Imidacloprid-resistant Populations Research Articles

Cross-Resistance to Imidacloprid in Brown Planthopper <i>Nilaparvata lugens</i> (Stal)

Cross-resistance pattern of imidacloprid resistance in brown planthopper, Nilaparvata lugens (Stål) to three insecticides viz., dinotefuran, pymetrozine and triflumezopyrim was evaluated with laboratory developed imidacloprid resistant (IMI-R) and susceptible populations (IMI-S). IMI-R population had a 6-fold resistance compared to IMI-S population. The LC50 values for dinotefuran in IMI-R and IMI-S populations were 466.6 and 129.2 mg/ L with a resistance ratio of 3.6-fold. Whereas, LC50 values for triflumezopyrim and pymetrozine in IMI-R and IMI-S populations were 0.27 and 0.17 mg/L; 49.07 and 31.5 mg/L with resistant ratios of 1.58 and 1.55, respectively. These findings clearly demonstrated cross-resistance between imidacloprid and dinotefuran but no obvious cross-resistance to triflumezopyrim and pymetrozine.

Glutathione S-transferase directly metabolizes imidacloprid in the whitefly, Bemisia tabaci

The whitefly Bemisia tabaci poses a significant threat to various crops and ornamental plants and causes severe damage to the agricultural industry. Over the past few decades, B. tabaci has developed resistance to several pesticides, including imidacloprid. Therefore, elucidating the mechanism that leads to insecticide detoxification is very important for controlling B. tabaci and managing whitefly resistance to neonicotinoid insecticides. Among insect detoxification enzymes, glutathione S-transferase (GST) is an important phase II detoxification enzyme that helps detoxify exogenous toxic substances. In this study, we cloned the BtGSTz1 gene and observed that its expression level was greater in imidacloprid-resistant populations than sensitive populations of B. tabaci. By silencing BtGSTz1 via RNA interference, we found a significant increase in the mortality of imidacloprid-resistant B. tabaci. Additionally, prokaryotic expression and in vitro metabolism studies revealed that the recombinant BtGSTz1 protein could metabolize 36.36% of the total imidacloprid, providing direct evidence that BtGSTz1 plays a crucial role in the detoxification of imidacloprid. Overall, our study elucidated the role of GSTs in physiological activities related to insecticide resistance, which helps clarify the resistance mechanisms conferred by GSTs and provides useful insights for sustainable integrated pest management.

Insecticide resistance reduces the profitability of insect-resistant rice cultivars

IntroductionPreventing crop yield loss caused by pests is critical for global agricultural production. Agricultural pest control has largely relied on chemical pesticides. The interaction between insecticide resistance and the adaptation of herbivorous pests to host plants may represent an emerging threat to future food security. ObjectivesThis study aims to unveil genetic evidence for the reduction in the profitability of resistant cultivars derived from insecticide resistance in target pest insects. MethodsAn experimental evolution system encompassing resistant rice and its major monophagous pest, the brown planthopper Nilaparvata lugens, was constructed. Whole genome resequencing and selective sweep analysis were utilized to identify the candidate gene loci related to the adaptation. RNA interference and induced expression assay were conducted to validate the function of the candidate loci. ResultsWe found that the imidacloprid-resistant population of N. lugens rapidly adapted to resistant rice IR36. Gene loci related to imidacloprid resistance may contribute to this phenomenon. Multiple alleles in the nicotinic acetylcholine receptor (nAChR)-7-like and P450 CYP4C61 were significantly correlated with changes in virulence to IR36 rice and insecticide resistance of N. lugens. One avirulent/susceptible genotype and two virulent/resistant genotypes could be inferred from the corresponding alleles. Importantly, we found that the virulent/resistant genotypes already exist in the wild in China, exhibiting increasing frequencies along with insecticide usage. We validated the relevance of these genotypes and the virulence to three more resistant rice cultivars. Knockdown of the above two genes in N. lugens significantly decreased both the resistance to imidacloprid and the virulence towards resistant rice. ConclusionOur findings provide direct genetic evidence to the eco-evolutionary consequence of insecticide resistance, and suggest an urgent need for the implementation of predictably sustainable pest management.

Changes in Transcriptome and Gene Expression in Sogatella furcifera (Hemiptera: Delphacidae) in Response to Cycloxaprid.

The white-backed planthopper, Sogatella furcifera (Horváth), causes substantial damage to crops by direct feeding or virus transmission, especially southern rice black-streaked dwarf virus, which poses a serious threat to rice production. Cycloxaprid, a novel cis-nitromethylene neonicotinoid insecticide, has high efficacy against rice planthoppers, including imidacloprid-resistant populations. However, information about the influence of cycloxaprid on S. furcifera (Hemiptera: Delphacidae) at the molecular level is limited. Here, by de novo transcriptome sequencing and assembly, we constructed two transcriptomes of S. furcifera and profiled the changes in gene expression in response to cycloxaprid at the transcription level. We identified 157,906,456 nucleotides and 131,601 unigenes using the Illumina technology from cycloxaprid-treated and untreated S. furcifera. In total, 38,534 unigenes matched known proteins in at least one database, accounting for 29.28% of the total unigenes. The number of coding DNA sequences was 28,546 and that of amino acid sequences in the coding region was 22,299. In total, 15,868 simple sequence repeats (SSRs) were identified. The trinucleotide repeats accounted for 45.1% (7,157) of the total SSRs and (AAG/CTT)n were the most frequent motif. There were 359 differentially expressed genes that might have been induced by cycloxaprid. There were 131 upregulated and 228 downregulated genes. Twenty-two unigenes might be involved in resistance against cycloxaprid, such as cytochrome P450, glutathione S-transferase (GST), acid phosphatase (ACP), and cadherin. Our study provides vital information on cycloxaprid-induced resistance mechanisms, which will be useful to analyze the molecular mechanisms of cycloxaprid resistance and may lead to the development of novel strategies to manage S. furcifera.

Low expression levels of nicotinic acetylcholine receptor subunits Boα1 and Boβ1 are associated with imidacloprid resistance in Bradysia odoriphaga.

Neonicotinoid insecticide imidacloprid acts on insect nicotinic acetylcholine receptors (nAChRs). The mechanisms of insect resistance to imidacloprid include target-site alteration and increased detoxification metabolism. In Bradysia odoriphaga, cytochrome P450 monooxygenase has been found involved in metabolic resistance to imidacloprid. However, the situation of target-site related resistance to imidacloprid in B. odoriphaga is still unknown. Nine field-collected B. odoriphaga populations showed various sensitivities to imidacloprid compared with the susceptible (SS) strain, including susceptibility, decreased susceptibility, low resistance, moderate resistance and high resistance. Seven nAChR subunit genes including α1, α2, α3, α7, α8, β1 and β3, were examined for site mutation and changes in transcription levels in field populations. No nAChR polymorphism potentially related to the resistant phenotypes was found. However, differential expression of nAChR subunit genes was found in imidacloprid resistant field population. In high imidacloprid resistant population LC-2 (93.14-fold resistance), the transcription levels of α1, α2 and β1 subunits were significantly down-regulated, while the transcription levels of α3 and α8 subunits were significantly up-regulated, compared with that in SS strain. In addition, imidacloprid acute exposure induced differential expression of nAChR subunit genes in B. odoriphaga. Furthermore, RNA interference (RNAi) suppressed the transcriptional expression of Boα1 and Boβ1, and decreased mortality of B. odoriphaga by 23.03% and 18.69%, respectively, when treated with imidacloprid. These results indicated that, although no target-site mutation was found in imidacloprid resistant B. odoriphaga population, the reduced expression of α1 and β1 subunits contributed to B. odoriphaga resistance to imidacloprid. © 2020 Society of Chemical Industry.

Cycloxaprid: a novel cis-nitromethylene neonicotinoid insecticide to control Bemisia tabaci.

Bemisia tabaci is a worldwide insect pest with high ability to develop resistance to many insecticides. Cycloxaprid is a novel cis-configuration neonicotinoid insecticide that is effective against a wide range of imidacloprid-resistant pests. The lethal and sublethal effects of cycloxaprid on B. tabaci B were estimated. Cycloxaprid showed higher toxicity against adult, nymph and egg of B. tabaci B than imidacloprid. And cycloxaprid provided better efficacies against B. tabaci than imidacloprid in fields. Moreover, cycloxaprid was similarly toxic to the relatively susceptible and imidacloprid-resistant populations. The LC50 of cycloxaprid against the resistant adult, nymph and egg were 6.2, 11.5 and 111.3mg L-1 , respectively, while they were 5.7, 8.9 and 129.4mg L-1 for the susceptible population. In addition, when resistant adult of B. tabaci B was treated with LC30 of cycloxaprid (3.2mg L-1 ), the net reproductive rate and female ratio of F1 generation B. tabaci were reduced. Moreover, the age-specific fecundity demonstrated the highest fecundity sharply decreased in cycloxaprid (7.0 offspring/ day) treated group compared with the control (26.0 offspring/ day). Our results indicated that cycloxaprid is a promising insecticide for the management of B. tabaci and insecticide-induced resurgence might not occur after exposure of B. tabaci to low sublethal concentrations of cycloxaprid. © 2019 Society of Chemical Industry.

Overexpression of a glutathione S-transferase (Mdgst) and a galactosyltransferase-like gene (Mdgt1) is responsible for imidacloprid resistance in house flies.

Neonicotinoids are the largest class of insecticides and are used for control of house fly populations at animal production facilities throughout the world. There have been several reports of neonicotinoid resistance in house fly populations, but identification of the factors involved in resistance has proven challenging. The KS8S3 population of house flies is highly resistant to the neonicotinoid insecticide imidacloprid due to two factors: one on chromosome 3 and one on chromosome 4. A comparative transcriptomic approach was used, followed by validation using transgenic Drosophila melanogaster to investigate the genes responsible for resistance in the KS8S3 strain. Overexpression of a microsomal glutathione S-transferase (Mdgst) was identified as the factor likely responsible for resistance on chromosome 3. Resistance on chromosome 4 appears to be due to an unidentified trans-regulatory gene which causes overexpression of a galactosyltransferase-like gene (Mdgt1). No single nucleotide polymorphisms were found that could be associated with imidacloprid resistance. Identification of the underlying processes that cause imidacloprid resistance is an important first step towards the development of novel and sensitive resistance monitoring techniques. It will be valuable to investigate if overexpression of Mdgst and Mdgt1 are found in other imidacloprid resistant populations. © 2018 Society of Chemical Industry.

The binding properties of cycloxaprid on insect native nAChRs partially explain the low cross-resistance with imidacloprid in Nilaparvata lugens.

Neonicotinoids, such as imidacloprid, are selective agonists of insect nicotinic acetylcholine receptors (nAChRs) used to control Nilaparvata lugens, a major rice insect pest. High imidacloprid resistance has been reported in N. lugens both in the laboratory and in the field. Cycloxaprid (CYC), an oxa-bridged cis-nitromethylene neonicotinoid, showed high insecticidal activity against N. lugens and low cross-resistance in imidacloprid-resistant strains and field populations. Binding studies demonstrated that imidacloprid has two binding sites with different affinities (Kd = 3.18 ± 0.43 pm and 1.78 ± 0.19 nm) in N. lugens nAChRs. CYC was poor at displacing [3 H]imidacloprid at its high-affinity binding site (Ki = 159.38 ± 20.43 nm), but quite efficient at the low-affinity binding site (Ki = 1.27 ± 0.35 nm). These data showed that CYC had overlapping binding sites with imidacloprid only at its low-affinity binding site. Therefore, the low displacement ability of CYC against imidacloprid binding at its high-affinity site could partially explain the low cross-resistance of CYC in imidacloprid-resistant populations. The high insecticidal activity, low cross-resistance and different binding properties on insect nAChRs of CYC show that it is a potential insecticide for the control of N. lugens and related insect pests, especially ones with high resistance to neonicotinoids. © 2018 Society of Chemical Industry.

Imidacloprid-susceptible Nilaparvata lugens individuals exceeded resistant individuals in a mixture population with density pressure.

Fitness costs associated with insecticide resistance in pest insects have mainly been studied under optimal laboratory conditions. However, resistant insects face more stressors than just insecticides in the field, and how the resistant population reacts to these stressors is of practical importance for the control of pest insects such as the brown planthopper Nilaparvata lugens. The aim of the present study was to explore the impact of population density on the competitiveness of resistant and susceptible individuals. Two isogenic N. lugens populations, a highly imidacloprid-resistant population (HZ-R) with a resistance ratio (RR) of 227.10 and a relatively susceptible population (HZ-S) with an RR of 2.99, were created from a field-resistant population (HZ; RR 62.51). The high resistance levels of HZ-R and HZ were mainly attributable to the overexpression of multiple cytochrome P450 (CYP) genes such as CYP6ER1, CYP6AY1, CYP6CW1 and CYP4CE1 compared with HZ-S, this being supported by piperonyl butoxide synergism. HZ-R was observed to be more resistant to thiacloprid and etofenprox compared with HZ and HZ-S. Most interestingly, in high population density treatments, HZ-S individuals were much more competitive than HZ-R individuals. Imidacloprid-resistant individuals of N. lugens are less competitive than their susceptible counterparts under density pressure. © 2017 Society of Chemical Industry.

Expression Differences of Resistance-Related Genes Induced by Cycloxaprid Using qRT-PCR in the Female Adult of Sogatella furcifera (Hemiptera: Delphacidae).

As a newer cis-nitromethylene neonicotinoid pesticide at present, cycloxaprid has good industrialization prospects, including the management of imidacloprid-resistant populations, because this chemical have an excellent efficiency against rice planthoppers. Sogatella furcifera (Horváth) is the most economically important pest of rice worldwide and has developed resistance to many insecticides. This study focused on the expression change of these resistance genes, induced by cycloxaprid, involved in metabolic detoxification and receptor protein. Twenty-two differentially expressed genes (DEGs) that may be related with the insecticide resistance were found in the transcriptome of S. furcifera, including 2 cytochrome P450 genes, 2 glutathione S-transferase (GST) genes, 1 acid phosphatase (ACP) gene, 12 decarboxylase genes, 2 glycolipid genes, 1 cadherin gene, and 2 glycosyltransferase genes, which were up- or downregulated in response to an exposure of cycloxaprid. Furthermore, two P450 genes (CYP4 and CYP6 family, respectively), two decarboxylase genes, and one glycosyltransferase gene were validated by qRT-PCR. Expression differences of these genes verified successfully by qRT-PCR in response to different concentrations and times treated with cycloxaprid could explain the insecticide resistance mechanism under cycloxaprid stress in S. furcifera.

RNA interference of three up-regulated transcripts associated with insecticide resistance in an imidacloprid resistant population of Leptinotarsa decemlineata

The Colorado potato beetle, Leptinotarsa decemlineata (Say), is a major agricultural pest of potatoes in the Central Sands production region of Wisconsin. Previous studies have shown that populations of L. decemlineata have become resistant to many classes of insecticides, including the neonicotinoid insecticide, imidacloprid. Furthermore, L. decemlineata has multiple mechanisms of resistance to deal with a pesticide insult, including enhanced metabolic detoxification by cytochrome p450s and glutathione S-transferases. With recent advances in the transcriptomic analysis of imidacloprid susceptible and resistant L. decemlineata populations, it is possible to investigate the role of candidate genes involved in imidacloprid resistance. A recently annotated transcriptome analysis of L. decemlineata was obtained from select populations of L. decemlineata collected in the Central Sands potato production region, which revealed a subset of mRNA transcripts constitutively up-regulated in resistant populations. We hypothesize that a portion of the up-regulated transcripts encoding for genes within the resistant populations also encode for pesticide resistance and can be suppressed to re-establish a susceptible phenotype. In this study, a discrete set of three up-regulated targets were selected for RNA interference experiments using a resistant L. decemlineata population. Following the successful suppression of transcripts encoding for a cytochrome p450, a cuticular protein, and a glutathione synthetase protein in a select L. decemlineata population, we observed reductions in measured resistance to imidacloprid that strongly suggest these genes control essential steps in imidacloprid metabolism in these field populations.

Cycloxaprid: A novel cis-nitromethylene neonicotinoid insecticide to control imidacloprid-resistant cotton aphid (Aphis gossypii)

Imidacloprid is a nicotinic acetylcholine receptor (nAChR) agonist with potent insecticidal activity. However, resistance to imidacloprid is a significant threat and has been identified in several pest species. Cycloxaprid with cis-configuration is a novel neonicotinoid insecticide, which shows high activity against imidacloprid-resistant pests. The LC50 of imidacloprid against the resistant Aphis gossypii was 14.33mgL−1 while it was only 0.70mgL−1 for the susceptible population, giving a resistance ratio of 20.47. In this imidacloprid-resistant population, a point mutation (R81T) located in the loop D region of the nAChR β1 subunit was found out. But this point mutation did not decrease the activity of cycloxaprid against A. gossypii. The LC50 of cycloxaprid was 1.05 and 1.36mgL−1 for the imidacloprid-susceptible and imidacloprid-resistant populations, respectively. In addition, cycloxaprid provided better efficacies against resistant A. gossypii than imidacloprid in the fields. Although cycloxaprid was highly toxic to A. gossypii, it showed high selective activity between A. gossypii and its predominant natural enemies, Harmonia axyridis and Chrysoperla sinica. These results demonstrate that cycloxaprid is a promising insecticide against imidacloprid-resistant A. gossypii and suitable for the integrated pest management.

Genetics and realized heritability of resistance to imidacloprid in a poultry population of house fly, Musca domestica L. (Diptera: Muscidae) from Pakistan

Imidacloprid, a post-synaptic, nicotinic insecticide, has been commonly used for the management of different pests including Musca domestica worldwide. Many pests have developed resistance to this insecticide. A 16-fold imidacloprid-resistant population of M. domestica infesting poultry was selected using imidacloprid for 13 continuous generations to study the inheritance and realized heritability of resistance. Toxicological bioassay at G14 showed that the imidacloprid-selected population developed 106-fold resistance when compared to the susceptible population. Reciprocal crosses of susceptible and resistant populations showed an autosomal trait of resistance to imidacloprid in M. domestica. There was incompletely recessive resistance in F1 (Imida-SEL ♂×Susceptible ♀) and F1† (Imida-SEL ♀×Susceptible ♂) having dominance value 0.53 and 0.31, respectively. Monogenic model of inheritance showed that imidacloprid resistance was controlled by multiple factors. The realized heritability value was 0.09 in the imidacloprid-selected population of M. domestica. It was concluded that imidacloprid resistance in M. domestica was autosomally inherited, incompletely recessive and polygenic. These findings should be helpful for better and more successful management of M. domestica.

Toxicities and sublethal effects of seven neonicotinoid insecticides on survival, growth and reproduction of imidacloprid‐resistant cotton aphid, Aphis gossypii

Imidacloprid has been a major neonicotinoid insecticide for controlling Aphis gossypii (Glover) (Homoptera: Aphididae) and other piercing-sucking pests. However, the resistance to imidacloprid has been recorded in many target insects. At the same time, cross-resistance of imidacloprid and other insecticides, especially neonicotinoid insecticides, has been detected. Results showed that the level of cross-resistance was different between imidacloprid and tested neonicotinoid insecticides (no cross-resistance: dinotefuran, thiamethoxam and clothianidin; a 3.68-5.79-fold cross-resistance: acetamiprid, nitenpyram and thiacloprid). In the study of sublethal effects, imidacloprid at LC(20) doses could suppress weight gain and honeydew excretion, but showed no significant effects on longevity and fecundity of the imidacloprid-resistant cotton aphid, A. gossypii. However, other neonicotinoid insecticides showed significant adverse effects on biological characteristics (body weight, honeydew excretion, longevity and fecundity) in the order of dinotefuran > thiamethoxam and clothianidin > nitenpyram > thiacloprid and acetamiprid. The results indicated that dinotefuran is the most effective insecticide for use against imidacloprid-resistant A. gossypii. To avoid further resistance development, the use of nitenpyram, acetamiprid and thiacloprid should be avoided on imidacloprid-resistant populations of A. gossypii.