Search

Abstract Brown marmorated stink bug (BMSB), Halyomorpha halys (Stål) (Hemiptera: Penta-tomidae), is a highly polyphagous and invasive insect pest with more than 300 plant species as hosts, including a wide range of economic crops. To date, several egg parasitoid species are reported to attack BMSB. In this study, two species of Trissolcus (Hymenoptera: Platygastridae) and one Anastatus (Hymenoptera: Eupelmidae) were recovered from field exposed BMSB sentinel eggs in Beijing, China. The wasps’ small size of only 1.5 mm and 4 mm in length make them difficult to identify morphologically and required taxonomist for identification of specific species. In addition, these parasitoids have morphology sibling species and have been misidentified several times in China. To overcome these problems, a molecular method with species-specific primers designed for the COI gene has been developed to identify Trissolcus, Anastatus and their host samples from field collected samples. After successful morphological confirmation with experts, DNA extractions were carried out from these samples. PCR amplification using published primers for T. japonicus, A. japonicus and BMSB specimens confirmed the species. As for T. cultratus, species specific primers TCYF and TCYR were developed which produce 340-bp PCR products length while no positive amplifications found in other wasps and host. Sensitivity analysis of markers revealed that TCYF and TCYR primers could detect as low a DNA template concentration as 0.00025 ng/μL. This indicates that PCR with these primers specifically and sensitively differentiates T. cultratus specimens from other similar wasp species. All the primers tested in this study could discriminate between parasitized and non-parasitized BMSB eggs. This molecular identification method shows promise for conveniently identifying Trissolcus and Anastatus species in host-parasitoid associations and accurately evaluating parasitism rates in the field.

The olfactory system of insects is crucial in modulating behaviors such as host seeking, mating, and oviposition. Odorant-binding proteins (OBPs) are involved in semiochemical recognition. OBPs recognize and bind odorants and transport them to odorant receptors located in olfactory neurons. Harmonia axyridis (Coleoptera: Coccinellidae) is a widely used predacious biological control agent for many agricultural and forestry pests. This study identified 19 OBPs in H. axyridis based on the antennal and whole-body transcriptomes of adults and obtained all the full-length open reading frames, including 11 ‘Classic’ OBPs, 7 ‘Minus-C’ OBPs and 1 ‘Plus-C’ OBP. They encoded 125 to 241 amino acid proteins with molecular weights ranging from 13.75 to 27.75 kDa and isoelectric points ranging from 4.15 to 8.80. Phylogenetic analyses were used to study the relationships between H. axyridis OBPs and OBPs from other species of Coleoptera. Quantitative real-time PCR (qPCR) analysis showed that HaxyOBP2, 3, 5, 8, 10, 12, 13, 14, and 15 were highly expressed in antennae of both adult females and males. Moreover, HaxyOBP2, 3, 5, 12, and 15 were more abundantly expressed in antennae than other body parts, while HaxyOBP13 and HaxyOBP14 were expressed predominantly, and at similar levels, in the head and antennae. The other OBP genes were highly expressed in non-olfactory tissues including the thorax, abdomen, legs, and wings. These results provide valuable information for further study of H. axyridis olfaction, which may ultimately enhance its use as a biocontrol agent.

Harmonia axyridis is an important natural enemy that consumes many agricultural and forestry pests. It relies on a sensitive olfactory system to find prey and mates. Odorant-binding proteins (OBPs) as the first-step of recognizing volatiles, transport odors through sensillum lymph to odorant receptors (ORs). However, little is known about the molecular mechanisms of H. axyridis olfaction. In this study, four H. axyridis antenna specific OBP genes, HaxyOBP3, 5, 12, and 15, were bacterially expressed and the binding features of the four recombinant proteins to 40 substances were investigated using fluorescence competitive binding assays. Three-dimensional structure modeling and molecular docking analysis predicted the binding sites between HaxyOBPs and candidate volatiles. Developmental expression analyses showed that the four HaxyOBP genes displayed a variety of expression patterns at different development stages. The expression levels of HaxyOBP3 and HaxyOBP15 were higher in the adult stage than in the other developmental stages, and HaxyOBP15 was significantly transcriptionally enriched in adult stage. Ligand-binding analysis demonstrated that HaxyOBP3 and HaxyOBP12 only combined with two compounds, β-ionone and p-anisaldehyde. HaxyOBP5 protein displayed binding affinities with methyl salicylate, β-ionone, and p-anisaldehyde (Ki = 18.15, 11.71, and 13.45 μM). HaxyOBP15 protein had a broad binding profile with (E)-β-farnesene, β-ionone, α-ionone, geranyl acetate, nonyl aldehyde, dihydro-β-ionone, and linalyl acetate (Ki = 4.33–31.01 μM), and hydrophobic interactions played a key role in the binding of HaxyOBP15 to these substances according to molecular docking. Taken together, HaxyOBP15 exhibited a broader ligand-binding spectrum and a higher expression in adult stage than HaxyOBP3, 5, and 12, indicating HaxyOBP15 may play a greater role in binding volatiles than other three HaxyOBPs. The results will increase our understanding of the molecular mechanism of H. axyridis olfaction and may also result in new management strategies (attractants/repellents) that increase the biological control efficacy of H. axyridis.

Anastatus japonicus Ashmead is an egg parasitoid wasp important for the biological control of fruit crop pests. The olfaction of parasitoids is crucial to searching for host pests in fruit crops. In this study, we sequenced and analyzed the antennal and abdominal transcriptomes of A. japonicus to better understand the olfactory mechanisms in this species. A total of 201 putative olfactory receptor genes were identified, including 184 odorant receptors (ORs) and 17 ionotropic receptors (IRs). Then, we assayed the tissue-specific and sex-biased expression profiles of those genes based on the transcriptional levels. In total, 165 ORs and 15 IRs had upregulated expression in the antennae. The expression levels of 133 ORs, including odorant receptor co-receptor (AjapORco), and 10 IRs, including AjapIR8a, were significantly different between the female and male antennae. Our results provide valuable information for further studies on the molecular mechanisms of the olfactory system in A. japonicus.

Abstract: (E)-β-Caryophyllene is a sesquiterpene compound widely distributed in plants and functions in plant defence. However, little is known about the sequence and function of (E)-β-caryophyllene synthase in lima bean (Phaseolus lunatus). Here, we report a new full-length cDNA (PlCAHS) encoding (E)-β-caryophyllene synthase, a possible key enzyme of plant defence. The cDNA of PlCAHS contains an open reading frame of 1 761 bp, encoding a protein of 586 amino acids with a predicted mass of 67.95 kDa. The deduced amino acid sequence shows 52% identity with sesquiterpene synthase MtCAHS of Medicago truncatula. Based on phylogenetic analysis, PlCAHS is classified as the terpene synthases (TPS)-a subfamily. The recombinant enzyme, expressed in Escherichia coli, catalysed the formation of a major product (E)-β-caryophyllene (82%) and a minor product α-humulene (18%) from farnesyl diphosphate. Real-time quantitative PCR (qRT-PCR) analysis found that the PlCAHS transcript was significantly up-regulated in leaves after treatment with spider mites and alamethicin (ALA), suggesting its ecological function in plant defence.

Abstract The sycamore lace bug, Corythucha ciliata, is a pest of sycamore trees. In China, it is found in the most northern border where it has been known to become dormant during harsh winters. But the molecular and metabolic basis for dormancy in this insect is still unknown. In this study, we analyzed the transcript and metabolite profiles of this bug to identify key genes and metabolites that are significantly regulated during dormancy in adult females and males. In total, 149 differentially expressed genes (DEGs) were significantly up-regulated and 337 DEGs were significantly down-regulated in dormant adults (both females and males). We found major differences in heat shock protein (HSPs), immunity-responsive genes, NAD-dependent deacetylase sirtuin-1 (SIRT1) and genes involved in the spliceosome pathway that is known to regulate stress. Among the 62 metabolites identified by GC-MS, 12 metabolites including glycerol, trehalose, and alanine were significantly increased during C. ciliata dormancy. By integrating the transcriptome and metabolite datasets, we found that the metabolites in glycolysis/gluconeogenesis and citrate cycle (TCA) were significantly reduced. This study is the first to report both transcript and metabolite profiles of the overwintering responses of C. ciliata to cold stress at the molecular level.

Insect cuticle is an apical extracellular matrix produced by the epidermis, tracheal, hind‐ and foregut epithelia during embryogenesis and renewed during molting and metamorphosis. However, the underlying regulatory mechanism for embryonic cuticle formation remains largely unclear. Here, we investigate the function of the transcription factor POUM2 in the embryonic cuticular formation in Bombyx mori, a model lepidopteran insect. Clustered regularly interspaced palindromic repeats (CRISPR)/CRISPR‐associated protein‐9‐mediated knockout of POUM2 resulted in the defect of cuticular deposition, pigmentation, and sclerotization in the embryos. Differentially expressed transcripts analysis of 7‐d‐old embryos identified 174 up‐ or downregulated cuticular protein transcripts, 8 upregulated chitin degradation transcripts, 2 downregulated chitin synthesis transcripts and 48 up‐ or downregulated transcription factor transcripts in the POUM2−/− embryos. The expression levels of the key factors of the tyrosine metabolic pathway, such as tyrosine hydroxylase (Th), Dopa decarboxylase (DDC), and arylalkylamine N‐acetyltransferase (aaNAT), were significantly decreased in the POUM2−/− embryos. POUM2 isoform POUM2‐L specifically bound the POU cis‐regulatory element (CRE) in the Th promoter and increased the transcription of Th, whereas POUM2‐S could not bind the POU CRE, although it also increased the transcription of Th. Heterogeneous nuclear ribonucleoprotein Squid‐1 directly bound the POUM2 pre‐mRNA (messenger RNA) and inhibited the alternative splicing of POUM2‐L to POUM2‐S mRNA. These results suggest that POUM2 participates in the cuticular formation by regulating the chitin and cuticular protein synthesis and metabolism, and the cuticular pigmentation and sclerotization by regulating tyrosine metabolism during embryogenesis. This study provides new insights into novel function of POUM2 in embryogenesis. [ABSTRACT FROM AUTHOR]

Long wavelength InAs/AlSb quantum cascade lasers (QCLs) emitting at 13 μm, based on a diagonal transition scheme design through band structure engineering, have been grown and fabricated. This band structure engineering focuses on enhancing transition efficiency and suppressing carrier leakage. Our 3-mm-long, 25-μm-wide InAs/AlSb QCL has achieved a slope efficiency of 210 mW/A and a maximum peak power of 515 mW, despite encountering a substantial waveguide loss of 27 cm−1 and a relatively high threshold of 4.8 kA/cm2, due to the elevated residual doping level. Our InAs/AlSb QCL devices have demonstrated record-breaking performance in terms of slope efficiency, maximum peak power, and injection efficiency. Cavity length analysis suggests that reducing the residual doping by half could pave the way for achieving continuous wave output power in the realm of hundreds of milliwatts at room temperature for our designed 13 μm QCLs.

BackgroundThe relationship between white matter hyperintensities (WMH) and the core features of Alzheimer’s disease (AD) remains controversial. Further, due to the prevalence of co-pathologies, the precise role of WMH in cognition and neurodegeneration also remains uncertain.MethodsHerein, we analyzed 1803 participants with available WMH volume data, extracted from the ADNI database, including 756 cognitively normal controls, 783 patients with mild cognitive impairment (MCI), and 264 patients with dementia. Participants were grouped according to cerebrospinal fluid (CSF) pathology (A/T profile) severity. Linear regression analysis was applied to evaluate the factors associated with WMH volume. Modeled by linear mixed-effects, the increase rates (Δ) of the WMH volume, cognition, and typical neurodegenerative markers were assessed. The predictive effectiveness of WMH volume was subsequently tested using Cox regression analysis, and the relationship between WMH/ΔWMH and other indicators such as cognition was explored through linear regression analyses. Furthermore, we explored the interrelationship among amyloid-β deposition, cognition, and WMH using mediation analysis.ResultsHigher WMH volume was associated with older age, lower CSF amyloid-β levels, hypertension, and smoking history (all p ≤ 0.001), as well as cognitive status (MCI, p

Spodoptera litura is a destructive agricultural pest in tropical and subtropical areas. Understanding the molecular mechanisms of S. litura adaptation to its preferred host plants may help identify target genes useful for pest control. We used high‐throughput sequencing to characterize the expression patterns of messenger RNAs (mRNAs) and microRNAs (miRNAs) in the midgut of S. litura fed on Brassica juncea for 6 h and 48 h. A total of 108 known and 134 novel miRNAs were identified, 29 miRNAs and 237 mRNAs were differentially expressed at 6 h of B. juncea feeding, 26 miRNAs and 433 mRNAs were differentially expressed at 48 h. For the mRNAs, the up‐regulated genes were mostly enriched in detoxification enzymes (cytochrome P450, esterase, glutathione S‐transferase, uridine diphosphate‐glucuronosyl transferase), while the down‐regulated genes were mostly enriched in proteinases and immune‐related genes. Furthermore, most detoxification enzymes begin to up‐regulate at 6 h, while most digestion and immune‐related genes begin to up‐ or down‐regulate at 48 h. Eighteen and 37 differently expressed transcription factors were identified at 6 h and 48 h, which may regulate the functional genes. We acquired 136 and 41 miRNA versus mRNA pairs at 6 h and 48 h, respectively. Some down‐regulated and up‐regulated miRNAs were predicted to target detoxification enzymes and proteinases, respectively. Real‐time quantitative polymerase chain reaction of nine randomly selected miRNAs and 28 genes confirmed the results of RNA‐seq. This analyses of miRNA and mRNA transcriptomes provides useful information about the molecular mechanisms of S. litura response to B. juncea. [ABSTRACT FROM AUTHOR]

Krüppel homolog 1 (Kr‐h1), a zinc finger transcription factor, is involved in the metamorphosis and adult reproduction of insects. However, the role of Kr‐h1 in reproduction of holometabolic insects remains to be elucidated. The regulation network of Kr‐h1‐associated genes in the reproduction in Bombyx mori was investigated in this study. The higher expression level of BmKr‐h1 in the ovaries was detected during the late pupal stage and adults. RNA interference (RNAi)‐mediated depletion of BmKr‐h1 in the female at day 6 of pupae resulted in abnormal oocytes at 48 h post‐double‐stranded RNA treatment, which showed less yolk protein deposition and partially transparent chorion. RNA‐seq and subsequent differentially expressed transcripts analysis showed that knockdown of BmKr‐h1 caused a decrease in the expression of 2882 genes and an increase in the expression of 2565 genes in the oocytes at day 8 of pupae. Totally, 27 genes coding for transcription factors were down‐regulated, while six genes coding for other transcription factors were up‐regulated. BmKr‐h1 bound to the Kr‐h1 binding site of the transcription factors AP‐1 (activating protein‐1) and FOXG1 to increase their messenger RNA transcripts in the BmN cells, respectively. Gene Ontology enrichment and Kyoto Encyclopedia of Genes and Genomes pathway analyses of that positively co‐expressed with AP‐1 and FOXG1 transcripts showed mainly enrichment in the metabolic‐related pathways, the nutrient absorption and the yolk protein absorption processes. These data suggested that BmKr‐h1 might directly regulate the metabolic‐related pathways, the nutrient absorption and the yolk protein absorption processes or probably through AP‐1 and /or FOXG1 to regulate oocyte development. [ABSTRACT FROM AUTHOR]

The fall armyworm Spodoptera frugiperda recently invaded China, ravaging crops in many provinces. Deciphering the possible genetic basics for its successful invasion is critical for innovative and specific control for this gluttonous pest. Here we generated comparative genomic analyses between S. frugiperda and its native relative, S. litura, which differs in host preference, locomotivity and production behavior. We demonstrated that S. frugiperda genes are enriched in taste sensory perception and nervous system, obviously different from those of S. litura. Potential host adaptation genes showed generally an elevated ratio of non‐synonymous substitution rate to synonymous substitution rate, suggesting a faster evolution during the divergence of the two species. Focusing on these sets of genes, we identified 23 genes being under positive selection in S. frugiperda. Among them are two notable genes involved in sensory perception, gustatory receptor (GR) and an acetaldehyde oxidase, which are important for host detection in invasion and expansion processes. Another two genes are mitochondrial adenosine triphosphate synthase β subunit and ferritin heavy chain, which may be associated with the enhanced locomotivity and resistance, which fascinated long‐distance migration needed for invasion and rapid expansion. Another interesting gene is chorion protein, in which positive selection sites in S. frugiperda were found and a replacement in one site is predicted to affect the protein function, which might be associated with competent reproductivity in S. frugiperda to ensure genetic resources for expansion. [ABSTRACT FROM AUTHOR]

Fusion of the testis occurs in most Lepidoptera insects, including Spodoptera litura, an important polyphagous pest. Testicular fusion in S. litura is advantageous for male reproduction, and the molecular mechanism of fusion remains unknown. Doublesex influences the formation of genitalia, the behavior of courtship, and sexually dimorphic traits in fruit‐fly and silkworm, and is essential for sexual differentiation. However, its purpose in the testis of S. litura remains unknown. The doublesex gene of S. litura (Sldsx) has male‐specific SldsxM and female‐specific SldsxF isoforms, and exhibits a higher expression level in the male testis. At the testicular fusion stage (L6D6), Sldsx attained the highest expression compared to the pre‐fusion and post‐fusion periods. Moreover, Sldsx had a higher expression in the peritoneal sheaths of testis than that of germ cells in the follicle. CRISPR/Cas9 (Clustered Regularly Interspaced Short Palindromic Repeats/Cas9) was applied to S. litura to determine the role of Sldsx. A mixture of single guide RNA messenger RNA and Cas9 protein (300 ng/μL each) was injected into eggs within 2 h following oviposition. CRISPR/Cas9 successfully induced genomic mutagenesis of Sldsx at G0 generation. The mutant males had smaller testis surrounded by less tracheae. Moreover, the mutant males had abnormal external genitalia and could not finish mating with wild‐type females. Additionally, testes were fused for almost all mutant males. The results showed that Sldsx was not related to testicular fusion, and is required for both testis development and the formation and function of external genitalia in S. litura. The main roles of doublesex on the male are similar to other insects. [ABSTRACT FROM AUTHOR]

Silkworm mutants are valuable resources for both transgenic breeding and gene discovery. PiggyBac‐based random insertional mutagenesis has been widely used in gene functional studies. In order to discover genes involved in silk synthesis, a piggyBac‐based random insertional library was constructed using Bombyx mori, and the mutants with abnormal cocoon were particularly screened. By this means, a "thin cocoon" mutant was identified. This mutant revealed thinner cocoon shell and shorter posterior silk gland (PSG) compared with the wild type. The messenger RNA (mRNA) levels of all the three fibroin genes, including Fib‐H, Fib‐L and P25, were significantly down‐regulated in the PSG of mutants. Four piggyBac insertion sites were identified in Aquaporin (AQP), Longitudinals lacking protein‐like (Lola), Glutamyl aminopeptidase‐like (GluAP) and Loc101744460. The mRNA levels of all the four genes were significantly altered in the silk gland of mutants. In particular, the mRNA amount of AQP, a gene responsible for the regulation of osmotic pressure, decreased dramatically immediately prior to the spinning stage in the anterior silk gland of mutants. The identification of the genes disrupted in the "thin cocoon" mutant in this study provided useful information for understanding silk production and transgenic breeding of silkworms in the future. [ABSTRACT FROM AUTHOR]