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Cytochrome P450 monooxygenase (P450 or CYP) plays an important role in the metabolism of insecticides and plant allelochemicals by insects. CYP321B1, a novel Spodoptera litura P450 gene, was identified and characterized. CYP321B1 contains a 1488 bp open reading frame (ORF) that encodes a 495 amino acid protein. In fourth instar larvae, the highest CYP321B1 expression levels were found in the midgut and fat body. In the tannin feeding test, tannin can significantly induce the expression of CYP321B1 in the midgut and fat body of 4th instar larvae. To verify the function of CYP321B1, RNA interference and metabolome analysis were performed. The results showed that silencing CYP321B1 significantly reduced the rate of weight gain under tannin induction. Metabolome analysis showed silencing affected 47 different metabolites, mainly involved in secondary metabolite biosynthesis and amino acid metabolism, including amino acids, lipid fatty acids, organic acids and their derivatives. Henoxyacetic acid and cysteamine are the most highly regulated metabolites, respectively. These findings demonstrate that CYP321B1 plays an important role in tannin detoxification and metabolism. Functional knowledge about metabolite detoxification genes in this major herbivorous insect pest can provide new insights into this biological process and provide new targets for agricultural pest control.

Propylea japonica has been regarded as one of the most remarkable natural enemies against aphid in China. However, the mechanism of juvenile hormone (JH) regulation of reproduction in P. japonica is still unclear. In this study, we investigated the JH titers of P. japonica and the development of the ovaries. We selected the six different developmental stages of ladybeetle females for transcriptome sequencing. We identified 583 genes involved in insect reproduction regulation, including 107 insect hormone synthesis signaling pathway-related genes and 476 nutrition-sensing signaling pathway-related genes. Transcriptome analysis indicated that a large number JH synthesis- and metabolism-related enzyme genes and some potential nutrient signal sensing- and transduction-related genes were significantly differentially expressed during P. japonica development. We investigated the effects of Met gene silencing on the reproduction of female adults and found that the ovarian maturation, vitellogenesis, and follicular epithelium development in the dsMet treatment group were significantly inhibited.

Background Aphis gossypii, a polyphagous and recurrent pest induced by pesticides, causes tremendous loss in the yield of crops each year. Previous studies of pesticide-induced sublethal effects mechanism mainly focus on gene level. The symbiotic bacteria are also important participants of this mechanism, but their roles in hormesis are still unclear. Results In this study, life table parameters and 16S rRNA sequencing were applied to evaluate the sublethal and transgenerational effects of sulfoxaflor on adult A. gossypii after 24-h LC20 (6.96 mg L-1 ) concentration exposure. The results indicated that the LC20 of sulfoxaflor significantly reduced the finite rate of increase (λ) and net reproductive rate (R0 ) of parent generation (G0), and significantly increased mean generation time (T) of G1 and G2, but not of G3 and G4. Both reproductive period and fecundity of G1 and G2 were significantly higher than those of the control. Furthermore, our sequencing data revealed that more than 95% bacterial communities were dominated by the phylum Proteobacteria, in which the maximum proportion genus was the primary symbiont Buchnera and the facultative symbiont Arsenophonus. Compared to those of the control, the abundance and composition of symbiotic bacteria of A. gossypii for three successive generations (G0-G2) were changed after G0 A. gossypii was exposed to sulfoxaflor: the diversity of the bacterial community was decreased, but the abundance of Buchnera was increased (G0), while the abundance of Arsenophonus was decreased. Contrary to G0, G1 and G2 cotton aphid exhibited an increased relative abundance of Arsenophonus in sublethal treatment group. Conclusion Taken together, our results provide an insight into the interactions among pesticide resistance, aphids, and symbionts, which will eventually help to better manage the resurgence of A. gossypii. This article is protected by copyright. All rights reserved.

With the aging population worldwide, osteoporosis, as an age-related bone metabolic disease, is becoming a hot issue in public health. However, it is still a great challenge to realize osteoporotic bone healing due to the alteration of the bone microenvironment in osteoporosis patients. In this study, a nano-structured akermanite (nAK) coating was in situ constructed on Ti–6Al–4V implants to improve osteoporotic bone repair. In vitro studies indicated that both the surface nano-topography and bioactive ions released from the nAK coatings promoted the proliferation, osteogenesis, angiogenesis and inhibited osteoclastogenesis of ovariectomy rabbit-derived bone marrow mesenchymal stem cells (OVX-rBMSCs). Furthermore, the nAK-coated Ti–6Al–4V implants improved new bone formation and osseointegration in an osteoporosis rabbit model in vivo. These results indicated that the AK coating with a nano-structured surface on the Ti–6Al–4V implant could synergistically promote bone formation and osseointegration for osteoporosis patients. This may be a promising strategy to improve the bone regeneration and osseointegration capability of orthopedic implants under osteoporosis conditions.

Osteoporosis (OP) is a major systemic bone disease leading to an imbalance in bone homeostasis which remains a challenge in the current treatment of bone defects. Our previous studies on strontium (Sr) doping apparently stimulated osteogenesis of bioceramics, which suggested a promising strategy for the treatment of bone defects. However, the potential effects and the underlying mechanisms of Sr-doping on osteoporotic bone defects still remain unclear. Autophagy is a conventional self-degradation process of cells involved in bone homeostasis and regeneration under physiological and pathological conditions. Therefore, it is essential to design appropriate biomaterials and investigate the associated osteogenic mechanisms via autophagy. Based on this hypothesis, Sr-doped 45S5 bioglass (Sr/45S5) was fabricated, and ovariectomy bone marrow-derived mesenchymal stem cells (OVX-BMSCs) were applied as the in vitro cell culture model. First, the optimal Sr-doping concentration of 10 mol% was screened by cell proliferation, ALP staining, alizarin red S staining and the real-time PCR assay. Then, the results of western blot (WB) analysis showed that Sr-induced osteogenic differentiation of OVX-BMSCs was associated with time-dependent modulation of autophagy and related to the AKT/mTOR signaling pathway. Meanwhile, the autophagy in Sr-induced osteogenic differentiation of OVX-BMSCs was detected by WB, immunofluorescence staining and transmission electron microscopy. Furthermore, the effect of osteogenic differentiation of OVX-BMSCs has been significantly inhibited by the administration of autophagy inhibitors and the AKT/mTOR pathway inhibitors, respectively, in the early and late periods of osteogenic differentiation. Finally, the results of the model of femoral condyle defects in OVX-rats indicated that Sr10/45S5 granules remarkably enhanced bone regeneration which provided the evidences in vivo. Our research indicates that Sr-doping provides a promising strategy to promote osteogenic differentiation of OVX-BMSCs and bone regeneration in osteoporotic bone defects via early improvement of autophagy and late activation of the Akt/mTOR signaling pathway.

Research on microplastic pollution of terrestrial soils is catching up with the aquatic environment, especially agricultural soil systems. Plastic residues have caused various environmental problems in mulch film extensively used agricultural areas. However, studies focusing specifically on the potential influence of mulch film residues on the metabolic cycle of soil systems have yet to be conducted. Here, high-throughput sequencing combined with metabolomics were first used to study the effects of residual mulch on soil microbial communities and related metabolic functions. Plastic film treatment did not significantly affect soil physicochemical properties including pH, organic matter and nitrogen, etc in short term. However, it did significantly changed overall community structure of soil bacteria, and interfered with complexity of soil bacterial symbiosis networks; exposure time and concentration of residues were particularly important factors affecting community structure. Furthermore, metabolomics analysis showed that film residue significantly changed soil metabolite spectrum, and interfered with basic carbon and lipid metabolism, and also affected basic cellular processes such as membrane transport and, in particular, interfered with the biosynthesis of secondary metabolites, as well as, biodegradation and metabolism of xenobiotics. Additionally, through linear discriminant and collinear analysis, some new potential microplastic degrading bacteria including Nitrospira, Nocardioidaceae and Pseudonocardiaceae have been excavated.

Parasitism usually causes considerable changes in lipids and fatty acids by redirecting the development of the host. In this study, changes in weight and in free fatty acid content of cotton aphids were recorded after aphids had been parasitized. Results showed that the weight of parasitized Aphis gossypii was increased compared to nonparasitized aphids, and significantly increased weights were detected at 1, 2, and 3 instars after parasitization by Lysiphlebia japonica. Free fatty acid test kits and GC-MS showed that the fatty acid content increased in the early stage of parasitization but decreased after 3 days of parasitization. Seven genes related to the fatty acid synthesis pathway were significantly upregulated in the parasitized aphids, where they were 1.96-10.97 times greater. Our data described the change that occurs in the fatty acid content of parasitized A. gossypii.

The predatory ladybird beetle, Harmonia axyridis, is a predominant natural enemy of pest insects in cotton fields. Commercialization of genetically modified crops has promoted the increased use of the herbicide glyphosate. In this study, to assess potential negative effects of glyphosate on beneficial non-target organisms in cotton fields, we first examined how glyphosate exposure affected the development and endosymbiotic bacterial community of H. axyridis. The results showed that the survival rate, development duration, pupation rate and emergence rate of H. axyridis under low and high concentrations of glyphosate exposure were not significantly changed, but glyphosate did significantly reduce the body weight of H. axyridis. Based on 16S rRNA sequencing, there were no significant differences in the diversity or richness of the endosymbiotic bacteria of H. axyridis before and after glyphosate exposure. The dominant bacterial phyla Firmicutes and Proteobacteria and genera Staphylococcus and Enterobacter remained the same regardless of treatment with glyphosate, however the abundance and copy number of these bacteria were altered. Glyphosate treatment significantly reduced the abundance and gene copy number of Staphylococcus and increased the abundance and gene copy number of Enterobacter. This is the first report demonstrating that glyphosate can reduce the body weight H. axyridis and alter the bacterial endosymbiont community by affecting the abundance and gene copy number of dominant bacteria.

Lysozymes are used in sterilisation, antisepsis, dairy additives, inflammation, and cancer. One transgenic goat line expressing high levels of human lysozyme (hLZ) in goat milk has been developed in China. Herein, we established an event-specific real-time polymerase chain reaction (real-time PCR) method to detect the transgenic hLZ goat line. The developed method has high specificity, sensitivity and accuracy, and a wide quantitative dynamic range. The limit of detection and limit of quantification was 5 and 10 copies per reaction, respectively. The practical sample analysis results showed that the method could identify and quantify transgenic lysozyme content in trace samples in routine lab analyses. Furthermore, the potential applicability in risk assessment, such as molecular characterisation and gene horizontal transfer, was confirmed. We believe that this method is suitable for the detection of transgenic hLZ goat line and its derivate.

Multiplex and high-throughput assays are becoming the main trends in the development of new nucleic acid detection and quantification methods, such as those for genetically modified organism (GMO) analysis. Here, we report a novel universal LNA probe-mediated droplet digital polymerase chain reaction (PCR) method (ULNA-ddPCR) for multiple DNA target quantification in GMOs. In ULNA-ddPCR, only one universal LNA probe is used for multiple DNA targets instead of using one to one TaqMan probe. The specificity, sensitivity, dynamic range, and accuracy of the ULNA-ddPCR method are determined by employing GM rice analysis as an example. Simplex and triplex ULNA-ddPCR assays for three GM rice events, T2A-1, T1C-19, and G6H1, are established and evaluated. All results indicate that the developed simplex and triplex ULNA-ddPCR assays are suitable for quantitative analysis of GM rice events with high sensitivity, accuracy, and low cost. The ULNA-ddPCR method also has the potential for multiple DNA target quantification in other research fields.

Spodoptera litura is a major insect with a cosmopolitan distribution and strong resistance to multiple insecticides. Determining the molecular basis and key candidate genes of the insecticide resistance of S. litura may help in managing this insect. In this study, fifth-instar S. litura larvae were subjected to transcriptome analysis at 6, 12, 24, 48, and 72 h after feeding on an LC20 dose of avermectin. The result showed that genes responding to avermectin changed dynamically with different gene counts and resistance mechanisms at the fifth instar based on a metabolic pathway map. These responses included degrading the insecticide by a series of P450 and glutathione-S-transferase enzymes starting at the 12 h time point, with subsequent increases in the number of genes involved and shifts to TOLL and immune deficiency (IMD) pathways at 48 h after feeding the insecticide. Weighted correlation network analysis (WGCNA) determined a co-expression module related to the avermectin response at 12 and 24 h (r = 0.403, p = 0.0371; r = 0.436, p = 0.023), in which a hub gene (LOC111358940) related to metalloproteinase activity was identified. In addition, Analysis of the genes in the co-expression module further revealed that eight genes encoding UDP-glucuronosyltransferases were directly associated with insecticide response in S. litura. These results provide better understanding of the avermectin response mechanism of S. litura and may be useful in developing improved control strategies for this species. SUPPLEMENTARY INFORMATION: The online version of this article (10.1007/s13205-021-02651-9) contains supplementary material, which is available to authorized users.

Simple Summary The Aphis gossypii is a global problem for its pesticide resistance with substantial economic and ecological cost and a wide host range, including cotton and cucurbits. The development of insecticide resistance is rapid and widespread and threatens crop productivity. Biopesticides have emerged as a better alternative for pest control. Cucurbitacin B (CucB) and epigallocatechin gallate (EGCG) are the major secondary metabolites of host plants cucurbits and cotton. In this study, we used cotton- and cucurbit-specialized aphids (CO and CU) as a study system to better understand the effects of CucB and EGCG on cotton aphid. Our study showed that CucB and EGCG can significantly reduce the population-level fitness of A. gossypii, affect their ability to adapt to nonhost plants and alter the levels of some detoxifying enzymes, which showed a potential to be developed into new biopesticides against the notorious aphids. Abstract Aphis gossypii (Glover) is distributed worldwide and causes substantial economic and ecological problems owing to its rapid reproduction and high pesticide resistance. Plant-derived cucurbitacin B (CucB) and epigallocatechin gallate (EGCG) are known to have insecticidal and repellent activities. However, their insecticidal activity on cotton- and cucurbit-specialized aphids (CO and CU), the two important host biotypes of A. gossypii, remains to be investigated. In the present study, we characterized, for the first time, the effects of these two plant extracts on the two host biotypes of A. gossypii. CucB and EGCG significantly reduced the A. gossypii population-level fitness and affected their ability to adapt to nonhost plants. Activities of important detoxification enzymes were also altered, indicating that pesticide resistance is weakened in the tested aphids. Our results suggest that CucB and EGCG have unique properties and may be developed as potential biopesticides for aphid control in agriculture.

Implantable self-powered generators (ISPGs) have been extensively explored as energy supplies for driving electronics and electrically stimulated therapeutics in vivo. However, some drawbacks arise, such as complicated architectonics, inescapability of wire connection, energy instability, and consumption. In this study, a host-coupling bio-nanogenerator (HCBG) is developed to configure a self-powered regional electrical environment for powerful bone regeneration. An HCBG consists of a porous electret nanofiber mat coupled with interstitial fluid and stimulated objects of the host after implantation, forming a host coupling effect. This bio-nanogenerator not only overcomes the disadvantages of general ISPGs, but also accomplishes both biomechanical energy scavenging and electrical stimulation therapeutics. The enhancement of osteogenesis differentiation of bone marrow mesenchymal stem cells in vitro and bone regeneration in vivo are remarkably achieved. Moreover, osteogenic ability is systematically evaluated by regulating the electrical performance of HCBGs. Osteogenic differentiation is activated by upregulating more cytosolic calcium ion, following to activate the calcium ion-induced osteogenic signal pathway, while applying electrical stimulation. As an implantable medical technology, the HCBG provides an explorative insight to facilitate the development of ISPG-based electrical medical therapeutics.

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 extraction of Mg2+ by di-(2-ethylhexyl) phosphoric acid (P204) was carried out to separate lithium and magnesium. To facilitate the direct electrodeposition of metallic magnesium from the magnesium-rich organic phase, different diluents and ionic liquids were investigated. It was found that tributylhexylphosphonium bis (trifluoromethyl sulfonyl) imide ionic liquid ([P4446][NTf2]) can serve as an excellent supporting electrolyte to solve the electroconductibility of the weak polarity organic phase. The electroconductivity was further increased by more than 1500 times by the use of methyl isobutyl ketone (MIBK) as a diluent. The single-stage extraction rate of Mg2+ at a concentration up to 76.87 g/L reached 68.4% using 40% P204 + 55% MIBK + 5% [P4446][NTf2] at R(O/A) = 6:1, and the electroconductivity of organic phase was 254.5 μS·cm−1 after extraction. The extraction mechanism and thermokinetics indicated that both [P4446][NTf2] and MIBK do not participate in the extraction process, and the extraction is an obvious endothermic process.

As a new-type liquid lithium resource, the recovery of lithium from geothermal water is of great significance due to the increasing importance of lithium in modern industry. In this work, different lithium manganese oxides (LMOs), including LiMn2O4, Li1.66Mn1.66O4, and Li4Mn5O12, were synthesized and applied for selective recovery of lithium from geothermal water. It was found that the powdery Li4Mn5O12 exhibits high performances for low concentrations of Li+ and was then successfully granulated using chitosan (CTS) at a mass ratio of CTS/LMO = 3:2. The hydrophily of CTS leaded to a serious dissolution loss of CTS/LMO under acidic conditions, but it was successfully solved by direct cross-link of the spherical material using ethylene glycol diglycidyl ether (EGDE), by which the dissolution loss in 0.25 mol/L HCl was decreased from 27.65% to no more than 0.6%. The equilibrium adsorption capacity of Li+ in geothermal water reached 8.98 mg/g at 303.15 K, while the attenuation of adsorption capacity was no more than 1.1% after five times adsorption-desorption cycles. The separation factors between Li+ and other ions were all higher than 51 with a distribution coefficient of Li+ reaching up to 1113.27 mL/g. All these properties demonstrate that the developed material has high selectivity and stability, and can be used as a candidate for selective recovery of lithium from geothermal water or other solutions.

DNA Phosphorothioate (PT), replacing a non-bridging phosphate oxygen atom with a sulfur atom, is one kind of common DNA modification in bacteria. Whole genome scale description of the location and frequency of PT modification is the key to understand its biological function. Herein we developed a novel method, named with iodine-induced cleavage quantitative real-time PCR (IC-qPCR), to evaluate the frequency of PT modification at a given site in bacterial DNA. The efficiency, dynamic range, sensitivity, reproducibility and accuracy of IC-qPCR were well tested and verified employing an E. coli B7A strain as example. The amplification efficiency of IC-qPCR assay ranged from 91% to 99% with a high correlation coefficient ≥0.99. The limit of quantification was determined as low as 10 copies per reaction for the 607710 and 1818096 sites, and 5 copies for the 302695 and 4120753 sites. Based on the developed IC-qPCR method, the modification frequency of four PTs in E. coli B7A was determined with high accuracy, and the results showed that the PT modification was partial and that the modification frequency varied among investigated PT sites. All these results showed that IC-qPCR was suitable for evaluating the PT modification, which would be helpful to further understand the biological function of PT modification.

The cotton‐melon aphid, Aphis gossypii Glover, is a worldwide‐spreading species, and pesticide‐resistant populations are increasing rapidly. In this study, investigations were performed based on Illumina HiSeq sequencing of the 16S rDNA V4 region for the bacterial communities embodied as intracellular symbionts under natural and in pesticide‐treated populations of A. gossypii. The results revealed that more than 82% of bacterial communities belonged to the phylum Proteobacteria in which the maximum proportion (53.24%) was of the genus Arsenophonus; Hamiltonella composed 22.31; and 1.37% was of the genus Acinetobacter. The relative abundance of Hamiltonella was obvious, vertically transmitted, divided into two groups, and its infection influenced the bacterial communities in A. gossypii. Symbiont density and composition were changed in samples tested on different days. Azadirachtin and phoxim influenced on the composition of bacterial communities. Different biomarkers were used for pesticide‐treated samples with LEfSe results. These findings will increase awareness regarding bacterial communities in naturally occurring populations of A. gossypii and pave the way to study the relationship between symbionts and pesticide resistance.

A new microbial α-amylase inhibitor gene was cloned and characterized. The encoded, recombinant, α-amylase inhibitor protein was induced and expressed by isopropyl β-d-1-thiogalactopyranoside (IPTG) in Escherichia coli M15 cells. The effects of the α-amylase inhibitor protein on Helicoverpa armigera larvae were studied. Compared to the control, the weight of H. armigera larvae fed the diet with recombinant α-amylase inhibitor protein added at a concentration of 20 μg/g was reduced by 49.8%. The total soluble protein of H. armigera larvae fed the diet with the α-amylase inhibitor protein added was also reduced by 36.8% compared to the control. The recombinant α-amylase inhibitor protein showed inhibition activity against α-amylase of H. armigera. These results suggested that this α-amylase inhibitor protein may be a promising bioinsecticide candidate for controlling H. armigera.