Substitution mapping and characterization of brown planthopper resistance genes from indica rice variety, 'PTB33' (Oryza sativa L.).

Rice (Oryza sativa L.) yield is severely reduced by the brown planthopper (BPH), Nilaparvata lugens Stål, in Asian countries. Increasing resistance in rice against BPH can mitigate yield loss. Previous reports indicated the presence of three BPH resistance genes, BPH2, BPH17-ptb, and BPH32, in durable resistant indica rice cultivar 'PTB33'. However, several important questions remain unclear; the genetic locations of BPH resistance genes on rice chromosomes and how these genes confer resistance, especially with relationship to three major categories of resistance mechanisms; antibiosis, antixenosis or tolerance. In this study, locations of BPH2, BPH17-ptb, and BPH32 were delimited using chromosome segment substitution lines derived from crosses between 'Taichung 65' and near-isogenic lines for BPH2 (BPH2-NIL), BPH17-ptb (BPH17-ptb-NIL), and BPH32 (BPH32-NIL). BPH2 was delimited as approximately 247.5 kbp between RM28449 and ID-161-2 on chromosome 12. BPH17-ptb and BPH32 were located between RM1305 and RM6156 on chromosome 4 and RM508 and RM19341 on chromosome 6, respectively. The antibiosis, antixenosis, and tolerance were estimated by several tests using BPH2-NIL, BPH17-ptb-NIL, and BPH32-NIL. BPH2 and BPH17-ptb showed resistance to antibiosis and antixenosis, while BPH17-ptb and BPH32 showed tolerance. These results contribute to the development of durable BPH resistance lines using three resistance genes from 'PTB33'.

Potential Risks of Poaceous Plants as Infectious Sources of Rice Black-Streaked Dwarf Virus Transmitted by the Small Brown Planthopper, Laodelphax striatellus.

The recent reemergence of rice black-streaked dwarf virus (RBSDV) has caused severe rice yield losses in several areas of East Asia. To identify the most important infectious sources of RBSDV, we compared the susceptibility of major poaceous plants to RBSDV infection and survival and the RBSDV acquisition efficiency of a vector insect, the small brown planthopper Laodelphax striatellus. RBSDV infection and survival rates of L. striatellus were significantly high in wheat (Triticum aestivum 'Norin61') and rice (Oryza sativa 'Reiho'), indicating that these crops can be important sources of RBSDV. Our results also showed that RBSDV can complete its infection cycle between Italian ryegrass (Lolium multiflorum 'Hataaoba') and L. striatellus. These results indicate that control of RBSDV and L. striatellus on winter-spring crops of wheat and Italian ryegrass may avoid an RBSDV epidemic on rice during the following summer. In addition to infections of wheat and Italian ryegrass, RBSDV infections were detected in Avena fatua, Avena sterilis subsp. ludoviciana, Cynosurus echinatus, Festuca arundinacea, Festuca pratensis, Lolium perenne, and Vulpia myuros var. megalura, although the infection efficiency varied.

Remote sampling mass spectrometry for dry samples: Sheath-flow probe electrospray ionization (PESI) using a gel-loading tip inserted with an acupuncture needle.

RATIONALE: Probe electrospray ionization (PESI) is only applicable to liquid or wet samples. In this study, a sheath-flow PESI method for remote sampling mass spectrometry that can be applied to dry samples was developed. METHODS: An acupuncture needle (0.12 mm outer diameter, 700 nm tip diameter) was inserted into a gel-loading tip with a 0.1 mm protrusion out of the tip. Analytes were extracted by filling the latter tip with solvent and softly touching the sample surface for a short time (<1 s). A high voltage was applied to the acupuncture needle, and mass spectra of analytes were obtained by self-aspirating electrospray. RESULTS: Dry samples, such as lines of ballpoint pen ink on paper, pharmaceutical tablets, instant coffee, brown rice, and narcotics, gave strong ion signals. The sample carryover was negligible. The sequential electrospray was observed to be similar to conventional PESI. The limits of detection (LODs) for morphine and rhodamine B were found to be of the order of picograms. CONCLUSIONS: Because of its simplicity and versatility, sheath-flow PESI is a promising technique for on-site and nondestructive profile analysis of dry samples with bulky and complicated shapes, with a spatial resolution of ~0.3 mm.

Quantitative Analysis of Southern rice black-streaked dwarf virus in Sogatella furcifera and Virus Threshold for Transmission.

We investigated Southern rice black-streaked dwarf virus (SRBSDV) accumulation in a vector insect, the whitebacked planthopper (Sogatella furcifera), to elucidate the association of virus accumulation in the vector with virus transmission efficiency. Real-time quantitative reverse transcription polymerase chain reaction analysis confirmed that this virus is transmitted in a persistent-propagative manner. SRBSDV was successfully transmitted by S. furcifera males in which RNA accumulation of the capsid protein gene of SRBSDV was >10(3) in the whole body of S. furcifera, indicating that the threshold accumulation of the virus RNA for virus transmission is 10(3) in an S. furcifera male. The SRBSDV detection rate in the immigrant population of S. furcifera was high in 2011 (39.5%); however, most of the insects contained fewer than 10(3) RNAs of the capsid protein gene. This result indicates that the risk of SRBSDV epidemics could be estimated from the proportion of virus-transmissible S. furcifera (i.e., S. furcifera that contained more than 10(3) RNAs of the virus capsid protein gene) rather than the SRBSDV detection rate in S. furcifera.

Genetic basis of multiple resistance to the brown planthopper (Nilaparvata lugens Stål) and the green rice leafhopper (Nephotettix cincticeps Uhler) in the rice cultivar 'ASD7' (Oryza sativa L. ssp. indica).

The rice cultivar ASD7 (Oryza sativa L. ssp. indica) is resistant to the brown planthopper (BPH; Nilaparvata lugens Stål) and the green leafhopper (Nephotettix virescens Distant). Here, we analyzed multiple genetic resistance to BPH and the green rice leafhopper (GRH; Nephotettix cincticeps Uhler). Using two independent F2 populations derived from a cross between ASD7 and Taichung 65 (Oryza sativa ssp. japonica), we detected two QTLs (qBPH6 and qBPH12) for resistance to BPH and one QTL (qGRH5) for resistance to GRH. Linkage analysis in BC2F3 populations revealed that qBPH12 controlled resistance to BPH and co-segregated with SSR markers RM28466 and RM7376 in plants homozygous for the ASD7 allele at qBPH6. Plants homozygous for the ASD7 alleles at both QTLs showed a much faster antibiosis response to BPH than plants homozygous at only one of these QTLs. It revealed that epistatic interaction between qBPH6 and qBPH12 is the basis of resistance to BPH in ASD7. In addition, qGRH5 controlled resistance to GRH and co-segregated with SSR markers RM6082 and RM3381. qGRH5 is identical to GRH1. Thus, we clarified the genetic basis of multiple resistance of ASD7 to BPH and GRH.

Genetic mapping of the rice resistance-breaking gene of the brown planthopper Nilaparvata lugens.

Host plant resistance has been widely used for controlling the major rice pest brown planthopper (BPH, Nilaparvata lugens). However, adaptation of the wild BPH population to resistance limits the effective use of resistant rice varieties. Quantitative trait locus (QTL) analysis was conducted to identify resistance-breaking genes against the anti-feeding mechanism mediated by the rice resistance gene Bph1. QTL analysis in iso-female BPH lines with single-nucleotide polymorphism (SNP) markers detected a single region on the 10th linkage group responsible for the virulence. The QTL explained from 57 to 84% of the total phenotypic variation. Bulked segregant analysis with next-generation sequencing in F2 progenies identified five SNPs genetically linked to the virulence. These analyses showed that virulence to Bph1 was controlled by a single recessive gene. In contrast to previous studies, the gene-for-gene relationship between the major resistance gene Bph1 and virulence gene of BPH was confirmed. Identified markers are available for map-based cloning of the major gene controlling BPH virulence to rice resistance.

The Spread of Southern Rice Black-Streaked Dwarf Virus Was Not Caused by Biological Changes in Vector Sogatella furcifera.

The pandemic of Southern rice black-streaked dwarf virus (SRBSDV) in and after the late 2000s caused serious yield losses in rice in Southeast and East Asia. This virus was first recorded in China in 2001, but its exclusive vector insect, Sogatella furcifera, occurred there before then. To clarify the evolutionary origin of SRBSDV as the first plant virus transmitted by S. furcifera, we tested virus transmission using three chronological strains of S. furcifera, two of which were established before the first report of SRBSDV. When the strains fed on SRBSDV-infected rice plants were transferred to healthy rice plants, those established in 1989 and 1999 transmitted the virus to rice similarly to the strain established in 2010. SRBSDV quantification by RT-qPCR confirmed virus accumulation in the salivary glands of all three strains. Therefore, SRBSDV transmission by S. furcifera was not caused by biological changes in the vector, but probably by the genetic change of the virus from a closely related Fijivirus, Rice black-streaked dwarf virus, as suggested by ecological and molecular biological comparisons between the two viruses. This result will help us to better understand the evolutionary relationship between plant viruses and their vector insects and to better manage viral disease in rice cropping in Asia.

Male killing caused by a Spiroplasma symbiont in the small brown planthopper, Laodelphax striatellus.

Spiroplasma-mediated late male killing was found in the small brown planthopper, Laodelphax striatellus. Female-biased colonies (maternal lines, N = 4) were established from planthoppers collected in Taiwan and Japan. This sex ratio distortion was maternally inherited (sex ratio of total number of progenies [female:male]: 488:0 in F1, 198:7 in F2, 407:0 in F3; likelihood ratio test of all generations, P < 0.0001) and caused by male death during nymphal stages. The female-biased colonies were doubly infected with Spiroplasma and Wolbachia, and the non-biased colonies were infected solely with Wolbachia. Antibiotic treatment resulted in a normal sex ratio, strongly suggesting that bacteria are manipulating host reproduction. Spiroplasma-singly-infected planthopper colonies created by the antibiotic treatment produced progeny with strongly female-biased sex ratios (181:2; likelihood ratio test, χ(2) = 231.6, P < 0.0001). This is the first report of Spiroplasma-mediated male killing in hemimetabolous insects.

Dynamics of Southern rice black-streaked dwarf virus in rice and implication for virus acquisition.

A novel viral disease of rice caused by Southern rice black-streaked dwarf virus (SRBSDV) has spread throughout East and Southeast Asia since the mid-2000s. Outbreaks of this viral disease occur yearly in southern parts of Japan concurrently with overseas migration of the planthopper vector Sogatella furcifera from southern China during the rainy season (from late June to early July). We examined the dynamics (changes in titer and localization) of SRBSDV on rice using reverse-transcription real-time polymerase chain reaction and determined the relationship between virus titer in plants and virus acquisition by S. furcifera. Under a constant temperature of 27°C, a substantial increase of SRBSDV titer in the leaf sheath together with typical symptoms (stunted growth and twisting of leaf tips) was observed at 20 days after the end of a 7-day exposure of viruliferous S. furcifera. Approximately 40% of S. furcifera acquired SRBSDV through feeding for 5 days on rice plants that were infected following exposure to viruliferous vectors for 10 to 15 days. These results suggest that rice infected by S. furcifera can be a source of SRBSDV before the next generation of S. furcifera emerges.

Efficacy of pulmonary vein isolation on left atrial function in paroxysmal and persistent atrial fibrillation and the dependency on its baseline function.

BACKGROUND: The effects of pulmonary vein (PV) isolation in atrial fibrillation (AF) on left atrial (LA) function or PV flow have not been well documented. METHODS: We examined the LA function and PV flow before and 3-6 months after PV isolation in 67 AF patients (34 paroxysmal [PAF] and 33 persistent [CAF]) using transesophageal echocardiography. RESULTS: AF recurred in 6/34 patients with PAF and in 6/33 patients with CAF 6 months after PV isolation. A larger LA dimension, a lower systolic PV flow velocity, and a lower ratio of systolic to diastolic PV flow velocity were related to a higher incidence of AF recurrence. The increment of left atrial appendage (LAA) flow velocity (55% vs. 22%) and systolic PV flow velocity (57% vs. 20%) after PV isolation tended to be greater in CAF than in PAF. The changes in LAA flow velocity had reverse correlations with the baseline values before PV isolation (PAF: r = -0.73, CAF: r = -0.58). The changes in mitral flow velocity during atrial contraction in PAF had reverse correlations with the baseline values before PV isolation (r = -0.84). The changes in systolic and diastolic PV flow velocity of PAF had reverse correlations with the baseline values before PV isolation (r = -0.56, r = -0.66). CONCLUSION: The baseline LA function may affect AF recurrence as well as the improvement of LA function, and the benefit of successful PV isolation might be greater in CAF than in PAF.

Mitochondrial cox sequences of Nilaparvata lugens and Sogatella furcifera (Hemiptera, Delphacidae): low specificity among Asian planthopper populations.

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.

Mapping and pyramiding of two major genes for resistance to the brown planthopper (Nilaparvata lugens [Stål]) in the rice cultivar ADR52.

The brown planthopper (BPH), Nilaparvata lugens (Stål), is one of the most serious and destructive pests of rice, and can be found throughout the rice-growing areas of Asia. To date, more than 24 major BPH-resistance genes have been reported in several Oryza sativa ssp. indica cultivars and wild relatives. Here, we report the genetic basis of the high level of BPH resistance derived from an Indian rice cultivar, ADR52, which was previously identified as resistant to the whitebacked planthopper (Sogatella furcifera [Horváth]). An F(2) population derived from a cross between ADR52 and a susceptible cultivar, Taichung 65 (T65), was used for quantitative trait locus (QTL) analysis. Antibiosis testing showed that multiple loci controlled the high level of BPH resistance in this F(2) population. Further linkage analysis using backcross populations resulted in the identification of BPH-resistance (antibiosis) gene loci from ADR52. BPH25 co-segregated with marker S00310 on the distal end of the short arm of chromosome 6, and BPH26 co-segregated with marker RM5479 on the long arm of chromosome 12. To characterize the virulence of the most recently migrated BPH strain in Japan, preliminary near-isogenic lines (pre-NILs) and a preliminary pyramided line (pre-PYL) carrying BPH25 and BPH26 were evaluated. Although both pre-NILs were susceptible to the virulent BPH strain, the pre-PYL exhibited a high level of resistance. The pyramiding of resistance genes is therefore likely to be effective for increasing the durability of resistance against the new virulent BPH strain in Japan.

Enhanced resistance to four species of Clypeorrhynchan pests in Neotyphodium uncinatum infected Italian ryegrass.

Particular alkaloids produced by Neotyphodium endophytes show toxicity to invertebrates. Italian ryegrass (Lolium multiflorum Lamarck) cultivars and strains that are symbiotic with Neotyphodium endophytes have been recently established in Japan. N. uncinatum-infected Italian ryegrass lines accumulate N-formylloline, a type of loline alkaloid (1-aminopyrrolizidine) showing neurotoxicity to herbivorous insects. This study investigated the toxicity of N-formylloline and resistance of N. uncinatum-infected Italian ryegrass to vascular-sap feeding Clypeorrhynchan pests. When four vascular-sap feeding insects: Laodelphax striatellus (Fallén) (Homoptera: Delphacidae), Sogatella furcifera (Horváth) (Homoptera: Delphacidae), Cicadulina bipunctata (Melichar) (Homoptera: Cicadellidae), and Nephotettix cincticeps (Uhler) (Homoptera: Cicadellidae) fed on N. uncinatum-infected Italian ryegrass, significant decreases in survival rate were observed for three phloem-sap feeders but not for a xylem-sap feeder, N. cincticeps. This result suggests an uneven distribution of N-formylloline among plant tissues. A potency assay for N-formylloline using a Parafilm feeding sachet and a quantitative analysis of N-formylloline in plant showed a concentration-dependent lethal effect of N-formylloline on all four tested vascular-sap feeders. Our results strongly suggest that N. uncinatum-infected plants can control some Clypeorrhynchan pests in crop fields.

Unraveling the hierarchical genetic structure of tea green leafhopper, Matsumurasca onukii, in East Asia based on SSRs and SNPs.

Matsumurasca onukii (Matsuda, R. (1952). Oyo-Kontyu Tokyo, 8(1): 19-21), one of the dominant pests in major tea production areas in Asia, currently is known to occur in Japan, Vietnam, and China, and severely threatens tea production, quality, and international trade. To elucidate the population genetic structure of this species, 1633 single nucleotide polymorphisms (SNPs) and 18 microsatellite markers (SSRs) were used to genotype samples from 27 sites representing 18 geographical populations distributed throughout the known range of the species in East Asia. Analyses of both SNPs and SSRs showed that M. onukii populations in Yunnan exhibit high-genetic differentiation and structure compared with the other populations. The Kagoshima (JJ) and Shizuoka (JS) populations from Japan were separated from populations from China by SNPs, but clustered with populations from Jinhua (JH), Yingde (YD), Guilin (GL), Fuzhou (FZ), Hainan (HQ), Leshan (CT), Chongqing (CY), and Zunyi (ZY) tea plantations in China and the Vietnamese Vinh Phuc (VN) population based on the SSR data. In contrast, CT, CY, ZY, and Shaanxi (SX) populations clustered together based on SNPs, but were separated by SSRs. Both marker datasets identified significant geographic differentiation among the 18 populations. Various environmental and anthropogenic factors, including geographical barriers to migration, human transport of hosts (Camellia sinesis [L.] O. Kuntze) and adaptation of M. onukii to various local climatic zones possibly account for the rapid spread of this pest in Asia. The results demonstrate that SNPs from high-throughput genotyping data can be used to reveal subtle genetic substructure at broad scales in r-strategist insects.

Two hidden taxa in the Japanese encephalitis vector mosquito, Culex tritaeniorhynchus, and the potential for long-distance migration from overseas to Japan.

The Culex vishnui subgroups, particularly Culex tritaeniorhynchus, are considered the primary vectors of the Japanese encephalitis virus (JEV) in Asia. Recent molecular phylogenetic analyses of JEV isolates from Asian countries have shown that JEVs with diverse genetic variants are present in Asia. Furthermore, some JEV strains have been found to have crossed the East China Sea and been introduced into Japan. In this study, the possibility of overseas migration of the JE vector mosquito, Cx. tritaeniorhynchus was examined from the genetic, physical, and meteorological perspectives. Molecular phylogenetic analysis was performed based on both whole coding sequences and on the barcoding region of the mitochondrial cytochrome c oxidase subunit I (COI) gene of Cx. vishnui subgroups collected from Asian countries. Culex tritaeniorhymchus was classified into two genetically independent taxa by COI sequences: the Japanese type (Ct-J), which inhabits Japan except for the Amami Islands of southern Japan, and the continental type (Ct-C), which inhabits the Asian region except for Japan. It was confirmed that approximately 10% of Cx. tritaeniorhynchus trapped during the summer in western Kyushu were Ct-C, and that they could fly for up to 38 h continuously. The meteorological analysis also confirmed that the atmospheric flow occurring over the continent coincided with the date of Ct-C capture. This is the first report showing the existence of two taxa in Cx. tritaeniorhynchus. Their physical and physiological characteristics suggest the possibility of long-distance migration from overseas regions to Japan across the East China Sea. Future efforts are expected to provide evidence to support the occurrence of long-distance migration of Cx. tritaeniorhynchus with JEV.

Role of hydroperoxide lyase in white-backed planthopper (Sogatella furcifera Horváth)-induced resistance to bacterial blight in rice, Oryza sativa L.

A pre-infestation of the white-backed planthopper (WBPH), Sogatella furcifera Horváth, conferred resistance to bacterial blight caused by Xanthomonas oryzae pv. oryzae (Xoo) in rice (Oryza sativa L.) under both laboratory and field conditions. The infestation of another planthopper species, the brown planthopper (BPH) Nilaparvata lugens Stål, did not significantly reduce the incidence of bacterial blight symptoms. A large-scale screening using a rice DNA microarray and quantitative RT-PCR revealed that WBPH infestation caused the upregulation of more defence-related genes than did BPH infestation. Hydroperoxide lyase 2 (OsHPL2), an enzyme for producing C(6) volatiles, was upregulated by WBPH infestation, but not by BPH infestation. One C(6) volatile, (E)-2-hexenal, accumulated in rice after WBPH infestation, but not after BPH infestation. A direct application of (E)-2-hexenal to a liquid culture of Xoo inhibited the growth of the bacterium. Furthermore, a vapour treatment of rice plants with (E)-2-hexenal induced resistance to bacterial blight. OsHPL2-overexpressing transgenic rice plants exhibited increased resistance to bacterial blight. Based on these data, we conclude that OsHPL2 and its derived (E)-2-hexenal play some role in WBPH-induced resistance in rice.

Effect of barley chromosome addition on the susceptibility of wheat to feeding by a gall-inducing leafhopper.

The maize orange leafhopper Cicadulina bipunctata is distributed widely in tropical and subtropical regions of the Old World and feeds on various Poaceae. The leafhopper is recognized as an important pest of maize in several countries. Adults as well as nymphs of C. bipunctata induce growth stunting and galls characterized by the severe swelling of leaf veins on many cereal crops including wheat, rice, and maize, but do not on barley. To clarify the mechanism of growth stunting and gall induction by C. bipunctata, we used six barley chromosome disomic addition lines of wheat (2H-7H) and investigated the effect of barley (cv. Betzes) chromosome addition on the susceptibility of wheat (cv. Chinese Spring) to feeding by the leafhopper. Feeding by C. bipunctata significantly stunted the growth in 2H, 3H, 4H, and 5H, but did not in 6H and 7H. The degree of gall induction was significantly weaker and severer in 3H and 5H than in Chinese Spring, respectively. These results suggest that barley genes resistant to growth stunting and gall induction exist in 6H and 7H, and 3H, respectively. 5H is considered to be useful for future assays investigating the mechanism of gall induction by this leafhopper because of the high susceptibility to the feeding by C. bipunctata. Significant correlation between the degrees of growth stunting and gall induction was not detected in the six chromosome addition lines and Chinese spring. This implies that these two symptoms are independent phenomena although both are initiated by the feeding of C. bipunctata.

Genome-wide assessment and development of molecular diagnostic methods for imidacloprid-resistance in the brown planthopper, Nilaparvata lugens (Hemiptera; Delphacidae).

BACKGROUND: The brown planthopper, Nilaparvata lugens (Stål), is one of the most notorious pests of rice throughout Asia. The brown planthopper has developed high resistance to imidacloprid, a member of neonicotinoid insecticides. Several genes and mutations conferring imidacloprid resistance in N. lugens, especially in eastern and southeastern Asia populations, have been reported. Thus, the key mechanisms of imidacloprid resistance need to be examined. RESULTS: RNA-seq analyses revealed that only one cytochrome P450 monooxygenase gene, CYP6ER1, was commonly upregulated in the five resistant strains tested. Sequences of CYP6ER1, which were highly expressed in the imidacloprid-resistant strains, contained a three-nucleotide deletion in the coding region, and amino acid substitutions and deletion, compared to that in an imidacloprid-susceptible strain. RNAi-mediated gene knockdown of CYP6ER1 increased imidacloprid susceptibility in a resistant strain. Further, we established two simple and convenient PCR-based molecular diagnostic methods to detect the CYP6ER1 locus with the three-nucleotide deletion. Using these methods, the resistance of F2 progenies derived from the crosses of F1 siblings from susceptible and resistant parents was analyzed, showing that the imidacloprid resistance had a relationship to the CYP6ER1 locus with the three-nucleotide deletion. CONCLUSION: The overexpression of a variant CYP6ER1 with amino acid substitutions and deletion was involved in imidacloprid resistance in N. lugens. Based on these findings, molecular diagnostic methods have been developed and are promising tools for monitoring imidacloprid resistance in paddy fields. © 2020 Society of Chemical Industry.

Dispersal of the Common Cutworm, Spodoptera litura, Monitored by Searchlight Trap and Relationship with Occurrence of Soybean Leaf Damage.

The common cutworm, Spodoptera litura Fabricius (Lepidoptera: Noctuidae) is a major pest of soybean. Pheromone traps are used to monitor male adults, but the catch peaks do not always predict leaf damage in soybean fields. Thus, there is no accurate means of forecasting soybean damage, and insecticide is applied on the basis of farmers' observations of actual damage in fields. To understand the occurrence of soybean leaf damage, the dispersal of S. litura in a soybean field in southwestern Japan was preliminarily investigated using a searchlight trap in comparison to a pheromone trap at one location, from August to mid-October in 2016-2018. To determine the relationship between pest arrival and crop damage, trap catch numbers and the number of soybean leaves damaged by 1st-instar larvae were examined by separately comparing raw numbers and cumulative numbers. The raw catch numbers of the two trap types in August and September 2016 and 2018 preceded subsequent damage peaks by an average of 5.3 days. This temporal difference coincided with the estimated duration of the egg stage plus an assumed mating period. Furthermore, the cumulative catch numbers of the two traps in August and September were linearly associated with cumulative damaged leaves in the same period in each year and in the three-year period. The coefficient of determination (R2) of linear regression between the cumulative catch numbers of the searchlight trap and the cumulative damaged leaves for the three-year period was much higher than that between the cumulative catch of the pheromone trap and cumulative damage. This suggests that soybean leaf damage is closely linked to the number of S. litura arrivals at the survey site. Thus, the searchlight trap captured S. litura arrivals better than the pheromone trap. As the linear regression function of the cumulative catch of the searchlight trap for the three-year period was tentatively correlated with a prefectural economic injury level for soybean fields, it might be feasible to predict S. litura-induced soybean damage using searchlight traps. The cumulative female catch number of the searchlight trap was also linearly associated with damaged leaves, but the coefficient of determination was generally lower than that with the cumulative total catch. The female ratio of searchlight trap catches in September was <0.5 in contrast to S. litura migrating overseas (>0.5). The advantages and disadvantages of the two trapping methods, as well as necessary further studies are discussed. Our findings provide a foundation for S. litura monitoring with searchlight traps.

Energy Reserve Compensating for Trade-Off Between Metabolic Resistance and Life History Traits in the Brown Planthopper (Hemiptera: Delphacidae).

Development of insecticide resistance often changes life history traits of insect pests, because metabolic detoxification of insecticides in insect bodies requires huge energetic reserves. The brown planthopper, Nilaparvata lugens (Stål), an important insect pest of rice crop in East and Southeast Asia, has developed strong resistance to imidacloprid from mid-2000s. The aim of this study was to examine the costs of life history traits and reveal changes in energy reserves with developing imidacloprid resistance. We compared the life history traits (survival time, fecundity, developmental time, and hatchability) and total lipid content between imidacloprid-resistant and imidacloprid-susceptible (control) brown planthopper strains. As compared to the control strains, adults' survival time of the resistant females was shorter, and their fecundity was lower; the other life history traits did not differ significantly between the resistant and control strains. As the results, net reproductive rates (R0) were lower in the resistant strains than in the susceptible strains. However, the amount of stored lipids was larger in resistant females than control ones. Our findings demonstrated a physiological trade-off between the development of imidacloprid resistance and the reproductive traits of brown planthopper. The imidacloprid-resistant strains are likely to store lipids for metabolic detoxification rather than consume them for reproduction.