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The cuticular hydrocarbon (CHC) biosynthetic pathways branches off from the synthesis of fatty acids. Fatty acid elongases (ELOs) are enzymes catalyzing the synthesis of long-chain fatty acids and thereby contribute to the diversification of CHCs. Based on bioinformatics analyses we identified 20 ELO genes in the brown planthopper, Nilaparvata lugens. RNA interference against these genes demonstrated that 9 NlELO genes were essential for the survival of N. lugens nymphs and adults. Indeed, knockdown of NlELO s 1, 3, 4, 7 , 8, 9, 10, 12 and 18 caused lethal phenotypes with a thin and wizened body and reduced lipids in the fat body. Surface analysis by scanning electron microscopy and CHC quantification indicated that knockdown of NlELO s 2, 3, 8 and 16 additionally resulted in a smooth body surface and a decrease in CHC amounts. Therefore, we speculate that long-chain CHCs are needed for CHC attachment to the cuticle surface. CHC deficiency, in turn, resulted in increased adhesion of water droplets and secreted honeydew to the animal surface and the inability of N. lugens to survive in paddy fields with varying humidity. Our present study provides an initial comprehensive analysis of ELO gene functions in an insect, and may serve to better understand the biology of CHCs. Image 1 • Roles of 20 fatty acid elongase (ELO) genes are examined in the brown planthopper Nilaparvata lugens. • NlELO s 2, 3, 8, and 16 function in the production of cuticular hydrocarbons. • NlELO s 2, 3, 8, and 16 are important for both cuticle hydrophobicity and for water retention. • NlELO s 1, 4, 7, 9, 10, 12, and 18 play roles in development and physiology of the brown planthopper. [ABSTRACT FROM AUTHOR]

Simple Summary: During insect mating, male insects trigger post-mating responses in females through seminal fluid proteins, inducing a physiological transition in females that is conducive to reproduction. Simultaneously, this process indirectly increases the male's chances of producing offspring through competition. Females, on the other hand, elicit short- and long-term post-mating responses through proteins and neural networks. Within the intricate regulatory network, seminal fluid proteins and octopamine play pivotal roles. The network spans from the female's reproductive organs and abdominal ganglion to the brain and operates by activating receptors for seminal fluid proteins and octopamine. This activation leads to the modulation of various signals, influencing the female's physiological behavior, organ structure, and hormone secretion, and facilitating enhanced nutrient intake, increased expression of reproductive-related genes, ovulation, and sperm storage. Ultimately, these processes aid in the successful oviposition of females. Following insect mating, females often exhibit a series of physiological, behavioral, and gene expression changes. These post-mating responses (PMRs) are induced by seminal fluid components other than sperm, which not only form network proteins to assist sperm localization, supplement female-specific protein requirements, and facilitate the formation of specialized functional structures, but also activate neuronal signaling pathways in insects. This review primarily discusses the roles of seminal fluid proteins (SFPs) and octopamine (OA) in various PMRs in insects. It explores the regulatory mechanisms and mediation conditions by which they trigger PMRs, along with the series of gene expression differences they induce. Insect PMRs involve a transition from protein signaling to neuronal signaling, ultimately manifested through neural regulation and gene expression. The intricate signaling network formed as a result significantly influences female behavior and organ function, contributing to both successful reproduction and the outcomes of sexual conflict. [ABSTRACT FROM AUTHOR]

Yeast-like symbiotes (YLS) are endosymbionts that are closely related to the growth, development and reproduction of their host, the brown planthopper (BPH), Nilaparvata lugens Stål (Hemiptera: Delphacidae). In order to understand the relationship between the population of YLS in BPH cells and the survival rate of BPH, eight different fungicides were applied to rice plants infested by BPH, and the number of YLS and mortality of BPH were determined. Three of the fungicides, 27% toyocamycin & tetramycin P & tetrin B & tetramycin A, 0.01% trichodermin, and 75% trifloxystrobin & tebuconazole WG, were found to significantly reduce the number of YLS in BPH, subsequently causing a high mortality of BPH. The three fungicides were each mixed with a commonly used insecticide-imidacloprid, and the fungicide/insecticide mixtures could cause a marked reduction in YLS number in BPH, resulting in a significantly higher mortality of BPH than did the imidacloprid alone. The mixture of 27% toyocamycin & tetramycin P & tetrin B & tetramycin A with imidacloprid showed the best inhibitory effect on BPH population. Our study demonstrated a high dependence of the BPH survival rate on the number of YLS harbored in BPH fat-body cells. It implies that using specific fungicides as an additive to imidacloprid for controlling BPH could be a novel way to enhance the efficacy of insecticide, minimizing the use of imidacloprid in paddy fields. [ABSTRACT FROM AUTHOR]

Insect cuticular hydrocarbons (CHCs) are organic compounds of the surface lipid layer, which function as a barrier against water loss and xenobiotic penetration, while also serving as chemical signals. Plasticity of CHC profiles can vary depending upon numerous biological and environmental factors. Here, we investigated potential sources of variation in CHC profiles of Nilaparvata lugens, Laodelphax striatellus and Sogatella furcifera, which are considered to be the most important rice pests in Asia. CHC profiles were quantified by GC/MS, and factors associated with variations were explored by conducting principal component analysis (PCA). Transcriptomes were further compared under different environmental conditions. The results demonstrated that CHC profiles differ among three species and change with different developmental stages, sexes, temperature, humidity and host plants. Genes involved in cuticular lipid biosynthesis pathways are modulated, which might explain why CHC profiles vary among species under different environments. Our study illustrates some biological and ecological variations in modifying CHC profiles, and the underlying molecular regulation mechanisms of the planthoppers in coping with changes of environmental conditions, which is of great importance for identifying potential vulnerabilities relating to pest ecology and developing novel pest management strategies. [ABSTRACT FROM AUTHOR]

Fatty acyl‐CoA reductase (FAR) is one of the key enzymes, which catalyses the conversion of fatty acyl‐CoA to the corresponding alcohols. Among the FAR family members in the brown planthopper (Nilaparvata lugens), NlFAR7 plays a pivotal role in both the synthesis of cuticular hydrocarbons and the waterproofing of the cuticle. However, the precise mechanism by which NlFAR7 influences the formation of the cuticle structure in N. lugens remains unclear. Therefore, this paper aims to investigate the impact of NlFAR7 through RNA interference, transmission electron microscope, focused ion beam scanning electron microscopy (FIB‐SEM) and lipidomics analysis. FIB‐SEM is employed to reconstruct the three‐dimensional (3D) architecture of the pore canals and related cuticle structures in N. lugens subjected to dsNlFAR7 and dsGFP treatments, enabling a comprehensive assessment of changes in the cuticle structures. The results reveal a reduction in the thickness of the cuticle and disruptions in the spiral structure of pore canals, accompanied by widened base and middle diameters. Furthermore, the lipidomics comparison analysis between dsNlFAR7‐ and dsGFP‐treated N. lugens demonstrated that there were 25 metabolites involved in cuticular lipid layer synthesis, including 7 triacylglycerols (TGs), 5 phosphatidylcholines (PCs), 3 phosphatidylethanolamines (PEs) and 2 diacylglycerols (DGs) decreased, and 4 triacylglycerols (TGs) and 4 PEs increased. In conclusion, silencing NlFAR7 disrupts the synthesis of overall lipids and destroys the cuticular pore canals and related structures, thereby disrupting the secretion of cuticular lipids, thus affecting the cuticular waterproofing of N. lugens. These findings give significant attention with reference to further biochemical researches on the substrate specificity of FAR protein, and the molecular regulation mechanisms during N. lugens life cycle. [ABSTRACT FROM AUTHOR]

Hundreds of insect genome sequences have been reported; however, most sequencing projects have not determined the sex chromosomes, and no Y chromosomes from a heterometabolous insect have been identified and characterized to date. The brown planthopper (Nilaparvata lugens Stål) is the most economically damaging pest to rice and is also an ideal research subject for paddy ecology and functional genomics. We previously assembled a draft female genome mainly using second‐generation sequencing technologies, with a contig N50 of only 24 kb, due to the large size and excessive repetitive regions in the N. lugens genome. Here, we utilize third‐generation sequencing technologies and Hi‐C data to generate a high‐quality male N. lugens assembly with a contig N50 of 1.01 Mb, a scaffold N50 of 69.96 Mb and more than 95.6% of the assembled bases located on 16 chromosomes. Fourteen autosomes and two sex chromosomes (X + Y) were identified, filling in the gap related to the Y chromosome in heterometabolous insects. A total of 18,021 protein‐coding genes and 6423 long‐noncoding RNAs were predicted with full‐length cDNA sequencing data. All 315 of the Y chromosome genes (Y‐genes) were derived from autosomal and X‐chromosome duplications. Large‐scale RNA interference (RNAi) experiments were conducted against the N. lugens Y‐genes, demonstrating that 7 Y‐genes were essential for normal BPH development or male organ development, suggesting the importance of Y‐genes. The first identified Y chromosome in heterometabolous insects will help gain more insight into sex determination, fertility and chromosome evolution. [ABSTRACT FROM AUTHOR]

Trichoid sensilla are the most common mechanoreceptors in insects; depending on their distribution, they can act as either exteroceptors or proprioceptors. In this study, the internal structure of the trichoid sensillum from Nilaparvata lugens was studied, using focused ion beam scanning electron microscopy (FIB-SEM). We reconstructed a three-dimensional (3D) model derived from the FIB-SEM data set. The model displayed characteristic mechanosensory sensilla components, including a hair inserted in the socket, a dendrite going through the laminated cuticle, and an electron-dense tubular body at the dendrite terminal. The detailed 3D model showed the relationship between the microtubules within the tubular body and those outside of the tubular body. We also found an autocellular junction in the tormogen cell, indicating that the tormogen cell grows around the dendrite sheath to form a hollow column shape during sensilla morphogenesis. [ABSTRACT FROM AUTHOR]

Abstract: In this paper, a new encapsulation carrier was applied in order to improve the survival of Lactobacillus bulgaricus (L. bulgaricus). L. bulgaricus was encapsulated in alginate–milk microspheres prepared by extrusion method. Around 100% encapsulation yield was achieved. The tolerance of encapsulated L. bulgaricus to adverse environments such as low pH (pH 2.0 and 2.5), high concentration of bile salt (1.0% and 2.0%) and long time storage (1month), was investigated. Release characteristic of encapsulated L. bulgaricus in Simulated Intestine Fluid (SIF) was also studied. The results showed that encapsulation could improve the tolerance of L. bulgaricus to adverse environments. The viability of encapsulated L. bulgaricus did not change after 120min incubation in Simulated Gastric Fluid (SGF) pH 2.5. The viability of encapsulated L. bulgaricus could be kept more than 8logCFU/g after 120min incubation in SGF pH 2.0. The viability of encapsulated L. bulgaricus in 1% bile salt solution was reduced from 9.98logCFU/g microspheres to 9.24 and 8.48logCFU/g microspheres after 1 and 2h incubation, respectively. Around 1.3 and 2.1logCFU/g microspheres were reduced after 1 and 2h exposure in 2% bile salt solution, respectively. Full viability of encapsulated L. bulgaricus could be preserved after 1month’s storage at 4°C. L. bulgaricus encapsulated in alginate–milk microspheres could be completely released in 1h. These studies demonstrated encapsulation of L. bulgaricus in alginate–milk microspheres is an effective protection technique against extreme simulated gastrointestinal environment. [Copyright &y& Elsevier]

Heat shock 70kDa proteins (HSP70s) and their cochaperones DNAJs are ubiquitous molecular chaperones, which function as the "HSP70/DNAJ machinery" in a myriad of biological processes. At present, a number of HSP70s have been classified in many species, but studies on DNAJs, especially in insects, are lacking. Here, we first systematically identified and characterized the HSP70 and DNAJ family members in the brown planthopper (BPH), Nilaparvata lugens, a destructive rice pest in Asia. A total of nine HSP70 and 31 DNAJ genes were identified in the BPH genome. Sequence and phylogenetic analyses revealed the high diversity of the NlDNAJ family. Additionally, spatio-temporal expression analysis showed that most NlHSP70 and NlDNAJ genes were highly expressed in the adult stage and gonads. Furthermore, RNA interference (RNAi) revealed that seven NlHSP70s and 10 NlDNAJs play indispensable roles in the nymphal development, oogenesis, and female fertility of N. lugens under physiological growth conditions; in addition, one HSP70 (NlHSP68) was found to be important in the thermal tolerance of eggs. Together, our results in this study shed more light on the biological roles of HSP70/DNAJ in regulating life cycle, coping with environmental stresses, and mediating the interactions within, or between, the two gene families in insects. [ABSTRACT FROM AUTHOR]

Graphical abstract Highlights • Knockdown of flightin gene resulted in male courtship songs loss. • The lack of male courtship songs prolonged copulation latency and caused female rejection. • The mute males exhibited greater competitiveness. Abstract In duet-based courtship, species- and sex-specific vibrational signals enable animals to identify the species and sex of the singer and also provide the necessary information with which to locate a partner. Substrate-borne communication has been described in a wide variety of insects. Here, we focus on the gene necessary for the emission of male vibrational signals and whether the male song fulfills such a functional role in the mating system of the brown planthopper (BPH, Nilaparvata lugens). We generated mute BPH adult males via RNA interference (RNAi) of the flightin gene, which encodes a myosin-binding protein expressed exclusively in the dorsal longitudinal muscle (DLM) in the basal two abdominal segments used for driving the vibration of the male-specific tymbal structure in short-winged (brachypterous) BPH adults. Transmission electron microscopy (TEM) observation showed that flightin knockdown disrupted the normal sarcomere structure of the abdominal DLM. No courtship song could be detected in the brachypterous males after RNAi treatment. Behavior and competition trials showed that the lack of male courtship songs prolonged copulation latency and even caused female rejection. Unexpectedly, the mute males exhibited greater competitiveness when competing against normal males. [ABSTRACT FROM AUTHOR]

[Display omitted] • CYP380C10 is the ancestor gene of the CYP4G subfamily in N. lugens. • CYP380C10 is involved in CHC biosynthesis. • Knockdown of CYP380C10 generated lethal defect. • Knockdown of CYP380C10 decreased the bodyweight and the amounts of CHCs. • CYP380C10 is an importantly functional gene in waterproof and water-retention. Cuticular hydrocarbons (CHCs) are important components in the integument of insects and are required for development and survival. Insect-specific CYP4G subfamily, of the P450 enzymes, catalyze the oxidative decarbonylation step in the biosynthesis of CHCs. Here, we characterized CYP380C10 gene function in a Hemiptera rice pest, Nilaparvata lugens. We used RNA interference–mediated expression silencing to reveal that NlCYP380C10 played a key role in waterproofing and water–retention in the integument of N. lugens. Knockdown of NlCYP380C10 significantly reduced body weight and caused mortality. Scanning electron microscopy showed the loss of the lipid layer on the surface of the abdominal cuticle of the ds NlCYP380C10 -injected adults. Furthermore, CHC profile analysis revealed that NlCYP380C10 knockdown significantly decreased the amounts of CHCs in adult females. This suggested that NlCYP380C10 was involved in CHC biosynthesis. Reduction of CHC content caused the loss of the intact lipid layer of the cuticle, which resulted in loss of the waterproofing and water-retention functions. This led to failure of molting and eclosion. Our findings expanded the knowledge of CHC biosynthesis in the insect integument and led to a better understanding of the functional roles of CYP450 genes involved in waterproofing and water–retention in insects. [ABSTRACT FROM AUTHOR]

Nicotinamide adenine dinucleotide phosphate (NADPH)-cytochrome P450 reductase (CPR) is an essential enzyme that transfers electrons from NADPH to cytochrome P450 monooxygenases. CPR is involved in cuticular hydrocarbon (CHC) synthesis in insects and is vital for insect development and survival. Here, we clarify the physiological function of a CPR gene in Nilaparvata lugens, an important rice pest, by using RNA interference. CPR gene knockdown leads to the functional loss of waterproofing and water retention in the integument of female adults, which causes significantly reduced body weight and a lethal phenotype. Scanning electron microscopy shows that the lipid layer on the outermost surface of the abdominal cuticle becomes thin in dsCPR-injected adults. Furthermore, CHC profile analysis reveals that CPR knockdown significantly decreases the contents of CHCs with a carbon chain length ≥ C27 in adult females. Moreover, we find that CPR knockdown generates a deficient phenotype in ovaries with deformed oocytes and a complete failure of egg-laying. These findings suggest that CPR plays multiple functional roles in CHC biosynthesis and embryo development in insects. [ABSTRACT FROM AUTHOR]