The brown planthopper (BPH), Nilaparvata lugens (Stål) (Hemiptera: Delphacidae), a major pest of rice in Asia, is able to successfully puncture sieve tubes in rice with its piercing stylet and then to ingest phloem sap. How BPH manages to continuously feed on rice remains unclear. Here, we cloned the gene NlSEF1, which is highly expressed in the salivary glands of BPH. The NlSEF1 protein has EF-hand Ca2+-binding activity and can be secreted into rice plants when BPH feed. Infestation of rice by BPH nymphs whose NlSEF1 was knocked down elicited higher levels of Ca2+ and H2O2 but not jasmonic acid, jasmonoyl-isoleucine (JA-Ile) and SA in rice than did infestation by control nymphs; Consistently, wounding plus the recombination protein NlSEF1 suppressed the production of H2O2 in rice. Bioassays revealed that NlSEF1-knockdown BPH nymphs had a higher mortality rate and lower feeding capacity on rice than control nymphs. These results indicate that the salivary protein in BPH, NlSEF1, functions as an effector and plays important roles in interactions between BPH and rice by mediating the plant’s defense responses.
ORCID IDs: 0000-0002-5714-7586 (J.Z.); 0000-0002-3262-6134 (Y.L.).The brown planthopper (BPH) Nilaparvata lugens is one of the most destructive insect pests on rice (Oryza sativa) in Asia. After landing on plants, BPH rapidly accesses plant phloem and sucks the phloem sap through unknown mechanisms. We discovered a salivary endo-b-1,4-glucanase (NlEG1) that has endoglucanase activity with a maximal activity at pH 6 at 37°C and is secreted into rice plants by BPH. NlEG1 is highly expressed in the salivary glands and midgut. Silencing NlEG1 decreases the capacity of BPH to reach the phloem and reduces its food intake, mass, survival, and fecundity on rice plants. By contrast, NlEG1 silencing had only a small effect on the survival rate of BPH raised on artificial diet. Moreover, NlEG1 secreted by BPH did not elicit the production of the defense-related signal molecules salicylic acid, jasmonic acid, and jasmonoyl-isoleucine in rice, although wounding plus the application of the recombination protein NlEG1 did slightly enhance the levels of jasmonic acid and jasmonoyl-isoleucine in plants compared with the corresponding controls. These data suggest that NlEG1 enables the BPH's stylet to reach the phloem by degrading celluloses in plant cell walls, thereby functioning as an effector that overcomes the plant cell wall defense in rice.
BackgroundThe brown planthopper (BPH), Nilaparvata lugens (Stål), a destructive rice pest in Asia, can quickly overcome rice resistance by evolving new virulent populations. Herbivore saliva plays an important role in plant–herbivore interactions, including in plant defense and herbivore virulence. However, thus far little is known about BPH saliva at the molecular level, especially its role in virulence and BPH–rice interaction.Methodology/Principal FindingsUsing cDNA amplification in combination with Illumina short-read sequencing technology, we sequenced the salivary-gland transcriptomes of two BPH populations with different virulence; the populations were derived from rice variety TN1 (TN1 population) and Mudgo (M population). In total, 37,666 and 38,451 unigenes were generated from the salivary glands of these populations, respectively. When combined, a total of 43,312 unigenes were obtained, about 18 times more than the number of expressed sequence tags previously identified from these glands. Gene ontology annotations and KEGG orthology classifications indicated that genes related to metabolism, binding and transport were significantly active in the salivary glands. A total of 352 genes were predicted to encode secretory proteins, and some might play important roles in BPH feeding and BPH–rice interactions. Comparative analysis of the transcriptomes of the two populations revealed that the genes related to ‘metabolism,’ ‘digestion and absorption,’ and ‘salivary secretion’ might be associated with virulence. Moreover, 67 genes encoding putative secreted proteins were differentially expressed between the two populations, suggesting these genes may contribute to the change in virulence.Conclusions/SignificanceThis study was the first to compare the salivary-gland transcriptomes of two BPH populations having different virulence traits and to find genes that may be related to this difference. Our data provide a rich molecular resource for future functional studies on salivary glands and will be useful for elucidating the molecular mechanisms underlying BPH feeding and virulence differences.
Long non-coding RNA (lncRNA) plays an important role in the development of human malignant tumours. Recently, an increasing number of lncRNAs have been identified and investigated in a variety of tumours. However, the expression pattern and biological function of lncRNAs in cervical cancer still remain largely unexplored. Differentially expressed lncRNAs in cervical cancer and para-carcinoma tissues were identified by screening using The Cancer Genome Atlas (TCGA), and candidate lncRNAs were verified by quantitative real-time PCR. We found that lncRNAC5orf66-AS1 was significantly upregulated in cervical cancer tissues and cells. Over-expression of C5orf66-AS1 promoted the proliferation of cervical cancer cells, while downregulation of C5orf66-AS1 promoted the apoptosis of cervical cancer cells. C5orf66-AS1 was identified as the sponge of miR-637 by RNA immunoprecipitation (RIP) and luciferase reporter assays. Exogenous miR-637 and RING1 interventions could reverse the proliferation ability mediated by C5orf66-AS1 in cervical cancer cells. In vivo experiments also confirmed that downregulation of C5orf66-AS1 inhibited the tumour growth. LncRNA C5orf66-AS1, as a competitive endogenous RNA (ceRNA), regulated the effect of RING1 on the proliferation, apoptosis and cell cycle of cervical cancer cells through adsorbing miR-637. Taken together, our findings provided a new theoretical and experimental basis for investigating the pathogenesis and exploring effective therapeutic targets for cervical cancer.
BackgroundIntake of ω-3 PUFAs have been demonstrated to have positive effects on pregnancy outcome, whose receptor, GPR120, regulates several cellular functions including differentiation, metabolism and immune reaction. However, whether GPR120 is involved in decidualization and pregnancy remains unknown.MethodsDecidua tissue from women with normal pregnancy and spontaneous abortion were collected to determine the expression profile of GPR120. Abortion mouse models and artificially induced deciduoma in mice were established to evaluate the effect of GPR120 on pregnancy outcome and in vivo decidualization. HESCs and primary DSCs were used to explore the roles of GPR120 in decidualization and mechanisms involved.FindingsWe found that GPR120 functioned to promote decidualization by upregulating glucose uptake and pentose-phosphate pathway (PPP) of human endometrial stromal cells. Firstly, the expression of GPR120 in decidua of spontaneous abortion was downregulated compared to normal decidua. Lack of GPR120 predisposed mice to LPS or RU486 induced abortion. Decidualization was augmented by GPR120 via improving GLUT1-mediated glucose uptake and G6PD- mediated PPP. FOXO1 was upregulated by GPR120 via activation of ERK1/2 and AMPK signaling and increased the expression of GLUT1. Furthermore, the expression of chemokines and cytokines in decidual stromal cells was enhanced by GPR120. Lastly, GPR120 agonist ameliorated LPS-induced abortion in the mice.InterpretationGPR120 plays significant roles in decidualization and the maintenance of pregnancy, which might be a potential target for diagnosis and treatment of spontaneous abortion.FundMinistry of Science and Technology of China, National Natural Science Foundation of China, the Program of Science and Technology Commission of Shanghai Municipality.
BackgroundThe brown planthopper (BPH), Nilaparvata lugens (Stål), one of the most serious rice insect pests in Asia, can quickly overcome rice resistance by evolving new virulent populations. The insect fat body plays essential roles in the life cycles of insects and in plant-insect interactions. However, whether differences in fat body transcriptomes exist between insect populations with different virulence levels and whether the transcriptomic differences are related to insect virulence remain largely unknown.Methodology/Principal FindingsIn this study, we performed transcriptome-wide analyses on the fat bodies of two BPH populations with different virulence levels in rice. The populations were derived from rice variety TN1 (TN1 population) and Mudgo (M population). In total, 33,776 and 32,332 unigenes from the fat bodies of TN1 and M populations, respectively, were generated using Illumina technology. Gene ontology annotations and Kyoto Encyclopedia of Genes and Genomes (KEGG) orthology classifications indicated that genes related to metabolism and immunity were significantly active in the fat bodies. In addition, a total of 339 unigenes showed homology to genes of yeast-like symbionts (YLSs) from 12 genera and endosymbiotic bacteria Wolbachia. A comparative analysis of the two transcriptomes generated 7,860 differentially expressed genes. GO annotations and enrichment analysis of KEGG pathways indicated these differentially expressed transcripts might be involved in metabolism and immunity. Finally, 105 differentially expressed genes from YLSs and Wolbachia were identified, genes which might be associated with the formation of different virulent populations.Conclusions/SignificanceThis study was the first to compare the fat-body transcriptomes of two BPH populations having different virulence traits and to find genes that may be related to this difference. Our findings provide a molecular resource for future investigations of fat bodies and will be useful in examining the interactions between the fat body and virulence variation in the BPH.
BACKGROUND Calcium (Ca2+)‐binding proteins in the saliva of herbivorous insects function as effectors to attenuate host plant defenses and thus improve insect feeding performance. Silencing these genes via transgenic plant‐mediated RNAi is thus a promising pest control strategy. However, their sequences and functions in the small brown planthopper Laodelphax striatellus (SBPH) remain to be investigated. RESULTS We identified a putative EF‐hand Ca2+‐binding protein (LsECP1) in SBPH watery saliva. LsECP1 was expressed extremely high in the salivary glands but at a low level during the egg stage. Transient LsECP1 expression in rice cells indicated its cytoplasm and nucleus localization. The bacterially expressed recombinant LsECP1 protein exhibited Ca2+‐binding activity. Rice plants fed by SBPH nymphs with knocked down LsECP1 exhibited higher levels of cytosolic Ca2+, jasmonic acid (JA), jasmonoyl‐isoleucine (JA‐Ile) and hydrogen peroxide (H2O2). Consistently, application of heterogeneously expressed LsECP1 protein suppressed wound‐induced JA, JA‐Ile and H2O2 accumulation in rice. Thus, LsECP1 knockdown by dsRNA injection resulted in reduced feeding, fecundity and survival rates of SBPH reared on rice plants. Transgenic rice plants constitutively expressing LsECP1 dsRNA were produced, and plant‐mediated LsECP1 knockdown enhanced rice resistance to SBPH. CONCLUSION SBPH LsECP1 acts as an effector to impair host rice defense responses and promotes SBPH performance. This discovery provides a potential gene target for plant‐mediated RNAi‐based pest management. © 2021 Society of Chemical Industry
Background and context: Surgical procedures are evolving toward less invasive and more tailored approaches to consider the specific pathology, morphology, and life habits of a patient. However, these new surgical methods require thorough preoperative planning and an advanced understanding of biomechanical behaviors. In this sense, patient-specific modeling is developing in the form of digital twins to help personalized clinical decision-making.Purpose: This study presents a patient-specific finite element model approach, focusing on tibial plateau fractures, to enhance biomechanical knowledge to optimize surgical trauma procedures and improve decision-making in postoperative management.Study design: This is a level 5 study.Methods: We used a postoperative 3D X-ray image of a patient who suffered from depression and separation of the lateral tibial plateau. The surgeon stabilized the fracture with polymethyl methacrylate cement injection and bi-cortical screw osteosynthesis. A digital twin of the patient’s fracture was created by segmentation. From the digital twin, four stabilization methods were modeled including two screw lengths, whether or not, to inject PMMA cement. The four stabilization methods were associated with three bone healing conditions resulting in twelve scenarios. Mechanical strength, stress distribution, interfragmentary strains, and fragment kinematics were assessed by applying the maximum load during gait. Repeated fracture risks were evaluated regarding to the volume of bone with stress above the local yield strength and regarding to the interfragmentary strains.Results: Stress distribution analysis highlighted the mechanical contribution of cement injection and the favorable mechanical response of uni-cortical screw compared to bi-cortical screw. Evaluation of repeated fracture risks for this clinical case showed fracture instability for two of the twelve simulated scenarios.Conclusion: This study presents a patient-specific finite element modeling workflow to assess the biomechanical behaviors associated with different stabilization methods of tibial plateau fractures. Strength and interfragmentary strains were evaluated to quantify the mechanical effects of surgical procedures. We evaluate repeated fracture risks and provide data for postoperative management.
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