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.     

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.     

Responses by the brown planthopper,Nilaparvata lugens,to conspecific density on resistant and susceptible rice varieties

This study examines the nature of intraspecific interactions among Nilaparvata lugens (Stål) (Hemiptera: Delphacidae) planthoppers feeding on resistant and susceptible rice varieties. Planthopper nymphs produced less honeydew and gained less weight when feeding on rice variety IR62 (resistant) compared to susceptible rice varieties. A series of bioassays was conducted that varied N. lugens nymph densities on IR62 and IR22 (susceptible). Increasing nymph density facilitated feeding by conspecifics; however, intraspecific competition increased mortality of nymphs on IR62 (but rarely on IR22). Furthermore, nymph weights declined with increasing conspecific density on IR22, and the effects were weak on IR62. More female nymphs than males survived on IR62 but this was not affected by density. Nitrogenous fertilizer increased competition among N. lugens on young plants of IR22, but not on IR62. Results indicate that nymphs have a low efficiency in accessing resources when feeding on IR62, even where the plants have received fertilizer. Female‐biased survival and biomass compensation for mortality may promote population recovery after development on the resistant plant and accelerate adaptation to the resistant variety.     

Biochar amendment to soils impairs developmental and reproductive performances of a major rice pest Nilaparvata lugens (Homopera: Delphacidae)

Biochar amendments to soils had aroused much interest for having potential for greenhouse gas mitigation, soil improvement and increased crop productivity. However, little attention had been focused on the influence of biochar amendments on herbivorous insect pests. This study investigated whether a biochar amendment affected developmental and reproductive performances of the rice brown planthopper (Nilaparvata lugens) feeding on rice plants. The biochar from the pyrolysis of wheat straw was used for treatments of soils (from a fallow rice field), and the treated soils were applied to grow rice seedlings in small vials, in which N. lugens life history was observed. The nymphal development time was delayed and nymph‐to‐adult survival decreased with a high level of 200 g/kg biochar application. Herbivore lifetime fecundity decreased with increasing amounts of biochar, from 256 eggs under the control down to 69 eggs under the high level (200 g/kg) of biochar application. Egg‐hatching rate significantly decreased at the highest biochar level (200 g/kg), compared to the other lower biochar levels. Our results suggest that biochar amendment to rice fields may have negative impacts on the rice brown planthoppers when applied at level of 200 g/kg of soil.     

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.     

PHYSIOLOGICAL EFECTS OF PAICHONGDING APPLIED TO RICE ON THE Nilaparvata lugens (STÅ L), THE BROWN PLANTHOPPER

Nilaparvata lugens (Stål) is a major rice pest in Asia. Paichongding is a novel neonicotinoid insecticide developed in 2008. The effects of this insecticide on the activity of detoxification enzymes of N. lugens and on rice resistance to the pest were examined in the laboratory. The results showed that paichongding could significantly decrease the acetylcholinesterase and GSHs transferase activities of N. lugens. The variation tendency of mixed function oxidase (MFO) activity was similar with that of the esterase. After 12 h treatment, there was no significance between the treatment and control. However, the activities of MFO and esterase increased after 24 and 48 h treatment, which suggested that MFO and esterase may play an important role in the detoxification of paichongding for N. lugens. Our results also demonstrated that treated with paichongding, damage levels of rice plants were significantly lower than those of control plants except 15 days after treatment. Compared with the control, injury indices decreased 70.22, 49.12, 34.44, and 23.23% at 3 , 6 , 9, and 12 days after paichongding treatment, respectively. The laboratory results suggested that paichongding may be effective for the control of brown planthopper.     

Mechanisms of resistance to the brown planthopperNilaparvata lugens in wild rice (Oryza spp.) cultivars

Three wild rice species and six cultivated rice varieties were evaluated to determine their mechanisms of resistance toNilaparvata lugens (Stal.). Wild rice species,Oryza officinalis, O. punctata, andO. latifolia and cultivated rices Rathu Heenati (Bph 3), Babawee (bph 4), ARC 10 550 (bph 5), Swarnalata (Bph 6), Ptb 33 (bph 2+Bph 3) and the susceptible Taichung Native (TN 1) (no resistance gene) were included in the study. In a free choice seedbox screening test, wild rice species maintained their high level of resistance through the 48 h exposure toN. lugens nymphs while plant damage ratings of cultivated rice varieties increased with time. Wild rices were non preferred and significantly more individuals settled on susceptible TN 1 followed by cultivated rices. The quantity of food ingested and assimilated byN. lugens on wild rices was less than on cultivated resistant varieties.N. lugens caged on resistant wild rices had slow nymphal development, reduced longevity, low fecundity, and low egg hatchability as compared toN. lugens on cultivated resistant varieties.     

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.     

Herbivore-induced rice resistance against rice blast mediated by salicylic acid

In agro-ecosystems,plants are important mediators of interactions between their associated herbivorous insects and microbes,and any change in plants induced by one species may lead to cascading effects on interactions with other species.Often,such effects are regulated by phytohormones such as jasmonic acid(JA)and salicylic acid(SA).Here,we investigated the tripartite interactions among rice plants,three insect herbivores(Chilo suppressalis,Cnaphalocrocis medinalis or Nilapai-vata lugens),and the causal agent of rice blast disease,the fungus Magnaporthe oryzae.We found that pre-infestation of rice by C.suppressalis or N.lugens but not by C.medinalis conferred resistance to M.oryzae.For C.suppressalis and N.lugens,insect infestation without fungal inoculation induced the accumulation of both JA and SA in rice leaves.In contrast,infestation by C.medinalis increased JA levels but reduced SA levels.The exogenous application of SA but not of JA conferred resistance against M.oryzae.These results suggest that preinfestation by C suppressalis or N.lugens conferred resistance against M.oryzae by increasing SA accumulation.These findings enhance our understanding of the interactions among rice plant,insects and pathogens,and provide valuable information for developing an ecologically sound strategy for controlling rice blast.     

Influence of rice black streaked dwarf virus on the ecological fitness of non-vector planthopper Nilaparvata lugens （Hemiptera： Delphacidae）

Rice black streak dwarf virus （RBSDV） is transmitted by the small brown planthopper （SBPH）, Laodelphax striatellus （Fallen）. Non-vector rice brown planthopper （BPH）, Nilaparvata lugens （Stal）, shares the same host rice plants with SBPH in paddy fields. The changes in nutritional composition of rice plants infected by RBSDV and the ecological fitness of BPH feeding on the infected plants were studied under both artificial climate chamber and field conditions. Contents of 16 detected amino acids and soluble sugar in RBSDV infected rice plants were higher than those in the healthy ones. On the diseased plants BPH had significantly higher nymphal survival rates, nymphal duration of the males, weight of the female adults, as well as egg hatchability compared to BPH being fed on healthy plants. However, there was no obvious difference in female nymph duration, longevity and fecundity. Defense enzymes （superoxidase dismutase, SOD and catalase, CAT） and detoxifying enzymes （carboxylesterase, CAE and glutathione S-transferase, GST） in BPH adults fed on diseased plants had markedly higher activities. The results indicate rice plants infected by RBSDV improved the ecological fitness of the brown planthopper, a serious pest but not a transmitter of the RBSDV virus.     

Study of the relationship between the content of the rare earth element Eu in rice plants and in Nilaparvata lugens (Hemiptera: Delphacidae)

Abstract The current study investigated changes in the content of the rare earth element Europium (Eu) in roots, shoots and leaves of rice plants and in Nilaparvata lugens Stål (Hemiptera: Delphacidae) using an Eu marker and hydroponic culture. The results showed that N. lugens infestation significantly reduced Eu content in roots, shoots and leaves of two rice varieties, Shenyou 1 and Xieyou 963. The Eu content in roots, shoots and leaves of the susceptible variety, Shenyou 1, was significantly higher than that in the resistant variety, Xieyou 963. The Eu content of N. lugens fed on Shenyou 1 was significantly higher than of those fed on Xieyou 963. In addition, the Eu level was elevated at a higher density of N. lugens infestation. Eu content in the bodies of N. lugens was related to their weight and honeydew excretion, with a significant positive correlation. Thus, Eu content in the bodies of N. lugens can be considered an index of the amount of phloem sap taken in by N. lugens because the amount of honeydew excretion is proportional to the amount of phloem sap consumed. The ratios of Eu content in N. lugens to that in roots, shoots and leaves of rice plants were elevated at a higher N. lugens infestation density. That ratio was maximal in leaves, was intermediate in shoots and was minimal in roots. There was no significant difference in ratios between the two plant varieties. An Eu marker may be useful in the screening of resistant varieties and in the study of the mechanisms of resistance.     

褐飞虱的迁人和翅型分化规律的研究

褐飞虱Nilaparvata lugens(Stal)属迁飞性水稻害虫,其成虫有短翅和长翅两种翅型。 褐飞虱迁入雌虫均未经过交配,其短翅型由一对显性等位基因控制。翅型分化同时受到遗传和内分泌系统的协调控制,外界条件如密度和寄主等因子通过内分泌系统来影响翅型的分化。褐飞虱翅型分化的敏感龄期雌虫为1～3龄,而雄虫为1-5龄;不同若虫蜜度处理对褐飞虱成虫的前翅形成有一定的影响。分蘖期水稻饲养的褐飞虱短翅型比例明显高于孕穗期水稻饲养的褐飞虱。不同密度下各生物型间的翅型分化差异不显著。迁入地的浙江各种群属温带型,其雌虫短翅率低且与密度呈显著负相关;菲律宾热带种群雌虫在不同密度下均为短翅型,而雄虫的短翅率随密度增加而上升。广西种群接近热带型,其雌虫短翅率高但不随密度而变化。试验各种群的雄虫在中等密度甚至高密度时其短翅率出现最高。     

Ectopic expression of the spike protein of Rice Ragged Stunt Oryzavirus in transgenic rice plants inhibits transmission of the virus to insects

Rice ragged stunt oryzavirus (RRSV) replicates in both its insect vector, Nilaparvata lugens, and its plant host, rice, and has a complex multi-component particle bearing spikes on its outer surface. Transgenic rice lines expressing the 39 kDa spike protein showed good resistance to infection by RRSV. Furthermore, N. lugens fed on these plants prior to feeding on RRSV-infected plants were significantly protected against RRSV infection. The viral titre in insects initially fed on transgenic plants and then on RRSV-infected plants was inversely proportional to the levels of the 39 kDa protein expressed in the transgenic plants. This suggests that the 39 kDa protein interferes with the interaction between the intact virus particles and insect cell receptors and that the spike protein of RRSV contributes to vector specificity. This approach would probably be a more environment-friendly and sustainable method of virus control than by actual eradication of insect vectors.     

THE VIRULENCE CHANGE AND DAMAGE CHARACTERISTICS OF VARIOUS GEOGRAPHIC POPULATIONS OF BROWN PLANTHOPPER*

Abstract The virulence change and damage characteristics of brown planthopper (BPH), Nilaparvata lugens Stal, populations collected from Guangxi, Yunnan and Zhejiang Provinces were studied by using various resistant rice varieties. The results indicated that the Zhejiang population could successfully infest varieties IR26 and Mudgo, showing the characters of BPH biotype 2 after 1989. The Guangxi population, however, could adapt the resistant varieties IR26, Mudgo and ASD7 after 1991. In 1997, the nymphal survival indices of Guangxi and Yunnan populations on Rathu Heenati (Bph3) were up to 70. 9 and 66. 7 respectively. This may imply that the BPH populations in Guangxi and Yunnan have shifted into a new biotype. In free-choice test, there was obvious difference in the feeding preference of nymphs and adults of Zhejiang population, the adult preferred to feeding on resistant variety IR26 and the nymphs chose the hosts randomly. Under 26 C and L12: D12, the nymphal duration of Zhejiang population was the longest in the three tested populations. For Guangxi population, the nymphal duration on ASD7 was significantly longer than on TN1, although it could infest resistant variety ASD7. The nymphal survival rate of Zhejiang population fed on IR26 was higher than those fed on TN1 and ASD7. On varieties IR26 and ASD7, an obviously higher nymphal survival rate was observed in Yunnan population as compared to those of Zhejiang and Guangxi populations. These results indicate that difference in virulence and damage characteristics could be found in different geographical populations of BPH.     

Effect of rice lines transformed with Bacillus thuringiensis toxin genes on the brown planthopper and its predator Cyrtorhinus lividipennis

Five transgenic rice lines, each containing an insecticidal toxin gene from Bacillus thuringiensis (Bt) under control of a different promoter, were tested for effects on two non-target insects: the brown planthopper, Nilaparvata lugens (Stål) (Homoptera: Delphacidae), and its predator Cyrtorhinus lividipennis (Hemiptera: Miridae). Bt toxin was detected by ELISA in the honeydew of N. lugens that fed on rice lines with the CaMV 35S and actin promoters. Nilaparvata lugens produced greater volumes of acidic honeydew (derived from xylem feeding) on all five Bt rice lines than on non-transgenic control lines. The amount of honeydew derived from phloem feeding did not differ between Bt and control lines. There were no differences between N. lugens reared on Bt and control lines in any of the five fitness parameters measured (survival to the adult stage, male and female weight, and male and female developmental time). There were no differences between C. lividipennis reared on N. lugens nymphs from Bt and control lines, in any of the three fitness parameters examined (survival to the adult stage and male and female developmental time). Our results indicate that N. lugens and its natural enemies will be exposed to Bt toxins from rice lines transformed with some Bt gene constructs, but that this exposure might not affect N. lugens and C. lividipennis fitness.     

Direct and feeding‐induced interactions between two rice planthoppers, Sogatella furcifera and Nilaparvata lugens: effects on dispersal capability and performance

Abstract. 1. A series of laboratory experiments was conducted to explore the effects of inter‐specific interactions, both direct interactive effects and those induced through previous feeding, on the dispersal capability (proportion of macropterous adults) and performance (development time and survival) of two wing‐dimorphic planthoppers, the whitebacked planthopper Sogatella furcifera and the brown planthopper Nilaparvata lugens, two pests of rice throughout Asia. 2. An asymmetric effect of inter‐specific crowding on dispersal capability was detected between the two planthoppers. With density controlled, the proportion of macropterous adults in N. lugens was higher when raised in mixed populations with S. furcifera than in pure cultures comprised of conspecifics, suggesting that interspecific effects on dispersal capability are stronger than intraspecific effects. In contrast, interspecific effects on macroptery (%) were weaker than intraspecific impacts for S. furcifera. 3. This trend was parallelled by adverse competitive effects on survival, with interspecific effects stronger than intraspecific impacts for N. lugens and intraspecific effects stronger for S. furcifera. 4. Crowding also affected the incidence of macroptery indirectly through feeding‐induced changes in plant physiology. Rearing N. lugens on plants fed on previously by heterospecifics resulted in a higher incidence of macroptery in females and protracted development than for N. lugens raised on plants fed on previously by conspecifics. By contrast, the intraspecific effect of previous feeding was stronger than the interspecific effect in S. furcifera, with higher macropter production and prolonged development occurring on plants exposed previously to conspecifics. 5. The results suggest that interspecific interactions between herbivorous insects, both direct and via feeding‐induced changes in plant physiology, can have negative consequences for performance and survival and promote the production of flight‐capable adults that can disperse.     

Feeding‐induced changes in defence enzymes and PR proteins and their implications in host resistance to Nilaparvata lugens

Six rice genotypes showing susceptible and resistant reactions to brown planthopper (BPH), Nilaparvata lugens were studied for feeding‐induced changes in defence enzymes and pathogenesis‐related (PR) proteins. The high resistant genotypes PTB 33, ADT 45 and ASD 7 and moderately resistant genotypes CO 43 and KAU 1661 recorded the greater expression of defence enzymes peroxidase, polyphenol oxidase, phenylalanine ammonia lyase, total phenol and β‐1,3 glucanase in response to N. lugens feeding at 1 day after infestation (DAI) compared with susceptible genotype TN1. The greater activity of chitinase was observed in resistant cultivars at 3 DAI and the activity was sustained for more than 1 week compared with susceptible TN1. In conclusion, the current study revealed that these defence enzymes and PR proteins might attribute to the resistance mechanisms in rice plants against BPH infestation.     

Metabolic Transformation and Accumulation of O-Ethyl S,S-Diphenyl Phosphorodithiolate (Hinosan®) in Rice Plants

³²P-labeled organophosphorus fungicide Hinosan was sprayed on rice plants at various growth stages, and metabolic fate of the pesticide in the rice plants was studied. Identification of Hinosan and its metabolites in the n-hexane and water extracts was conducted by TLC and GLC (FPD).

The rate of hydrolysis of Hinosan in rice plants seemed to be slower than that of other organophosphorus pesticides. Hexane-soluble components, which were detected throughout the experimental period, consisted mainly of Hinosan. Among the water-soluble metabolites identified were O-ethyl S-phenyl phosphorothioic acid and S,S-diphenyl phosphoro- dithioic acid, which were detected one to four days after the application, and ethyl phosphate and phosphoric acid which increased with the lapse of time.

Upon examination of the radioactivity of Hinosan and its metabolites in rice grains, a certain level was detected in husk, but very little in hulled rice and polished rice.     

Electrical penetration graphs indicate that tricin is a key secondary metabolite of rice,inhibiting phloem feeding of brown planthopper,Nilaparvata lugens

Tricin (5,7,4′‐trihydroxy‐3′,5′‐dimethoxyflavone) is a valuable secondary metabolite which is widely present in gramineous plants, including cultivated rice (Oryza sativa L.) (Poaceae). It can defend the rice plant against damage by the brown planthopper (BPH), Nilaparvata lugens (Stål) (Hemiptera: Delphacidae), one of the most important pests of rice. This study was conducted to elucidate the mechanisms of action of tricin on BPH feeding behavior. BPH feeding behavior in resistant (Rathu Heenati, RHT) and susceptible (Taichuang native 1, TN1) rice varieties and artificial diets was monitored using the electrical penetration graph (EPG) technique. Tricin concentrations in leaves of varieties RHT and TN1 were quantitatively analyzed by liquid chromatography, coupled to tandem mass spectrometric techniques. Six (NP and N1‐5) and four (NP, N1, N2, and N4) types of waveforms occurred during feeding on rice plants and artificial diets, respectively. The tricin concentration of rice varieties was correlated with total and average durations of N4. Moreover, EPG data indicated that tricin significantly increased the duration of non‐probing and pathway periods and strongly inhibited phloem ingestion (N4). The inhibition was strongly dose dependent, resulting in complete suppression of activity in the phloem region when the tricin concentration was increased to 1 g l^?1. This study revealed that tricin disturbed the feeding behavior of BPH mainly by increasing the non‐probe period and inhibiting phloem ingestion. We confirmed the hypothesis that tricin is a ‘stylet probing stimulant’ of rice planthoppers as proposed in previous studies. The information on the ecological effect of tricin from this study may be useful to clarify the resistance mechanism against BPH of RHT and other tricin‐containing rice varieties.