Brummer‐dependent lipid mobilization regulates starvation resistance in Nilaparvata lugens

Energy homeostasis is an essential characteristic of all organisms, requiring fluctuation in energy accumulation, mobilization, and exchange with the external environment. In insects, energy mobilization is under control of the lipase brummer (bmm), which regulates nutritional status by hydrolyzing the ester bonds in triacylglycerol (TAG). In the present study, we investigated the role of bmm in the lipid mobilization and starvation resistance in the brown planthopper (BPH; Nilaparvata lugens), which is economically one of the most important rice pests in Asia. A severe decrease in TAG and glyceride contents was observed in the starved BPHs, while there was a partial rescue after refeeding. The starvation condition caused a significant increase in the expression levels of Nlbmm, and supplement of food after starvation dramatically reduced the Nlbmm expression. Sucrose rescue after starvation significantly suppressed the expression of Nlbmm, while caused an accumulation of TAG and glyceride. Knockdown of Nlbmm by double‐stranded RNA treatment extended the lifespan to starvation, whereas it increased the level of TAG and glyceride in the BPHs. The decreased lipolysis rate by dsNlbmm‐treated BPHs eventually resulted in increase of starvation resistance. These data demonstrated that the regulation of energy homeostasis by Nlbmm affects starvation resistance, probably through lipid mobilization control in N. lugens.     

Enhancement of the geomagnetic field reduces the phototaxis of rice brown planthopper Nilaparvata lugens associated with frataxin down‐regulation

The geomagnetic field (GMF) is an environmental cue that provides directional information for animals. The intensity of GMF is varied over space and time. Variations in the GMF intensity affect the navigation of animals and their physiology. In this study, the phototaxis of the migratory insect rice planthopper Nilaparvata lugens (N. lugens) and frataxin in N. lugens (Nl‐fh), which is a mitochondrial protein required for cellular iron homeostasis and iron‐sulfur cluster assembly, were investigated by using different intensities of magnetic field. From the results, individuals of N. lugens showed decreased phototaxis when reared and tested in a behavioral arena under a strong magnetic field. Besides the reduction in performance, an accompanying effect of the strong magnetic field condition was a reduced level of Nl‐fh‐messenger RNA, and a Nl‐fh knockdown indeed impaired the phototactic behavior in a tested sample of insects. This leads to the conclusion that the expression of frataxin is dependent on the strength of the surrounding magnetic field and that functional frataxin facilitates phototactic behavior in N. lugens.     

Brummer lipase is an evolutionary conserved fat storage regulator in Drosophila

Energy homeostasis, a fundamental property of all organisms, depends on the ability to control the storage and mobilization of fat, mainly triacylglycerols (TAG), in special organs such as mammalian adipose tissue or the fat body of flies. Malregulation of energy homeostasis underlies the pathogenesis of obesity in mammals including human. We performed a screen to identify nutritionally regulated genes that control energy storage in the model organism Drosophila. The brummer (bmm) gene encodes the lipid storage droplet-associated TAG lipase Brummer, a homolog of human adipocyte triglyceride lipase (ATGL). Food deprivation or chronic bmm overexpression depletes organismal fat stores in vivo, whereas loss of bmm activity causes obesity in flies. Our study identifies a key factor of insect energy homeostasis control. Their evolutionary conservation suggests Brummer/ATGL family members to be implicated in human obesity and establishes a basis for modeling mechanistic and therapeutic aspects of this disease in the fly.     

Identification of a sugar gustatory receptor and its effect on fecundity of the brown planthopper Nilaparvata lugens

In insects, the gustatory system plays a crucial role in multiple physiological behaviors, including feeding, toxin avoidance, courtship, mating and oviposition. Gustatory stimuli from the environment are recognized by gustatory receptors. To date, little is known about the function of gustatory receptors in agricultural pest insects. In this study, we cloned a sugar gustatory receptor gene, N/Gr11, from the brown planthopper (BPH), Nilaparvata lugens (Stal), a serious pest of rice in Asia;we then identified its ligands, namely, fructose, galactose and arabinose, by calcium imaging assay. After injection of N/Gr11 doublestranded RNA, we found that the number of eggs laid by BPH decreased. Moreover, we found that N/Gr11 inhibited the phosphorylation of adenosine monophosphate-activated protein kinase (AMPK) and promoted the phosphorylation of protein kinase B (AKT). These findings demonstrated that N/Gr11 could accelerate the fecundity of BPH through AMPK- and AKT-mediated signaling pathways. This is the first report to indicate that a gustatory receptor modulates the fecundity of insects and that the receptor could be a potential target for pest control.     

The role of chemosensory protein 10 in the detection of behaviorally active compounds in brown planthopper,Nilaparvata lugens

Chemosensory proteins (CSPs) play important roles in insects’ chemoreception, although their specific functional roles have not been fully elucidated. In this study, we conducted the developmental expression patterns and competitive binding assay as well as knock‐down assay by RNA interference both in vitro and in vivo to reveal the function of NlugCSP10 from the brown planthopper (BPH), Nilaparvata lugens (Stål), a major pest in rice plants. The results showed that NlugCSP10 messenger RNA was significantly higher in males than in females and correlated to gender, development and wing forms. The fluorescence binding assays revealed that NlugCSP10 exhibited the highest binding affinity with cis‐3‐hexenyl acetate, eicosane, and (+)‐β‐pinene. Behavioral assay revealed that eicosane displayed attractant activity, while cis‐3‐hexenyl acetate, similar to (+)‐β‐pinene significantly repelled N. lugens adults. Silencing of NlugCSP10, which is responsible for cis‐3‐hexenyl acetate binding, significantly disrupted cis‐3‐hexenyl acetate communication. Overall, findings of the present study showed that NlugCSP10 could selectively interrelate with numerous volatiles emitted from host plants and these ligands could be designated to develop slow‐release mediators that attract/repel N. lugens and subsequently improve the exploration of plans to control this insect pest.     

The regulation of lipid metabolism by a hypothetical P-loop NTPase and its impact on fecundity of the brown planthopper

BackgroundInsect fecundity can be regulated by multiple genes in several important signaling pathways which form an extremely complicated regulatory network. However, there are still many genes that have significant impact on insect fecundity but their action mode are still unknown.MethodsQuantitative real-time PCR (qRT-PCR), immunofluorescence and western blot were used to study the expression profile of Nl23867 in the brown planthopper, Nilaparvata lugens. RNA interference (RNAi), RNA-seq and isobaric tags for relative and absolute quantification (iTRAQ) were performed to investigate the action mode of Nl23867 in the regulation of fecundity. High performance liquid chromatography (HPLC) analysis was performed to detect the fatty acid contents.ResultsWe show that knockdown of Nl23867, a gene encoding a hypothetical P-loop NTPase, significantly decreased fecundity of N. lugens. Underdeveloped ovaries, fewer eggs laid and reduction in vitellogenin (Vg) protein expression were observed after RNAi knockdown of Nl23867, and most of the affected genes and pathways are fatty acid metabolism-related. We further determined that Nl23867 directly impacts the palmitic acid biosynthesis by regulating the expression of palmitoyl-protein thioesterase (PPT), subsequently affecting the content of total lipids in N. lugens.ConclusionsNl23867 regulates the fecundity of N. lugens by modulating the biosynthetic pathway of palmitic acid and affecting lipid metabolism during vitellogenesis and oocyte development.General significanceThe presented study pioneers the exploration into how a function-unknown gene takes part in the regulation of fecundity in an insect, and will contribute to the construction of gene regulatory network for insect fecundity.     

Chromosomal‐level genomes of three rice planthoppers provide new insights into sex chromosome evolution

The brown planthopper Nilaparvata lugens, white‐backed planthopper Sogatella furcifera, and small brown planthopper Laodelphax striatellus are three major insect pests of rice. They are genetically close; however, they differ in several ecological traits such as host range, migration capacity, and in their sex chromosomes. Though the draft genome of these three planthoppers have been previously released, the quality of genome assemblies need to be improved. The absence of chromosome‐level genome resources has hindered in‐depth research of these three species. Here, we performed a de novo genome assembly for N. lugens to increase its genome assembly quality with PacBio and Illumina platforms, increasing the contig N50 to 589.46 Kb. Then, with the new N. lugens genome and previously reported S. furcifera and L. striatellus genome assemblies, we generated chromosome‐level scaffold assemblies of these three planthopper species using HiC scaffolding technique. The scaffold N50s significantly increased to 77.63 Mb, 43.36 Mb and 29.24 Mb for N. lugens, S. furcifera and L. striatellus, respectively. To identify sex chromosomes of these three planthopper species, we carried out genome re‐sequencing of males and females and successfully determined the X and Y chromosomes for N. lugens, and X chromosome for S. furcifera and L. striatellus. The gene content of the sex chromosomes showed high diversity among these three planthoppers suggesting the rapid evolution of sex‐linked genes, and all chromosomes showed high synteny. The chromosome‐level genome assemblies of three planthoppers would provide a valuable resource for a broad range of future research in molecular ecology, and subsequently benefits development of modern pest control strategies.     

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

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 Gβγ 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 Gβγ when bound to galactose. Thus, our study provides novel insights into the signaling pathways regulated by the sugar gustatory receptors in insects.     

Synergistic roles of acyl‐CoA binding protein (ACBP1) and sterol carrier protein 2 (SCP2) in Toxoplasma lipid metabolism

Toxoplasma gondii relies on apicoplast‐localised FASII pathway and endoplasmic reticulum‐associated fatty acid elongation pathway for the synthesis of fatty acids, which flow through lipid metabolism mainly in the form of long‐chain acyl‐CoA (LCACoAs) esters. Functions of Toxoplasma acyl‐CoA transporters in lipid metabolism remain unclear. Here, we investigated the roles of acyl‐CoA‐binding protein (TgACBP1) and a sterol carrier protein‐2 (TgSCP2) as cytosolic acyl‐CoA transporters in lipid metabolism. The fluormetric binding assay and yeast complementation confirmed the acyl‐CoA binding activities of TgACBP1 and TgSCP2, respectively. Disruption of either TgACBP1 or TgSCP2 caused no obviously phenotypic changes, whereas double disruption resulted in defects in intracellular growth and virulence to mice. Gas chromatography coupled with mass spectrometry (GC–MS) results showed that TgACBP1 or TgSCP2 disruption alone led to decreased abundance of C18:1, whereas double disruption resulted in reduced abundance of C18:1, C22:1, and C24:1. ¹³C labelling assay combined with GC–MS showed that double disruption of TgACBP1 and TgSCP2 led to reduced synthesis rates of C18:0, C22:1, and C24:1. Furthermore, high performance liquid chromatography coupled with high resolution mass spectrometry (HPLC‐HRMS) was used for lipidomic analysis of parasites and indicated that loss of TgACBP1 and TgSCP2 caused serious defects in production of glycerides and phospholipids. Collectively, TgACBP1 and TgSCP2 play synergistic roles in lipid metabolism in T. gondii.     

Characterization of neutral lipases revealed the tissue-specific triacylglycerol hydrolytic activity in Nilaparvata lugens

The lipid metabolism plays an essential role in the development and reproduction of insects, and lipases are important enzymes in lipid metabolism. In Nilaparvata lugens, an important insect pest on rice, triacylglycerol hydrolytic activities were different among tissues, with high activity in integument, ovary, and fat body, but low activity in intestine. To figure out the tissue-specific triacylglycerol hydrolytic activity, we identified 43 lipases in N. lugens. Of these 43 lipases, 23 belonged to neutral lipases, so this group was selected to perform further experiments on triacylglycerol hydrolysis. The complete motifs of catalytic triads, β9 loop, and lid motif, are required for the triacylglycerol hydrolytic activity in neutral lipases, which were found in some neutral lipases with high gene expression levels in integument and ovary, but not in intestine. The recombinant proteins of 3 neutral lipases with or without 3 complete motifs were obtained, and the activity determination confirmed the importance of 3 motifs. Silencing XM_022331066.1, which is highly expressed in ovary and with 3 complete motifs, significantly decreased the egg production and hatchability of N. lugens, partially through decline of the lipid metabolism. In summary, at least one-third of important motifs were incomplete in all neutral lipases with high gene expression in intestine, which could partially explain why the lipase activity in intestine was much lower than that in other tissues. The low activity to hydrolyze triacylglycerol in N. lugens intestine might be associated with its food resource and nutrient components, and the ovary-specific neutral lipases were important for N. lugens reproduction.     

Elevated acetyl‐CoA by amino acid recycling fuels microalgal neutral lipid accumulation in exponential growth phase for biofuel production

Microalgal neutral lipids [mainly in the form of triacylglycerols (TAGs)], feasible substrates for biofuel, are typically accumulated during the stationary growth phase. To make microalgal biofuels economically competitive with fossil fuels, generating strains that trigger TAG accumulation from the exponential growth phase is a promising biological approach. The regulatory mechanisms to trigger TAG accumulation from the exponential growth phase (TAEP) are important to be uncovered for advancing economic feasibility. Through the inhibition of pyruvate dehydrogenase kinase by sodium dichloroacetate, acetyl‐CoA level increased, resulting in TAEP in microalga Dunaliella tertiolecta. We further reported refilling of acetyl‐CoA pool through branched‐chain amino acid catabolism contributed to an overall sixfold TAEP with marginal compromise (4%) on growth in a TAG‐rich D. tertiolecta mutant from targeted screening. Herein, a three‐step α loop‐integrated metabolic model is introduced to shed lights on the neutral lipid regulatory mechanism. This article provides novel approaches to compress lipid production phase and heightens lipid productivity and photosynthetic carbon capture via enhancing acetyl‐CoA level, which would optimize renewable microalgal biofuel to fulfil the demanding fuel market.     

Cover Caption

RNA interference (RNAi) has been considered as a potential strategy for insect pest control especially for sucking insects, for example, planthoppers, aphids and psyllids. Metamorphosis is a unique developmental model of insects. Therefore, using RNAi technology, regulating the key genes during insect growth and development will not affect other organisms. This strategy probably realizes biological and ecological safety of insect pest control. Photo in cover shows a brown planthopper Nilaparvata lugens molting. Photo provided by Prof. Xue‐Xia Miao.     

On the functional significance of symbiotic microorganisms in the Homoptera: a comparative study of Acyrthosiphon pisum and Nilaparvata lugens

All phloem‐feeding Homoptera possess symbiotic microorganisms. Although the phylogenetic position and anatomical location of the micro‐ organisms differ, the underlying theme of the symbiosis is the same; the microorganisms improve the nutritional quality of the diet through the provision of essential amino acids. The symbiosis has been well documented in aphids, but little information is available from other homopteran groups. The impact of the loss of bacterial symbionts in the pea aphid Acyrthosiphon pisum Harris and eukaryotic yeast‐like symbionts in the Asian rice brown planthopper Nilaparvata lugens Stål was examined in parallel. The weight and relative growth rate of aphids and planthoppers was significantly reduced by symbiont loss, and characteristic features of aposymbiotic pea aphids, so‐called ‘metabolic signatures’, were, for the first time, observed in aposymbiotic N. lugens. For example, the amount of protein per unit fresh weight was reduced by 26 and 10%, and the free amino acid levels increased 1.8‐ and 1.4‐fold, in aposymbiotic A. pisum and N. lugens, respectively. In addition, the concentration of the amino acid glutamine was elevated in the tissues of aposymbiotic insects. The data are discussed in the context of our current understanding of the nutritional role of the symbiosis and the mechanisms of nitrogen metabolism in the two insect species. It is concluded that the metabolic adjustments of the insects to symbiont loss are broadly equivalent.