Protection of rice against Nilaparvata lugens by direct toxicity of sodium selenate

Nilaparvata lugens is one of the most notorious pest insects of cultured rice, and outbreaks of N. lugens cause high economic losses each year. While pest control by chemical pesticides is still the standard procedure for treating N. lugens infections, excessive use of these insecticides has led to the emergence of resistant strains and high pesticide residues in plants for human consumption and the environment. Therefore, novel and environment-friendly pest control strategies are needed. In previous studies, selenium was shown to protect selenium-accumulating plants from biotic stress. However, studies on nonaccumulator (crop) plants are lacking. In this study, rice plants (Oryza sativa, Nipponbare) were treated with sodium selenate by seed priming and foliar spray and then infested with N. lugens. Brown planthoppers feeding on these plants showed increased mortality compared to those feeding on control plants. Treatment of the plants with sodium selenate did not affect the enzymes involved in the biosynthesis of the plant stress hormones jasmonic acid and salicylic acid, suggesting that the observed insect mortality cannot be attributed to the activation of these hormonal plant defenses. Feeding assays using an artificial diet supplemented with sodium selenate revealed direct toxicity toward N. lugens. With a low concentration of 6.5 ± 1.5 µM sodium selenate, half of the insects were killed after 3 days. In summary, sodium selenate treatment of plants can be used as a potential alternative pest management strategy to protect rice against N. lugens infestation through direct toxicity.     

THE ROLES OF DETOXIFYING ENZYMES AND AChE INSENSITIVITY IN METHAMIDOPHOS RESISTANCE DEVELOPMENT AND DECLINE IN NILAPARVATA LUGENS

Methamidophos resistance of brown planthopper (Nilaparvata lugens Stål, BPH) was selected in laboratory. After successive selection for 9 generations, the selection was ceased by rearing BPH without contact with any insecticide for 9 generations. In the full course, the successive changes of esterase activity, MFO activity, GSTs activity and AChE insensitivity were analyzed. The results showed that the change of esterase activity was high correlated with that of methamidophos in the full course, which indicated that esterase played very important role both in the resistance development and in the resistance decline. However, the change of AChE insensitivity only significantly correlated with that of resistance in the development stage, and the change of MFO activity or GSTs activity only significantly correlated with that of the resistance in the decline stage, which indicated the changes of AChE insensitivity, MFO activity or GSTs activity only played some roles in different stages of the resistance change.     

Biochemical mechanism of rice nitrogen promoting the resistance development of Nilaparvata lugens (Stål) to buprofezin

Overusing nitrogen fertilizer not only pollute farmlands, but also make Nilaparvata lugens (Stål) less susceptible to pesticides. The current study investigated the relationship between nitrogen fertilizer application and buprofenzin resistance in N. lugens in rice by conducting a series of field efficacy tests and laboratory bioassays. The results showed that the efficacy of buprofezin decreased significantly with high level of nitrogen fertilizer application, compared to those with low or no nitrogen application in rice fields. In the laboratory, the resistance ratios of N. lugens to buprofezin, assayed at three concentrations of nitrogen (0, 1.0, 3.0 mmol/L), have developed to 14.05-, 18.40- and 25.81-fold after 20 generations of resistance selection, respectively. Meanwhile, the resistance ratios of N. lugens without buprofezin treated were 1.00-, 3.30- and 4.67-fold after 20 generations as the nitrogen concentrations increased. The enzymatic assays revealed that the activities of P450 and esterase increased significantly after resistance selection with buprofezin and nitrogen, especially on the esterase activity in N. lugens under the condition of nitrogen treatment. These results suggested that nitrogen fertilizer application contributed to the resistance development in N. lugens to buprofezin by enhancing the activity of esterase.     

Carboxylesterase genes in nitenpyram-resistant brown planthoppers,Nilaparvata lugens

Carboxylesterases (CarEs) represent one of the major detoxification enzyme families involved in insecticide resistance. However, the function of specific CarE genes in insecticide resistance is still unclear in the insect Nilaparvata lugens (Stål), a notorious rice crop pest in Asia. In this study, a total of 29 putative CarE genes in N. lugens were identified, and they were divided into seven clades; further, the β-esterase clade was significantly expanded. Tissue-specific expression analysis found that 17 CarE genes were abundantly distributed in the midgut and fat body, while 12 CarE genes were highly expressed in the head. The expression of most CarE genes was significantly induced in response to the challenge of nitenpyram, triflumezopyrim, chlorpyrifos, isoprocarb and etofenprox. Among these, the expression levels of NlCarE2, NlCarE4, NlCarE9, NlCarE17 and NlCarE24 were increased by each insecticide. Real-time quantitative polymerase chain reaction and RNA interference assays revealed the NlCarE1 gene to be a candidate gene mainly involved in nitenpyram resistance, while simultaneously silencing NlCarE1 and NlCarE19 produced a stronger effect than silencing either one individually, suggesting a cooperative relationship in resistance formation. These findings lay the foundation for further clarification of insecticide resistance mediated by CarE in N. lugens.     

Establishing the role of detoxifying enzymes in field‐evolved resistance to various insecticides in the brown planthopper (Nilaparvata lugens) in South India

The brown planthopper (BPH), Nilaparvata lugens (Stål), is one of the major pests of rice throughout Asia. Extensive use of insecticides for suppressing N. lugens has resulted in the development of insecticide resistance leading to frequent control failures in the field. The aim of the present study was to evaluate resistance in the field populations of N. lugens from major rice growing states of South India to various insecticides. We also determined the activity of detoxifying enzymes (esterases [ESTs], glutathione S‐transferases [GSTs], and mixed‐function oxidases [MFOs]). Moderate levels of resistance were detected in the field populations to acephate, thiamethoxam and buprofezin (resistance factors 1.05–20.92 fold, 4.52–14.99 fold, and 1.00–18.09 fold, respectively) as compared with susceptible strain while there were low levels of resistance to imidacloprid (resistance factor 1.23–6.70 fold) and complete sensitivity to etofenoprox (resistance factor 1.05–1.66 fold). EST activities in the field populations were 1.06 to 3.09 times higher than the susceptible strain while for GST and MFO the ratios varied from 1.29 to 3.41 and 1.03 to 1.76, respectively. The EST activity was found to be correlated to acephate resistance (r = 0.999, P ≥ 0.001). The high selection pressure of organophosphate, neonicotinoid, and insect growth regulator (IGR) in the field is likely to be contributing for resistance in BPH to multiple insecticides, leading to control failures. The results obtained will be beneficial to IPM recommendations for the use of effective insecticides against BPH.     

Imidacloprid‐induced transference effect on some elements in rice plants and the brown planthopper Nilaparvata lugens (Hemiptera: Delphacidae)

Abstract The widespread use of imidacloprid against insect pests has not only increased the rate of the development of target pest resistance but has also resulted in various negative effects on rice plants and Nilaparvata lugens resurgence. However, the effect of imidacloprid on elements in rice plants and the transference of these element changes between rice and N. lugens are currently poorly understood. The present study investigated changes of Cu, Fe, Mn, Zn, Ca, K, Mg and Na contents in rice plants following imidacloprid foliar sprays in the adult female of N. lugens that develops from nymphs that feed on treated plants and honeydew produced by females. The results indicated that imidacloprid foliar spray significantly increased Fe and K contents in leaf sheaths. Generally, Fe, Mn, K and Na contents in leaf blades were noticeably decreased, but Ca contents in leaf blades for 10 and 30 mg/kg imidacloprid treatments were significantly increased. The contents of most elements except K and Mg in the adult females and honeydew were significantly elevated. Multivariate statistical analysis showed that Fe, Mn and Na in leaf blades and Fe and Mn in leaf sheaths could be proportionally transferred to N. lugens. The relationship between most elements in adult female bodies and in the honeydew showed a positive correlation coefficient. There were significant differences in the contents of some elements in rice plants and N. lugens from different regions.     

斜纹夜蛾抗药性及其防治对策的研究进展

斜纹夜蛾Spodopteralitura是多种作物的重要害虫 ,对有机氯、有机磷、氨基甲酸酯类、拟除虫菊酯类以及Bt制剂等杀虫剂均产生抗药性。本文对斜纹夜蛾抗药性的形成与发展、抗性机理及其防治对策等方面进行了综述     

Pesticide-induced susceptibility of rice to brown planthopper Nilaparvata lugens

The effects of rice plants treated with various pesticides (jingganmycin, bisultap and methamidophos) on feeding, survival rates and population growth of Nilaparvata lugens Stål (Homoptera: Delphacidae), susceptibility of the treated rice plants and amounts of free amino acids and sucrose were studied. Experiments indicated that the effects of the tested pesticides were dependent on nymphal age, pesticide and their dose and time after application. Jingganmycin at 75 g a.i. ha^–1 significantly increased the N. lugens population. Both jingganmycin and bisultap increased the survival rate of N. lugens nymphs. The feeding rate of the insects was also affected by the pesticide application, but the effect varied between nymphal age and time after application and lasted no longer than 15 d. Results clearly indicated that pesticide application increased the susceptibility of rice plants to N. lugens. Although the free amino acids in rice plants did not change with the pesticide treatments, the concentration of sucrose significantly decreased 5 d after application and the C/N ratio significantly decreased in jingganmycin treated plants 5 d and 10 d after application.     

Ethiprole resistance in Nilaparvata lugens (Hemiptera: Delphacidae): possible mechanisms and cross-resistance

This article reports the current status of ethiprole resistance in Nilaparvata lugens Stål in the central region of Thailand, together with the associated resistance mechanisms. A resistance survey found that a field population had developed 308.5-fold resistance to ethiprole. Further selection with ethiprole for nine generations in the laboratory led to 453.1-fold ethiprole resistance. However, following this selection procedure, the resistance of N. lugens to other insecticides decreased to about one-third of its original resistance. This result implies that there is no cross-resistance between ethiprole and other kinds of insecticides in this pest. In an in vivo study of synergisms, triphenyl phosphate (TPP) exhibited a strong synergism (SR 4.2) with ethiprole in the resistant hoppers, piperonyl butoxide (PBO) also showed significant synergistic effects with ethiprole (1.6), but diethyl maleate (DEM) did not show any obvious synergism with ethiprole (1.2). An in vitro biochemical study indicated that esterase activity increased with ethiprole resistance in N. lugens, that P450 monooxygenase activity also increased significantly with high resistance, but that glutathione S-transferase activity did not. These results reveal that increases in esterase activity and P450 monooxygenase activity cause the ethiprole resistance observed in the field populations of N. lugens. Whether the mechanisms for ethiprole resistance involve target-site sensitivity is not yet known; further molecular analysis is required. However, an analysis of insecticide cross-resistance and the insecticide application history of the resistant populations indicated that target resistance was present and that rotation between insecticides with different modes of action will provide a key countermeasure to maintain the efficacy of ethiprole.     

Transgenic plants expressing the AaIT/GNA fusion protein show increased resistance and toxicity to both chewing and sucking pests

The adoption of pest‐resistant transgenic plants to reduce yield losses and decrease pesticide use has been successful. To achieve the goal of controlling both chewing and sucking pests in a given transgenic plant, we generated transgenic tobacco, Arabidopsis, and rice plants expressing the fusion protein, AaIT/GNA, in which an insecticidal scorpion venom neurotoxin (Androctonus australis toxin, AaIT) is fused to snowdrop lectin (Galanthus nivalis agglutinin, GNA). Compared with transgenic tobacco and Arabidopsis plants expressing AaIT or GNA, transgenic plants expressing AaIT/GNA exhibited increased resistance and toxicity to one chewing pest, the cotton bollworm, Helicoverpa armigera. Transgenic tobacco and rice plants expressing AaIT/GNA showed increased resistance and toxicity to two sucking pests, the whitefly, Bemisia tabaci, and the rice brown planthopper, Nilaparvata lugens, respectively. Moreover, in the field, transgenic rice plants expressing AaIT/GNA exhibited a significant improvement in grain yield when infested with N. lugens. This study shows that expressing the AaIT/GNA fusion protein in transgenic plants can be a useful approach for controlling pests, particularly sucking pests which are not susceptible to the toxin in Bt crops.     

Impacts of nitrogen fertilizer on major insect pests and their predators in transgenic Bt rice lines T2A‐1 and T1C‐19

Nitrogen is a critical factor for plant development and nitrogen input is one of the important tactics to enhance the development and yield of crops. Nevertheless, nitrogen input could influence the occurrence of insects positively or negatively. Nitrogen is also one of the main elements composing the insecticidal crystal (Cry) protein. Cry protein production could affect nitrogen partitioning in Bt plants and as such nitrogen input may influence insect pest management in transgenic Bt rice, Oryza sativa L. (Poaceae). To test this possibility, we evaluated the impacts of nitrogen regimes on the main insect pests and their predators on two Bt rice lines, T2A‐1 and T1C‐19, expressing Cry2A and Cry1C, respectively, and their non‐transgenic parental counterpart MH63. The results showed that Cry proteins with different nitrogen regimes have enough insecticidal activity on rice leaffolder, Cnaphalocrocis medinalis Guenée (Lepidoptera: Crambidae), in both laboratory and field experiments. Laboratory studies indicated that relevant parameters of ecological fitness in brown planthopper, Nilaparvata lugens (Stål) (Hemiptera: Delphacidae), a non‐target insect pest, were significantly affected by nitrogen input both on Bt and MH63 rice lines. Nymphal survival, female adult longevity, and egg hatchability in N. lugens differed significantly among rice varieties. The experiments conducted in rice fields also demonstrated that nitrogen was positively correlated with the abundance of N. lugens on Bt rice, similar to that on MH63 rice. The abundances of two predators – the wolf spider Pirata subpiraticus (Boesenberg & Strand) (Araneae: Lycosidae) and the bug Cyrthorhinus lividipennis Reuter (Hemiptera: Miridae) – were significantly affected by rice growth stages but not by nitrogen input and rice varieties. In conclusion, the above results indicate that high nitrogen regimes for Bt rice (T2A‐1 and T1C‐19) and non‐Bt rice (MH63) cannot facilitate the management of insect pests.     

Evidence of sibling species between two host-associated populations of brown planthopper, N. lugens (stål) (Homoptera: Delphacidea) complex based on morphology and host–plant relationship studies

Morphological and host–plant relationship studies were conducted to differentiate two sympatric populations of brown planthopper (BPH), Nilaparvata lugens, one from rice (Oryza sativa) and the other from Leersia hexandra, a weed grass. In morphometric studies based on esterase activities, an UPGMA dendrogram using 17 quantitative morphological characters, including stridulatory organs (courtship signal-producing organs) between two sympatric populations of N. lugens, one from rice and the other from L. hexandra, a weed grass revealed that both populations were separated from each other. An out-group, N. bakeri, was found to be completely different from the two sympatric populations of N. lugens. Rice plants were best suited for the establishment of the rice-infesting population, and L. hexandra was a favourable host for the Leersia-infesting population. The individuals derived from one host did not thrive on the other host, as shown by a significant reduction in survival and nymphal development, ovipositional preferences, ovipositional response, and egg hatchability. Therefore, morphological and host–plant relationship studies indicate that rice-associated population with high esterase activities and L. heaxandra-associated population with low esterase activities are two closely related sibling species.     

The Insect Ecdysone Receptor is a Good Potential Target for RNAi-based Pest Control

RNA interference (RNAi) has great potential for use in insect pest control. However, some significant challenges must be overcome before RNAi-based pest control can become a reality. One challenge is the proper selection of a good target gene for RNAi. Here, we report that the insect ecdysone receptor (EcR) is a good potential target for RNAi-based pest control in the brown planthopper Nilaparvata lugens, a serious insect pest of rice plants. We demonstrated that the use of a 360 bp fragment (NlEcR-c) that is common between NlEcR-A and NlEcR-B for feeding RNAi experiments significantly decreased the relative mRNA expression levels of NlEcR compared with those in the dsGFP control. Feeding RNAi also resulted in a significant reduction in the number of offspring per pair of N. lugens. Consequently, a transgenic rice line expressing NlEcR dsRNA was constructed by Agrobacterium- mediated transformation. The results of qRT-PCR showed that the total copy number of the target gene in all transgenic rice lines was 2. Northern blot analysis showed that the small RNA of the hairpin dsNlEcR-c was successfully expressed in the transgenic rice lines. After newly hatched nymphs of N. lugens fed on the transgenic rice lines, effective RNAi was observed. The NlEcR expression levels in all lines examined were decreased significantly compared with the control. In all lines, the survival rate of the nymphs was nearly 90%, and the average number of offspring per pair in the treated groups was significantly less than that observed in the control, with a decrease of 44.18-66.27%. These findings support an RNAi-based pest control strategy and are also important for the management of rice insect pests.     

Flight duration of the brown planthopper, Nilaparvata lugens (Homoptera: Delphacidae)

ABSTRACT.

• 1 The brown planthopper, Nilaparvata lugens (Stål), is a major pest of rice in Asia. It is known to make wind-assisted migratory flights each year to colonize the summer rice growing areas of China, Japan and Korea.
• 2 Modelling windborne displacements between rice growing areas in Asia requires migratory behaviour and flight duration to be established for this insect.
• 3 Field and laboratory observations suggest that N. lugens take-off at dusk and that some continue flying for up to 24–26 h if the temperature is ≥ 17°C.
• 4 Trajectories for 10 m above ground level and 1.5 km above mean sea level are used to identify possible sources and, hence, to estimate the flight times of N.lugens caught in nets on ships on the East China Sea in 1973 and 1981.
• 5 Estimated flight times between the sources and the ships ranged from about 9 to 30 h.
• 6 Results suggest that long-distance migration can occur in surface winds, when they are strong, but that long-distance migration is more likely at 1.5 km.
• 7 When simulating windborne displacements of N.lugens, it can be assumed that in areas and at heights where the temperature is ≥ 17°C, some migrants will fly downwind for up to 30 h after a dusk take-off. Others will fly for shorter periods, giving the population as a whole the opportunity to colonize all the rice crops flown over.

     

Bt rice could provide ecological resistance against nontarget planthoppers

Genetically engineered (GE) rice lines expressing Lepidoptera‐active insecticidal cry genes from the bacterium Bacillus thuringiensis (Bt) have been developed in China. Field surveys indicated that Bt rice harbours fewer rice planthoppers than non‐Bt rice although planthoppers are not sensitive to the produced Bt Cry proteins. The mechanisms underlying this phenomenon remain unknown. Here, we show that the low numbers of planthoppers on Bt rice are associated with reduced caterpillar damage. In laboratory and field‐cage experiments, the rice planthopper Nilapavata lugens had no feeding preference for undamaged Bt or non‐Bt plants but exhibited a strong preference for caterpillar‐damaged plants whether Bt or non‐Bt. Under open‐field conditions, rice planthoppers were more abundant on caterpillar‐damaged non‐Bt rice than on neighbouring healthy Bt rice. GC–MS analyses showed that caterpillar damage induced the release of rice plant volatiles known to be attractive to planthoppers, and metabolome analyses revealed increased amino acid contents and reduced sterol contents known to benefit planthopper development. That Lepidoptera‐resistant Bt rice is less attractive to this important nontarget pest in the field is therefore a first example of ecological resistance of Bt plants to nontarget pests. Our findings suggest that non‐Bt rice refuges established for delaying the development of Bt resistance may also act as a trap crop for N. lugens and possibly other planthoppers.     

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

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

Molecular characterization and gene functional analysis of Dicer‐2 gene from Nilaparvata lugens (Hemiptera: Geometroidea)

Abstract Nilaparvata lugens (Stål) (Hemiptera: Geometroidea), a serious rice pest in many countries of Asia, causes a great loss in rice production every year. RNA interference (RNAi) is a powerful technology for gene function study in insects and a potential tool for pest control. As a core component of RNAi pathway, Dicer‐2 (Dcr‐2) protein determines the production of small interfering RNA (siRNA) and is crucial for the efficiency of RNAi. In this study, the full‐length complementary DNA (cDNA) of N. lugens Dcr‐2 (NlDcr‐2) was first cloned and analyzed, and then the RNAi experiment was conducted to explore the function of NlDcr‐2 gene. The complete Dcr‐2 cDNA of N. lugens was 4 971 bp in length with an open reading frame (ORF) of 1,656 amino acids. Phylogenetic and protein domain analysis showed that the predicted NlDcr‐2 protein was similar to Tribolium castaneum. In the RNAi experiment, the messenger RNA level of NlDcr‐2 was significantly reduced by NlDcr‐2 double‐stranded RNA (dsRNA) (dsDcr‐2). Fifty‐five per cent decrease of NlDcr‐2 was found after 4 days of unremitting feeding. No significant effect was observed on the development of N. lugens after dsRNA ingestion.