Small Brown Planthopper Research Articles

Roles of LsCYP4DE1 in wheat adaptation and ethiprole tolerance in Laodelphax striatellus

The cytochrome P450 monooxygenase (P450) gene family has an important role in detoxifying host plant allelochemicals and pesticides. In this study, we screened differentially expressed genes of the small brown planthopper (Laodelphax striatellus, SBPHs) that were reared for more than ten generations on rice and wheat plants, and found that only a few P450s were associated with host adaptation. LsCYP4DE1, whose expression was 9.5-fold higher in the wheat-adapted SBPH (wSBPH) than in the rice-adapted SBPH (rSBPH), appeared to have an important role in the colonization of wheat plants. Knocking down the expression of LsCYP4DE1 led to increased mortality, as well as decreased performance of SBPHs reared on wheat. However, no significant difference was found in dsLsCYP4DE1-treated SBPHs on rice plants. In addition, LsCYP4DE1 was potentially associated with pesticide tolerance, and suppression of its expression led to increased sensitivity to the pesticide ethiprole. Our results revealed potential roles of LsCYP4DE1 in wheat adaptation and ethiprole tolerance, and provide useful information for pest management.

Olfactory co-receptor Orco stimulated by Rice stripe virus is essential for host seeking behavior in small brown planthopper.

Laodelphax striatellus, the small brown planthopper (SBPH), is an economically important pest, besides sucking damage, which transmits rice viruses to cause severe damages to rice. In the process of virus transmission, the host orientation behavior of insect is mainly driven by olfaction. In this context, the molecular basis of olfaction in SBPH is of particular interest. Here, we identified the gene that encodes olfactory receptor co-receptor (Orco) and analyzed its expression profiles in Rice stripe virus (RSV)-infected and RSV-free SBPH. It was found that LstrOrco shared high identity with other Orcos from different order insects. LstrOrco was mainly expressed in the head of SBPH, and its expression was significantly stimulated by RSV-infection. The behavioral bioassay revealed that viruliferous SBPH might have a stronger olfactory and seeking ability for rice than RSV-free insect. After silencing of LstrOrco expression, the olfaction and seeking behavior of nymphs for rice seedlings was significantly inhibited, mainly in the increase of the 'no response' percent and the prolongation of the response time. These results suggested that Orco played an important role in olfactory signaling and seeking behavior of SBPH, which provided a basic for future development of olfactory-based agriculture pest management strategies. © 2018 Society of Chemical Industry.

Fiprole insecticide resistance of Laodelphax striatellus: electrophysiological and molecular docking characterization of A2'N RDL GABA receptors.

Phenylpyrazole (fiprole) insecticides, including ethiprole, fipronil and flufiprole with excellent activity on rice planthoppers, are very important in Asia but resistance has developed after decades of use. The molecular mechanism of fipronil- but not ethiprole-resistance has been previously studied in rice planthoppers. In our laboratory, a small brown planthopper Laodelphax striatellus strain with ethiprole-resistance was cultured and the molecular mechanisms of ethiprole resistance and of cross-resistance among fiprole insecticides were investigated. Ethiprole-resistant L. striatellus has >5000-fold resistance compared to the susceptible strain, and exhibits around 200-fold cross-resistance with fipronil and flufiprole. RDL genes were isolated from susceptible and ethiprole-resistant L. striatellus and expressed in Xenopus oocytes. Electrophysiological studies showed fiprole insecticides inhibited γ-aminobutyric acid (GABA)-induced current with IC50 = 0.1-1.4 μM to LsRDL-S homomers. In LsRDL-R with A2'N mutation, only 1-13% inhibition was observed on treatment with 10 μM ethiprole, fipronil or flufiprole. Homology models indicate A2'N mutation allows crosslinking hydrogen bonding between Asn sidechains at the 2' position around the channel pore, blocking insecticides from interacting near this position. In contrast, insecticides showed favorable binding near A2' in wild-type L. striatellus. Cross-resistance is increasing for fiprole insecticides in L. striatellus and management strategies are necessary to minimize resistance. © 2018 Society of Chemical Industry.

Transmission Characteristics of Barley Yellow Striate Mosaic Virus in Its Planthopper Vector Laodelphax striatellus.

The most economically important plant viruses are specifically transmitted by phytophagous insects that significantly affect viral epidemiology. Barley yellow striate mosaic virus (BYSMV), a member of the genus Cytorhabdovirus, is transmitted by the small brown planthopper (SBPH, Laodelphax striatellus) in a persistent-propagative manner. However, the infection route of BYSMV in SBPHs is poorly understood. In this study, immunofluorescence confocal laser scanning microscopy (iCLSM) was performed to investigate the route of BYSMV in SBPHs. We unexpectedly found that BYSMV initially infected the hindgut epithelium of SBPHs, instead of the midgut epithelium initially infected by other persistent-propagative viruses. Subsequently, BYSMV disseminated to the hindgut visceral muscles and spread to other parts of alimentary canals, hemolymph, and salivary glands. Comparative analysis of gene expression on viral mRNAs and the BYSMV nucleoprotein by using different molecular detection and immunohistochemistry further demonstrated that BYSMV initially infected and replicated in the hindgut epithelial cells of SBPHs. Collectively, our study provides the first insight into that hindgut is initial infection site of BYSMV that represents a new dissemination route of persistent-propagative viruses.

NS3 Protein from Rice stripe virus affects the expression of endogenous genes in Nicotiana benthamiana

BackgroundRice stripe virus (RSV) belongs to the genus Tenuivirus. It is transmitted by small brown planthoppers in a persistent and circulative-propagative manner and causes rice stripe disease (RSD). The NS3 protein of RSV, encoded by the viral strand of RNA3, is a viral suppressor of RNA silencing (VSR). NS3 plays a significant role in viral infection, and NS3-transgenic plants manifest resistance to the virus.MethodsThe stability and availability of NS3 produced by transgenic Nicotiana benthamiana was investigated by northern blot analysis. The accumulation of virus was detected by western blot analysis. Transcriptome sequencing was used to identify differentially expressed genes (DEGs) in NS3-transgenic N. benthamiana.ResultsWhen the host plants were inoculated with RSV, symptoms and viral accumulation in NS3-transgenic N. benthamiana were reduced compared with the wild type. Transcriptome analysis identified 2533 differentially expressed genes (DEGs) in the NS3-transgenic N. benthamiana, including 597 upregulated genes and 1936 downregulated genes. These DEGs were classified into three Gene Ontology (GO) categories and were associated with 43 GO terms. KEGG pathway analysis revealed that these DEGs were involved in pathways associated with ribosomes (ko03010), photosynthesis (ko00195), photosynthesis-antenna proteins (ko00196), and carbon metabolism (ko01200). More than 70 DEGs were in these four pathways. Twelve DEGs were selected for RT-qPCR verification and subsequent analysis. The results showed that NS3 induced host resistance by affecting host gene expression.ConclusionNS3, which plays dual roles in the process of infection, may act as a VSR during RSV infection, and enable viral resistance in transgenic host plants. NS3 from RSV affects the expression of genes associated with ribosomes, photosynthesis, and carbon metabolism in N. benthamiana. This study enhances our understanding of the interactions between VSRs and host plants.

Molecular characterization and evolution of a chemosensory receptor gene family in three notorious rice planthoppers, Nilaparvata lugens, Sogatella furcifera and Laodelphax striatellus, based on genome and transcriptome analyses.

The white-backed planthopper (WBPH) Sogatella furcifera, the brown planthopper (BPH) Nilaparvata lugens, and the small brown planthopper (SBPH) Laodelphax striatellus (Hemiptera: Delphacidae) are rice pests that damage rice plants by sap-sucking and by transmitting viruses. Host-seeking behavior involves chemosensory receptor genes that include odorant receptors (ORs), ionotropic receptors (IRs) and gustatory receptors (GRs). We used genome and transcriptome data to identify 141 ORs, 28 GRs and 25 IRs in BPH; 135 ORs, 18 GRs and 16 IRs in WBPH; and 37 ORs, 14 GRs and 6 IRs in SBPH. A phylogenetic analysis identified several specific OR clades of rice planthoppers, the results indicating that these OR members might be used to respond to specific host volatiles. OR co-receptor (Orco) is the most conserved and essential OR gene among these species and RNA interference (RNAi) can decrease their mRNA expression level to <50%. RNAi knockdown rice planthoppers were anosmia and were unable to seek or locate rice plants in behavioral tests. The results demonstrate the importance of the planthopper Orco genes in locating rice plants. This information may aid in the development of RNAi-based transgenic rice and other pest management technologies. © 2018 Society of Chemical Industry.

Long chain fatty acid coenzyme A ligase (FACL) regulates triazophos-induced stimulation of reproduction in the small brown planthopper (SBPH), Laodelphax striatellus (Fallen)

The small brown planthopper (SBPH), Laodelphax striatellus (Fallen) is a major pest insect of rice, wheat, and maize in China and other countries. SBPH not only damage rice plants through sucking plant sap, but also transmits rice virus diseases, for example, striped virus disease (RSV), black streaked dwarf, and maize rough disease virus. Therefore, understanding of pesticide-induced stimulation of reproduction in SBPH is of great significance for the pest management. Our previous study discovered that triazophos (TZP) increased reproduction of SBPH. But the molecular mechanisms are unclear. Here, by using proteomic analysis, we screened and cloned the gene of long chain fatty acid coenzyme A ligase (FACL), and silenced FACL to examine influences of TZP on reproduction and glycerin content in SBPH females. In TZP-treated females vs control females, there were 41 differential proteins in 18 pathways related to reproduction, of which 8 were up-regulated and 33 were down-regulated. TZP + dsFACL eliminated TZP-induced stimulation of reproduction of SBPH females (↓about 73.92%) and decreased glycerin content and body weight (↓about 19.93% and 13.62%). TZP + dsFACL treatment led to reduced expression of FACL (↓about 61.88%). FACL is a key gene of TZP-induced increase of reproduction of SBPH.

Differential Expression of P450 Genes and nAChR Subunits Associated With Imidacloprid Resistance in Laodelphax striatellus (Hemiptera: Delphacidae).

Imidacloprid is a key insecticide used for controlling sucking insect pests, including the small brown planthopper (Laodelphax striatellus, Fallén) (Hemiptera: Delphacidae), an important agricultural pest of rice. A strain of L. striatellus (YN-ILR) developed 21-fold resistance when selected with imidacloprid on a susceptible YN strain. An in vitro study on piperonyl butoxide synergism indicated that enhanced detoxification mediated by cytochrome P450s contributed to imidacloprid resistance to some extent, and multiple P450 genes showed altered expression in the imidacloprid-resistant YN-ILR strain compared with the susceptible YN strain (CYP425B1-CYP6BD10 had 1.51- to 11.45-fold higher expression, CYP4CE2-CYP4DD1V2 had 0.12- to 0.57-fold lower expression). While there were no mutations in target nicotinic acetylcholine receptor (nAChR) genes, subunits of Lsα1, Lsβ1, and Lsβ3 in the YN-ILR strain showed 3.86-, 4.39-, and 2.59-fold higher expression and Lsa8 displayed 0.38-fold lower expression than the YN strain. Moreover, 21-fold moderate imidacloprid resistance in individuals of L. striatellus did not produce a fitness cost. The findings suggest that L. striatellus has the capacity to develop resistance to imidacloprid through P450 detoxification and potential target nAChR expression changes, and moderate imidacloprid resistance was not associated with a fitness cost.

Comparative proteomics analysis of maize (Zea mays) leaves infected by small brown planthopper (Laodelphax striatellus)

Maize rough dwarf disease (MRDD) is a viral disease caused by brown planthopper infestation, and leads to great yield loss, especially in China. Comparative proteomics was performed using maize inbred line Zheng 58 and LN 287. MRDD pathogen was detected as rice black-streaked dwarf virus (RBSDV) by quantitative real time PCR (qRT-PCR) in Shandong Province, China. The modified trichloroacetic acid (TCA)/acetone method was used for soluble protein extraction from leaves. Two-dimensional electrophoresis (2-DE) analysis was performed on 24-cm long, pH 4–7 linear immobilized pH gradient (IPG) strips, and gels were stained with silver and coomassie brilliant blue. We identified 944 proteins expressed in RBSDV infected maize leaves by proteomics approaches. Among these, 44 protein spots that revealed a 1.5-fold difference in intensity were identified by mass spectrometry between mock-inoculated and RBSDV infected samples. Among these, 17 and 26 spots were up-regulated, and 27 and 18 spots were down-regulated in the virus infected samples of Zheng 58 and LN 287, respectively. Differential protein spots were analyzed by mass spectrometry identification, which could be divided into six categories. Furthermore, the expression of stress-related proteins was detected and confirmed by qRT-PCR. This study lays the foundation for further investigations, enabling the enhancement of MRDD resistance in maize.

Variations in Endosymbiont Infection Between Buprofezin-Resistant and Susceptible Strains of Laodelphax striatellus (Fallén).

The endosymbionts Wolbachia and Rickettsia have been shown to be correlated with the insecticide resistance of mosquito and whitefly. The small brown planthopper (SBPH), Laodelphax striatellus, harbours many species of endosymbionts, and has developed a high resistance to buprofezin in China. In this study, we examined the species and the infection incidences of endosymbionts in a buprofezin-resistant (BR) strain, a buprofezin-susceptible (BS) strain, and the BR strain after exposure to buprofezin, and we also investigated the change in buprofezin susceptibility after removal of Wolbachia from the BR strain. The results showed that Wolbachia infection incidences were 100% in both the BR and BS strains, but the Wolbachia density in the BR strain was significantly higher than that in the BS strain. There were no significant differences in Arsenophonus infection incidence between the two strains. However, the infection incidence of Serratia and double infection incidence of Serratia + Wolbachia in the BR strain were significantly higher than that in the BS strain. After the BR strain was exposed to 1200mg/L buprofezin, the infection incidence of Arsenophonus in the surviving individuals increased, and the infection rate of Serratia did not differ, but the double infection incidence of Serratia + Wolbachia decreased. And when a Wolbachia-infected line originating from the BR strain was cleared of Wolbachia, its susceptibility to buprofezin increased. The results suggest that Serratia and Wolbachia infection might improve the buprofezin resistance of SBPH.

Two Fungicides Alter Reproduction of the Small Brown Planthopper Laodelphax striatellus by Influencing Gene and Protein Expression

Aside from their intended actions, fungicides can drive pest insect outbreaks due to virtually continuous use and pest evolution. Small brown planthopper (SBPH), Laodelphax striatellus, outbreaks occurred recently in many provinces in China, with devastating rice losses. Because exposure to the fungicide jinggangmycin (JGM) increased reproduction of the brown plant hopper, Nilaparvata lugens, via its influence on fatty acid synthase, we posed the hypothesis that JGM and carbendazim (CBM) influence SBPH reproduction via their influence on enzymes involved in other aspects of lipid metabolism. Exposure to the fungicide CBM stimulated SBPH reproduction (egg-laying up by 78%) and to another fungicide, JGM, led to decreased egg-laying (down by 47.3%). These inverse effects are mediated by down-regulated expression of l-3-hydroxyacyl-coenzyme A dehydrogenase (LCHAD) in JGM-treated females and up-regulated expression of hydroxysteroid dehydrogenase-like protein 2-like (HSD) in CBM-treated females. RNAi knockdown of, separately, LCHAD and HSD led to reduced egg-laying (down by 52% for dsLCHAD and by 73% for dsHSD). dsLCHAD, dsHSD, and JGM treatments also led to severely reduced ovarian development in experimental SBPH, with shorted and thinned valvula and lack of egg cells in ovaries. Valvula of CBM-treated females enlarged, with banana-shaped eggs in ovaries. These data strongly support our hypothesis.

Fluxametamide: A novel isoxazoline insecticide that acts via distinctive antagonism of insect ligand-gated chloride channels

Fluxametamide is a novel wide-spectrum insecticide that was discovered and synthesized by Nissan Chemical Industries, Ltd. To identify the mode of action of fluxametamide, we first performed [3H]4′-ethynyl-4-n-propylbicycloorthobenzoate (EBOB) binding assays. Fluxametamide potently inhibited the specific binding of [3H]EBOB to housefly-head membranes, suggesting that fluxametamide affects insect γ-aminobutyric acid (GABA)-gated chloride channels (GABACls). Next, the antagonism of housefly GABACls and glutamate-gated chloride channels (GluCls) was examined using the two-electrode voltage clamp (TEVC) method. Fluxametamide inhibited agonist responses in both ion channels expressed in Xenopus oocytes in the nanomolar range, indicating that this insecticide is a ligand-gated chloride channel (LGCC) antagonist. The insecticidal and LGCC antagonist potencies of fluxametamide against fipronil-susceptible and fipronil-resistant strains of small brown planthoppers and two-spotted spider mites, which are insensitive to fipronil, were evaluated. Fluxametamide exhibited similar levels of both activities in these fipronil-susceptible and fipronil-resistant arthropod pests. These data indicate that fluxametamide exerts distinctive antagonism of arthropod GABACls by binding to a site different from those for existing antagonists. In contrast to its profound actions on the arthropod LGCCs, the antagonistic activity of fluxametamide against rat GABACls and human glycine-gated chloride channels was nearly insignificant, suggesting that fluxametamide has high target-site selectivity for arthropods over mammals. Overall, fluxametamide is a new type of LGCC antagonist insecticide with excellent safety for mammals at the target-site level.

Ribosomal protein L18 is an essential factor that promote rice stripe virus accumulation in small brown planthopper

Rice stripe virus (RSV) transmitted by the vector, small brown planthopper (SBPH), can cause a severe rice disease. The nucleocapsid (N) protein is the major component of RSV ribonucleoprotein particles (RNPs), and it plays important roles in viral persistent-propagative transmission by SBPH. To gain further insights into the vector components enabling RSV transmission, a GAL4-based yeast two-hybrid system was utilized to find unknown vector factors that interact with the N protein. Thirteen different proteins were identified as factors that interact with the N protein. The interaction between 60S ribosomal protein L18 (RPL18) and the N protein was further studied. Although the expression of RPL18 was not altered in insects during RSV infection, RPL18 was validated to bind directly to RSV RNPs and interact with RSV N protein. Knockdown of RPL18 dramatically reduced viral RNA and protein levels, especially viral protein expression, indicating a requirement for RPL18 in RSV translation and replication. Our results provide evidence that RPL18 is a critical factor required for RSV accumulation in SBPH, which suggests that the vector factor RPL18 may be as a potential target to develop for controlling the transmission of rice virus.

Effects of agricultural practices and fine-scale landscape factors on spiders and a pest insect in Japanese rice paddy ecosystems

We examined the effects of environmentally friendly (EF) farming and landscape factors on the abundances of major spider guilds and a rice pest, the small brown planthopper (SBPH), Laodelphax striatellus (Fallen), in a Japanese rice paddy ecosystem. The abundances of all spider guilds increased by EF farming, whereas different spider guilds showed contrasting responses to the size of the forest within 200 m of the fields. The abundances of ground spiders and horizontal web weavers increased with increasing forest area, unlike small ground spiders, whose abundance decreased. The abundances of SBPH nymphs and adults decreased by EF farming but responded differently to landscape: nymphs decreased with increasing forest area, but adults did not. Further analysis indicated a potential negative relationship between web-weaving spiders and SBPH nymphs. Our findings suggest that the size of the forest adjacent to fields is an important determinant of spider guild composition and pest abundance.

Cry64Ba and Cry64Ca, Two ETX/MTX2-Type Bacillus thuringiensis Insecticidal Proteins Active against Hemipteran Pests.

Genetically modified crops that express insecticidal Bacillus thuringiensis (Bt) proteins have become a primary approach for control of lepidopteran (moth) and coleopteran (beetle) pests that feed by chewing the plants. However, the sap-sucking insects (Hemiptera) are not particularly susceptible to Bt toxins. In this study, we describe two Cry toxins (Cry64Ba and Cry64Ca) from Bt strain 1012 that showed toxicity against two important hemipteran rice pests, Laodelphax striatellus and Sogatella furcifera Both of these proteins contain an ETX/MTX2 domain and share common sequence features with the β-pore-forming toxins. Coexpression of cry64Ba and cry64Ca genes in the acrystalliferous Bt strain HD73- resulted in high insecticidal activity against both hemipteran pests. No toxicity was observed on other pests such as Ostrinia furnacalis, Plutella xylostella, or Colaphellus bowringi Also, no hemolytic activity or toxicity against cancer cells was detected. Binding assays showed specific binding of the Cry64Ba/Cry64Ca toxin complex to brush border membrane vesicles isolated from L. striatellus Cry64Ba and Cry64Ca are Bt Cry toxins highly effective against hemipteran pests and could provide a novel strategy for the environmentally friendly biological control of rice planthoppers in transgenic plants.IMPORTANCE In Asia, rice is an important staple food, whose production is threatened by rice planthoppers. To date, no effective Bacillus thuringiensis (Bt) protein has been shown to have activity against rice planthoppers. We cloned two Bt toxin genes from Bt strain 1012 that showed toxicity against small brown planthoppers (Laodelphax striatellus) and white-backed planthoppers (Sogatella furcifera). To our knowledge, the proteins encoded by the cry64Ba and cry64Ca genes are the most efficient insecticidal Bt Cry proteins with activity against hemipteran insects reported so far. Cry64Ba and Cry64Ca showed no toxicity against some lepidopteran or coleopteran pests. These two proteins should be able to be used for integrated hemipteran pest management.

Incidence and occurrence profiles of the small brown planthopper (Laodelphax striatellus Fallén) in Korea in 2011–2015

The small brown planthopper (SBPH, Laodelphax striatellus Fallén) is an important pest that causes severe yield losses in rice by transmitting viral plant diseases. For the proper management of this pest, therefore, it is necessary to understand its temporal and spatial dynamics by establishing a periodical monitoring system. A dataset, including the number of SBPHs by location, collection method [aerial collection net (AeCN) or light trap (LT)] and period (May–Aug.) for five years (2011–2015) was provided by the Rural Developmental Administration, and missing values were imputed using multiple imputation methods. Of the 15,848 individuals collected, approximately 47% and 52.9% were collected using the AeCN and LT methods, respectively. Large numbers of SBPHs were generally collected from western coastal regions using AeCNs but not LTs. A high incidence of migratory SBPHs was observed during Julian days 144–166 using the AeCN method, with slightly different migration periods in each year. Generally, the migratory SBPHs made up 39.4% of the total populations of SBPHs during those periods. According to clustering analysis, the migratory region was located along the western coastal regions. Putative migration paths were estimated by trajectory cluster analyses using meteorological data. Interestingly, an L-shaped trajectory path emerged as a potentially important route for migratory SBPHs, passing through major wheat cultivation areas in Henan, Anhui, Jiangsu, and Zhejiang provinces in China, where high densities of SBPHs occur from late May to early June. These results would provide valuable information to predict the incidence period of migratory SBPHs and establish a proactive management system against SBPH, including the basis for the detection of hazardous factors and decisions regarding appropriate pesticide treatment periods.

Comparing the genetic diversity and structure of indigenous Korean and Chinese populations of Laodelphax striatellus Fallén using mitochondrial haplotypes

The small brown planthopper, Laodelphax striatellus Fallén, is a major migratory pest from China to Korea and transmits viral diseases of rice plants. In this study, we compared the genetic structures of overwintered indigenous Korean and Chinese populations. The eight Korean populations consisted of 33 haplotypes, and 16 of these were newly identified. The genetic diversity of the Korean population revealed high haplotype diversity (Hd) and low nucleotide diversity (π) of 0.86±0.017 and 0.0024±0.0001 on average, respectively. These values for the Chinese population were Hd=0.92±0.016 and π=0.0026±0.0002. Overall, genetic distances by pairwise FST were generally low, ranging from −0.022 to 0.089. Moreover, the Korean population revealed its ongoing population expansion by showing negative values in the neutrality test (Tajima's D=−1.4, Fu's Fs=−15.8) and a unimodal pattern in the mismatch distribution analysis. The genetic structures and population relationships were determined using 8 indigenous Korean and 30 Chinese populations by Bayesian approaches and population tree construction, respectively. The genetic structure was inferred as five (K=5), and all indigenous Korean populations as well as 10 Chinese populations from the Shanghai city, Anhui, Henan, Jiangsu, Liaoning, Shandong, and Zhejiang provinces were grouped into ‘G3’. These results provide partial support for unidirectional gene flow by migratory Chinese populations into Korea. Further analysis conducted using specimens from China and Korea collected in similar periods is necessary to clarify the migration population's origin and its effect on the genetic structure of the indigenous populations.

Resistance to cycloxaprid in Laodelphax striatellus is associated with altered expression of nicotinic acetylcholine receptor subunits.

Cycloxaprid is a new oxabridged cis-configuration neonicotinoid insecticide, the resistance development potential and underlying resistance mechanism of which were investigated in the small brown planthopper, Laodelphax striatellus (Fallén), an important agricultural pest of rice. A cycloxaprid-resistant strain (YN-CPD) only achieved 10-fold higher resistance, in contrast to 106-fold higher resistance to buprofezin and 332-fold higher resistance to chlorpyrifos achieved after exposure to similar selection pressure, and the cycloxaprid selected line showed no cross-resistance to the buprofezin and chlorpyrifos-selected resistance strains. Moreover, we identified 10 nicotinic acetylcholine receptor (nAChR) subunits from the transcriptome of L. striatellus, and six segments had open reading frames (ORFs). While we did not find mutations in the nAChR genes of L. striatellus, subunits Lsα1 and Lsβ1 exhibited, respectively, 9.60-fold and 3.36-fold higher expression in the resistant strain, while Lsα8 exhibited 0.44-fold lower expression. Suppression of Lsα1 through ingestion of dsLsα1 led to an increase in susceptibility to cycloxaprid. The findings indicate that resistance to cycloxaprid develops slowly compared with resistance to other chemicals and without cross-resistance to chlorpyrifos or buprofezin; over-expressed Lsα1 is associated with low cycloxaprid resistance levels, but the importance of over-expressed Lsβ1 and reduced expression of Lsα8 could not be excluded. © 2017 Society of Chemical Industry.

Identification of virus-derived siRNAs and their targets in RBSDV-infected rice by deep sequencing.

RNA interference (RNAi) is a conserved mechanism against viruses in plants and animals. It is thought to inactivate the viral genome by producing virus-derived small interfering RNAs (vsiRNAs). Rice black-streaked dwarf virus (RBSDV) is transmitted to plants by the small brown planthopper (Laodelphax striatellus), and seriously threatens production of rice in East Asia, particularly Oryza sativa japonica subspecies. Through deep sequencing, genome-wide comparisons of RBSDV-derived vsiRNAs were made between the japonica variety Nipponbare, and the indica variety 9311. Four small RNA libraries were constructed from the leaves and shoots of each variety. We found 659,756 unique vsiRNAs in the four samples, and only 43,485 reads were commonly shared. The size distributions of vsiRNAs were mostly 21- and 22-nt long, and A/U bias (66-68%) existed at the first nucleotide of vsiRNAs. Additionally, vsiRNAs were continuously but heterogeneously distributed along S1-S10 segments of the RBSDV genome. Distribution profiles of vsiRNA hotspots were similar in different hosts and tissues, and the 5'- and 3'-terminal regions of S4, S5, and S8 had more hotspots. Distribution and abundance of RBSDV vsiRNAs could be useful in designing efficient targets for exploiting RNA interference for virus resistance. Degradome analysis found 25 and 11 host genes appeared to be targeted by vsiRNAs in 9311 and Nipponbare. We report for the first time vsiRNAs derived from RBSDV-infected rice.

Spread and yield loss mechanisms of rice stripe disease in rice paddies

Rice stripe disease is an economically important disease of rice caused by the Rice stripe virus (RSV), which is transferred by the small brown planthopper (SBPH). The recent rapid increase in damage to rice crops throughout Japan caused by this disease makes it imperative to develop control methods as soon as possible. To obtain basic data for developing such methods, we studied how the disease causes damage and spreads within paddy fields. Our investigations revealed that diseased plants first appear in mid-June to early July, after which the disease spreads from affected plants to adjacent plants. This suggests that SBPH carrying RSV enter paddy fields, where they infect plants as they move about and lay eggs. Subsequently, hatched viruliferous nymphs infect surrounding plants, thereby spreading the disease. Our analysis of the damage caused by rice stripe disease showed that the earlier the onset of disease, the more extensive the damage caused, and that the disease reduces yield by reducing the number of healthy panicles. This suggests that to reduce damage caused by this disease, it is necessary to ensure the growth of a sufficient number of healthy panicles by controlling the vector insect during the crop’s early growth period. To be most effective, pest control efforts should be timed to target either the first-generation adults that colonize the paddy fields or the second-generation nymphs and adults that cause the rapid increase in the number of diseased plants within a field.