The brown planthopper (BPH), Nilaparvata lugens (Stal) is a resurgent pest of rice crops throughout Asia. We recently discovered that octopamine (OA) and OA2B2 operate in the BPH mating system, where it mediates a wide range of molecular, physiological and behavioural changes. Here, we report on outcomes of experiments designed to test the hypothesis that OA/OA2B2 signalling mediates responses to three abiotic stressors, starvation, high temperature (37 degrees C), and induced oxidative stress. We found per os RNAi-mediated OA2B2 silencing led to significantly decreased survival, measured in days, following exposure to each of these stressors. We selected a biologically costly process, reproductive biology, as a biotic stressor. Silencing of OA2B2 led to decreased total protein content in ovaries and fat bodies, downregulated expression of vitellogenin (Vg) and Vg receptor (VgR), inhibited fat body Vg protein synthesis, shortened the oviposition period, prolonged the preoviposition period, reduced the number of laid eggs, body weight and female longevity. In addition, the silencing treatments also led to inhibited ovarian development, and ovarian Vg uptake, reduced numbers of egg masses and offspring and lower hatching rates and population growth index. These data support our hypothesis that OA2B2 acts in mediating BPH resistance to biotic and abiotic stressors.

Mating triggers physiological and behavioral changes in female insects. In many species, females experience postmating behavioral and physiological changes that define a post-mated state. These changes are comprised of several conditions, including long-term refractoriness to re-mating and increased production and laying of eggs. Here, we report that mating led to several changes in brown planthopper (BPH) females, including increased octopamine (OA), cAMP concentrations, and activities of several enzymes. Mating also led to changes in the expression of several genes acting in female physiology, including those in the cAMP/PKA signal transduction pathway. OA injections into virgin females led to similar changes. RNAi silencing of the gene encoding tyramine beta-hydroxylase, involved in the final step in OA synthesis, led to decreased expression of these genes, and reduced the cAMP/PKA signaling. At the whole-organism level, the RNAi treatments led to reduced fecundity, body weights, and longevity. RNAi silencing of genes acting in OA signaling led to truncated ovarian development, egg maturation, and ovarian vitellogenin (Vg) uptake. The impact of these decreases is also registered at the population level, seen as decreased population growth. We infer that OA signaling modulates the postmating state in female BPH and possibly other hemipterans.

In Asian rice systems, Cyrtorhinus lividipennis Reuter is an important predator that preys on rice planthopper eggs and young nymphs, as a primary food source. Alanine aminotransferase (ALT) acts in many physiological and biochemical processes in insects. We cloned the full-length complementary DNA of C. lividipennis ClALT. Expression analysis showed higher expression in the fat body and midgut compared to other tissues. It is expressed in all C. lividipennis developmental stages and at least four organs. Silencing of ClALT by RNA interference significantly decreased the ClALT enzyme activity and ClALT expression compared to dsGFP-treated controls at 2 days after emergence (DAE). Silencing of ClALT influenced free hemolymph amino acid compositions, resulting in a reduction of Aspartic acid (Asp) and Alanine (Ala) proportions, and increased Cysteine (Cys) and Valine (Val) proportions in females at 2 DAE. dsClALT treatments led to decreased soluble total protein concentrations in ovary and fat body, and to lower reduced vitellogenin (Vg) expression, body weight, and the numbers of laid eggs. The double-stranded RNA viruse treatments also led to prolonged preoviposition periods and hindered ovarian development. Western blot analysis indicated that silencing ClALT also led to reduced fat body Vg protein abundance at 2 DAE. These data support our hypothesis that ClALT influences amino acid metabolism and fecundity in C. lividipennis.

BACKGROUND Triflumezopyrim (TFM), a novel mesoionic insecticide, has high efficiency at a low dosage, and is mainly used to control hopper species. A previous study demonstrated that seed dressing with TFM effectively controlled rice planthopper populations in mechanically transplanted rice fields; however, mode of action for control was unclear.RESULTS The study shows that seed dressing with TFM resulted in elevated levels of oxalic acid, flavonoids, phenolic substances, callose and other compounds associated with Nilaparvata lugens resistance in rice plants, and low TFM residue content in rice plant stem and grain. Host choice behavioral experiments showed that N. lugens females prefer feeding on untreated rice plants. Electrical penetration graph (EPG) data showed that seed dressing with TFM at medium and high doses significantly prolonged the non-probing period and inhibited phloem ingestion in N. lugens females. These changes led to a significant decrease in female secretion of honeydew, expression of genes encoding vitellogenin and juvenile hormone acid methyltransferase, body weight and longevity, and significantly influenced several physiological parameters resulting in impaired oocyte growth, fecundity and population. Field survey data showed that seed dressing with TFM was efficacious and relatively durable in protecting rice plants from infestation by planthoppers.CONCLUSION This study revealed that seed dressing with TFM enhances rice plant resistance to N. lugens by limiting phloem ingestion and increasing the N. lugens non-probing period; this leads to reduced fecundity of females and lowers N. lugens numbers in the field.

The predatory mirid bug, Cyrtorhinus lividipennis Reuter, feeds on brown planthopper (BPH) eggs that are deposited on rice and gramineous plants surrounding rice fields. The development and reproduction of C. lividipennis are inhibited by feeding on BPH eggs from gramineous species, and the underlining regulatory mechanism for this phenomenon is unclear. In the present study, HPLC-MS/MS analysis revealed that the concentrations of six amino acids (AAs:Ala, Arg, Ser, Lys, Thr, and Pro) were significantly higher in rice than in five gramineous species. When C. lividipennis fed on gramineous plants with BPH eggs, expression of several genes in the target of rapamycin (TOR) pathway (Rheb, TOR, and S6K) were significantly lower than that in the insects fed on rice plants with BPH eggs. Treatment of C. lividipennis females with rapamycin, dsRheb, dsTOR, or dsS6K caused a decrease in Rheb, TOR, and S6K expression, and these effects were partially rescued by the juvenile hormone (JH) analog, methoprene. Dietary dsTOR treatment significantly influenced a number of physiological parameters and resulted in impaired predatory capacity, fecundity, and population growth. This study indicates that these six AAs play an important role in the mediated-TOR pathway, which in turn regulates vitellogenin (Vg) synthesis, reproduction, and population growth in C. lividipennis.

The mirid bug,Cyrtorhinus lividipennisReuter, is an important predator of rice planthoppers in Asia. In a previous study,C. lividipennisfed on gramineous weeds with brown planthopper (BPH) eggs had reduced development compared to those fed on rice with BPH eggs. In the current study, the concentrations of selected amino acids (AAs) were higher in rice than five gramineous species, which might explain the enhanced growth ofC. lividipennison rice. WhenC. lividipenniswas fed on AA-deprived artificial diets, the Wnt/beta-catenin pathway was inhibited. Furthermore,C. lividipennisfemales silenced for expression ofFrizzled 2(Fz2) showed a significant reduction in the Wnt/beta-catenin and target of rapamycin (TOR) pathways. SilencingFz2led to decreased expression of the vitellogenin gene (Vg), lower Vg accumulation in oocytes, reduced soluble protein in ovaries and fat bodies, reduced titers of juvenile hormone, prolonged preoviposition periods, and lower predation capacity, body weight, and egg numbers as controlled to controls.Fz2silencing resulted in undeveloped ovaries and the inhibition of oocyte growth in the ovarioles, resulting in decreased numbers of offspring and reduced hatching rates. The silencing ofFz2also resulted in aberrant embryos with undeveloped eyespots and organs, suggesting thatFz2is an essential gene for embryonic development, oogenesis, and egg maturation. In summary, this study established a potential link between Wnt and TOR pathways, which interact synergistically to regulateC. lividipennisreproduction in response to AA signals. These results provide valuable new information that can be applied to large-scale rearing ofC. lividipennispredators, which is important for reducing planthopper damage in rice fields.

The brown planthopper (BPH), Nilaparvata lugens, is a resurgent pest with an unexpected response to jinggangmycin (JGM), a broadly applied antibiotic used to control rice sheath blight disease. JGM stimulates BPH fecundity, but the underlining molecular mechanisms remain unclear. Here we report that JGM sprays led to increased glucose concentrations, photosynthesis and gene expression, specifically Rubsico, sucrose phosphate synthase, invertase 2 (INV2) and INV3 in rice plants. JGM sprays led to high-glucose rice plants. Feeding BPH on these plants led to increased insulin-like signaling and vitellogenin synthesis. Treating BPH with metformin, a gluconeogenesis inhibitor, reversed the influence of feeding on high-glucose rice, which was rescued by glucose injections. Silencing insulin-like peptide 2 using per os dsRNA led to reduction in juvenile hormone (JH) III titers and other fecundity parameters, which were reversed by topical applications of the JH analog, methoprene. We infer that JGM acts via two broad mechanisms, one through increasing rice plant sugar concentrations and a second by upregulating BPH insulin-like signaling. (C) 2020 Elsevier Ltd. All rights reserved.

The antibiotic jinggangmycin (JGM) is broadly applied in Chinese rice producing regions to control rice blight, a fungal disease. Aside from protecting rice plants from the disease, JGM leads to the unexpected action of stimulating brown planthopper (BPH; Nilaparvata lugens; Hemiptera: Delphacidae) reproduction to the extent it can influence population sizes. The JGM-induced BPH population growth has potential for severe agricultural problems and we are working to understand and mitigate the mechanisms of the enhanced reproduction. UDP-glucuronosyltransferases (UGTs) are multifunctional detoxification enzymes responsible for biotransformation of diverse lipophilic compounds. The biological significance of this enzyme family in insect fecundity is not fully understood, however, upregulated UGT12 in JGM-treated BPH, may influence fecundity through metabolism of developmental hormones. This idea prompted our hypothesis that NIUGT12 is a positive modulator of BPH reproductive biology. JGM treatment led to significant increases in accumulations of mRNA encoding NIUGT12, numbers of eggs laid, oviposition period, juvenile hormone Ill titers, and fat body, and ovarian protein contents. dsUGT12 treatment suppressed NIUGT12 expression and reversed JGM-enhanced effects, resulting in under-developed ovaries and reduced expression of juvenile hormone acid methyltransferase and the JH receptor, methoprene tolerant. Application of the JH analog, methoprene, on dsUGT12 treated-females partially reversed the dsUTG12 influence on vitellogenin synthesis and on NIUGT12 expression. These results represent an important support for our hypothesis.

Hexokinase is a rate-limiting enzyme that plays pivotal roles in glucose homeostasis and energy metabolism via glucose (Glc) phosphorylation and Glc signaling mediation. Previous investigations have revealed the modulatory role of Hexokinase (Hex) genes involved in proper glucose regulation during insect diapause and embryo development, whereas whether it functions in insect fecundity remains largely unknown. We aimed to explore the relationship between Triazophos (TZP)-induced Hex-1 and fecundity of female Nilaparvata lugens. In this study, Hex-1 expression were characterized at different developmental stages and in various tissues of N. lugens, with the highest expression registered in brain tissues and 5th instar nymph. The present findings indicated that TZP + dsHex-1 silencing significantly reduced protein synthesis, including the fat body and ovarian protein content of female adults. Meanwhile, the glycometabolism with respect to the soluble sugar, trehalose and glucose content in female adults were strikingly influenced as a result of Hex-1 knockdown. The relative transcript level of Hex-I, vitellogenin (NlVg) and vitellogenin receptor (NlVgR) considerably decreased in TZP + dsHex-1 treated females compared to TZP and TZP + dsGFP-treated groups. More importantly, TZP + dsHex-1 silencing led to reduced number of eggs laid and vitellogenin (Vg) accumulation as well as retarded ovary development compared with TZP-treated and TZP + dsGFP-treated groups. Taken together, it is proposed that Hex-1 implicates in N. lugens fecundity by exerting profound effects on glycometabolism, protein sythesis and NlVg expression.