Chilo Suppressalis Research Articles

High temperature-induced Cscaspase-8 disrupts the developmental relationship between Chilo suppressalis and its endoparasitoid

Host hemolymph is an important place of growth and development for most endoparasitoids. Immunofluorescence assay showed that parasitism induced Chilo suppressalis larvae to produce large numbers of granulocytes, but high temperatures led to granulocytes apoptosis and loss of phagocytosis. In addition, high temperatures activated the endoplasmic reticulum apoptotic pathway, leading to apoptosis of prohemocytes. In the present study, the initiator Cscaspase-8 was obtained from the rice pest C. suppressalis. The results of real-time PCR showed that Cscaspase-8 expression was highest in hemocytes; furthermore, transcription was most highly in female adults. Cscaspase-8 was significantly induced when larvae were exposed to 39 °C for a 2-h period. Cscaspase-8 expression was significantly elevated after 2 d of parasitism. Results of the interference test showed that the survival rate of C. suppressalis larvae is not affected by Cscaspase-8 gene silencing under high temperature and parasitism stress. However, developmental delays were observed in Cotesia chilonis larvae when the host Cscaspase-8 gene was knocked down. These results contribute to the current knowledge on the regulatory mechanisms of apoptosis in insects subjected to high temperature and parasitism stress.

(E)-β-Farnesene Protects Rice from Rice Striped Stem Borer Attack by Inhibiting Its Beneficial Gut Microbes.

Herbivores harbor gut microbes that affect their development and contribute to their nutrition, reproduction, and survival. Plant defenses could target the herbivore's beneficial gut microbes, but this has not been well studied in rice. In this context, we identified a rice terpene synthase gene, Os04g0341500. It was strongly induced after feeding by rice striped stem borers (SSB, Chilo suppressalis), and it can catalyze the (E)-β-farnesene (Eβf) synthesis. When added to artificial diets, Eβf impaired the development and survival of SSB larvae. High-throughput amplicon sequencing revealed that SSB fed on Eβf were decreased in beneficial gut microbes, compared to those feeding on the corresponding control feed. In vitro feeding of Eβf suggested that this antimicrobial sesquiterpene directly inhibited the growth of SSB gut microbes. The present study suggested that the Eβf-induced decrease of relative abundance of gut microbes potentially impairs larval development and survival in SSB.

The Threat of the Fall Armyworm to Asian Rice Production Is Amplified by the Brown Planthopper.

The recent invasion of the fall armyworm (FAW) into Asia not only has had a major impact on maize yield but is feared to also pose a risk to rice production. We hypothesized that the brown planthopper (BPH) may aggravate this risk based on a recently discovered mutualism between the planthopper and the rice striped stem borer. Here we show that BPH may indeed facilitate a shift of FAW to rice. FAW females were found to strongly prefer to oviposit on BPH-infested rice plants, which emitted significantly elevated levels of five volatile compounds. A synthetic mixture of these compounds had a potent stimulatory effect on ovipositing females. Although FAW caterpillars exhibited relatively poor growth on both uninfested and BPH-infested rice, a considerable portion completed their development on young plants. Moreover, FAW were found to readily pupate and survive in exceedingly moist soils typical for rice cultivation, further highlighting FAW's potential to switch to rice. We conclude that BPH, by changing the bouquet of volatiles emitted by rice plants, may greatly facilitate this switch. These findings, together with a current increase of nonflooded upland rice in Asia, warrant careful monitoring and specific control measures against FAW to safeguard Asian rice production.

Expression of a novel NaD1 recombinant antimicrobial peptide enhances antifungal and insecticidal activities

This study aimed to increase the antifungal and insecticidal activities of NaD1, as an antimicrobial peptides (AMP), by improving its interaction with the fungal cell wall and chitin monomeric units in insect midguts. Hence, the chitin-binding domains (CBDs) of wheat germ agglutinin protein (WGA) were fused to either N- or C-terminus of NaD1 generating transgenic Nicotiana tabacum hairy roots (HRs). Molecular assessments confirmed the integration of NaD1 transgenes, their transcription and production of recombinant peptides in the HR lines. Total protein of (CBD)4-NaD1 and NaD1-(CBD)4 transgenic lines inhibited the growth of Pyricularia oryzae mycelium, suggesting that fusion of CBD to NaD1 can increase NaD1 half-life, leading to higher affinity toward cell wall chitin. Furthermore, feeding the third-instar larvae of Chilo suppressalis with both (CBD)4-NaD1 and NaD1-(CBD)4 extracts exhibited a higher mortality rate. Both NaD1-CBDs caused a significant decrease in trypsin (TRY) and chymotrypsin (CTR) activities in the larvae, while enhancing the activity of antioxidant enzymes CAT, POD, APX, and SOD. Therefore, feeding the larvae by total extract of NaD1-(CBD)4 and (CBD)4-NaD1 HR lines probably increased affinity to midgut chitin in C. suppressalis, enhancing insecticidal activities. Overall, the results indicate that recombinant peptides are effective in enhancing fungal and insect resistance.

Functional and evolutionary analysis of key enzymes triacylglycerol lipase, glycogen hydrolases in the glycerol and glucose biosynthesis pathway and cellular chaperones for freeze-tolerance of the Rice stem borer, Chilo suppressalis

Freeze-tolerance is an important physiological trait for terrestrial environmental adaptation and intraspecific geographic-lineage diversification in ectothermic animals, yet there remains a lack of systematic studies on its underlying genetic mechanisms and evolution. To address this problem, we employed the widely distributed rice pest, the Chilo suppressalis, as a model to explore the genetic mechanisms and evolutionary history of freeze-tolerance. First, we systematically characterized its antifreeze mechanisms by performing functional validation of potential key genes in laboratory-reared lines. This revealed the functional roles of glycerol biosynthesis in freeze-tolerance, including the triacylglycerol-originated pathway via triacylglycerol lipase (Tgl) hydrolysis and the glycogen-originated pathway via α-amylase (Aa) and maltase (Ma) hydrolysis, as well as the roles of the cellular chaperones Hsc70 and Hsf1. Then, we investigated the evolution of freeze-tolerance by collecting representative geographical samples and performing population genetic analyses, which suggested differentiated strategies of cold adaptation in different geographic populations. Taken together, our findings demonstrate the functional basis of cold resistance in Chilo suppressalis and reveal the evolutionary history of freeze-tolerance in natural populations, providing insights into organismal freeze-tolerance and clues for pest control.

Resistance Realized Heritability and Fitness Cost of Cyproflanilide in Rice Stem Borer, Chilo suppressalis (Lepidoptera: Pyralidae)

The rice stem borer (RSB) Chilo suppressalis is a devastating rice pest with resistance to a number of insecticides. Recently, the new meta-diamide insecticide cyproflanilide has been considered an effective insecticide to control RSB. However, its resistance risk has not been reported. In the present study, we aimed to assess the resistance risk and evaluate the fitness cost after the RSB was exposed to cyproflanilide. After five generations of selection, the resistance level of RSB increased by 1.5-fold. When h2 was 0.125, a 10-fold resistance increase in the LD50 values was expected in fourteen and thirty-one generations at the selection intensity of 90% and 50%, respectively. The selected population (RSB-SEL) had significant differences in the developmental duration of eggs, 1st, 2nd, 3rd, and 6th instar larvae, and female pupae compared to the unselected population (RSB-UNSEL). Besides, the adult longevity was shortened, and the average pupal weight of males, the emergence rate, the sex ratio, the oviposition, the mean fecundity, and the full life cycle rate were decreased in RSB-SEL. The intrinsic rate of increase (r), the net reproductive rate (R0), and the finite rate of increase (λ) of RSB-SEL were significantly lower than those of RSB-UNSEL, while the mean generation time (T) of RSB-SEL was significantly longer than that of RSB-UNSEL. Based on the results of the prediction of the generations required for a 10-fold resistance increase in the LD50, a potential risk of resistance development exists in RSB after continuous and excessive use of cyproflanilide. These results will be useful in designing the dose of cyproflanilide to control C. suppressalis in field.

OPTIMIZATION OF SEX PHEROMONE DOSE AND TRAP DENSITY FOR MATING DISRUPTION OF CHILO SUPPRESSALIS (LEPIDOPTERA; PYRILADAE)

The rice stem borer, Chilo suppressalis (Walker) is an oligophagous pest that mainly feeds on rice and currently is largely controlled through chemicals, leading to several ecological problems and often low control efficiency due to the larval feeding habits. Therefore, the alternative and eco-friendly control strategies is needed to manage this noxious insect pests, applications of trapping devices specifically sex pheromone traps are one of the most substitute control methods to monitoring and control moths by killing or mating disruption on large scale. The aim of the current study was to optimize the appropriate pheromone dose, trap density and trap placement height for mating disruption of C. suppressalis. During first year (2021), a mixture of two pheromones (Z11-16: Ald and Z9-16: Ald) in equal ratio (50:50) were applied at different doses (50, 100, 200, 300 and 500µg/20µL), with various trap heights (3, 4.5 and 6 feet) and compared with insecticide (fipronil G 0.2% 15 kg/ha). Results revealed that, 200 and 300µg/20µL caught significantly maximum (119.63±15.86) moths of C. suppressalis and resulted in lowest (4.96%) damage (white ear and dead hearts) as compared to other doses and treatments. The second year´s trial was designed to optimize the trap density in which 8, 12, 18 and 25 traps/ha were tested with a median dosage 250µg/20µl, based on first year’s results. It exhibited that 18 traps/ha captured maximum number (121.95±19.77) moths and reduced the damage symptoms significantly followed by other 25, 12 and 8 traps/ha with significant difference (P ≤0.001). Moreover, results of best treatment (appropriate sex pheromone dosage, trap placement height and trap density/ha) revealed highest (3:52) cost benefit ratio with minimum (7.90 $) expenditure also highest yield 7400 kg/ha was recorded as compared to other treatments. Therefore, installation of 18 traps/ha at 4.5 feet height with a dose of 250µg/20µL can manage C. suppressalis in rice-growing areas and consequently reduce management cost as compared to the insecticides. Thus, it is recommended as an important component for the eco-friendly management of rice stem borer to ensure food security. Key words: Chilo suppressalis, Pheromone dosage, Trap density,Trap height and mating disruption.

The irregular developmental duration mainly caused by the broad-complex in Chilo suppressalis

Chilo suppressalis, a critical rice stem borer pest, poses significant challenges to rice production due to its overlapping generations and irregular developmental duration. These characteristics complicate pest management strategies. According to the dynamic analysis of the overwintering adults of C. suppressalis in fields, it indicates that the phenomenon of irregular development of C. suppressalis exists widely and continuously. This study delves into the potential role of the Broad-Complex (Br-C) gene in the developmental duration of C. suppressalis. Four isoforms of Br-C, named CsBr-C Z1, CsBr-C Z2, CsBr-C Z4, and CsBr-C Z7, were identified. After CsBr-Cs RNAi, the duration of larva development spans extended obviously. And, the average developmental duration of dsCsBr-Cs feeding individuals increased obviously. Meanwhile, the average developmental duration of the dsCsBr-C Z2 feeding group was the longest among all the RNAi groups. After dsCsBr-Cs feeding continuously, individuals pupated at different instars changed obviously: the proportion of individuals pupated at the 5th instar decreased and pupated at the 7th instar or higher increased significantly. Moreover, the pupation rate of dsCsBr-Cs (except dsCsBr-C Z7) were significantly lower than that of dsGFP. The same results were obtained from the mutagenesis in CsBr-C genes mediated by CRISPR/Cas9. The average developmental duration of CsBr-Cs knockout individuals was significantly prolonged. And, the instar of pupation in knockout individuals was also delayed significantly. In conclusion, this work showed that CsBr-Cs played a crucial role in pupal commitment and affected the developmental duration of C. suppressalis significantly.

Oral secretions from striped stem borer (Chilo suppressalis) induce defenses in rice.

The striped stem borer (SSB, Chilo suppressalis) is one of the most destructive insect pests on rice. As a chewing insect, SSB larval feeding causes a dramatic increase in rice defense responses. However, the effects of oral secretions (OSs) during SSB feeding on rice defense remain largely unexplored. In this study, based on transcriptome analysis results, treatment with SSB OSs regulated the expression of genes involved in the plant defense-related pathways of calcium, mitogen-activated protein kinases, reactive oxygen species, jasmonic acid (JA), herbivore-induced plant volatiles (HIPVs), and protease inhibitors. Unsurprisingly, treatment with SSB OSs elicited the accumulation of JA and JA-isoleucine in rice. The defense mechanisms activated by the cascade not only induced the expression of trypsin inhibitors, inhibiting the normal growth of SSB larvae but also induced HIPVs emission, rendering rice attractive to a common larval parasitoid. High-throughput proteome sequencing of SSB OSs led to 534 proteins being identified and 343 proteins with two or more unique peptides being detected. The study demonstrates that SSB OSs trigger both direct and indirect defense mechanisms in rice, akin to the effects of SSB feeding. It identifies specific proteins in SSB OSs that may influence the interactions between SSB and rice during feeding, providing valuable insights for effectors research. © 2024 Society of Chemical Industry.

Infestation of Rice Striped Stem Borer (Chilo suppressalis) Larvae Induces Emission of Volatile Organic Compounds in Rice and Repels Female Adult Oviposition.

Plants regulate the biosynthesis and emission of metabolic compounds to manage herbivorous stresses. In this study, as a destructive pest, the pre-infestation of rice striped stem borer (SSB, Chilo suppressalis) larvae on rice (Oryza sativa) reduced the subsequent SSB female adult oviposition preference. Widely targeted volatilomics and transcriptome sequencing were used to identify released volatile metabolic profiles and differentially expressed genes in SSB-infested and uninfested rice plants. SSB infestation significantly altered the accumulation of 71 volatile organic compounds (VOCs), including 13 terpenoids. A total of 7897 significantly differentially expressed genes were identified, and genes involved in the terpenoid and phenylpropanoid metabolic pathways were highly enriched. Correlation analysis revealed that DEGs in terpenoid metabolism-related pathways were likely involved in the regulation of VOC biosynthesis in SSB-infested rice plants. Furthermore, two terpenoids, (-)-carvone and cedrol, were selected to analyse the behaviour of SSB and predators. Y-tube olfactometer tests demonstrated that both (-)-carvone and cedrol could repel SSB adults at higher concentrations; (-)-carvone could simultaneously attract the natural enemies of SSB, Cotesia chilonis and Trichogramma japonicum, and cedrol could only attract T. japonicum at lower concentrations. These findings provide a better understanding of the response of rice plants to SSB and contribute to the development of new strategies to control herbivorous pests.

Extreme genetic signatures of local adaptation in a notorious rice pest, Chilo suppressalis

ABSTRACT Climatic variation stands as a significant driving force behind genetic differentiation and the evolution of adaptive traits. Chilo (C.) suppressalis, commonly known as the rice stem borer, is a highly destructive pest that crucially harms rice production. The lack of natural population genomics data has hindered a more thorough understanding of its climate adaptation, particularly the genetic basis underlying adaptive traits. To overcome this obstacle, our study employed completely resequenced genomes of 384 individuals to explore the population structure, demographic history, and gene flow of C. suppressalis in China. This study observed that its gene flow occurred asymmetrically, moving from central populations to peripheral populations. Using genome-wide selection scans and genotype-environment association studies, we identified potential loci that may be associated with climatic adaptation. The most robust signal was found to be associated with cold tolerance, linked to a homeobox gene, goosecoid (GSC), whose expression level was significantly different in low and high latitudes. Moreover, downregulating the expression of this gene by RNAi enhances its cold tolerance phenotypes. Our findings have uncovered and delved into the genetic foundation of the ability of C. suppressalis to adapt to its environment. This is essential in ensuring the continued effectiveness and sustainability of novel control techniques.

Enhancing bioactivity of Callistemon citrinus (Curtis) essential oil through novel nanoemulsion formulation

The main focus of this study was to create a stable and efficient nanoemulsion (NE) using Callistemon citrinus essential oil (EO). Various factors affecting the NE’s stability were optimized including oil %, Tween 80%, time of sonication, and its accelerated stability was examined. The research also considered the antibacterial, antifungal, and larvicidal effects of the optimized NE (B10). The optimum NE stood out for its stability, featuring a particle size of 33.15 ± 0.32 nm. Analysis via IR spectroscopy confirmed successful EO encapsulation in B10. The formulation remained stable for six months, with B10 showing significantly higher antibacterial and antifungal potency compared to the pure oil. When samples were subjected to tests against Fusarium oxysporum, B10 exhibited a MIC value of 62.5 mg/mL, whereas the pure oil showed a MIC value of 250 mg/mL. This indicates that the B10 formulation was 50 times more effective than the EO. In terms of antibacterial activity against Escherichia coli, the MIC value was 0.256 mg/mL for B10 and 4 mg/mL for the EO. Also, pure oil and B10 displayed larvicidal effects against Chilo suppressalis (Walker) larvae, with B10 eliminating 95.2% of larvae in 48 h. Overall, stable and optimum C. citrinus NE with its strong antimicrobial qualities, shows promise as an effective fungicide and insecticide.

MiRNA-mediated insect-resistant transgenic rice poses no risk to a non-target parasitoid, Cotesia chilonis, via direct feeding or through its target host.

MicroRNAs (miRNAs) have started to play an important role in pest control, and novel miRNA-based transgenic insect-resistant plants are now emerging. However, an environmental risk assessment of these novel transgenic plants expressing insect miRNAs must be undertaken before promoting their application. Here, transgenic miR-14 rice, which has high resistance to the rice stem borer Chilo suppressalis, was used as an example for evaluation in this study. Taking the tier1 risk assessment method in Bacillus thuringiensis (Bt) crops as a reference, the effects of the direct exposure of a non-target parasitoid, Cotesia chilonis, to a high concentration of miRNA were evaluated. The results showed that direct feeding with miR-14 at high concentration had no significant effects on the biological parameters of Co.chilonis, whereas when miR-14 was injected into Ch.suppressalis-parasitized larvae, the development duration of Co.chilonis was significantly affected. In combination with the real conditions of the rice paddy field, it could be inferred that transgenic miR-14 rice has no significant negative effects on the important non-target parasitoid, Co.chilonis. These results will provide a foundation for the establishment of a new safety evaluation system for novel RNAi-based transgenic plants.

The jasmonate pathway and water loss in rice leaves induced by a stem-borer inhibit leaf-feeder growth.

Plant-mediated interactions between herbivores play an important role in regulating the composition of herbivore community. The fall armyworm (FAW), Spodoptera frugiperda, which has become one of the most serious pests on corn in China since it invaded in 2018, has been found feeding rice in the field. However, how FAW interacts with native rice insect pests remains largely unknown. Here, we investigated the interaction between FAW and a resident herbivore, striped stem borer (SSB, Chilo suppressalis) on rice. The infestation of rice leaf sheaths (LSs) by SSB larvae systemically enhanced the level of jasmonic acid (JA), abscisic acid (ABA), and trypsin proteinase inhibitors (TPIs), reduced relative water content (RWC) in leaf blades (LBs), and suppressed the growth of FAW larvae. In contrast, because FAW larvae infested LBs and did not affect defence responses in LSs, they did not influence the performance of SSB larvae. Using different mutants, together with bioassays and chemical analysis, we revealed that SSB-induced suppression of FAW larvae growth depended on both the SSB-activated JA pathway and RWC in LBs, whereas the ABA pathway activated by SSB larvae promoted the growth of FAW larvae by impeding water loss. These results provide new insights into mechanisms underlying plant-mediated interactions between herbivores.

A parasitic wasp-releasing engineering to promote ecosystem services in paddy systems

In East Asia, rice is a key crop that is subject to significant insect driven crop damage often leading to wide scale reliance on insecticides. We assessed the effects of releasing conditions (including density, frequency and distribution points) of a parastic wasp (i.e., Trichogramma japonicum) on the control efficacy it provided against two rice pest herbivores (i.e., Chilo suppressalis and Cnaphalocrocis medinalis). Optimal control was achieved by three releases of 150,000 wasps per hectare across 75 release points within a hectare rice field. A following 2-year large-scale field experiment demonstrated that the release of parasitic wasps allowed a 60.0 % reduction in insecticide use when compared to the insecticide treatment alone. Importantly, there was also a 3.7 % increase in pest control, 2.5 % increase in rice yield, and an overall 4.5 % increase in net economic benefit when the release of parasitic wasps was combined with insecticide application. This was an improvement on the use of either parasitic wasps alone or insecticide application alone. Release of parasitic wasps had co-benefits for other invertebrate predators whose abundance was increased by 65.8 % relative to the insecticide only treatment, however, this fell to a 26.4 % increase when parasitic wasps release was combined with insecticide use. Additionally, the combined treatment group of parasitic wasps and insecticides exhibited the highest net economic benefit (40488.9±593.2 Chinese Yuan (CNY) ha−1), significantly surpassing both the parasitic wasps-only treatment by 8.9 % and the treatment of no parasitic wasps and no insecticide by 21.8 %. These findings suggest that parasitic wasps releasing is a feasible approach to ecological engineering to promote multiple ecosystem services linked to greater crop yield in agroecosystems.

Metabolomic Profiling Reveals the Anti-Herbivore Mechanisms of Rice (Oryza sativa).

The use of secondary metabolites of rice to control pests has become a research hotspot, but little is known about the mechanism of rice self-resistance. In this study, metabolomics analysis was performed on two groups of rice (T1, with insect pests; T2, without pests), indicating that fatty acids, alkaloids, and phenolic acids were significantly up-regulated in T1. The up-regulated metabolites (p-value < 0.1) were enriched in linoleic acid metabolism, terpene, piperidine, and pyridine alkaloid biosynthesis, α-linolenic acid metabolism, and tryptophan metabolism. Six significantly up-regulated differential metabolites in T1 were screened out: N-trans-feruloyl-3-methoxytyramine (1), N-trans-feruloyltyramine (2), N-trans-p-coumaroyltyramine (3), N-cis-feruloyltyramine (4), N-phenylacetyl-L-glutamine (5), and benzamide (6). The insect growth inhibitory activities of these six different metabolites were determined, and the results show that compound 1 had the highest activity, which significantly inhibited the growth of Chilo suppressalis by 59.63%. Compounds 2-4 also showed a good inhibitory effect on the growth of Chilo suppressalis, while the other compounds had no significant effect. RNA-seq analyses showed that larval exposure to compound 1 up-regulated the genes that were significantly enriched in ribosome biogenesis in eukaryotes, the cell cycle, ribosomes, and other pathways. The down-regulated genes were significantly enriched in metabolic pathways, oxidative phosphorylation, the citrate cycle (TCA cycle), and other pathways. Eighteen up-regulated genes and fifteen down-regulated genes from the above significantly enriched pathways were screened out and verified by real-time quantitative PCR. The activities of detoxification enzymes (glutathione S-transferase (GST); UDP-glucuronosyltransferase (UGT); and carboxylesterase (CarE)) under larval exposure to compound 1 were measured, which indicated that the activity of GST was significantly inhibited by compound 1, while the activities of the UGT and CarE enzymes did not significantly change. As determined by UPLC-MS, the contents of compound 1 in the T1 and T2 groups were 8.55 ng/g and 0.53 ng/g, respectively, which indicated that pest insects significantly induced the synthesis of compound 1. Compound 1 may enhance rice insect resistance by inhibiting the detoxification enzyme activity and metabolism of Chilo suppressalis, as well as promoting cell proliferation to affect its normal growth and development process. The chemical-ecological mechanism of the insect resistance of rice is preliminarily clarified in this paper.

Functional Analysis of Insecticide Inhibition and Metabolism of Six Glutathione S-Transferases in the Rice Stem Borer, Chilo suppressalis.

The glutathione S-transferases (GSTs) are important detoxifying enzymes in insects. Our previous studies found that the susceptibility of Chilo suppressalis to abamectin was significantly increased when the CsGST activity was inhibited by glutathione (GSH) depletory. In this study, the potential detoxification mechanisms of CsGSTs to abamectin were explored. Six CsGSTs of C. suppressalis were expressed in vitro. Enzymatic kinetic parameters including Km and Vmax of recombinant CsGSTs were determined, and results showed that all of the six CsGSTs were catalytically active and displaying glutathione transferase activity. Insecticide inhibitions revealed that a low concentration of abamectin could effectively inhibit the activities of CsGSTs including CsGSTd1, CsGSTe4, CsGSTo2, CsGSTs3, and CsGSTu1. However, the in vitro metabolism assay found that the six CsGSTs could not metabolize abamectin directly. Additionally, the glutathione transferase activity of CsGSTs in C. suppressalis was significantly increased post-treatment with abamectin. Comprehensive analysis of the results in present and our previous studies demonstrated that CsGSTs play an important role in detoxification of abamectin by catalyzing the conjugation of GSH to abamectin in C. suppressalis, and the high binding affinities of CsGSTd1, CsGSTe4, CsGSTo2, CsGSTs3, and CsGSTu1 with abamectin might also suggest the involvement of CsGSTs in detoxification of abamectin via the noncatalytic passive binding and sequestration instead of direct metabolism. These studies are helpful to better understand the detoxification mechanisms of GSTs in insects.

Comparative assessment of the gut bacterial diversity associated with field population of three rice stem borers and their in vitro insecticide degradation ability

Insects harbour diverse range of microbes in the digestive tract. An investigation was carried out to isolate and characterize the culturable gut bacterial fauna of three stem borers attacking rice (Scirpophaga incertulus, Sesamia inferens and Chilo suppressalis) morphologically, biochemically and genetically (based on 16S rDNA sequences). Their ability to degrade two commonly used insecticides, i.e., chlorpyrifos and chlorantraniliprole was quantified in vitro through minimal media study using GC–MS and HPLC, respectively. There was a great variation in the gut bacterial composition of these three borers in spite of belonging to same guild and being collected from the same host (Varshadhan: CRLC-899) at the same time from ICAR-NRRI farm, Cuttack, India. Bacillus was prominent in S. incertulus and C. suppressalis whereas Stenotrophomonas was dominant in S. inferens. It is the first report of encountering B. pumilus, B. cereus and Acinetobacter calcoaceticus from the gut of S. incertulus and Paraclostridium from the gut of S. inferens. Chlorpyrifos and chlorantraniliprole was degraded to the tune of 44.87–92.02 % and 10.52 to 74.38 %, respectively. B. subtilis most efficiently degraded chlorpyrifos, while Klebsiella sp. and A. calcoaceticus were superior in degrading Chlorantraniliprole. The gut bacterial consortium of S. inferens was the most efficient in degrading the insecticides. The differential abilities of the gut bacterial isolates to degrade the insecticides provide insights for their possible involvement in xenobiotic detoxification in their host and further study on this aspect may improve management strategies of these pests. These insects can also serve as novel sources for profiling insecticide-degrading microbes for bioremediation programmes.

Knockdown of MAPK p38-linked genes increases the susceptibility of Chilo suppressalis larvae to various transgenic Bt rice lines

Bacillus thuringiensis (Bt) toxins have provided exceptional control of agricultural insect pests, however, over reliance on the proteins would potentially contribute to the development of field tolerance. Developing new sustainable insect pest control methods that target the mechanisms underlying Bt tolerance can potentially support the Bt control paradigm while also providing insights into basic insect physiology. The MAPK p38 pathway is strongly associated with Bt tolerance in Chilo suppressalis, a major pest of rice. To gain insights into how this pathway impacts tolerance, high-throughput screening of C. suppressalis larval midguts initially identified eight novel target genes. Increased larval sensitivity to the transgenic cry1Ca rice strain T1C-19 was observed following RNA interference-mediated knockdown of four of the genes, Cscnc, Csgcp, Cszfp26 and CsZMYM1. Similar enhanced sensitivity to the TT51 (expressing Cry1Ab/1Ac) and T2A-1 (expressing Cry2Aa) transgenic rice lines occurred when Cszfp26 and CsZMYM1 were knocked down. All four target genes are downstream of the MAPK p38 pathway but do not participate in negative feedback loop of the pathway. These results implicate Cscnc, Csgcp, Cszfp and CsZMYM1 in the C. suppressalis transgenic cry1Ca rice tolerance mechanism regulated by MAPK p38. These findings further enhance our understanding of the MAPK p38-dependent molecular mechanisms underlying Bt tolerance in C. suppressalis and open new avenues of tolerance management to develop.

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-partof 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 hormonelevels. 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.