ESM1 May Be Used as a New Indicator for the Diagnosis and Prognosis of Early and Advanced Stage Digestive Tract Cancers.

Background: The biological function and prognostic significance of endothelial cell specific molecule 1 (ESM1) in various cancers have been validated. This study aimed to explore the expression and clinical diagnosis values in patients with stomach adenocarcinoma (STAD) and esophageal carcinoma (ESCA). Methods: Online database Gene Expression Omnibus was used to screen for abnormally expressed genes in STAD and ESCA. Besides, 36 STAD and 36 ESCA patients were enrolled, and their corresponding control groups were also 36 people each. Reverse transcription-quantitative polymerase chain reaction and Western blot were performed to analyze the expression of ESM1. Overall survival (OS) curve and receiver operating characteristics curve (ROC) analysis were used to assess the prognosis, and the sensitivity and specificity of ESM1 for the diagnosis of STAD and ESCA, respectively. Additionally, the effects of ESM1 on cell viability, migration, and invasion were analyzed by cell counting kit-8, transwell migration and invasion assays. Results: The results showed that the poor OS of STAD and ESCA patients was correlated with high ESM1. Besides, ESM1 was increased in ESCA and STAD in in vivo and in vitro studies. ESM1 has a high accuracy [area under the curve (AUC) > 0.79] at stage I and IV of STAD and ESCA. Knockdown of ESM1 suppressed the cell viability, migration, and invasion and increased the apoptosis rate of AGS and TE1 cells. Conclusion: Our study suggested that ESM1 might be used as a new indicator for the diagnosis and prognosis of early and advanced stage digestive tract cancers.

Alanine metabolism mediates energy allocation of the brown planthopper to adapt to resistant rice.

INTRODUCTION: During the adaptation to host plant resistance, herbivorous insects faced the challenge of overcoming plant defenses while ensuring their own development and reproductive success. To achieve this, a strategic allocation of energy resources for detoxification and ecological fitness maintenance became essential. OBJECTIVE: This study aimed to elucidate the intricate energy allocation mechanisms involved in herbivore adaptation that are currently poorly understood. METHODS: The rice Oryza sativa and its monophagous pest, the brown planthopper (BPH), Nilaparvata lugens were used as a model system. An integrated analysis of metabolomes and transcriptomes from different BPH populations were conducted to identify the biomarkers. RNA interference of key genes and exogenous injection of key metabolites were performed to validate the function of biomarkers. RESULTS: We found that alanine was one of the key biomarkers of BPH adaptation to resistant rice variety IR36. We also found that alanine flow determined the adaptation of BPH to IR36 rice. The alanine aminotransferase (ALT)-mediated alanine transfer to pyruvate was necessary and sufficient for the adaptation. This pathway may be conserved, at least to some extent, in BPH adaptation to multiple rice cultivars with different resistance genes. More importantly, ALT-mediated alanine metabolism is the foundation of downstream energy resource allocation for the adaptation. The adapted BPH population exhibited a significantly higher level of energy reserves in the fat body and ovary when fed with IR36 rice, compared to the unadapted population. This rendered the elevated detoxification in the adapted BPH and their ecological fitness recovery. CONCLUSION: Overall, our findings demonstrated the crucial role of ALT-mediated alanine metabolism in energy allocation during the adaptation to resistant rice in BPH. This will provide novel knowledge regarding the co-evolutionary mechanisms between herbivores and their host plants.

Development of a prognostic model for personalized prediction of colon adenocarcinoma (COAD) patient outcomes using methylation-driven genes.

The objective of this study was to identify methylation-driven genes and explore their prognostic value in colon adenocarcinoma (COAD). The Cancer Genome Atlas (TCGA) database was used to acquire collated COAD transcriptome gene expression matrix (containing 59,427 transcripts), transcriptome gene methylation level matrix (containing 29,602 methylated modified genes), which included 517 samples containing 41 samples of normal tissue (NT) & 476 samples of COAD, and patient clinical information files (including patient survival time, survival status, age, gender and tumor stage, etc.), for all COAD samples. A total of 9807 differentially expressed genes (DEGs) were obtained by DEG analysis of the COAD transcriptional expression matrix, of which 5874 were up-regulated and 3933 were down-regulated. And 46 methylation-driven DEGs (MD-DEGs) in COAD were obtained by DEG analysis, differential analysis of gene methylation levels, and correlation analysis between them. Next, three prognostic associated MD-DEGs (PMD-DEGs) (IDUA, ZBTB18 and C5orf38) were identified by Cox regression analysis, and a prognostic model composed of the three PMD-DEGs was constructed by least absolute shrinkage and selection operator (LASSO) regression analysis and cross-validation analysis. In addition, survival analysis, the receiver operating characteristics (ROC) curve analysis and independent prognostic analysis were used to evaluate and verify that the prognostic model we constructed could accurately and independently predict the prognosis of COAD patients. Finally, we constructed a nomogram based on the prognosis model to accurately and personalized predict the survival prognosis of COAD patients. In conclusion, we identified the methylation driver gene of COAD and constructed a prognostic model and nomogram to personalized predict the prognosis of patients, which opened a new prospect for accurate diagnosis and treatment in clinical practice.

Poaceae-specific ß-1,3;1,4-d-glucans link jasmonate signalling to OsLecRK1-mediated defence response during rice-brown planthopper interactions.

Brown planthopper (BPH, Nilaparvata lugens), a highly destructive insect pest, poses a serious threat to rice (Oryza sativa) production worldwide. Jasmonates are key phytohormones that regulate plant defences against BPH; however, the molecular link between jasmonates and BPH responses in rice remains largely unknown. Here, we discovered a Poaceae-specific metabolite, mixed-linkage ß-1,3;1,4-d-glucan (MLG), which contributes to jasmonate-mediated BPH resistance. MLG levels in rice significantly increased upon BPH attack. Overexpressing OsCslF6, which encodes a glucan synthase that catalyses MLG biosynthesis, significantly enhanced BPH resistance and cell wall thickness in vascular bundles, whereas knockout of OsCslF6 reduced BPH resistance and vascular wall thickness. OsMYC2, a master transcription factor of jasmonate signalling, directly controlled the upregulation of OsCslF6 in response to BPH feeding. The AT-rich domain of the OsCslF6 promoter varies in rice varieties from different locations and natural variants in this domain were associated with BPH resistance. MLG-derived oligosaccharides bound to the plasma membrane-anchored LECTIN RECEPTOR KINASE1 OsLecRK1 and modulated its activity. Thus, our findings suggest that the OsMYC2-OsCslF6 module regulates pest resistance by modulating MLG production to enhance vascular wall thickness and OsLecRK1-mediated defence signalling during rice-BPH interactions.

Oxalic Acid Inhibits Feeding Behavior of the Brown Planthopper via Binding to Gustatory Receptor Gr23a.

Plants produce diverse secondary compounds as natural protection against microbial and insect attack. Most of these compounds, including bitters and acids, are sensed by insect gustatory receptors (Grs). Although some organic acids are attractive at low or moderate levels, most acidic compounds are potentially toxic to insects and repress food consumption at high concentrations. At present, the majority of the reported sour receptors function in appetitive behaviors rather than aversive taste responses. Here, using two different heterologous expression systems, the insect Sf9 cell line and the mammalian HEK293T cell line, we started from crude extracts of rice (Oryza sativa) and successfully identified oxalic acid (OA) as a ligand of NlGr23a, a Gr in the brown planthopper Nilaparvata lugens that feeds solely on rice. The antifeedant effect of OA on the brown planthopper was dose dependent, and NlGr23a mediated the repulsive responses to OA in both rice plants and artificial diets. To our knowledge, OA is the first identified ligand of Grs starting from plant crude extracts. These findings on rice-planthopper interactions will be of broad interest for pest control in agriculture and also for better understanding of how insects select host plants.

Ligand dose-dependent activation of signaling pathways through the gustatory receptor NlGr11 linked to feeding efficacy in Nilaparvata lugens.

Insects often face both conditions with sufficient nutrients and conditions of undernutrition in the field. Through gustatory receptors, insects sense nutrients and regulate their physiological functions such as feeding and reproduction. However, it remains unclear whether signaling pathways activated by gustatory receptors depend on the concentration of nutrients and whether the difference in signaling pathways directly affects insects' physiological functions. Herein, we found that a sugar gustatory receptor, NlGr11, from the brown planthopper (BPH), Nilaparvata lugens, activated G protein-coupled signaling and ionotropic pathways when bound to high galactose concentration. BPHs subsequently demonstrated longer feeding times, feeding loads, and higher vitellogenin (NlVg) expression than BPHs exposed to high galactose concentrations, which only activated the ionotropic pathway. For the first time, our findings link plant nutrient conditions, signaling pathways activated by nutrients, and their gustatory receptors, and nutrient dose-dependent feeding efficacy and vitellogenin (Vg) expression in an insect. This will help us to better understand the molecular mechanism for insect feeding strategies on plants at different stages of nutritional conditions.

Population density modulates insect progenitive plasticity through the regulation of dopamine biosynthesis.

Insect fecundity is a quantitative phenotype strongly affected by genotypes and the environment. However, interactions between genotypes and environmental factors in modulating insect fecundity remain largely unknown. This study investigated the impact of population density on the fecundity of Nilaparvata lugens (brown planthopper; BPH) carrying homozygous high- (HFG) or low- (LFG) fecundity homozygous genotypes. Under low population densities, the fecundity and population growth rate of both genotypes showed similar increasing trends across generations, while the trends between HFG and LFG under high population densities were opposite. Through a combination of temporal analysis and weighted gene co-expression network analyses on RNA-seq data of HFG and LFG under low and high population densities in the 1st, 3rd, and 5th generations, we identified 2 gene modules that were associated with these density-dependent progenitive phenotypes. Four pathways related to the neural system were simultaneously enriched by the 2 gene modules. Furthermore, Nlpale, which encodes a tyrosine hydroxylase, was identified as a key gene. The RNA interference of this gene and manipulation of its downstream product dopamine significantly affected the basic and density-dependent progenitive phenotypes of BPH. These findings indicated that dopamine biosynthesis is the key regulatory factor that determines fecundity in response to density changes in different BPH genotypes. Thus, this study provides insights into the interaction of a typical environmental factor and insect genotype during the process of population regulation.

Bacterial distribution on the ocular surface of patients with primary Sjögren's syndrome.

Many studies have shown that gut microbial dysbiosis is a major factor in the etiology of autoimmune diseases but none have suggested that the ocular surface (OS) microbiome is associated with Sjögren's syndrome (SS). In this prospective study, we analyzed bacterial distribution on the OS in patients with primary SS. Among the 120 subjects included in this study, 48 patients (group A) had primary SS, whereas 72 subjects (group B) had dry eye symptoms that were unrelated to SS. We evaluated clinical dry eye parameters such as the OS disease index, ocular staining score (OSS), Schirmer's I test, and tear break-up time (TBUT). Conjunctival swabs were used to analyze the microbial communities from the two groups. Bacterial 16S rRNA genes were sequenced using the Illumina MiSeq platform, and the data were analyzed using the QIIME 1.9.1 program. The Shannon index was significantly lower in group A than in group B microbiota (p < 0.05). An analysis of similarity using the Bray-Curtis distance method found no difference in beta-diversity between the two groups (p > 0.05). In group A, Actinobacteria at the phylum level and Corynebacteria at the genus level exhibited low abundance than group B, but the differences were not statistically significant (p > 0.05). SS apparently decreases the diversity of the OS microbial community. These observations may be related to the pathophysiology of SS and should be investigated in future studies.

Dietary guanidineacetic acid supplementation ameliorated meat quality and regulated muscle metabolism of broilers subjected to pre-slaughter transport stress by metabolomics analysis.

Pre-slaughter transport stress could induce multiple comprehensive variations in physiological and metabolic parameters of broilers. However, the entire metabolomics of pre-slaughter transport stress and supplementation of exogenous energy regulatory substances on broilers is still poorly understood. The metabolome characteristics of broilers subjected to 3 h pre-slaughter transport stress combined with 1,200 mg/kg guanidinoacetic acid (GAA1,200) supplementation were analyzed using gas chromatography-mass spectrometry (GC-MS) in this study. The results showed that, compared to the control group (no transport), 3 h pre-slaughter transport stress (T3h) decreased creatine (Cr), phosphocreatine (PCr) and adenosine triphosphate (ATP), and increased adenosine diphosphate (ADP), adenosine monophosphate (AMP) and the ratio of AMP to ATP in pectoralis muscle (PM) of broilers by high performance liquid chromatography (HPLC) analysis. However, GAA1,200 supplementation reversed the negative effects induced by 3 h pre-slaughter transport stress. Besides, GAA1,200 supplementation elevated mRNA expression of creatine transporter in PM. Our metabolomics approaches demonstrated that 38 and 48 significant metabolites were separately identified between the control group and T3h group, and T3h group and 3 h pre-slaughter transport stress combined with GAA1,200 supplementation group using the standard of variable importance in the projection values >1 and P < 0.05. Among these, the metabolites involved in amino acid metabolism (alanine, glycine, serine, threonine, cysteine , methionine, phenylalanine, tyrosine, and tryptophan), oxidative stress (3-methylhistidine, 1-methylhistidine and glutathione), non-protein amino acid (citrulline) metabolism, and energy metabolism (Cr, PCr, sarcosine, and glycocyamine) were confirmed through pathway enrichment analysis, which could be chosen as suitable candidate targets for further analysis of the effects of exogenous energy substances on broilers subjected to transport stress.

Genome-wide Analysis of Alternative Gene Splicing Associated with Virulence in the Brown Planthopper Nilaparvata lugens (Hemiptera: Delphacidae).

Alternative splicing of protein coding genes plays a profound role in phenotypic variation for many eukaryotic organisms. The development of high-throughput sequencing and bioinformatics algorithms provides the possibility of genome-wide identification of alternative splicing events in eukaryotes. However, for the brown planthopper Nilaparvata lugens, a destructive pest of rice crops, whole-genome distribution of alternative splicing events and the role of alternative splicing in the phenotypic plasticity of virulence have not previously been estimated. Here, we developed an analysis pipeline to identify alternative splicing events in the genome of N. lugens. Differential expression analysis and functional annotation were performed on datasets related to different virulence phenotypes. In total, 27,880 alternative splicing events corresponding to 9,787 multi-exon genes were detected in N. lugens. Among them, specifically expressed alternative splicing transcripts in the virulent Mudgo population were enriched in metabolic process categories, while transcripts in the avirulent TN1 population were enriched in regulator activity categories. In addition, genes encoding odorant receptor, secreted saliva protein and xenobiotic metabolic P450 monooxygenase showed different splicing patterns between Mudgo population and TN1 population. Host change experiment also revealed that an isoform of a P450 gene could be specially induced by the stimulation of resistant rice variety Mudgo. This research pioneered a genome-wide study of alternative gene splicing in the rice brown planthopper. Differences in alternative splicing between virulent and avirulent populations indicated that alternative splicing might play an important role in the formation of virulence phenotypes in N. lugens.

Therapeutic Efficacy of Autologous Platelet Concentrate Injection on Macular Holes with High Myopia, Large Macular Holes, or Recurrent Macular Holes: A Multicenter Randomized Controlled Trial.

We aimed to evaluate the anatomical and functional outcomes of pars-plana vitrectomy (PPV) with or without autologous platelet concentrate (APC) injection in patients with recurrent macular holes (MHs), large MHs, or MHs with high myopia. This multicenter, prospective, interventional randomized controlled trial was conducted from March 2017 to April 2020. Participants were randomly allocated to a PPV group or a PPV+APC group. All participants underwent standard 25-gauge PPV, and eyes in the PPV+APC group underwent PPV with intravitreal APC injection before air-gas exchange. A total of 117 patients were enrolled (PPV group: n = 59, PPV+APC group: n = 58). Hole closure was achieved in 47 participants (79.7%) in the PPV group and 52 participants (89.7%) in the PPV+APC group. There were no between-group differences in the anatomical closure rate or functional outcomes including best-corrected visual acuity, metamorphopsia, pattern-reversal visual evoked potential, or Visual Function Questionnaire-25 score. The use of APC injection does not improve the anatomical and functional outcomes of surgery for large MHs, recurrent MHs, or MHs with high myopia. The adjunctive use of APC can be considered in selected cases because it is not inferior to conventional MH surgery, is relatively simple to perform, and is not affected by the surgeon's skill.

Machine learning prediction of pathologic myopia using tomographic elevation of the posterior sclera.

Qualitative analysis of fundus photographs enables straightforward pattern recognition of advanced pathologic myopia. However, it has limitations in defining the classification of the degree or extent of early disease, such that it may be biased by subjective interpretation. In this study, we used the fovea, optic disc, and deepest point of the eye (DPE) as the three major markers (i.e., key indicators) of the posterior globe to quantify the relative tomographic elevation of the posterior sclera (TEPS). Using this quantitative index from eyes of 860 myopic patients, support vector machine based machine learning classifier predicted pathologic myopia an AUROC of 0.828, with 77.5% sensitivity and 88.07% specificity. Axial length and choroidal thickness, the existing quantitative indicator of pathologic myopia only reached an AUROC of 0.758, with 75.0% sensitivity and 76.61% specificity. When all six indices were applied (four TEPS, AxL, and SCT), the discriminative ability of the SVM model was excellent, demonstrating an AUROC of 0.868, with 80.0% sensitivity and 93.58% specificity. Our model provides an accurate modality for identification of patients with pathologic myopia and may help prioritize these patients for further treatment.

FoxO directly regulates the expression of TOR/S6K and vitellogenin to modulate the fecundity of the brown planthopper.

As a conserved transcription factor, FoxO plays a crucial role in multiple physiological processes in vivo, including stress resistance, longevity, growth and reproduction. Previous studies on FoxO have focused on human, mouse, Drosophila melanogaster and Caenorhabditis elegans, while there are few reports on agricultural pests and little is known about how FoxO modulates insect fecundity. In Asia, the brown planthopper (BPH) Nilaparvata lugens (Stål) is one of the most serious pests in rice production and high fecundity is the basis of the outbreak of BPH. Here, using the genome-wide ChIP-seq of NlFoxO in BPH, we found that NlFoxO binds to the promoters of ribosomal proteinS6 kinase (NlS6K) and serine/threonine-protein kinase mTOR (NlTOR) and increases their expression levels. We also found that NlFoxO directly binds to the exon of vitellogenin (NlVg) and has a specific inhibitory effect on its expression. In addition, the number of eggs laid and their hatching rate decreased significantly after injection of NlFoxO double-stranded RNA into BPH adults. Our findings provide direct evidence that FoxO modulates insect fecundity through binding to the promoters of NlS6K, NlTOR and the exon of NlVg and affecting their gene expression in the Vg network.

Galactose-NlGr11 inhibits AMPK phosphorylation by activating the PI3K-AKT-PKF-ATP signaling cascade via insulin receptor and Gßγ.

As ligands of the sugar gustatory receptors, sugars have been known to activate the insulin/insulin-like growth factor signaling pathway; however, the precise pathways that are activated by the sugar-bound gustatory receptors in insects remain unclear. In this study, we aimed to investigate the signaling cascades activated by NlGr11, a sugar gustatory receptor in the brown planthopper Nilaparvata lugens (Stål), and its ligand. Galactose-bound NlGr11 (galactose-NlGr11) activated the -phosphatidylinositol 3-kinase (PI3K)-AKT signaling cascade via insulin receptor (InR) and Gßγ in vitro. In addition, galactose-NlGr11 inhibited the adenosine monophosphate-activated protein kinase (AMPK) phosphorylation by activating the AKT-phosphofructokinase (PFK)-ATP signaling cascade in vitro. Importantly, the InR-PI3K-AKT-PFK-AKT signaling cascade was activated and the AMPK phosphorylation was inhibited after feeding the brown planthoppers with galactose solution. Collectively, these findings confirm that NlGr11 can inhibit AMPK phosphorylation by activating the PI3K-AKT-PFK-ATP signaling cascades via both InR and Gßγ when bound to galactose. Thus, our study provides novel insights into the signaling pathways regulated by the sugar gustatory receptors in insects.

Effects of Different Nutritional Conditions on the Growth and Reproduction of Nilaparvata lugens (Stål).

Growth and reproduction are the two most basic life processes of organisms and the distribution of energy in these processes is a core issue of the life history of organisms. Nilaparvata lugens (Stål), the brown planthopper (BPH), is a single-feeding rice pest. In the present study, this species was used as a model for testing the effects of nutritional conditions on various growth and reproduction indicators. First, the third-instar nymphs were fed with three different concentrations (100, 50, and 25%) of artificial diet until the second day of adulthood. The results showed that as the nutrient concentration decreased, the body development and oviposition of BPH were hindered. The total lipid content in the fat bodies was also significantly reduced. RT-PCR analysis showed compared to the 100% concentration group, the expression levels of vitellogenin (Vg) genes in the fifth-instar nymphs, adults, and in different tissues (ovary, fat body, and other tissues) were significantly decreased in the 50 and 25% treatment groups. Western blot analysis showed that Vg protein expression was highest in the 100% group, followed by the 50% group, with no expression in the 25% group. These results indicate that growth and reproduction in the BPH are regulated by, or correlated with, nutrient concentration. This study is of great significance as it reveals the adaptive strategies of the BPH to nutritional deficiencies and it also provides valuable information for the comprehensive control of this pest.

SMRT sequencing of the full-length transcriptome of the white-backed planthopper Sogatella furcifera.

The white-backed planthopper Sogatella furcifera is an economically important rice pest distributed throughout Asia. It damages rice crops by sucking phloem sap, resulting in stunted growth and plant virus transmission. We aimed to obtain the full-length transcriptome data of S. furcifera using PacBio single-molecule real-time (SMRT) sequencing. Total RNA extracted from S. furcifera at various developmental stages (egg, larval, and adult stages) was mixed and used to generate a full-length transcriptome for SMRT sequencing. Long non-coding RNA (lncRNA) identification, full-length coding sequence prediction, full-length non-chimeric (FLNC) read detection, simple sequence repeat (SSR) analysis, transcription factor detection, and transcript functional annotation were performed. A total of 12,514,449 subreads (15.64 Gbp, clean reads) were generated, including 630,447 circular consensus sequences and 388,348 FLNC reads. Transcript cluster analysis of the FLNC reads revealed 251,109 consensus reads including 29,700 high-quality reads. Additionally, 100,360 SSRs and 121,395 coding sequences were identified using SSR analysis and ANGEL software, respectively. Furthermore, 44,324 lncRNAs were annotated using four tools and 1,288 transcription factors were identified. In total, 95,495 transcripts were functionally annotated based on searches of seven different databases. To the best of our knowledge, this is the first study of the full-length transcriptome of the white-backed planthopper obtained using SMRT sequencing. The acquired transcriptome data can facilitate further studies on the ecological and viral-host interactions of this agricultural pest.

The Shape of Posterior Sclera as a Biometric Signature in Open-angle Glaucoma: An Intereye Comparison Study.

PURPOSE: To characterize intereye differences in posterior segment parameters and determine their significance in open-angle glaucoma patients with unilateral damage. METHODS: Both eyes from 65 subjects without any nerve damage and 43 patients undergoing treatment for unilateral open-angle glaucoma were included in this study. A 12.0×9.0×2.6 mm volume of the posterior segment in each eye was scanned with swept-source optical coherence tomography. Coronally reconstructed optical coherence tomography images were analyzed to determine the deepest point of the eye (DPE), which we then calculated the distance (Disc-DPE distance), depth (Disc-DPE depth), angle (Disc-DPE angle) from the optic disc center to the DPE. Posterior pole shape was analyzed measuring the posterior pole-cross-sectional area, posterior pole-horizontal width (PP-HW), and posterior pole-vertical width) of the posterior pole. These measurements and their intereye absolute difference (IAD; absolute difference in measurements between the right and left eyes) values were compared between the healthy and unilateral glaucomatous patients. RESULTS: The posterior sclera measurements, including the Disc-DPE distance, Disc-DPE depth, and posterior pole-cross-sectional area, were significantly different between the unilateral glaucoma eyes and contralateral healthy eyes (P=0.043, P=0.035, and P=0.049, respectively). By contrast, none of the intereye differences in optic nerve head parameters were significant in the unilateral glaucoma patients. In comparison with the IAD values, the baseline intraocular pressure and PP-HW of the posterior segment showed significant differences between the healthy and the unilateral glaucoma patients (P=0.019 and P=0.036, respectively). A multivariate analysis showed that a larger baseline intraocular pressure IAD [odds ratio (OR), 1.381; P=0.009)] and larger PP-HW IAD (OR, 1.324; P=0.032) were significantly associated with the presence of glaucoma. CONCLUSIONS: Compared with the fellow healthy eyes, glaucomatous eyes had larger and more steeply curved posterior poles, which represent a structural variation of the posterior sclera that might be associated with glaucomatous optic neuropathy.

Effect of glycogen synthase and glycogen phosphorylase knockdown on the expression of glycogen- and insulin-related genes in the rice brown planthopper Nilaparvata lugens.

Nilaparvata lugens is a serious threat to rice growth. Glycogen metabolism is one of the important physiological processes of insects, which is mainly regulated by glycogen synthase (GS) and glycogen phosphorylase (GP). In the present study, trehalose content was significantly reduced at 72 h after NlGP and NlGS knockdown, whereas glucose content was significantly increased at both 48 h and 72 h after GS knockdown. RNAi combined with RNA-Seq was used to identify NlGP- and NlGS-related pathways and genes in N. lugens. A total of 593 genes were up-regulated and 5969 genes were down-regulated after NlGP and NlGS knockdown, respectively. Moreover, the NlGS-knockdown group was mapped to 10,967 pathways, whereas the NlGP-knockdown group was mapped to 7948 pathways, and the greatest differences between the groups were associated with carbohydrate, lipid, amino acid and energy metabolism. Meanwhile, 1800, 1217, and 1211 transcripts in the NlGP-knockdown group and 2511, 1666, and 1727 transcripts in the NlGS-knockdown group were involved in bioprocess, cellular ingredients and molecular function, respectively. Almost all these genes were down-regulated by either NlGP or NlGS knockdown, with significant down-regulation of the 6-trehalose phosphate synthase (TPS), trehalase (TRE), GS, GP, phosphoacetylglucosamine mutase (PGM, n = 2), Insulin receptors (InRs) and insulin-like peptides (Ilps) genes. These results have demonstrated that RNAi-mediated NlGP and NlGS knockdown could lead to content of trehalose and glucose out of balance, but have no obvious effect on glycogen content, and have suggested that GS plays more complex role in other metabolism pathway of N. lugens.

Two alternative splicing variants of a sugar gustatory receptor modulate fecundity through different signalling pathways in the brown planthopper, Nilaparvata lugens.

Insect gustatory receptors play crucial roles in multiple physiological behaviours. Although the alternative splicing of some gustatory receptors has been observed in insect species, differences in their ligands and functions have rarely been reported. Here, we cloned NlGr10a and NlGr10b, two alternative splicing variants of a sugar gustatory receptor gene in the brown planthopper (BPH), Nilaparvata lugens (Stål), and found that their ligands were different by calcium imaging assay. The ligands of NlGr10a were fructose and cellobiose, and the ligand of NlGr10b was arabinose. Subsequently, the RNAi results showed that knockdown of both splicing variants decreased the number of eggs laid by BPH females, and the egg hatching rate after knockdown of NlGr10a was significantly lower than that after knockdown of NlGr10b. Furthermore, NlGr10a promoted the fecundity of BPH through the AMPK- and AKT-NlVg/NlVgR signalling pathways, whereas NlGr10b promoted the fecundity only through the AMPK- and AKT-NlVg signalling pathways. These findings broaden our understanding of the difference in the ligands and functions of alternative splicing variants of gustatory receptors in insects.

Comparative metabolomics analysis of different resistant rice varieties in response to the brown planthopper Nilaparvata lugens Hemiptera: Delphacidae.

INTRODUCTION: The brown planthopper (BPH, Nilaparvata lugens Stål, Hemiptera: Delphacidae) is one of the most devastating insect pests of the crucially important cereal crop, rice (Oryza sativa L.). Currently, multiple BPH-resistant rice varieties have been cultivated and generalized to control BPH. However, the defence metabolic responses and their modes of action against BPH in different rice cultivars remain uncharacterized. OBJECTIVE: We used a non-biased metabolomics approach to explore the differences in metabolite profiles in response to BPH infestation in the susceptible TN1 rice cultivar and two resistant cultivars (IR36 and IR56). METHODS: The metabolomic detection based on gas chromatography-mass spectrometry (GC-MS) and liquid chromatography-mass spectrometry (LC-MS) was performed to investigate the content changes of identified metabolites in TN1, IR36 and IR56 rice varieties at various time points (0 h, 24 h, 48 h and 96 h) post BPH feeding. The differentially expressed metabolites were screened and the corresponding metabolic pathways were further enriched. RESULTS: The results showed that compared to that in TN1, the content changes of most primary metabolites were more stable, but the concentration alterations of some defence-related metabolites were more acute and persistent in IR36 and IR56. Furthermore, the differentially expressed pathways analysis revealed that cyanoamino acids and lipids metabolism was persistently induced in IR36, but changes in thiamine, taurine and hypotaurine metabolism were more significant in IR56 during BPH infestation. Besides, the contents of quercetin and spermidine which were harmful to BPH fitness, were significantly elevated by BPH in TN1 and IR36, and the quercetin level was significantly decreased during BPH feeding in IR56. CONCLUSION: The results of the differences in metabolite profiles in response to BPH infestation in different rice cultivars were useful to clarify the metabolic mechanism of rice plants during BPH infestation and to provide new resources to control this insect pest.