Dispersal kernel of green rice leafhopper estimated from truncated data

Characterizing dispersal kernels from truncated data is important for managing and predicting population dynamics. We used mark-recapture data from 10 previously published replicated experiments at three host plant development stages (seedling, tillering, and heading) to estimate parameters of the normal and exponential dispersal kernels for green rice leafhopper, Nephotettix cincticeps (Uhler). We compared classic statistical methods for estimating untruncated distribution parameters from truncated data with maximum likelihood (MLE) and the method of statistical moments for simulated and empirical data. Simulations showed that both methods provided accurate parameter estimates with similar precision. The method of moments is algebraically complex, but simple to calculate, while the MLE methods require numerical solutions of nonlinear equations. Simulations also showed that accurate, precise estimates of the parameters of the untruncated distributions could be attained even under severe truncation with sufficient numbers of recaptures. Both diffusivity and the exponential mean were higher with later plant growth stage, showing that insects moved farther and faster at the heading stage. Precision of the estimates was not strongly related to percent capture, size of the experimental field, or the number of leafhoppers captured. The leptokurtic exponential kernel fit the data better than the normal kernel for all the experiments. These results support an alternative explanation for the strong density-dependent population regulation of this species at the heading stage. Instead of leafhopper density per se, the increase in movement at this stage could integrate the populations in the separate fields, leveling densities throughout the landscape.     

Electronic measurement of probing activities of the green leafhopper of rice

The electronic measurement system developed for aphids has been modified to measure leafhopper salivation and ingestion activities. Several distinct waveforms were recorded when the rice green leafhopper, Nephotettix cincticeps (Uhler) probed rice plants and into sucrose solutions. These waveforms were correlated with ingestion and salivation activities based on chemical analysis of the excreta.

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Elektronische messung des probiersaugens der grünen reiszikade

Zusammenfassung Das für Blattläuse entwickelte Messystem wurde abgeändert, um die Probiersaugaktivität von Zikaden zu messen. Es wurden mehrere unterscheidbare Wellenformen registriert, wenn die Grüne Reiszikade, Nephotettix cincticeps, Reispflanzen und Rohrzuckerlösungen probierte. Basierend auf der chemischen Analyse der Ausscheidungen konnten diese Wellenformen mit der Futteraufnahme-und Speichelbildungsaktivität korreliert werden.

     

A rice protein library: a data-file of rice proteins separated by two-dimensional electrophoresis

Proteins extracted from embryos, endosperms and leaves of rice were separated by two-dimensional electrophoresis and relative molecular weights and isoelectric points were determined. The separated proteins were electroblotted onto a polyvinylidene difluoride membrane and 85 electroblotted proteins were analyzed by a gas-phase protein sequencer. The N-terminal amino-acid sequences of 27 out of 85 proteins were determined in this manner. The N-terminal regions of the remaining proteins could not be sequenced and they were inferred to have a blocking group at the N-terminus. Among proteins, 11 could be sequenced after deblocking by in situ treatment with pyroglutamyl peptidase. The internal amino-acid sequences of 23 proteins were determined by sequence analysis of peptides obtained by Cleveland peptide mapping. The amino-acid sequences determined here were compared with those of known plant and animal proteins. The concanavalin A-peroxidase method was used to determine whether the 85 proteins were glycosylated and the diagonal electrophoresis method was used to determine whether they contained disulphide bonding. Finally, we constructed a data-file of rice proteins including information on relative molecular weight, isoelectric point, amino-acid sequence, sequence homology, glycosylation, and the presence of disulphide bonding.     

Aberrations caused by neem oil odour in green leafhopper feeding on rice plants

Using an electronic device and a lignin-specific dye, the feeding behaviour of the green leafhopper,Nephotettix virescens (Distant), (Homoptera: Cicadellidae) was monitored on rice plants kept in an arena permeated with the odour of neem (Azadirachta indica A. Juss) seed oil. The garlicky odour of neem oil disrupted the normal feeding behaviour of the leafhopper. Phloem feeding byN. virescens on rice plants kept in an arena permeated with odour of 6, 12, or 25% neem oil was significantly reduced compared with that on control plants kept in an arena with paraffin oil or 1.66% aqueous Teepol solution. Reduced intake from phloem was associated with a significant increase in the insect's probing frequency and an increase in durations of salivation and xylem ingestion.

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Alterations du comportement sur riz de Nephotettix virescens, dues à l'odeur d'essence de neem (Azadirachta indica)

Résumé De nombreux essais en plein champ à l'I.R.R.I. ont révélé une réduction du T.V. (tungro virus) transmis par (Nephotettix virescens) sur des parcelles de riz sensible non-traitées, entourées de parcelles pulvérisées périodiquement avec de l'essence de graines de neem (Azadirachta indica).L'essence de neem a une forte odeur d'ail, perceptible à distance. Nous avons examiné l'effet de l'odeur d'essence de neem sur le comportement alimentaire deN. verescens, grâce à un dispositif électronique et un colorant spécifique de la lignine. Sur du riz maintenu dans une enceinte imprégnée d'odeur d'essence de neem, nous avons constaté une diminution de la consommation de phloème, une augmentation significative de la fréquence des piqûres et une augmentation de temps de salivation et d'absorption de xylème. Ces altérations du comportement alimentaire provoquées par l'odeur d'essence de neem peuvent expliquer la diminution de la fréquence du T.V. spécifique du phloème dans des champs de riz non-traités, entourés de champs pulvérisés avec de l'essence de neem.

     

Influence of pymetrozine on feeding behaviors of three rice planthoppers and a rice leafhopper using electrical penetration graphs

Pymetrozine reportedly inhibits feeding of plant sap-sucking insects, such as aphids and brown planthopper (Nilaparvata lugens (St?l)). By using electrical penetration graph (EPG), this study was conducted to investigate any differential effect of pymetrozine on the feeding behaviors of four major rice sap-sucking insect species, 1) N. lugens, 2) white-backed planthopper (Sogatella furcifera (Horváth)), 3) small brown planthopper (Laodelphax striatellus (Fallen)), and 4) green rice leafhopper (Nephotettix cincticeps Uhler). On pymetrozine-free TN1 rice plants, white-backed planthopper and small brown planthopper showed a significantly less activity in the phloem phases than brown planthopper or green rice leafhopper while green rice leafhopper engaged in relatively more xylem ingestion than brown planthopper, white-backed planthopper, and small brown planthopper. On the plants treated with 100 mg liter(-1) of pymetrozine, all four insect species showed significant increases, in total duration of nonprobing and significant decreases in the activities in phloem tissue, while all species showed similar feeding behavior during the pathway and xylem phases. This study revealed that, regardless of whether the insects on untreated plants spent more time feeding on phloem than xylem (brown planthopper) or more time on xylem than phloem (green rice leafhopper) or similar times on phloem and xylem (white-backed planthopper and small brown planthopper), their feeding behavior was disturbed by pymetrozine and exhibited similar patterns of sharp decline in activity in the phloem tissue and a significant increase the nonprobing.     

Fine mapping of Grh1, a major gene associated with antibiosis to green rice leafhopper in rice

Green rice leafhopper (GRH, Nephotettix cincticeps Uhler) is one of the insect pests that damage cultivated rice in East Asia. GRH also transmits viruses such as rice dwarf virus. The mortality of GRH nymphs is high in rice cultivar Shingwang, indicating that Shingwang is resistant to GRH. Genetic analyses were performed to map GRH resistance in Shingwang using F2 and F3 populations derived from a cross between a GRH-resistant near-isogenic line (NIL-IS60) from Shingwang and recurrent parent Ilpum. Resistance to GRH in Shingwang was found to be controlled by a single dominant gene (Grh1) mapped within an approximately 670-kb region between 8.10 and 8.77 Mb on the short arm of chromosome 5. Genotypes with three simple sequence repeat markers (RM18166, RM516, and RM18171) and one indel marker (Indel 15040) co-segregated with GRH resistance controlled by the Grh1 locus. A detailed map of the Grh1 locus will facilitate marker-assisted selection of resistance to GRH in rice breeding.     

A systems model and computer simulation of the green rice leafhopper populations in control programmes

A simulation model of the green rice leafhopper-spiders system was presented. The validity of this model for simulation purpose was tested by comparing the calculated values with observed ones (Fig. 4). The effectiveness of various control measures against leafhoppers was evaluated by computer simulations. The computer simulation demonstrated that the wrong use of selective insecticide, contrary to expectation, brought an increase in the pest density, i. e., that the egg densities of leafhoppers in the 2nd and 3rd generations are increased by the insecticidal application in February, while they are decreased by the July application (Table 2). To obtain satisfactory control by sterile-male release, 320,000 sterile-male per square kilometer should be released even in the combined use with insecticides (Fig. 5). The escape of leafhoppers from predation by spiders was demonstrated by the simulation. It is suggested that spiders are able to suppress the leafhopper populations at a low density when there is a very favorable balance between spiders and leafhoppers, and this condition may be realized by sophisticated use of selective insecticides (Fig. 6). Factors and/or processes which have to be involved in a more improved systems model are discussed.     

Predation ofCyrtorhinus lividipennis Reuter on eggs of the green leafhopper and brown planthopper in rice

The functional response of Cyrtorhinus lividipennis feeding on brown planthopper (BPH) and green leafhopper (GLH) eggs was found to be Holling's Type II. The Random Predator Equation fitted the data satisfactorily. Using Manly's preference index, α, both the male and female C. lividipennis were found to prefer BPH eggs. There was, however, no evidence of switching and the ratio of the respective searching efficiencies could account for the preference.     

Successional leafhopper assemblages: Pattern and process

Auchenorrhyncha (leafhopper) faunas of a series of experimental plots of different successional age were recorded by vacuum sampling. There were clear successional trends in the assemblages. The life-history strategy of the dominant species changed during succession; the mobile and multivoltine or bivoltine species, which overwinter as adults or nymphs, are replaced by less active, univoltine or bivoltine species which overwinter as eggs. Leafhopper assemblages clearly responded to the changes in plant species composition and vegetation structure during succession; however, disentangling the relative importance of the various factors requires further experimental manipulation.     

The outbreak mechanisms of the green rice leafhopper,Nephotettix cincticeps Uhler,in northern Japan

Population Ecology - Intraspecific regulatory processes keep the population ofNephotettix cincticeps stable at a low density in southern Japan. In northern Japan, however, the yearly population...     

High mortality caused by high dose of dsRNA in the green rice leafhopper Nephotettix cincticeps (Hemiptera: Cicadellidae)

RNA interference (RNAi) is a useful tool for gene functional studies in non-model insects and pest insects. Establishment of experimental conditions for RNAi, which differ from insect to insect, is important for evaluating the effect of dsRNA injection of relevant genes. When injecting dsRNA into the green rice leafhopper Nephotettix cincticeps (Uhler), high mortality was observed. Therefore, the adverse effects of injection of dsRNA on leafhopper development were examined to assess the suitable conditions for RNAi in this species. Injection manipulation by using a glass capillary did not affect leafhopper survival but delayed the molt of the corresponding instar. High mortality was observed when large amounts of dsRNA were administered. This adverse effect of dsRNA was examined in 2 genes, exogenous EGFP gene and endogenous peptidoglycan recognition protein gene (NcPGRP12). Injection of a high dose (60 ng/insect) caused high mortality in all stages tested: 4th instar, 5th instar, and female adult. A relatively low dose (6 ng/insect) did not cause high mortality, retaining a high potential for gene silencing. Since RNAi is highly effective in this species, the deleterious effect of large amounts of dsRNA could be avoided by administering a low dsRNA dose.     

Laccase-type phenoloxidase in salivary glands and watery saliva of the green rice leafhopper, Nephotettix cincticeps

The activity and composition of leafhopper saliva are important in interactions with the host rice plant, and it may play a physiological role in detoxifying toxic plant substances or ingesting sap. We have characterized diphenoloxidase in the salivary glands of Nephotettix cincticeps, its activity as a laccase, and its presence in the watery saliva with the objective of understanding its function in feeding on rice plants. Nonreducing SDS-PAGE of salivary gland homogenates with staining by the typical laccase substrate 2,2'-azino-bis (3-ethylbenzthiazoline-6-sulfonic acid) (ABTS), hydroquinone or syringaldazine revealed a band at a molecular mass of approximately 85 kDa at pH 5. A band also appeared at a molecular mass of approximately 200 kDa when the gels were treated with dopamine, L-3,4-dihydroxyphenylalanine (DOPA) or catechol at pH 7. The ABTS-oxidizing activity of the homogenates was drastically inhibited by N-hydroxyglycine, a specific inhibitor of laccase. However, the dopamine-oxidizing activity was not inhibited by N-hydroxyglycine, while it was inhibited by phenylthiourea (PTU). Thus, the salivary glands of N. cincticeps contain two types of phenoloxidases: a laccase (85 kDa) and a phenoloxidase (200 kDa). Laccase activity was detected in a holidic sucrose diet that was fed on for 16 h by two females, but only a trace of catechol oxidase activity was observed, suggesting that the laccase-type phenoloxidase was the predominant phenoloxidase secreted in watery saliva. The laccase exhibited an optimum pH of 4.75-5 in McIlvaine buffer and had a PI of 4.8. Enzyme activity was histochemically localized in V cells of the posterior lobe of the salivary glands. It remained at the same level throughout the adult stage from 2 days after eclosion. A possible function of N. cincticeps salivary laccase may be rapid oxidization of potentially toxic monolignols to nontoxic polymers during feeding on the rice plant. This is the first report proving that laccase occurs in the salivary glands of Hemiptera species and is secreted in the watery saliva.     

Life table analysis of the green rice leafhopper,Nephotettix virescens (distant) (Hemiptera: Cicadellidae), and efficient vector of rice tungro disease in asynchronous rice fields in Indonesia

Population dynamics of Nephotettix virescens was studied in 17 paddy fields transplanted at intervals of about 1 month in 1988–1990. The adult density was highest either in the immigrant or the 1st generation and sharply decreased to the 2nd generation. The survival rate of the 1st generation was lowest in the transition season when areal population density increased. Key factor analysis revealed that the nymphal and adult mortality of the 1st generation (kn) was the principal source of population fluctuations. No significant correaltion was found between kn and natural enemy density, natural enemy density/healthy egg density, or the precipitation during the nymphal period. On these bases adult emigration was suspected to be the key factor. Areal population build-up of N. virescens in the transition season was considered to occur as a result of increasing immigration to young stages of rice.     

Pathogenesis-related proteins in somatic hybrid rice induced by bacterial blight

Rice bacterial blight, caused by Xanthomonasoryzae pv. Oryzae (Xoo), is one of the most serious rice diseases worldwide. The bacterial blight resistance trait from Oryza meyeriana, a wild rice species, was introduced into an elite japonica rice cultivar using asymmetric somatic hybridization. This study was carried out with the intention of understanding the molecular mechanism of incompatible interaction between Xoo and the stable somatic hybrids by using proteomic analyses. Proteins were extracted from leaves at 24, 48, and 72 h after Xoo inoculation and separated by 2-DE. A total of 77 protein spots changed their intensities significantly (p<0.05) by more than 1.5-fold at least at one time point. Sixty-four protein spots were successfully identified by MS analysis. Among them, 51 were known to be involved in photosynthesis. Up-regulation of Rubisco large subunit (RcbL) small fragments and down-regulation of RcbL big fragments indicated that intact RcbL and RcbL big fragments degraded following Xoo attack, which was further confirmed by Western blot analysis. The differential expression of proteins related to signal transduction, antioxidant defense, photosynthesis, metabolism, and protein turnover during the Xoo infection, suggests the existence of a complex regulatory network in the somatic hybrid rice that increases resistance toward Xoo infection and damage.     

Iron deficiency in rice shoots: identification of novel induced genes using RDA and possible relation to leaf senescence

Rice plants are highly susceptible to Fe-deficiency. Under nutrient deprivation, plant cells undergo extensive metabolic changes for their continued survival. To provide further insight into the pathways induced during Fe-deficiency, rice seedlings were grown for 3, 6 and 9 days in the presence or absence of Fe. Using RDA (Representational Difference Analysis), sequences of 32 induced genes in rice shoots under Fe-deficiency were identified. About 30% of the sequences found have been previously reported as responsive to other abiotic and even biotic stresses. However, this is the first report that indicates their relation to Fe deprivation. Differential expression of selected genes was confirmed by semi-quantitative RT-PCR analysis. The identification of classical senescence-related sequences, such as lipase EC 3.1.1.-, ubiquitin-conjugating enzyme EC 6.3.2.19, beta-Glucosidase EC 3.2.1.21 and cysteine synthase EC 2.5.1.47, besides the higher accumulation of total soluble sugars prior to the decrease of total chlorophyll content in Fe-deficient leaves, indicate that sugar accumulation may be one of the factors leading to premature leaf senescence induced by Fe-deficiency.     

Absence of protein polymorphism attributable to insecticide-insensitivity of acetylcholinesterase in the green rice leafhopper, Nephotettix cincticeps

The cDNA sequence of acetylcholinesterase (AChE) from the green rice leafhopper, Nephotettix cincticeps, was amplified, based on conserved peptide sequences of AChEs. A 2.3 kb contiguous sequence, containing an ORF encoding an AChE precursor with 677 amino acid residues was obtained. The deduced protein sequence showed the most similarity to that of AChE in the Colorado potato beetle, having common features in the primary AChE structure. cDNA sequences of individual leafhoppers from an insecticide susceptible strain and the resistant strain Nakagawara, whose methylcarbamate-insensitive AChEs show 10(2) or more I(50) ratio for propoxur, were compared. No fixed inter-strain difference was identified in the protein sequence, though amino acid substitution polymorphism was found at one position in the susceptible strain. Insecticide-insensitivity of leafhopper AChE does not result from changes in the protein primary structure that is encoded by the AChE gene sequence isolated in this study.     

Artificial rearing of the rice green leafhopper, Nephotettix cincticeps, on a holidic diet

The rice green leafhopper, Nephotettix cincticeps, was reared from the first instar to the adult stage on a holidic diet. Adult emergence from the diet-reared nymphs was ca. 53.3%, similar to rice seedlingreared; nymphal period being 22.5 days at 25°. The diet-reared females did not lay eggs under artificial feeding conditions although they had normal eggs in their ovarioles and could oviposit on rice seedlings in a test tube. The diet containing 1.68% (w/v) amino acid mixture and 5% (w/v) sucrose was best for growth and development.

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Résumé Nephotettix cincticeps a été élevé des larves venant d'éclore à l'adulte sur un régime holidique. Le taux d'adulte obtenus (53,3%) est voisin de celui obtenu avec élevage sur pousses de riz. Le développement larvaire demande 22,5±0,4 jours à 25°. Les femelles élevées avec ce régime ne pondent pas d'ufs en alimentation artificielle bien qu'elles aient un nombre normal d'ufs dans les ovarioles et pondent sur pousses de riz dans un tube témoin. Un régime contenant 1,68% d'acides aminés mélangés et 5% de sucrose assure la meilleure croissance et le développement le plus rapide.