In the present paper, four cDNAs encoding the alpha and gamma subunits of elongation factor 1 (EF-1) were cloned and sequenced from Nilaparvata lugens, named NlEF-1α, NlEF-1γ, and its yeast-like symbiont (YLS), named YsEF-1α and YsEF-1γ, respectively. Comparisons with sequences from other species indicated a greater conservation for EF-1α than for EF-1γ. NlEF-1α has two identical copies. The deduced amino acid sequence homology of NlEF-1α and NlEF-1γ is 96 and 64%, respectively, compared with Homalodisca vitripennis and Locusta migratoria. The deduced amino acid sequence homology of YsEF-1α and YsEF-1γ is 96 and 74%, respectively, compared with Metarhizium anisopliae and Ophiocordyceps sinensis. Reverse transcription-quantitative polymerase chain reaction (RT–qPCR) analysis revealed that the expression level of NlEF-1α and NlEF-1γ mRNA in hemolymph, ovary, fat body and salivary glands were higher than the midgut and leg tissue. YsEF-1α and YsEF-1γ was highly expressed in fat body. The expression level of NlEF-1α was higher than that of NlEF-1γ. Through RNA interference (RNAi) of the two genes, the mortality of nymph reached 92.2% at the 11th day after treatment and the ovarian development was severely hindered. The RT–qPCR analysis verified the correlation between mortality, sterility and the down-regulation of the target genes. The expression and synthesis of vitellogenin (Vg) protein in insects injected with NlEF-1α and NlEF-1γ double-stranded RNA (dsRNA) was significantly lower than control groups. Attempts to knockdown the YsEF-1 genes in the YLS was unsuccessful. However, the phenotype of N. lugens injected with YsEF-1α dsRNA was the same as that injected with NlEF-1α dsRNA, possibly due to the high similarity (up to 71.9%) in the nucleotide sequences between NlEF-1α and YsEF-1α. We demonstrated that partial silencing of NlEF-1α and NlEF-1γ genes caused lethal and sterility effect on N. lugens. NlEF-1γ shares low identity with that of other insects and therefore it could be a potential target for RNAi-based pest management.

The brown planthopper Nilaparvata lugens is a serious phloem-feeding pest of rice in China. The current study focuses on a saccharopine dehydrogenase (SDH) that catalyzes the penultimate reaction in biosynthesis of the amino acid lysine (Lys), which plays a role in insect growth and carnitine production (as a substrate). The protein, provisionally designated as NlylsSDH [a SDH derived from yeast-like symbiont (YLS) in N. lugens], had a higher transcript level in abdomens, compared with heads, wings, legs and thoraces, which agrees with YLS distribution in N. lugens. Ingestion of Nlylssdh targeted double-stranded RNA (dsNlylssdh) for 5, 10 and 15 days decreased the mRNA abundance in the hoppers by 47, 70 and 31%, respectively, comparing with those ingesting normal or dsegfp diets. Nlylssdh knockdown slightly decreased the body weights, significantly delayed the development of females, and killed approximately 30% of the nymphs. Moreover, some surviving adults showed two apparent phenotypic defects: wing deformation and nymphal cuticles remained on tips of the legs and abdomens. The brachypterours/macropterours and sex ratios (female/male) of the adults on the dsRNA diet were lowered compared with the adults on diets without dsRNA. These results suggest that Nlylssdh encodes a functional SDH protein. The adverse effect of Nlylssdh knockdown on N. lugens implies the importance of Lys in hopper development. This study provides a proof of concept example that Nlylssdh could serve as a possible dsRNA-based pesticide for planthopper control.

The brown planthoppers (BPH) Nilaparvata lugens (Stål) and the white-backed planthoppers (WBPH) Sogatella furcifera (Horváth) annually migrate from tropical and subtropical regions to temperate regions in Asia, including Japan, Korea and northern China. To elucidate the genetic divergence based on geography of planthoppers and to estimate their migration route on the basis of molecular data, we analysed a part of their mitochondrial genome sequences. Sequences of cytochrome oxidase subunit I (cox1) – transfer RNA for Leu (trnL2) – cox2 were determined for 579 BPH (1,928 bp) and 464 WBPH (1,927 bp) individuals collected from 31 and 25 locations, respectively, in East and Southeast Asia. Thirty and 20 mitochondrial haplotypes were detected for BPH and WBPH, respectively. Single populations of both planthoppers included multiple haplotypes, and many haplotypes were shared in some populations and areas. The most frequently detected haplotypes accounted for approximately 50% of all BPH and WBPH individuals. To evaluate gene flow among planthoppers in different regions in Asia, pairwise fixation index (Fst) values were calculated. For BPH, high Fst values (0.580–0.926) were shown between planthoppers in Papua New Guinea (PNG) and the other areas and moderate Fst values (0.176–0.362) were observed between those in southern Philippines and other areas. For WBPH, the Fst value was the highest between Taiwan and southern Vietnam (0.236), and low among the other areas. AMOVA indicated no genetic structure among eight areas, excluding southern Philippines and PNG, for BPH, and among ten areas for WBPH. These data indicate that both planthoppers do not show much differentiation of local populations and/or have genetically intermixed Asian populations. These data also indicate that it may be difficult to distinguish regional planthopper populations on the basis of differences in mitochondrial sequences.

Brown Planthopper (BPH, Nilaparvata lugens (Stål)) is one of the most serious pests of rice in both temperate and tropical regions of East and South Asia and has become especially problematic over the past few years. In order to analyze the effect of the change of rice cropping system on the population dynamics of BPH, field surveys of the occurrence and distribution of BPH were performed and other relevant data, including light trap data and ovary dissection data were collected in nearly 40 Chinese counties encompassing six provinces (or municipalities), including Hainan, Guangxi, Anhui, Shanghai, Fujian and Guangdong from April to October in 2007.

The mixed planting areas of single- and double-cropping rice in China include Hubei, South and Central Anhui, North Hunan, and North Jiangxi. In these areas, double-cropping rice has now been greatly reduced and single-cropping rice has been rapidly increasing since 1997. The surveys revealed that when the immigration peak of BPH occurred in June and July, the single-cropping rice was at the tillering to booting stage and fit for BPH, but early rice had already matured and most of late rice had not yet been transplanted. BPH immigrants from southern rice areas prefer to inhabit and breed in single-cropping rice paddies. Moreover, farming activities between early rice and late rice interrupted the continuous growth of BPH populations in double-cropping rice paddies. As a result, in comparison with data collected 30 years ago, the spatiotemporal dynamics and migration patterns of BPH have dramatically changed in the lower-middle reaches of the Yangtze River. In the mixed planting areas, due to their high suitability, the BPH population in single-cropping rice grew so quickly that it caused serious local damage and there was mass emigration of macropterous progeny to the Yangtze River Delta in late August and early September.

Global warming may also affect BPH populations, where results suggest steadily warmer autumns have occurred from the 1990s on, with such conditions gradually the norm. The combination of ‘cooler summer’ and ‘warmer autumn’ are conditions known to promote outbreaks of BPH in the lower-middle reaches of the Yangtze River. Immigrant BPH arrivals in late August and September now cause serious damage to late-maturing mid-season rice and late rice in the lower-middle reaches of the Yangtze River.

The tropical fire ant Solenopsis geminata (Fabricius) often nests very abundantly in the earthen banks (bunds) around irrigated rice fields in the tropics. Where some farmers habitually drain fields to the mud for about 3–4 days, the ants can quickly spread up to about 20 m into the fields where they collect food, including pest prey such as the eggs and young of the apple snail Pomacea caniculata (Lamarck) and insects such as lepidopterous larvae and hoppers, notably Nilaparvata lugens (Stäl) the brown planthopper (Bph) and green leafhoppers Nephotettix spp. Even in drained fields, the activity of S. geminata is restricted by rainfall in the wet season. The relatively few ant workers that forage characteristically into drained fields and on to the transplanted clumps of rice plants (hills) kill the normally few immigrant Bph adults but are initially slower acting than other species of the natural enemy complex. However, larger populations of Bph are fiercely attacked and effectively controlled by rapidly recruited ant workers; whereas, in the absence of the ant, the other natural enemies are inadequate. In normal circumstances, there is no ant recruitment in response to initially small populations of immigrant Bph and no evidence of incompatibility between ant foragers and other natural enemies such as spiders. However, when many ants are quickly and aggressively recruited to attack large populations of Bph, they temporarily displace some spiders from infested hills. It is concluded that, in suitable weather conditions and even when insecticides kill natural enemies within the rice field, periodic drainage that enables S. geminata to join the predator complex is valuable for ant-based control of pests such as snails and Lepidoptera, and especially against relatively large populations of Bph. Drainage practices to benefit ants are fully compatible with recent research, which shows that periodic drainage combats problems of ‘yield decline’ in intensively irrigated tropical rice and is also needed in South East Asia to make better use of seriously declining water supplies for irrigation.

Nilaparvata lugens Stål, the Brown Planthopper (BPH), is a major pest in rice. The lycosid Pardosa pseudoannulata (Bösenberg & Strand) and the linyphiid Atypena formosana (Oi) are the early dominant predators in irrigated rice. Later, predatory bugs, including the mirid Cyrtorhinus lividipennis Reuter, become dominant. In unsprayed rice, BPH numbers normally remain low. While P. pseudoannulata is known to be a key natural enemy of BPH, the contribution to BPH reduction by the smaller A. formosana is less well known. Due to the size difference, A. formosana may also be an intraguild prey of P. pseudoannulata. To investigate predation on BPH by the two spider species alone, together and in combination with C. lividipennis, two cage experiments were conducted, the first with adult spiders, the second with immature/unmated spiders. In the latter, spiders were introduced with a delay to allow a better establishment of C. lividipennis. In both experiments, BPH numbers were lowest in treatments with more than one predator species present. Intermediate BPH reduction was obtained with P. pseudoannulata followed by A. formosana and C. lividipennis. While P. pseudoannulata can prey upon large nymphs and adult BPH, A. formosana favour early instar BPH, leading to a more even control across instars in treatments with both spider species. Pardosa pseudoannulata numbers increased more with A. formosana or A. formosana and C. lividipennis present. In contrast, P. pseudoannulata reduced A. formosana and C. lividipennis numbers. The presence of C. lividipennis apparently lessened intraguild predation on A. formosana and may be an intraguild prey of A. formosana. Competition for prey may have added to the antagonistic interactions found. From a biological control perspective, the presence of both spiders in early rice is an advantage for the biological control of BPH in rice. In early rice, results show that A. formosana is an important predator of BPH and, like C. lividipennis, can complement control by P. pseudoannulata and serve as intraguild prey for the latter species.

The brown planthopper Nilaparvata lugens Stål is one of the major insect pests of rice Oryza sativa L. The host resistance exhibits profound effects on growth, development and propagation of N. lugens. To investigate the molecular response of N. lugens to host resistance, a cDNA-amplified fragment length polymorphism (cDNA-AFLP) technique was employed to identify the differentially expressed genes in the nymphs feeding on three rice varieties. Of the 2800 cDNA bands analysed, 54 were up-regulated and seven down-regulated qualitatively in N. lugens when the ingestion sources were changed from susceptible rice plants to resistant ones. Sequence analysis of the differential transcript-derived fragments showed that the genes involved in signalling, stress response, gene expression regulation, detoxification and metabolism were regulated by host resistance. Four of the transcript-derived fragments corresponding to genes encoding for a putative B subunit of phosphatase PP2A, a nemo kinase, a cytochrome P450 monooxygenase and a prolyl endopeptidase were further characterized in detail. Northern blot analysis confirmed that the expression of the four genes was enhanced in N. lugens feeding on resistant rice plants. The roles of these genes in the defensive response of N. lugens to host plant resistance were discussed.

Random amplified polymorphic DNA (RAPD) amplified by the polymerase chain reaction (PCR) was used to estimate the relatedness of three biotypes of brown planthopper, Nilaparvata lugens (Stål), from the Philippines. Each individual brown planthopper had a unique set of RAPD bands, but no bands were diagnostic for any one biotype. Brown planthopper types 1, 2 and 3 were genetically homogeneous. This supports results of other studies which conclude that brown planthopper biotypes are not sub-specific categories, but merely represent individual variation for virulence to rice cultivars.

Buprofezin treatment of brown planthopper, Nilaparvata lugens Stal. at sub-lethal doses resulted in the survival of individuals with varying reproductive potentials. From this reproductive potential of different stages, population build up of second generation was calculated. Logistic function was found to fit more closely the data on population growth at different concentrations. It could thus serve as a mathematical model for predicting the second generation population build up in a buprofezin treated population.

Pure line populations of brown planthopper, Nilaparvata lugens (Ståi), are needed for identifying new genetic sources of resistance. Among the known N. lugens biotypes in the Philippines, Biotype 1 can infest rice cultivare which lack genes for resistance. Biotype 2 can damage cultivars carrying Bph1 resistance gene, in addition to those that are susceptible to Biotype 1. Biotype 3 can infest rice cultivars carrying bph2 resistance gene, in addition to those susceptible to Biotype 1. In the greenhouse Biotypes 1, 2, and 3 are maintained on TN1 (no resistance gene), Mudgo (Bph1 gene) and ASD7 (bph2 gene) rice plants, respectively. Because Biotype 1 population is reared on TN1 plants, susceptible to all the three biotypes, accidental infestations of Biotype 1, Biotype 2 and Biotype 3 cannot be ruled out.

N. lugens biotypes were separated from each other by their differential ingestion and assimilation of food on different susceptible and resistant rice cultivars. A homogeneous population of Biotype 1 of N. lugens was produced from single pairs of males and females selected on the basis of their low food intake on rice cultivars Mudgo and ASD7. The purity of such Biotype 1 population was further demonstrated for three generations based on differential nymphal growth and development and adult longevity and fecundity on resistant Mudgo and ASD7 and on the susceptible TN1 rice cultivars. The technique can also be useful in identifying biotypes of other sucking insect pests.

In order to evaluate the migratory potential of the brown planthopper, Nilaparvata lugens, under as near to normal environmental conditions as possible, adult macropterous insects were captured from the field whilst in the process of flight initiation at the times of their natural take-off peaks. The insects were then flight-mounted and flown in the field. Average flight durations following the dusk take-off were 4 hr 55 min for males and 6 hr 17 min for females. However, many insects flew throughout the night and there was a high frequency of flight cessation at dawn. Following a dawn take-off both sexes flew for much shorter periods, an average of 2 hr 00 min for males and 2 hr 04 min for females. Female insects were reluctant to initiate a second major flight. The role of mating and ovary development in female flight willingness is discussed.

Resistant rice varieties have proved to be highly effective against the brown planthopper Nilaparvata lugens (Stål), but their long-term stability is threatened because of the evolution of prolific biotypes. At present, identification of biotypes is based principally on the differential reactions of host rice varieties to pest in greenhouse screenings and in field plantings of test nurseries at many locations. So far, three N. lugens biotypes that attack different rice varieties have been identified in the Philippines. Biotype 1 can survive on and damage only those varieties that do not carry any genes for resistance. Biotype 2 can thrive on varieties carrying Bph 1 resistance gene and on those susceptible to biotype 1. Biotype 3 can infest and destroy varieties having bph 2 resistance gene and those susceptible to biotype 1. However, none of these biotypes is capable of damaging varieties with Bph 3 and bph 4 genes for resistance, and the variety Ptb 33 carrying two unidentified genes is resistant to almost all known biotypes. Recently, a population of N. lugens was found thriving on the common weed grass Leersia hexandra Swartz on the IRRI farm and in some locations in Luzon, Philippines. This biotype does not survive on rice.

Several varieties which are resistant in the Philippines are susceptible in India and Sri Lanka because the South Asian biotypes of N. lugens are more virulent than the Southeast Asian biotypes. To monitor the pest biotypes in different geographical regions and to identify new sources of resistance, an International Rice Brown Planthopper Nursery (IRBPHN) has been established in many cooperating Asian countries.

Biotypes can also be differentiated by studying their behavioural and physiological responses to plants of known genotypes. Distinctive differences can be observed in insect feeding, growth, longevity, fecundity, oviposition and population increase on differential varieties and relative susceptibility or tolerance for hosts' allelochemics. Recent investigations of the existence of subtle morphological, cytological, and electrophoretic differences among N. lugens biotypes and results of hybridization experiments strongly complement other methods of biotype identification.

The evolution of biotypes is a complex process that is governed by the interactions of the genetic and other biological characteristics of the pest populations and the genetic makeup of cultivated varieties. While the strategy of sequential release of varieties with major resistance genes has been fairly successful, the monogenic nature of those varieties makes them vulnerable to the development of virulent pest biotypes. Therefore, present breeding endeavors envisage use of both major and minor resistance genes for effective pest suppression.

Nilaparvata lugens (Stål) biotype 1, 2 and 3 populations maintained as stock cultures for several years showed distinct morphological segregation based on multiple discriminant analysis using Wilk's Lambda method. Morphological characters of the rostrum, legs and antennae of both brachypterous and macropterous males and females provided a significant degree of certainty in classifying the three biotypes. The predicted biotype composition of the brown planthopper populations formed separate clusters.

Changes in the physiological status of the host rice plant had a profound effect on wing morphism in the brown planthopper, Nilaparvata lugens (Stål). Significant increases in macroptery in the hopper progenies reared on senescent or hopperburned hosts occurred mainly because of a general decline in the host's nutritional status and allelochemic factors. Starvation of young nymphs on nutritionally-depleted hosts probably inhibits the secretion of juvenile hormone by the corpus allatum, suppressing brachyptery and expressing macroptery. Also, any juvenilizing effect of the host's allelochemics would tend to dissipate with the host's senescence. Therefore, the proportion of macroptery in the hopper progeny developing on the rice host plant is determined by the extent of depletion of the host's nutritional allelochemic factors. The observed significant increase in macroptery at a density of 50 or more hoppers per plant may have also been mediated through the host plant. Brown planthopper progenies of macropterous and brachypterous Biotypes 1, 2, 3 and the field population differed in their expression of macroptery probably because of their inherent differences in sensitivity to macropteryinducing stresses.