358’ and NC358 × NC350) showed the same FAW susceptibility as the non-Bt susceptible check ‘DKC69-72’. Both parents contributed similarly to FAW resistance, or no maternal/cytoplasmic effect was detected in the experimental hybrids. Full article

Target pests of genetically engineered crops producing both defensive allelochemicals and Bacillus thuringiensis (Bt) toxins often sequentially or simultaneously uptake allelochemicals, Bt toxins, and/or insecticides. How the three types of toxins interact to kill pests remains underexplored. Here we investigated the interactions of Bt toxin Vip3A, plant allelochemical flavone, and insecticide emamectin benzoate in Spodoptera frugiperda. Simultaneous administration of flavone LC₂₅ + Vip3A LC₂₅, emamectin benzoate LC₂₅ + Vip3A LC₂₅, and flavone LC₁₅ + emamectin benzoate LC₁₅ + Vip3A LC₁₅ but not flavone LC₂₅ + emamectin LC₂₅ yielded a mortality significantly higher than their expected additive mortality (EAM). One-day pre-exposure to one toxin at LC₅ followed by six-day exposure to the same toxin at LC₅ plus another toxin at LC₅₀ showed that the mortality of flavone LC₅ + Vip3A LC₅₀, emamectin benzoate LC₅ + Vip3A LC₅₀, and Vip3A LC₅ + emamectin benzoate LC₅₀ were significantly higher than their EAM, while that of flavone LC₅ + emamectin benzoate LC₅₀ was significantly lower than their EAM. No significant difference existed among the mortalities of Vip3A LC₅ + flavone LC₅₀, emamectin benzoate LC₅ + flavone LC₅₀, and their EAMs. The results suggest that the interactions of the three toxins are largely synergistic (inductive) or additive, depending on their combinations and doses. Full article

To achieve high-quality corporate development, it is essential to undergo green transformation. Unlike previous literature, this paper explores relevant mechanisms affecting firms’ green transformation from a novel perspective of green credit guidelines (GCG). Using A-share listed industrial enterprises in China from 2010 to 2020 as a sample, we combine the generalized moment estimation model (GMM) with the difference-in-difference model (DID) and demonstrate that (1) GCG significantly promotes enterprises’ green transformation measured by the method of super-efficient Slacks-Based Measure and Data Envelopment Analysis (SBM-DEA). (2) GCG tends to improve green technology innovation, which further facilitates firms’ green transformation, and corporate social responsibility (CSR) reinforces the positive relationship between GCG and firms’ green transformation. Correspondingly, we provide policy recommendations for China and other developing countries. Full article

To better understand the geothermal system of the Gonghe Basin, we deployed 471 magnetotelluric survey points with an average distance of 2~3 km, covering the eastern and southern areas of the Basin. We used ModEM inversion software to carry out 3D inversion of 431 survey points and established a 3D-electrical model at a depth of 50 km in the area. The resistivity model shows that the low resistivity in the shallow part of the basin is related to the Cenozoic loose sedimentary cover, while the resistivity values of the mountains around the basin and the magmatic rock uplift zone are higher. The electrical model also shows that the high-conductivity layer is widely distributed in the middle and lower crust (15~35 km) of the basin, and direction of the high-conductivity layer is consistent with that of NW–SE fault in the basin. These high-conductivity layers may be the principal reason for the high heat flow values in the Gonghe Basin. Our resistivity model also shows that there is an obvious discontinuity between high- and low-resistivity blocks at different depths in the middle and upper crust. These discontinuities are consistent with the faults observed on the surface, which are related to the strong topographic relief. Our electrical model shows that these faults in the middle and upper crust are connected with the high-conductivity layer as the channel of heat transfer to the shallow part. Finally, the heat energy is enriched in the Triassic granite to form dry hot rock (HDR). The 3D-magnetotelluric imaging results depict the 3D-distribution characteristics of the geothermal system in the eastern and southern parts of the Gonghe Basin. Full article

Flavone is widely found in plants and plays an important role in plant defense against pests. Many pests, such as Helicoverpa armigera, use flavone as a cue to upregulate counter-defense genes for detoxification of flavone. Yet the spectrum of the flavone-inducible genes and their linked cis-regulatory elements remains unclear. In this study, 48 differentially expressed genes (DEGs) were found by RNA-seq. These DEGs were mainly concentrated in the retinol metabolism and drug metabolism-cytochrome P450 pathways. Further in silico analysis of the promoter regions of 24 upregulated genes predicted two motifs through MEME and five previously characterized cis-elements including CRE, TRE, EcRE, XRE-AhR and ARE. Functional analysis of the two predicted motifs and two different versions of ARE (named ARE1 and ARE2) in the promoter region of the flavone-inducible carboxylesterase gene CCE001j verified that the two motifs and ARE2 are not responsible for flavone induction of H. armigera counter-defense genes, whereas ARE1 is a new xenobiotic response element to flavone (XRE-Fla) and plays a decisive role in flavone induction of CCE001j. This study is of great significance for further understanding the antagonistic interaction between plants and herbivorous insects. Full article

In this work, we study the effect of substituents in cobalt(II) and iron(II) phthalocyanines (CoPcR₄ and FePcR₄ with R = H, F, Cl, tBu) on the structural features of their films, and their chemi-resistive sensor response to a low concentration of nitric oxide. For the correct interpretation of diffractograms of phthalocyanine films, structures of CoPcCl₄ and FePcCl₄ single crystals were determined for the first time. Films were tested as active layers for the determination of low concentrations of NO (10–1000 ppb). It was found that the best sensor response to NO was observed for the films of chlorinated derivatives MPcCl₄ (M = Co, Fe), while the lowest response was in the case of MPc(tBu)₄ films. FePcCl₄ films exhibited the maximal response to NO, with a calculated limit of detection (LOD) of 3 ppb; the response and recovery times determined at 30 ppb of NO were 30 s and 80 s, respectively. The LOD of a CoPcCl₄ film was 7 ppb. However, iron phthalocyanine films had low stability and their sensitivity to NO decreased rapidly over time, while the response of cobalt phthalocyanine films remained stable for at least several months. In order to explain the obtained regularities, quantum chemical calculations of the binding parameters between NO and phthalocyanine molecules were carried out. It was shown that the binding of NO to the side atoms of phthalocyanines occurred through van der Waals forces, and the values of the binding energies were in direct correlation with the values of the sensor response to NO. Full article

Transgenic crops producing insecticidal proteins from the bacterium Bacillus thuringiensis (Bt) control some important insect pests. However, evolution of resistance by pests reduces the efficacy of Bt crops. Here we review resistance to Bt cotton in the pink bollworm, Pectinophora gossypiella, one of the world’s most damaging pests of cotton. Field outcomes with Bt cotton and pink bollworm during the past quarter century differ markedly among the world’s top three cotton-producing countries: practical resistance in India, sustained susceptibility in China, and eradication of this invasive lepidopteran pest from the United States achieved with Bt cotton and other tactics. We compared the molecular genetic basis of pink bollworm resistance between lab-selected strains from the U.S. and China and field-selected populations from India for two Bt proteins (Cry1Ac and Cry2Ab) produced in widely adopted Bt cotton. Both lab- and field-selected resistance are associated with mutations affecting the cadherin protein PgCad1 for Cry1Ac and the ATP-binding cassette transporter protein PgABCA2 for Cry2Ab. The results imply lab selection is useful for identifying genes important in field-evolved resistance to Bt crops, but not necessarily the specific mutations in those genes. The results also suggest that differences in management practices, rather than genetic constraints, caused the strikingly different outcomes among countries. Full article

Ultra-high performance liquid chromatography-quadrupole-time of flight tandem mass spectrometry (UHPLC-Q-TOF-MS/MS) was used to study the diversity of tea polysaccharides and the dynamic changes in the physicochemical indexes of tea samples. FT-IR spectra and the free radical scavenging ability of tea polysaccharides, during pile-fermentation of post-fermented tea, were analyzed. The results showed that 23 saccharide co mponents in tea polysaccharides were identified: these belonged to 11 monosaccharides, 5 oligosaccharides, and 6 derivatives of monosaccharides and oligosaccharides. The abundance of oligosaccharides decreased gradually, while monosaccharides, and derivatives of monosaccharides and oligosaccharides increased gradually with the development of pile-fermentation. According to the differences in polysaccharide composition and their abundance, the tea polysaccharide samples extracted from different pile-fermentation stages could be clearly classed into three groups, W-0, W-1~W-4 and W-5~C-1. The pile-fermentation process affected the yield, the content of each component, FT-IR spectra, and the DPPH free radical scavenging ability of tea polysaccharides. Correlation analysis showed that microorganisms were directly related to the changes in composition and the abundance of polysaccharides extracted from different pile-fermentation stages. The study will further help to reveal the function of tea polysaccharides and promote their practical application as a functional food. Full article

Contamination by ochratoxigenic fungi and its prevention during the pile-fermentation of post-fermented tea have always been a concern. The present study aimed to elucidate the anti-fungal effect and mechanism of polypeptides produced by B. brevis DTM05 (isolated from post-fermented tea) on ochratoxigenic fungi, and to to evaluate their use in the pile-fermentation process of post-fermented tea. The results showed that polypeptides (produced by B. brevis DTM05) with a strong antifungal effect against A. carbonarius H9 mainly had a molecular weight between 3 and 5 kDa. The Fourier-transform infrared spectra of this polypeptide extract showed that it was a mixture consisting mainly of polypeptides and small amounts of lipids and other carbohydrates. The polypeptide extracts significantly inhibited the growth of A. carbonarius H9, and its minimum inhibitory concentration (MIC) was 1.6 mg/L, which significantly reduced the survival rate of spores. The polypeptides also effectively controlled the occurrence and ochratoxin A (OTA) production of A. carbonarius H9 on the tea matrix. The lowest concentration of polypeptides that significantly inhibited the growth of A. carbonarius H9 on the tea matrix was 3.2 mg/L. The enhancement of the fluorescence staining signal in the mycelium and conidiospore showed that the polypeptides with a concentration of more than 1.6 mg/L increased the permeability of the mycelium membrane and conidial membrane of A. carbonarius H9. The significant increase in the extracellular conductivity of mycelia suggested the outward leakage of intracellular active substances, and also further indicated an increase in cell membrane permeability. Polypeptides with a concentration of 6.4 mg/L significantly down-regulated the expression level of the polyketide synthase gene related to OTA production (acpks) in A. carbonarius H9, which may be the fundamental reason why polypeptides affect OTA production. In conclusion, reasonable use of the polypeptides produced by B. brevis can destroy the structural integrity of the cell membrane, make the intracellular active substances leak outward, accelerate the death of fungal cells and down-regulate the expression level of the polyketide synthase gene in A. carbonarius; thus, they can effectively control the contamination of ochratoxigenic fungi and OTA production during the pile-fermentation of the post-fermented tea. Full article

During the messenger RNA (mRNA) maturation process, RNA polyadenylation is a key step, and is coupled to the termination of transcription. Various cis-acting elements near the cleavage site and their binding factors would affect the process of polyadenylation, and AAUAAA, a highly conserved hexamer, was the most important polyadenylation signal (PAS). PAS usage is one of the critical modification determinants targeted at mRNA post-transcription. The full-length transcriptome has recently generated a massive amount of sequencing data, revealing poly(A) variation and alternative polyadenylation (APA) in Spodoptera frugiperda. We identified 50,616 polyadenylation signals in Spodoptera frugiperda via analysis of full-length transcriptome combined with expression Sequence Tags Technology (EST). The polyadenylation signal usage in Spodoptera frugiperda is conserved, and it is similar to that of flies and other animals. AAUAAA and AUUAAA are the most highly conserved polyadenylation signals of all polyadenylation signals we identified. Additionally, we found the U/GU-rich downstream sequence element (DSE) in the cleavage site. These results demonstrate that APA in Spodoptera frugiperda plays a significant role in root growth and development. This is the first polyadenylation signal usage analysis in agricultural pests, which can deepen our understanding of Spodoptera frugiperda and provide a theoretical basis for pest control. Full article