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.

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.

Long-term field insecticide susceptibility data and laboratory experiments reveal evidence for cross resistance to other neonicotinoids in the imidacloprid-resistant brown planthopper Nilaparvata lugens.

BACKGROUND: Long-term monitoring data is helpful to understand the fluctuation of susceptibility and pattern of cross resistance in insecticide resistance management. After the occurrence of imidacloprid resistance, the brown planthopper (BPH) has gradually developed resistance to thiamethoxam and clothianidin since 2010, but not to dinotefuran and nitenpyram. Here, we analyzed susceptibilities data of five neonicotinoids during 2005-2017 in East Asia and Vietnam to conduct cross-resistance patterns among neonicotinoids. To determine the factors of development of cross resistance in laboratory bioassays, we used the imidacloprid resistant and control strains that were selected from filed populations in the Philippines and Vietnam. RESULTS: The Linear Mixed Models (LMM) analyses of insecticide susceptibility data showed that the slope values of imidacloprid resistance effects were 0.68 and 1.09 for resistance to thiamethoxam and clothianidin, respectively. Laboratory bioassay results showed that the LD50 values for thiamethoxam and clothianidin in resistant strains (1.4-5.5 µg g-1 ) were 3.2-16.4 times higher than those in the control strains (0.28-1.5 µg g-1 ). However, the increase in the LD50 values for imidacloprid was not related to that for dinotefuran and nitenpyram based on the results of the LMM analysis and laboratory bioassay. CONCLUSION: Our results demonstrate that the development of imidacloprid resistance result in strong-cross resistance to some neonicotinoids, thiamethoxam and clothianidin, but not to others, dinotefuran and nitenpyram. We anticipate that our findings will be a starting point for understanding mechanism of the different trend of cross resistance by analyzing long-term susceptibility data and laboratory bioassays in insect pests. © 2019 Society of Chemical Industry.

Selection for imidacloprid resistance and mode of inheritance in the brown planthopper, Nilaparvata lugens.

BACKGROUND: Strong resistance to imidacloprid in Nilaparvata lugens (Stål) has developed in Southeast and East Asia. Although the mode of inheritance for resistance is very useful information for pest control, this information is unknown in N. lugens. Here, we established two resistant strains that were selected from field populations in Vietnam and the Philippines, and conducted crossing experiments to determine the inheritance pattern. RESULTS: The resistance ratio of 50% lethal dose (LD50 ) values for the two resistance-selected strains, i.e., resistant strains originating from Vietnam (VT-Res) and the Philippines (PH-Res), to their control strains were ∼ 8- and 157-fold, respectively. Reciprocal cross experiments between VT-Res and the susceptible strain (S-strain), and between PH-Res and the S-strain showed that the degree of dominance was 0.81 and 0.82, and 0.95 and 0.96, respectively. Analysis of the F2 populations and backcrosses to the parental strains indicated that resistance is a major-gene trait following Mendelian inheritance. The strength of the resistance was suppressed by pre-treatment with piperonyl butoxide, an inhibitor of cytochrome P450-monooxygenases. CONCLUSION: Our results suggest that imidacloprid resistance in N. lugens is autosomal and an almost completely dominant major-gene trait that is likely manifested by high expression levels of a detoxification enzyme. © 2019 Society of Chemical Industry.