Transcriptome-wide N6-methyladenosine profiling reveals growth-defense trade-offs in the response of rice to brown planthopper (Nilaparvata lugens) infestation.

BACKGROUND: N6-Methyladenosine (m6A) is a common messenger RNA (mRNA) modification that affects various physiological processes in stress responses. However, the role of m6A modifications in plants responses to herbivore stress remains unclear. RESULTS: Here, we found that an infestation of brown planthopper (Nilaparvata lugens) female adults enhanced the resistance of rice to N. lugens. The m6A methylome analysis of N. lugens-infested and uninfested rice samples was performed to explore the interaction between rice and N. lugens. The m6A methylation mainly occurred in genes that were actively expressed in rice following N. lugens infestation, while an analysis of the whole-genomic mRNA distribution of m6A showed that N. lugens infestation caused an overall decrease in the number of m6A methylation sites across the chromosomes. The m6A methylation of genes involved in the m6A modification machinery and several defense-related phytohormones (jasmonic acid and salicylic acid) pathways was increased in N. lugens-infested rice compared to that in uninfested rice. In contrast, m6A modification levels of growth-related phytohormone (auxin and gibberellin) biosynthesis-related genes were significantly attenuated during N. lugens infestation, accompanied by the down-regulated expression of these transcripts, indicating that rice growth was restricted during N. lugens attack to rapidly optimize resource allocation for plant defense. Integrative analysis of the differential patterns of m6A methylation and the corresponding transcripts showed a positive correlation between m6A methylation and transcriptional regulation. CONCLUSION: The m6A modification is an important strategy for regulating the expression of genes involved in rice defense and growth during rice-N. lugens interactions. These findings provide new ideas for formulating strategies to control herbivorous pests. © 2024 Society of Chemical Industry.

The dynamics of N6-methyladenine RNA modification in resistant and susceptible rice varieties responding to rice stem borer damage.

N6-methyladenosine (m6A) is the most prevalent modification in cellular RNA which orchestrates diverse physiological and pathological processes during stress response. However, the differential m6A modifications that cope with herbivore stress in resistant and susceptible crop varieties remain unclear. Here, we found that rice stem borer (RSB) larvae grew better on indica rice (e.g., MH63, IR64, Nanjing 11) than on japonica rice varieties (e.g., Nipponbare, Zhonghua 11, Xiushui 11). Then, transcriptome-wide m6A profiling of representative resistant (Nipponbare) and susceptible (MH63) rice varieties were performed using a nanopore direct RNA sequencing approach, to reveal variety-specific m6A modifications against RSB. Upon RSB infestation, m6A methylation occurred in actively expressed genes in Nipponbare and MH63, but the number of methylation sites decreased across rice chromosomes. Integrative analysis showed that m6A methylation levels were closely associated with transcriptional regulation. Genes involved in herbivorous resistance related to mitogen-activated protein kinase, jasmonic acid (JA), and terpenoid biosynthesis pathways, as well as JA-mediated trypsin protease inhibitors, were heavily methylated by m6A, and their expression was more pronounced in RSB-infested Nipponbare than in RSB-infested MH63, which may have contributed to RSB resistance in Nipponbare. Therefore, dynamics of m6A modifications act as the main regulatory strategy for expression of genes involved in plant-insect interactions, which is attributed to differential responses of resistant and susceptible rice varieties to RSB infestation. These findings could contribute to developing molecular breeding strategies for controlling herbivorous pests.

Investigation on the Nanopore Heterogeneity in Coals and its Influence on Methane Adsorption: A Multifractal Theory Study.

Coalbed methane (CBM) reservoirs constitute a distinct class of dense organic rocks characterized by extremely low porosity and permeability. Conducting an in-depth investigation into pore heterogeneity assumes paramount importance for the exploration and development of CBM. This study focuses on the multifractal analysis of the pores with diameters below 300 nm in six coal samples sourced from the Junggar Basin and the Qinshui Basin in China. The analysis is based on a series of experiments involving CO2 adsorption, low-temperature N2 adsorption/desorption, and CH4 isothermal adsorption. This work delves into the influence of pore heterogeneity on gas adsorption capacity by linking the structural parameters to CH4 adsorption properties. The results indicate that both the micropores, as assessed through CO2 adsorption, and mesopores to macropores, measured via N2 adsorption, exhibit multifractal behavior. In contrast to micropores, the mesopores and macropores display stronger heterogeneity and lower connectivity. Generally, uniform and well-connected nanopores are anticipated to positively contribute to gas adsorption. However, there is a positive correlation between the Langmuir volume and the heterogeneity degree of micropores. This phenomenon is ascribed to the fact that the greater surface complexity in micropores involves a larger specific surface area and a higher abundance of adsorption sites. This research contributes to a more profound and precise comprehension of the heterogeneous pore structure within CBM reservoirs, thereby establishing a theoretical foundation for the sustainable exploitation of CBM.

New insight into the role of the self-assembly of heteroatom compounds in heavy oil viscosity enhancement.

Unveiling the role of heteroatom compounds in heavy oil viscosity is pivotal for finding targeted viscosity reduction methods to improve oil recovery. This research investigates the impact of heteroatoms in asphaltene molecules by utilizing quantum chemical calculations and molecular dynamics simulations to analyze their electrostatic potential characteristics, pairwise interactions, and dynamic behavior within realistic reservoirs. Heteroatom compounds can influence the molecular-level properties of asphaltenes and thus impact the macroscopic behavior of heavy oils. Research results suggest that the presence of ketone and aromatic rings in asphaltene molecules leads to the unrestricted movement of pi electrons due to their collective electronegativity. Two distinct configurations of asphaltene dimers, face-to-face, and edge-to-face, were observed. Intermolecular interactions were predominantly governed by van der Waals forces, highlighting their significant role in stabilizing asphaltene aggregates. The distribution of asphaltene molecules in the oil phase can be summarized as the "rebar-cement" theory. In the heteroatom-free system, the face-to-face peaks in the radial distribution function exhibit significantly reduced magnitudes compared to those in the heteroatom-containing system. This emphasizes the pivotal function of heteroatoms in connecting molecular components to form a more compact asphaltene structure, which may result in a higher viscosity of heavy oil. These findings give insight into the significance of heteroatoms in bridging molecular components and shaping the intricate structure of asphaltene and advance our understanding of heavy oil viscosity properties.

Tyrosine hydroxylase plays crucial roles in larval cuticle formation and larval-pupal tanning in the rice stem borer, Chilo suppressalis.

The striped stem borer, Chilo suppressalis (Walker), a notorious pest infesting rice, has evolved a high level of resistance to many commonly used insecticides. In this study, we investigate whether tyrosine hydroxylase (TH), which is required for larval development and cuticle tanning in many insects, could be a potential target for the control of C. suppressalis. We identified and characterized the full-length cDNA (CsTH) of C. suppressalis. The complete open reading frame of CsTH (MW690914) was 1683 bp in length, encoding a protein of 560 amino acids. Within the first to the sixth larval instars, CsTH was high in the first day just after molting, and lower in the ensuing days. From the wandering stage to the adult stage, levels of CSTH began to rise and reached a peak at the pupal stage. These patterns suggested a role for the gene in larval development and larval-pupal cuticle tanning. When we injected dsCsTH or 3-iodotyrosine (3-IT) as a TH inhibitor or fed a larva diet supplemented with 3-IT, there were significant impairments in larval development and larval-pupal cuticle tanning. Adult emergence was severely impaired, and most adults died. These results suggest that CsTH might play a critical role in larval development as well as larval-pupal tanning and immunity in C. suppressalis, and this gene could form a potential novel target for pest control.

A chromosome-level genome assembly of yellow stem borer (Scirpophaga incertulas).

The yellow stem borer Scirpophaga incertulas is the dominant pest of rice in tropical Asia. However, the lack of genomic resources makes it difficult to understand their invasiveness and ecological adaptation. A high-quality chromosome-level genome of S. incertulas, a monophagous rice pest, was assembled by combining Illumina short reads, PacBio HiFi long sequencing, and Hi-C scaffolding technology. The final genome size was 695.65 Mb, with a scaffold N50 of 28.02 Mb, and 93.50% of the assembled sequences were anchored to 22 chromosomes. BUSCO analysis demonstrated that this genome assembly had a high level of completeness, with 97.65% gene coverage. A total of 14,850 protein-coding genes and 366.98 Mb of transposable elements were identified. In addition, comparative genomic analyses indicated that chemosensory processes and detoxification capacity may play critical roles in the specialized host preference of S. incertulas. In summary, the chromosome-level genome assembly of S. incertulas provides a valuable genetic resource for understanding the biological characteristics of its invasiveness and developing an efficient management strategy.

Defence and nutrition synergistically contribute to the distinct tolerance of rice subspecies to the stem borer, Chilo suppressalis.

Damage caused by the rice striped stem borer (SSB), Chilo suppressalis (Walker) (Lepidoptera: Pyralidae), is much more severe on indica/xian rice than on japonica/geng rice (Oryza sativa) which matches pest outbreak data in cropping regions of China. The mechanistic basis of this difference among rice subspecies remains unclear. Using transcriptomic, metabolomic and genetic analyses in combination with insect bioassay experiments, we showed that japonica and indica rice utilise different defence responses to repel SSB, and that SSB exploited plant nutrition deficiencies in different ways in the subspecies. The more resistant japonica rice induced patterns of accumulation of methyl jasmonate (MeJA-part of a defensive pathway) and vitamin B1 (VB1-a nutrition pathway) distinct from indica cultivars. Using gene-edited rice plants and SSB bioassays, we found that MeJA and VB1 jointly affected the performance of SSB by disrupting juvenile hormone levels. In addition, genetic variants of key biosynthesis genes in the MeJA and VB1 pathways (OsJMT and OsTH1, respectively) differed between japonica and indica rice and contributed to performance differences; in indica rice, SSB avoided the MeJA defence pathway and hijacked the VB1 nutrition-related pathway to promote development. The findings highlight important genetic and mechanistic differences between rice subspecies affecting SSB damage which could be exploited in plant breeding for resistance.

Constitutively expressed small heat shock protein LsHsp21.5 not only enhances heat tolerance but also helps to maintain reproduction in female Laodelphax striatellus.

Coping with stressful conditions and maintaining reproduction are two key biological processes that affect insect population dynamics. Small heat shock proteins (sHSPs) are involved in the stress response and the development of insects. The sHsp gene Laodelphax striatellus (Hemiptera: Delphacidae) sHsp 21.5 (LsHsp21.5) showed constitutive, stage- and organ-specific expression in L. striatellus, a pest that damages cultivated rice in east Asia. The expression of LsHsp21.5 was highest in the ovary, with 43.60, 12.99 and 1.45 time higher expression here than in the head, gut and female fat bodies, respectively. The expression of this gene was weakly affected by heat or cold shock. The gene provided in vitro protection against heat damage to malate dehydrogenase and in vivo protection against heat stress in Escherichia coli (Enterobacteriales: Enterobacteriaceae) BL21(DE3) and L. striatellus. Moreover, L. striatellus reproduction decreased by 1.85 times when the expression of LsHsp21.5 was inhibited by RNA interference. The expression of some genes related to reproduction, such as the homologous gene of chorion protein, also declined. These results suggest that LsHsp21.5 expression not only protects other proteins against stress but also helps maintain the stable expression of some reproduction-related genes under non-stressful conditions, with impacts on L. striatellus fecundity.

Egg-associated secretions from the brown planthopper (Nilaparvata lugens) activate rice immune responses.

The brown planthopper (BPH, Nilaparvata lugens) is a notorious sap-sucking insect pest that damages rice (Oryza sativa) plants throughout Asia. During BPH feeding, saliva enters rice plant tissues, whereas during oviposition egg-associated secretions (EAS) are deposited in damaged plant tissue. Dynamic changes in rice to planthopper salivary effectors have been widely reported. However, the effects of EAS from planthopper on rice immunity remains largely unexplored. In this study, we found that both infestation of rice by gravid BPH female adults and treatment with the EAS elicited a strong and rapid accumulation of jasmonic acid (JA), JA-isoleucine, and hydrogen peroxide in rice. EAS enhanced plant defenses not only in rice but also in tobacco, and these impaired the performance of BPH on rice, as well as the performance of aphids and whiteflies on tobacco. High-throughput proteome sequencing of EAS led to 110 proteins being identified and 53 proteins with 2 or more unique peptides being detected. Some proteins from BPH EAS were also found in the salivary proteome from herbivores, suggesting potential evolutionary conservation of effector functions across feeding and oviposition; however, others were only identified in EAS, and these are likely specifically related to oviposition. These findings point to novel proteins affecting interactions between planthoppers and rice during oviposition, providing an additional source of information for effector studies.

Evolution of Wolbachia reproductive and nutritional mutualism: insights from the genomes of two novel strains that double infect the pollinator of dioecious Ficus hirta.

Wolbachia is a genus of maternally inherited endosymbionts that can affect reproduction of their hosts and influence metabolic processes. The pollinator, Valisia javana, is common in the male syconium of the dioecious fig Ficus hirta. Based on a high-quality chromosome-level V. javana genome with PacBio long-read and Illumina short-read sequencing, we discovered a sizeable proportion of Wolbachia sequences and used these to assemble two novel Wolbachia strains belonging to supergroup A. We explored its phylogenetic relationship with described Wolbachia strains based on MLST sequences and the possibility of induction of CI (cytoplasmic incompatibility) in this strain by examining the presence of cif genes known to be responsible for CI in other insects. We also identified mobile genetic elements including prophages and insertion sequences, genes related to biotin synthesis and metabolism. A total of two prophages and 256 insertion sequences were found. The prophage WOjav1 is cryptic (structure incomplete) and WOjav2 is relatively intact. IS5 is the dominant transposon family. At least three pairs of type I cif genes with three copies were found which may cause strong CI although this needs experimental verification; we also considered possible nutritional effects of the Wolbachia by identifying genes related to biotin production, absorption and metabolism. This study provides a resource for further studies of Wolbachia-pollinator-host plant interactions.

Small Brown Planthopper Nymph Infestation Regulates Plant Defenses by Affecting Secondary Metabolite Biosynthesis in Rice.

The small brown planthopper (SBPH, Laodelphax striatellus) is one of the most destructive insect pests in rice (Oryza sativa), which is the world's major grain crop. The dynamic changes in the rice transcriptome and metabolome in response to planthopper female adult feeding and oviposition have been reported. However, the effects of nymph feeding remain unclear. In this study, we found that pre-infestation with SBPH nymphs increased the susceptibility of rice plants to SBPH infestation. We used a combination of broadly targeted metabolomic and transcriptomic studies to investigate the rice metabolites altered by SBPH feeding. We observed that SBPH feeding induced significant changes in 92 metabolites, including 56 defense-related secondary metabolites (34 flavonoids, 17 alkaloids, and 5 phenolic acids). Notably, there were more downregulated metabolites than upregulated metabolites. Additionally, nymph feeding significantly increased the accumulation of seven phenolamines and three phenolic acids but decreased the levels of most flavonoids. In SBPH-infested groups, 29 differentially accumulated flavonoids were downregulated, and this effect was more pronounced with infestation time. The findings of this study indicate that SBPH nymph feeding suppresses flavonoid biosynthesis in rice, resulting in increased susceptibility to SBPH infestation.

The microbiome stabilizes circadian rhythms in the gut.

The gut microbiome is well known to impact host physiology and health. Given widespread control of physiology by circadian clocks, we asked how the microbiome interacts with circadian rhythms in the Drosophila gut. The microbiome did not cycle in flies fed ad libitum, and timed feeding (TF) drove limited cycling only in clockless per01 flies. However, TF and loss of the microbiome influenced the composition of the gut cycling transcriptome, independently and together. Moreover, both interventions increased the amplitude of rhythmic gene expression, with effects of TF at least partly due to changes in histone acetylation. Contrary to expectations, timed feeding rendered animals more sensitive to stress. Analysis of microbiome function in circadian physiology revealed that germ-free flies reset more rapidly with shifts in the light:dark cycle. We propose that the microbiome stabilizes cycling in the host gut to prevent rapid fluctuations with changing environmental conditions.

Identification of a fatty acid synthase gene (FAS1) from Laodelphax striatellus planthoppers contributing to fecundity.

Fatty acid synthase (FAS) is a multifunctional enzyme that plays an important role in the formation of fatty acids. The fatty acids take part in many processes, such as cell signaling and energy metabolism, and in insects they are important in both cuticular hydrocarbon (CHC) formation and reproduction. Here we characterized the sequence structure and function of an FAS from the small brown planthopper (SBPH), Laodelphax striatellus. The full-length open reading frame (ORF) sequence of LsFAS1 was 7122 bp, encoding a predicted protein of 2373 amino acid residues. There were 7 functional domains in the LsFAS1 protein sequence. Gene expression screening by real-time quantitative polymerase chain reaction (RT-qPCR) showed that LsFAS1 was expressed in all developmental stages. Relative expression was highest at the 4th-instar and female adult stages. Among different tissues, the expression level of LsFAS1 in the ovary was the highest. Phylogenetic analysis showed that LsFAS1 clustered in a clade with 2 FASs from Nilaparvata lugens. Furthermore, these 3 FASs are related to cockroach BgFAS and locust LmFAS. After RNA interference-mediated knock-down, most treated insects died at eclosion. In addition, the lifespan of dsFAS1-treated female adults was shorter than that of the dsGFP-injected control, and offspring production decreased. Also, the expression of vitellogenin (Vg) and vitellogenin receptor (VgR) genes decreased. Virgin females dissected at days 2 and 4 post-eclosion showed many matured oocytes in planthoppers treated with dsGFP but not with dsFAS1. These data highlight the importance of LsFAS1 in SBPH, including a role in reproduction.

Time-Series Transcriptomic Analysis Reveals the Molecular Profiles of Diapause Termination Induced by Long Photoperiods and High Temperature in Chilo suppressalis.

Survival and adaptation to seasonal changes are challenging for insects. Many temperate insects such as the rice stem borer (Chilo suppressalis) overcome the adverse situation by entering diapause, wherein development changes dynamically occur and metabolic activity is suppressed. The photoperiod and temperature act as major environmental stimuli of diapause. However, the physiological and molecular mechanisms that interpret the ecologically relevant environmental cues in ontogenetic development during diapause termination are poorly understood. Here, we used genome-wide high-throughput RNA-sequencing to examine the patterns of gene expression during diapause termination in C. suppressalis. Major shifts in biological processes and pathways including metabolism, environmental information transmission, and endocrine signalling were observed across diapause termination based on over-representation analysis, short time-series expression miner, and gene set enrichment analysis. Many new pathways were identified in diapause termination including circadian rhythm, MAPK signalling, Wnt signalling, and Ras signalling, together with previously reported pathways including ecdysteroid, juvenile hormone, and insulin/insulin-like signalling. Our results show that convergent biological processes and molecular pathways of diapause termination were shared across different insect species and provided a comprehensive roadmap to better understand diapause termination in C. suppressalis.

SoxC is Required for Ecdysteroid Induction of Neuropeptide Genes During Insect Eclosion.

In insects, the shedding of the old exoskeleton is accomplished through ecdysis which is typically followed by the expansion and tanning of the new cuticle. Four neuropeptides, eclosion hormone (EH), ecdysis triggering hormone (ETH), crustacean cardioactive peptide (CCAP) and bursicon (Bur) are known to control ecdysis. However, the regulation of these neuropeptide genes is still poorly understood. Here, we report that in the red flour beetle (RFB) Tribolium castaneum and the fall armyworm (FAW) Spodoptera frugiperda, knockdown or knockout of the SoxC gene caused eclosion defects. The expansion and tanning of wings were not complete. In both RFB and FAW, the knockdown or knockout of SoxC resulted in a decrease in the expression of EH gene. Electrophoretic mobility shift assays revealed that the SfSoxC protein directly binds to a motif present in the promoter of SfEH. The luciferase reporter assays in Sf9 cells confirmed these results. These data suggest that transcription factor SoxC plays a key role in ecdysteroid induction of genes coding for neuropeptides such as EH involved in the regulation of insect eclosion.

Terpene Synthases in Rice Pan-Genome and Their Responses to Chilo suppressalis Larvae Infesting.

Terpene synthase (TPS) catalyzes the synthesis of terpenes and plays an important role in plant defense. This study identified 45 OsTPS genes (32 core genes and 13 variable genes) based on the high-quality rice gene-based pan-genome. This indicates limitations in OsTPS gene studies based on a single reference genome. In the present study, through collinearity between multiple rice genomes, one OsTPS gene absent in the reference (Nipponbare) genome was found and two TPS genes in the reference genome were found to have atypical structures, which would have been ignored in single genome analysis. OsTPS genes were divided into five groups and TPS-b was lost according to the phylogenetic tree. OsTPSs in TPS-c and TPS-g were all core genes indicating these two groups were stable during domestication. In addition, through the analysis of transcriptome data, some structural variations were found to affect the expression of OsTPS genes. Through the Ka/Ks calculation of OsTPS genes, we found that different OsTPS genes were under different selection pressure during domestication; for example, OsTPS22 and OsTPS29 experienced stronger positive selection than the other OsTPS genes. After Chilo suppressalis larvae infesting, 25 differentially expressed OsTPS genes were identified, which are involved in the diterpene phytoalexins precursors biosynthesis and ent-kaurene biosynthesis pathways. Overall, the present study conducted a bioinformatics analysis of OsTPS genes using a high-quality rice pan-genome, which provided a basis for further study of OsTPS genes.

Caterpillar-Induced Rice Volatile (E)-ß-Farnesene Impairs the Development and Survival of Chilo suppressalis Larvae by Disrupting Insect Hormone Balance.

Significant research progress has recently been made on establishing the roles of tps46 in rice defense. (E)-ß-farnesene (Eßf) is a major product of tps46 activity but its physiological functions and potential mechanisms against Chilo suppressalis have not yet been clarified. In the present study, C. suppressalis larvae were artificially fed a diet containing 0.8 g/kg Eßf and the physiological performance of the larvae was evaluated. In response to Eßf treatment, the average 2nd instar duration significantly increased from 4.78 d to 6.31 d while that of the 3rd instar significantly increased from 5.70 d to 8.00 d compared with the control. There were no significant differences between the control and Eßf-fed 4th and 5th instars in terms of their durations. The mortalities of the 2nd and 3rd Eßf-fed instars were 21.00-fold and 6.39-fold higher, respectively, than that of the control. A comparative transcriptome analysis revealed that multiple differentially expressed genes are involved in insect hormone biosynthesis. An insect hormone assay on the 3rd instars disclosed that Eßf disrupted the balance between the juvenile hormone and ecdysteroid levels. Eßf treatment increased the juvenile hormones titers but not those of the ecdysteroids. The qPCR results were consistent with those of the RNA-Seq. The foregoing findings suggested that Eßf impairs development and survival in C. suppressalis larvae by disrupting their hormone balance. Moreover, Eßf altered the pathways associated with carbohydrate and xenobiotic metabolism as well as those related to cofactors and vitamins in C. suppressalis larvae. The discoveries of this study may contribute to the development and implementation of an integrated control system for C. suppressalis infestations in rice.

Planthopper-Secreted Salivary Calmodulin Acts as an Effector for Defense Responses in Rice.

The brown planthopper (Nilaparvata lugens, BPH) and small brown planthopper (Laodelphax striatellus, SBPH) are major pests of rice (Oryza sativa) in Asia. These piercing-sucking insects secrete saliva into the host during feeding. Nevertheless, it is largely unknown how planthoppers use salivary effectors to enable continuous feeding on rice. Here, we screened their salivary proteomes and selected eight salivary proteins conserved between SBPH and BPH as candidate effectors. Silencing calmodulin (CaM) impeded BPH and SBPH from penetrating the phloem. Hence, their food intake, survival, and fecundity on rice plants were reduced. By contrast, CaM silencing had a small effect on the survival rate of BPH and SBPH raised on artificial diet. The CaM amino acid sequences were the same for both BPH and SBPH. CaM was highly expressed in their salivary glands and secreted into the rice plants during feeding. Bacterially expressed recombinant CaM protein exhibited calcium-binding activity. In planta expression disclosed that CaM was localized to the plant cytoplasms and nuclei and suppressed plant defenses such as hydrogen peroxide (H2O2) accumulation and callose deposition. CaM-silenced BPH and SBPH nymphs elicited relatively high levels of H2O2 and callose accumulation in rice plants. The foregoing results reveal that CaM is an effector as it enables the planthopper to reach the phloem by suppressing callose deposition and H2O2 accumulation in rice.

Vitellogenin from planthopper oral secretion acts as a novel effector to impair plant defenses.

Vitellogenin (Vg) is a well-known nutritious protein involved in reproduction in nearly all oviparous animals, including insects. Recently, Vg has been detected in saliva proteomes of several piercing-sucking herbivorous arthropods, including the small brown planthopper (Laodelphax striatellus, SBPH). Its function, however, remains unexplored. We investigated the molecular mechanism underlying SBPH orally secreted Vg-mediated manipulation of plant-insect interaction by RNA interference, phytohormone and H2 O2 profiling, protein-protein interaction studies and herbivore bioassays. A C-terminal polypeptide of Vg (VgC) in SBPH, when secreted into rice plants, acted as a novel effector to attenuate host rice defenses, which in turn improved insect feeding performance. Silencing Vg reduced insect feeding and survival on rice. Vg-silenced SBPH nymphs consistently elicited higher H2 O2 production, a well-established defense mechanism in rice, whereas expression of VgC in planta significantly hindered hydrogen peroxide (H2 O2 ) accumulation and promoted insect performance. VgC interacted directly with the rice transcription factor OsWRKY71, a protein which is involved in induction of H2 O2 accumulation and plant resistance to SBPH. These findings indicate a novel effector function of Vg: when secreted into host rice plants, this protein effectively weakened H2 O2 -mediated plant defense through its association with a plant immunity regulator.

A salivary calcium-binding protein from Laodelphax striatellus acts as an effector that suppresses defense in rice.

BACKGROUND: Calcium (Ca2+ )-binding proteins in the saliva of herbivorous insects function as effectors to attenuate host plant defenses and thus improve insect feeding performance. Silencing these genes via transgenic plant-mediated RNAi is thus a promising pest control strategy. However, their sequences and functions in the small brown planthopper Laodelphax striatellus (SBPH) remain to be investigated. RESULTS: We identified a putative EF-hand Ca2+ -binding protein (LsECP1) in SBPH watery saliva. LsECP1 was expressed extremely high in the salivary glands but at a low level during the egg stage. Transient LsECP1 expression in rice cells indicated its cytoplasm and nucleus localization. The bacterially expressed recombinant LsECP1 protein exhibited Ca2+ -binding activity. Rice plants fed by SBPH nymphs with knocked down LsECP1 exhibited higher levels of cytosolic Ca2+ , jasmonic acid (JA), jasmonoyl-isoleucine (JA-Ile) and hydrogen peroxide (H2 O2 ). Consistently, application of heterogeneously expressed LsECP1 protein suppressed wound-induced JA, JA-Ile and H2 O2 accumulation in rice. Thus, LsECP1 knockdown by dsRNA injection resulted in reduced feeding, fecundity and survival rates of SBPH reared on rice plants. Transgenic rice plants constitutively expressing LsECP1 dsRNA were produced, and plant-mediated LsECP1 knockdown enhanced rice resistance to SBPH. CONCLUSION: SBPH LsECP1 acts as an effector to impair host rice defense responses and promotes SBPH performance. This discovery provides a potential gene target for plant-mediated RNAi-based pest management. © 2021 Society of Chemical Industry.