Brown Planthopper Nilaparvata Lugens Research Articles

Growth of Beauveria bassiana Combined with MIPC Insecticide and Its Efficacy to Control the Brown Planthopper Nilaparvata lugens

The objectives of this study were to evaluate the efficacy of MIPC to control the brown planthopper Nilaparvata lugens, one of the most damaging pests of rice in the field and determine the effect of various of its doses on the growth of the fungus B. bassiana. The research was carried out in a rice field and in the Biological Agents Development Laboratory, Department of Pests and Plant Diseases, Faculty of Agriculture, Brawijaya University, Indonesia. The field efficacy test of the MIPC insecticide was conducted by applying 0.5-, 1.0-, 1.5-, and 2.0-kg/ha doses. The insecticidal compatibility test of MIPC with B. bassiana was conducted in 0.5-, 1.0-, 1.5-, and 2.0-g/L doses of MIPC. Application of MIPC insecticide at 0.5-, 1.0-, 1.5-, and 2.0-kg/ha doses had no significant difference in reducing N. lugens population in the vegetative phase (28 to 70 days after planting). The MIPC doses of 1.0, 1.5, and 2.0 g/L were highly toxic or incompatible with the growth of B. bassiana. Meanwhile, a sublethal concentration of MIPC insecticide of 0.5 g/L was non-toxic to B. bassiana. Therefore, this study recommends the use of 0.5 g/L MIPC combined with B. bassiana fungi.

Host plants alter their volatiles to help a solitary egg parasitoid distinguish habitats with parasitized hosts from those without.

When attacked by herbivores, plants emit volatiles to attract parasitoids and predators of herbivores. However, our understanding of the effect of plant volatiles on the subsequent behaviour of conspecific parasitoids when herbivores on plants are parasitized is limited. In this study, rice plants were infested with gravid females of the brown planthopper (BPH) Nilaparvata lugens for 24 hr followed by another 24 hr in which the BPH eggs on plants were permitted to be parasitized by their egg parasitoid, Anagrus nilaparvatae; volatiles from rice plants that underwent such treatment were less attractive to subsequent conspecific parasitoids compared to the volatiles from plants infested with gravid BPH females alone. Chemical analysis revealed that levels of JA and JA-Ile as well as of four volatile compounds-linalool, MeSA, α-zingiberene and an unknown compound-from plants infested with BPH and parasitized by wasps were significantly higher than levels of these compounds from BPH-infested plants. Laboratory and field bioassays revealed that one of the four increased chemicals-α-zingiberene-reduced the plant's attractiveness to the parasitoid. These results suggest that host plants can fine-tune their volatiles to help egg parasitoids distinguish host habitats with parasitized hosts from those without.

Expression Profiles and Biochemical Analysis of Chemosensory Protein 3 from Nilaparvata lugens (Hemiptera: Delphacidae).

Insects have evolved highly sensitive olfactory sensory systems to detect plant hosts and mates, with plant volatiles playing an important role in informing insect behavior. Chemosensory proteins (CSPs) are thought to play a key role in this process, but in this respect, there is limited information on brown planthopper Nilaparvata lugens, one of the most destructive pests of rice. To expand our understanding of CSP function in N. lugens we explored expression profiles and binding characteristics of NlugCSP3. The ligands with higher binding affinity were also validated by molecular docking and behavioral assays. NlugCSP3 mRNA was expressed at relatively higher levels in antennae and abdomen of 3-day-old unmated macropterous males as well as in antennae of 3-day mated macropterous and brachypterous females. Fluorescence competitive binding assays revealed that 5 out of 25 candidate volatiles are strong binders (Ki < 10μM). Behavioral assays revealed that nonadecane and 2-tridecanone, which have high binding affinities in fluorescence competition-binding assays, displayed strong attractiveness to N. lugens. Pursuing this further, molecular docking analysis identified key amino acid residues involved in binding volatile compounds. Overall, our data provide a base for further investigation of the potential physiological functions of CSP3 in Nilaparvata lugens, and extend the function of NlugCSP3 in chemoreception of N. lugens.

An Overview of Embryogenesis: External Morphology and Transcriptome Profiling in the Hemipteran Insect Nilaparvata lugens

During embryogenesis of insects, the morphological and transcriptional changes are important signatures to obtain a better understanding of insect patterning and evolution. The brown planthopper Nilaparvata lugens is a serious insect pest of rice plants, but its embryogenesis has not uncovered. Here, we described embryonic development process of the pest and found it belongs to an intermediate-germ mode. The RNA-seq data from different times (6, 30, 96, and 150 h, after egg laying) of embryogenesis were then analyzed, and a total of 10,895 genes were determined as differentially expressed genes (DEGs) based on pairwise comparisons. Afterward, 1,898 genes, differentially expressed in at least two comparisons of adjacent embryonic stages were divided into 10 clusters using K means cluster analysis (KMCA). Eight-gene modules were established using a weighted gene co-expression network analysis (WGCNA). Gene expression patterns in the different embryonic stages were identified by combining the functional enrichments of the stage-specific clusters and modules, which displayed the expression level and reprogramming of multiple developmental genes during embryogenesis. The “hub” genes at each embryonic stage with possible crucial roles were identified. Notably, we found a “center” set of genes that were related to overall membrane functions and might play important roles in the embryogenesis process. After parental RNAi of the MSTRG.3372, the hub gene, the embryo was observed as abnormal. Furthermore, some homologous genes in classic embryonic development processes and signaling pathways were also involved in embryogenesis of this insect. An improved comprehensive finding of embryogenesis within the N. lugens reveals better information on genetic and genomic studies of embryonic development and might be a potential target for RNAi-based control of this insect pest.

RNA interference of tyrosine hydroxylase caused rapid mortality by impairing cuticle formation in Nilaparvata lugens (Hemiptera: Delphacidae).

The application of RNA interference (RNAi) technique in controlling agricultural insect pests has been receiving much attention since the discovery of RNAi. The brown planthopper (BPH) Nilaparvata lugens, a notorious pest of rice, has evolved a high level of resistance to many kinds of insecticides. Tyrosine hydroxylase (Th) is an indispensable survival gene in holometabolous insects, playing key roles in cuticle tanning and immunity. In this study, we investigated whether Th could be used as a potential target in controlling N. lugens. Here, we demonstrated that NlTh had a periodical expression pattern during molting with the highest level observed in epidermis. Dysfunction of NlTH by dsNlTh microinjection or 3-IT feeding similarly caused rapid death of N. lugens. Compared with dsGFP control BPHs, dsNlTh injected BPHs (i) had cuticle pigmentation and sclerotizaton defects; (ii) had less endocuticle lamella in tergum integument; (iii) showed higher mortality during the molting process as a result of defective cuticle shedding; (iv) showed feeding disorders indicated by a low number of probe wound dots on rice; (v) had more vulnerable cuticle. This study demonstrated that TH orthologues play a conservative and crucial role for exocuticle tanning in both holometabolous and hemimetabolous insects, and NlTh could be targeted for RNAi-mediated BPH control. The rapid lethal phenotype of NlTH dysfunction BPHs partly induced by cuticle formation defects. © 2020 Society of Chemical Industry.

Stable Establishment of Cardinium spp. in the Brown Planthopper Nilaparvata lugens despite Decreased Host Fitness.

The brown planthopper Nilaparvata lugens (Hemiptera) is a major pest of rice crops in Asia. Artificial transinfections of Wolbachia have recently been used for reducing host impacts, but transinfections have not yet been undertaken with another important endosymbiont, Cardinium This endosymbiont can manipulate the reproduction of hosts through phenotypes such as cytoplasmic incompatibility (CI), which is strong in the related white-backed planthopper, Sogatella furcifera (Hemiptera). Here, we stably infected N. lugens with Cardinium from S. furcifera and showed that it exhibits perfect maternal transmission in N. lugens The density of Cardinium varied across developmental stages and tissues of the transinfected host. Cardinium did not induce strong CI in N. lugens, likely due to its low density in testicles. The infection did decrease fecundity and hatching rate in the transinfected host, but a decrease in fecundity was not apparent when transinfected females mated with Wolbachia-infected males. The experiments show the feasibility of transferring Cardinium endosymbionts across hosts, but the deleterious effects of Cardinium on N. lugens limit its potential to spread in wild populations of N. lugens in the absence of strong CI.IMPORTANCE In this study we established a Cardinium-infected N. lugens line that possessed complete maternal transmission. Cardinium had a widespread distribution in tissues of N. lugens, and this infection decreased the fecundity and hatching rate of the host. Our findings emphasize the feasibility of transinfection of Cardinium in insects, which expands the range of endosymbionts that could be manipulated for pest control.

Krüppel homologue 1 interacts directly with Hairy and regulates ecdysis in the brown planthopper.

Juvenile hormone (JH) plays important roles in the growth and development of insects. JH and its receptor methoprene-tolerant (Met) regulate the expression of transcription factors to control the transcription of downstream genes. The expression of Hairy (Hry) and Krüppel homologue 1 (Kr-h1) is regulated by JH and JH receptors. Hry and Kr-h1 are both crucial in mediating JH signalling. However, whether they interact at the gene level in regulating metamorphosis and whether they interact physically at the protein level remain unknown. We used co-immunoprecipitation, glutathione S-transferase pull-down and RNA interference (RNAi) approaches to study the genetic and biochemical interactions of the two proteins Hry and Kr-h1. The results showed that brown planthopper (Nilaparvata lugens) Hry and Kr-h1 interact directly: Hry binds to the N-terminal of Kr-h1, which includes five zinc-finger domains. The RNAi experiment showed that downregulation of Hry reduced the ratio of ecdysis failure caused by knockdown of Kr-h1, indicating that the downregulation of Hry might mitigate ecdysis failure via the downregulation of Kr-h1. The expression of Hry increased significantly when Kr-h1 was downregulated, whereas it did not change significantly when both were downregulated. Our results suggest that the binding of Hry protein with Kr-h1 prevents the N-terminal five zinc-finger domains from binding with DNA, which in turn inactivates the transcription activator or inhibitor function of Kr-h1. Hry could possibly be used as a target for pesticide applications in the future.

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.

Insecticides and resurgence of rice brown plant hopper nilaparvata lugens (Stal)

Effect of certain insecticides in rice ecosystem was studied for their role in resurgence of brown plant hopper (BPH) during kharif and rabi 2016-17. Among fourteen insecticides evaluated, seven insecticides viz., chlorpyriphos, profenophos, deltamethrin, cypermethrin, bifenthrin, lambdacyhalothrin and imidacloprid were observed to significantly increase the incidence of BPH over untreated check after 1st, 2nd and 3rdsprays consistently and resulted in induced resurgence. Acephate, monocrotophos, cartap hydrochloride, fipronil, chlorantraniliprole, buprofezin and pymetrozine effectively controlled the BPH and did not induce resurgence. Thus, these can be used judiciously, with periodic monitoring of resistance and resurgence.

Fine mapping, candidate genes analysis, and characterization of a brown planthopper (Nilaparvata lugens Stål) resistance gene in the rice variety ARC5984

The brown planthopper (Nilaparvata lugens Stal; BPH) is the most destructive rice pests worldwide. Identification and application of major resistance genes and cultivation of resistant varieties are the most efficient methods to control this insect. In this study, the rice variety, ARC5984, was detected to be highly resistant to BPH due to the presence of a major resistance gene, which has primarily been mapped in a 6-cM region on the long arm of chromosome 4. Fine mapping of the resistance gene was performed using the recombinants derived from BC1F2 individuals; the target gene was described to be a 46-kb region in the Nipponbare genome. Based on the information on the genomic sequences of the varieties Nipponbare, ZS97 and MH63, six, seven and 11 candidate genes were identified in the finally located region, respectively. The relative expression of the candidate genes Os04g35190 and Os04g35210 was significantly different between the resistant and susceptible plants after BPH infestation. The sequencing results indicated the coding domain sequence (CDS) of Os04g35210 from ARC5984 to be identical to that of Bph6. But the CDS of Os04g35190 was different between in ARC5984 and 9311. Moreover, the preliminary-near-isogenic lines (pre-NILs; the lines carrying resistance genes) with genetic background of the 9311 showed significant antixenosis to BPH, and thus, restricted its growth and development when compared with that in the susceptible line of 9311. Finally, a marker-assisted selection (MAS) technique was developed for the markers, RM5635 and BF9; its efficiency has been confirmed by applying it in screening of backcrossed individuals. The development in the fine mapping of the major gene will facilitate efforts of cloning this important resistance gene and to apply it in breeding of BPH-resistance rice varieties.

Distinct functional properties of sodium channel variants are associated with usage of alternative exons in Nilaparvata lugens

Voltage-gated sodium channels (Nav) are essential for electrical signaling in the nervous system. They are also the primary targets of several classes of insecticides including pyrethroids. There is only one sodium channel gene in most insect species, whereas mammals possess at least nine sodium channel genes. Extensive alternative splicing and RNA editing of sodium channel transcripts have been documented in many insect species. However, the functional consequences of these post-transcriptional events have been evaluated only in DmNav and BgNav from Drosophila melanogaster and Blattella germanica, respectively. In this study, we isolated 41 full-length cDNA clones encoding 34 sodium channel (NlNav) variants from a major rice pest, the brown planthopper (Nilaparvata lugens Stål). The 34 NlNav variants represent 24 distinct splicing types based on the usage of nine alternative exons, six of which, including exon b, have been previously reported in other insect species. When expressed in Xenopus oocytes, NlNav variants lacking exon b generated significantly larger sodium currents than variants possessing exon b, suggesting an inhibitory effect of exon b on sodium current expression. A similar effect has been reported for exon b from BgNav. Mutational analysis showed that three conserved amino acid residues encoded by exon b are critical for its inhibitory effect. In addition, mutually exclusive exons k/l contribute to distinct functional properties and channel sensitivity to pyrethroids. Altogether, these results show that alternative splicing generates functional diversity of sodium channels in this insect species and that the role of exon b in regulating neuronal excitability is likely conserved among insect species.

Genetics of novel brown planthopper Nilaparvata lugens (Stål) resistance genes in derived introgression lines from the interspecific cross O. sativa var. Swarna × O. nivara

The brown planthopper (BPH) Nilaparvata lugens (Stål) (Homoptera: Delphacidae) is considered a threat to rice (Oryza sativa ssp.) crop in many parts of the world including India. Among the BPH-resistance (R) genes so far reported in rice, most of them are ineffective against BPH biotype 4 predominant in the Indian sub-continent. In this study, we show the introgression line RPBio4918-230S was identified as BPH resistant after five years of rigorous screening at seedling stage and two years at tillering and reproductive stages. The inheritance of resistance indicated that two recessive genes are involved at seedling and reproductive stages. The allelic relation with known genes using linked reported markers suggested that the genes present in RPBio4918-230S are different. We report here the genetics of the two newly introgressed BPH resistance genes from O. nivara in the background of Swarna which are effective at all the important growth stages. The genes have been tentatively named as bph39(t) and bph40(t). The honeydew area (feeding rate) and days to wilt parameters observed at 30 days after sowing in BC1F3 indicated that newly introgressed genes have both antibiosis and tolerance mechanisms for resistance. The BPH resistance genes identified in this study would facilitate the breeding of broad spectrum and durable resistance in rice against BPH biotype 4.

Predicting Rice Pest Population Occurrence with Satellite-Derived Crop Phenology, Ground Meteorological Observation, and Machine Learning: A Case Study for the Central Plain of Thailand

The brown planthopper Nilaparvata lugens (BPH) is one of the most harmful insect pests in rice paddy fields, which causes considerable yield loss and consequent economic problems, particularly in the central plain of Thailand. Accurate and timely forecasting of pest population incidence would support farmers in planning effective mitigation. In this study, artificial neural network (ANN), random forest (RF) and classic linear multiple regression (MLR) analyses were applied and compared to forecast the BPH population using weather and host-plant phenology factors during the crop dry season from 2006 to 2016 in the central plain of Thailand. Data from satellite earth observation was used to monitor crop phenology factors affecting BPH population density. An ANN model with integrated ground-based meteorological variables and satellite-derived host plant variables was more accurate for short-term forecasting of the peak abundance of BPH when compared with RF and MLR, according to a reasonably validating dataset (RMSE of natural log-transformed (ln) BPH light trap catches = 1.686, 1.737, and 2.015, respectively). This finding indicates that the utilization of ground meteorological observations, satellite-derived NDVI time series, and ANN have the potential to predict BPH population density in support of integrated pest management programs. We expect the results from this study can be applied in conjunction with the satellite-based rice monitoring system developed by the Geo-Informatic and Space Technology Development Agency of Thailand (GISTDA; http://rice.gistda.or.th) to support an effective pest early warning system.

Sublethal effects of abamectin on the development, fecundity, and wing morphs of the brown planthopper Nilaparvata lugens

Abstract The brown planthopper Nilaparvata lugens negatively affects rice yield by sucking nutrients from the rice stem and spreading viruses. In this study, the effects of sublethal concentrations of abamectin on development, fecundity, and wing morphs of N. lugens were investigated. Treatment with sublethal concentrations of abamectin showed no significant effects on the developmental period of F0 nymphs. Although LC10 and LC25 treatments resulted in 10% and 25% mortality, the LC25 treatment significantly prolonged the lifespan of macropterous and brachypterous females. Abamectin-treated brachypterous females showed significantly lower fecundity than control females. Furthermore, abamectin treatment showed no significant effects on the developmental period of F1 nymphs. However, LC25 treatment significantly inhibited the fecundity of brachypterous F1 females and significantly reduced the proportion of macropterous F1 females and brachypterous F1 males. LC25 treatment showed no significant effects on the fecundity of macropterous F1 females. Considering the proportion of macropterous and brachypterous F1 females, the relative fitness of the macropterous F1 nymphs in the control, LC10, and LC25 treatment groups was 1, 1.02, and 0.84, respectively, and that of brachypterous F1 nymphs was 1, 0.79, and 0.93, respectively. Sublethal concentrations of abamectin inhibited N. lugens emergence. The present findings indicate the potential of abamectin for N. lugens field control.

Classification of rice planthoppers based on shape descriptors

Here, classification of rice planthopper (RPH) based on shape descriptors was addressed to solve the low semantics problem of shape features in traditional RPH (mainly including the whiteback planthopper (Sogatella furcifera (Horvath)), the brown planthopper (Nilaparvata lugens (Stal)), and the small brown planthopper (Laodelphax striatellus (Fallen))) image classification research. Images of RPH were obtained from rice field by an automatic insect image acquisition device made by ourselves and insect images were divided into single images based on OTSU threshold segmentation algorithm. In terms of the images of RPH after segmentation, Fourier descriptors and Hu moments, which are from two aspects of contour curve and shape area, were extracted to describe shape features of RPH. Then, random forest (RF), an ensemble learning algorithm, was used as the classifier to distinguish RPH efficiently. The optimal number of trees and prediction variables of RF are chosen to be 150 and 4, respectively, by minimising the out-of-bag error. Experimental results show that classification accuracy of RPH based on shape descriptors reaches up to 93.93%. Therefore, it has been verified that the classification with the method presented here is accurate and semantic.

Mutations in NlInR1 affect normal growth and lifespan in the brown planthopper Nilaparvata lugens

The brown planthopper (BPH) Nilaparvata lugens contains two insulin receptor homologues, designated NlInR1 and NlInR2. NlInR1 is strikingly homologous to the typical InR in insects and vertebrates, containing a ligand-activated intracellular tyrosine kinase catalytic domain. Herein, we report an optimized CRISPR/Cas9 system to induce mutations in the NlInR1 locus in BPH, consisting of a Cas9 plasmid that is specifically expressed in the germline via the Nlvasa promoter and versatile sgRNA expression plasmids under the control of the U6 promoter. We systematically evaluated the efficiency of injection mix compositions and demonstrated an appropriate combination of Cas9/sgRNA to target essential genes. Furthermore, we showed that homozygous mutants for the NlInR1 gene are early embryonic lethal, whereas heterozygous mutants grow more slowly, exhibit a severe reduction in body weight and wing size and live longer than the wild type. Interestingly, the severity of the mutant phenotype was different when targeting distinct important domains of the NlInR1 locus. The severity of the mutant phenotype is similar to that of insulin/insulin-like growth factor (IGF) signaling pathway deficiencies in vertebrates, suggesting a conserved function of NlInR1 in the regulation of development and longevity. Global expression profiling suggests that NlInR1 regulates many cellular processes in BPH, including insulin resistance, phototransduction, metabolism, endocytosis, longevity, biosynthesis and protein processing. Our results also pave the way for understanding the precise molecular mechanism of insulin signaling in wing polyphenism in insects.

Functional Analysis of an Epsilon-Class Glutathione S-Transferase From Nilaparvata lugens (Hemiptera: Delphacidae).

Glutathione conjugation is a crucial step in xenobiotic detoxification. In the current study, we have functionally characterized an epsilon-class glutathione S-transferase (GST) from a brown planthopper Nilaparvata lugens (nlGSTE). The amino acid sequence of nlGSTE revealed approximately 36–44% identity with epsilon-class GSTs of other species. The recombinant nlGSTE was prepared in soluble form by bacterial expression and was purified to homogeneity. Mutation experiments revealed that the putative substrate-binding sites, including Phe107, Arg112, Phe118, and Phe119, were important for glutathione transferase activity. Furthermore, inhibition study displayed that nlGSTE activity was affected by insecticides, proposing that, in brown planthopper, nlGSTE could recognize insecticides as substrates.

Involvement of mind the gap in the organization of the tracheal apical extracellular matrix in Drosophila and Nilaparvata lugens.

The tracheal apical extracellular matrix (aECM) is vital for expansion of the tracheal lumen and supports the normal structure of the lumen to guarantee air entry and circulation in insects. Although it has been found that some cuticular proteins are involved in the organization of the aECM, unidentified factors still exist. Here, we found that mind the gap (Mtg), a predicted chitin-binding protein, is required for the normal formation of the apical chitin matrix of airway tubes in the model holometabolous insect Drosophila melanogaster. Similar to chitin, the Mtg protein was linearly arranged in the tracheal dorsal trunk of the tracheae in Drosophila. Decreased mtg expression in the tracheae seriously affected the viability of larvae and caused tracheal chitin spiral defects in some larvae. Analysis of mtg mutant showed that mtg was required for normal development of tracheae in embryos. Irregular taenidial folds of some mtg mutant embryos were found on either lateral view of tracheal dorsal trunk or internal view of transmission electron microscopy analysis. These abnormal tracheae were not fully filled with gas and accompanied by a reduction in tracheal width, which are characteristic phenotypes of tracheal aECM defects. Furthermore, in the hemimetabolous brown planthopper (BPH) Nilaparvata lugens, downregulation of NlCPAP1-N (a homolog of mtg) also led to the formation of abnormal tracheal chitin spirals and death. These results suggest that mtg and its homolog are involved in the proper organization of the tracheal aECMs in flies and BPH, and that this function may be conserved in insects.

OsMKK3, a Stress-Responsive Protein Kinase, Positively Regulates Rice Resistance to Nilaparvata lugens via Phytohormone Dynamics.

Plants undergo several but very precise molecular, physiological, and biochemical modulations in response to biotic stresses. Mitogen-activated protein kinase (MAPK) cascades orchestrate multiple cellular processes including plant growth and development as well as plant responses against abiotic and biotic stresses. However, the role of MAPK kinases (MAPKKs/MKKs/MEKs) in the regulation of plant resistance to herbivores has not been extensively investigated. Here, we cloned a rice MKK gene, OsMKK3, and investigated its function. It was observed that mechanical wounding, infestation of brown planthopper (BPH) Nilaparvata lugens, and treatment with methyl jasmonate (MeJA) or salicylic acid (SA) could induce the expression of OsMKK3. The over-expression of OsMKK3 (oe-MKK3) increased levels of jasmonic acid (JA), jasmonoyl-L-isoleucine (JA-Ile), and abscisic acid (ABA), and decreased SA levels in rice after BPH attack. Additionally, the preference for feeding and oviposition, the hatching rate of BPH eggs, and BPH nymph survival rate were significantly compromised due to over-expression of OsMKK3. Besides, oe-MKK3 also augmented chlorophyll content but impaired plant growth. We confirm that MKK3 plays a pivotal role in the signaling pathway. It is proposed that OsMKK3 mediated positive regulation of rice resistance to BPH by means of herbivory-induced phytohormone dynamics.

Knockdown of glycogen phosphorylase and glycogen synthase influences expression of chitin synthesis genes of rice brown planthopper Nilaparvata lugens

Abstract Glycogen synthase (GS) and glycogen phosphorylase (GP) are two key enzymes in the glycogen synthesis pathway, which catalyze trehalose and glucose transformation in insects. GS and GP can be regulated by trehalose metabolism, which plays an important role in insect growth. However, it is not known whether these genes can be targeted for pest control through regulation of chitin metabolism. We studied the function of Nilaparvata lugens GS and GP (NLGS and NLGP, respectively) using RNA interference, and reported that trehalose and the chitin biosynthesis pathways are regulated by GP and GS, especially TPS3, TRE1-1, and G6PI1, which decreased following knockdown of these two genes. The expression levels of TPS1, TPS2, and several chitin synthesis pathway family genes were significantly increased following dsNlGP injection. Additionally, despite there being no apparent change to the chitin content, an abnormal molting phenotype and wing deformity appeared, and close to 25% insects died. These results demonstrate that silencing of NLGP or NLGS can lead to molting deformities and an elevated mortality rate through the regulation of chitin pathway genes and chitinase genes. NLGP may play a key role in chitin synthesis due to the number of genes regulated, and higher deformity and mortality rates resulting from its knockdown.