Susceptibility To Imidacloprid Research Articles

Transcription factors, cap ‘n’ collar isoform C regulates the expression of CYP450 genes involving in insecticides susceptibility in Locusta migratoria

BackgroundThe cap ‘n’ collar (Cnc) belongs to the Basic Leucine Zipper (bZIP) transcription factor super family. Cap ‘n’ collar isoform C (CncC) is highly conserved in the animal kingdom. CncC contributes to the regulation of growth, development, and aging and takes part in the maintenance of homeostasis and the defense against endogenous and environmental stress. Insect CncC participates in the regulation of various kinds of stress-responsive genes and is involved in the development of insecticide resistance. ResultsIn this study, one full-length CncC sequence of Locusta migratoria was identified and characterized. Upon RNAi silencing of LmCncC, insecticide bioassays showed that LmCncC played an essential role in deltamethrin and imidacloprid susceptibility. To fully investigate the downstream genes regulated by LmCncC and further identify the LmCncC-regulated genes involved in deltamethrin and imidacloprid susceptibility, a comparative transcriptome was constructed. Thirty-five up-regulated genes and 73 down-regulated genes were screened from dsLmCncC-knockdown individuals. We selected 22 LmCncC-regulated genes and verified their gene expression levels using RT-qPCR. Finally, six LmCYP450 genes belonging to the CYP6 family were selected as candidate detoxification genes, and LmCYP6FD1 and LmCYP6FE1 were further validated as detoxification genes of insecticides via RNAi, insecticide bioassays, and metabolite identification. ConclusionsOur data suggest that the locust CncC gene is associated with deltamethrin and imidacloprid susceptibility via the regulation of LmCYP6FD1 and LmCYP6FE1, respectively.

Novel_miR-1517 mediates CYP6CM1 to regulate imidacloprid resistance in Bemisia tabaci (Hemiptera: Gennadius)

Bemisia tabaci (Hemiptera: Gennadius) is a notorious pest that is capable of feeding on >600 kinds of agricultural crops. Imidacloprid is critical in managing pest with sucking mouthparts, such as B. tabaci. However, the field population of B. tabaci has evolved resistance because of insecticide overuse. The overexpression of the detoxification enzyme cytochrome P450 monooxygenase is considered the main mechanism of imidacloprid resistance, but the mechanism underlying gene regulation remains unclear. MicroRNAs are a type of endogenous small molecule compounds that is fundamental in regulating gene expression at the post-transcriptional level. Whether miRNAs are related to the imidacloprid resistance of B. tabaci remains unknown. To gain deep insight into imidacloprid resistance, we conducted on miRNAs expression profiling of two B. tabaci Mediterranean (MED) strains with 19-fold resistance through deep sequencing of small RNAs. A total of 8 known and 1591 novel miRNAs were identified. In addition, 16 miRNAs showed significant difference in expression levels between the two strains, as verified by quantitative reverse transcription PCR. Among these, novel_miR-376, 1517, and 1136 significantly expressed at low levels in resistant samples, decreasing by 36.9%, 60.2%, and 15.6%, respectively. Moreover, modulating novel_miR-1517 expression by feeding with 1517 inhibitor and 1517 mimic significantly affected B. tabaci imidacloprid susceptibility by regulating CYP6CM1 expression. In this article, miRNAs related to imidacloprid resistance of B. tabaci were systematically screened and identified, providing important information for the miRNA-based technological innovation for this pest management.

The gut symbiont Sphingomonas mediates imidacloprid resistance in the important agricultural insect pest Aphis gossypii Glover

BackgroundNeonicotinoid insecticides are applied worldwide for the control of agricultural insect pests. The evolution of neonicotinoid resistance has led to the failure of pest control in the field. The enhanced detoxifying enzyme activity and target mutations play important roles in the resistance of insects to neonicotinoid resistance. Emerging evidence indicates a central role of the gut symbiont in insect pest resistance to pesticides. Existing reports suggest that symbiotic microorganisms could mediate pesticide resistance by degrading pesticides in insect pests.ResultsThe 16S rDNA sequencing results showed that the richness and diversity of the gut community between the imidacloprid-resistant (IMI-R) and imidacloprid-susceptible (IMI-S) strains of the cotton aphid Aphis gossypii showed no significant difference, while the abundance of the gut symbiont Sphingomonas was significantly higher in the IMI-R strain. Antibiotic treatment deprived Sphingomonas of the gut, followed by an increase in susceptibility to imidacloprid in the IMI-R strain. The susceptibility of the IMI-S strain to imidacloprid was significantly decreased as expected after supplementation with Sphingomonas. In addition, the imidacloprid susceptibility in nine field populations, which were all infected with Sphingomonas, increased to different degrees after treatment with antibiotics. Then, we demonstrated that Sphingomonas isolated from the gut of the IMI-R strain could subsist only with imidacloprid as a carbon source. The metabolic efficiency of imidacloprid by Sphingomonas reached 56% by HPLC detection. This further proved that Sphingomonas could mediate A. gossypii resistance to imidacloprid by hydroxylation and nitroreduction.ConclusionsOur findings suggest that the gut symbiont Sphingomonas, with detoxification properties, could offer an opportunity for insect pests to metabolize imidacloprid. These findings enriched our knowledge of mechanisms of insecticide resistance and provided new symbiont-based strategies for control of insecticide-resistant insect pests with high Sphingomonas abundance.

Insecticide resistance in field populations of the pear psyllids Cacopsylla permixta and Cacopsylla bidens in Iran

AbstractCacopsylla permixta and Cacopsylla bidens are major pests of pears in Iran. They are commonly sprayed with various insecticides up to 12 times a year but control by these means is no longer reliable, suggesting the evolution of resistance. Here, we report about 60‐fold variation in diazinon susceptibility both among five geographic populations of C. bidens and among three geographic populations of C. permixta, as well as 12‐ to 14‐fold variations in imidacloprid susceptibility in these populations. The levels of resistance to the two compounds were highly correlated in the different populations. Esterase, cytochrome P450, and glutathione S‐transferase activities all varied by about two‐fold among the different populations and were also highly correlated with population resistance to the two insecticides, implying differences in metabolism contribute to the differences in resistance. Bioassays with inhibitors of each class of enzyme indicated all three, but particularly the esterases, contributed to the variation in diazinon resistance, while esterase and, in particular, cytochrome P450 activities contributed to the variation in imidacloprid resistance. The most resistant populations of each species also showed some resistance to chlorpyrifos and acetamiprid, suggesting there may be wide cross resistance to organophosphates and neonicotinoids in these populations. The sampled psyllid populations showed lower levels of resistance to representatives of two other classes of insecticides currently used in Iran, spirotetramat and abamectin, though resistance to spirotetramat was detectable and was correlated with diazinon and imidacloprid resistance across populations. We conclude that metabolic resistance to a range of insecticides is now evolving in field populations of both species in Iran, warranting timely development of resistance management plans.

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

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. 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. 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.

UV-irradiation and leaching in water reduce the toxicity of imidacloprid-contaminated leaves to the aquatic leaf-shredding amphipod Gammarus fossarum.

Systemic neonicotinoid insecticides such as imidacloprid are increasingly applied against insect pest infestations on forest trees. However, leaves falling from treated trees may reach nearby surface waters and potentially represent a neonicotinoid exposure source for aquatic invertebrates. Given imidacloprid's susceptibility towards photolysis and high water solubility, it was hypothesized that the leaves' toxicity might be modulated by UV-irradiation during decay on the forest floor, or by leaching and re-mobilization of the insecticide from leaves within the aquatic ecosystem. To test these hypotheses, the amphipod shredder Gammarus fossarum was fed (over 7 d; n = 30) with imidacloprid-contaminated black alder (Alnus glutinosa) leaves that had either been pre-treated (i.e., leached) in water for up to 7 d or UV-irradiated for 1 d (at intensities relevant during autumn in Central Europe) followed by a leaching duration of 1 d. Gammarids' feeding rate, serving as sublethal response variable, was reduced by up to 80% when consuming non-pretreated imidacloprid-contaminated leaves compared to imidacloprid-free leaves. Moreover, both leaching of imidacloprid from leaves (for 7 d) as well as UV-irradiation reduced the leaves' imidacloprid load (by 46 and 90%) thereby mitigating the effects on gammarids' feeding rate to levels comparable to the respective imidacloprid-free controls. Therefore, natural processes, such as UV-irradiation and re-mobilization of foliar insecticide residues in water, might be considered when evaluating the risks systemic insecticide applications in forests might pose for aquatic organisms in nearby streams.

Temporal patterns of imidacloprid resistance throughout a growing season in Leptinotarsa decemlineata populations.

BACKGROUNDThe Colorado potato beetle, Leptinotarsa decemlineata (Say), is a major agricultural pest of commercial potatoes. Pest managers use a combination of control tactics to limit populations, including multiple insecticides. Finding a window of insecticide susceptibility and understanding genetic responses to insecticide exposure during a growing season may provide novel management recommendations for L. decemlineata.RESULTSWe examined temporal changes (during one growing season) in phenotypic response between a susceptible population and an imidacloprid‐resistant population. Beetles remained more susceptible to imidacloprid in the susceptible population throughout the growing season. Estimated mean LC50 values varied throughout the growing season in the resistant population, with increased susceptibility among overwintered and recently emerged adult beetles compared with a heightened level of resistance in the second generation. RNA transcript abundance was compared among multiple time points through the growing season, showing that cuticular proteins and cytochrome p450s were highly upregulated during peaks of measured resistance.CONCLUSIONTemporal variation in imidacloprid susceptibility of L. decemlineata was observed, which included early time points of susceptibility and later peaks in resistance. Heightened resistance occurred during the second generation and correlated to increased transcript abundance of multiple mechanisms of resistance, including multiple cuticular protein and cytochrome p450 transcripts. © 2016 The Authors. Pest Management Science published by John Wiley & Sons Ltd on behalf of Society of Chemical Industry.

Biochemical mechanisms of imidacloprid resistance in Nilaparvata lugens: Over-expression of cytochrome P450 CYP6AY1

Imidacloprid is a key insecticide extensively used for control of Nilaparvata lugens, and its resistance had been reported both in the laboratory selected strains and field populations. A target site mutation Y151S in two nicotinic acetylcholine receptor subunits and enhanced oxidative detoxification have been identified in the laboratory resistant strain, contributing importantly to imidacloprid resistance in N. lugens. To date, however, imidacloprid resistance in field population is primarily attributable to enhanced oxidative detoxification by over-expressed P450 monooxygenases. A resistant strain (Res), originally collected from a field population and continuously selected in laboratory with imidacloprid for more than 40 generations, had 180.8-fold resistance to imidacloprid, compared to a susceptible strain (Sus). Expression of different putative P450 genes at mRNA levels was detected and compared between Res and Sus strains, and six genes were found expressed significantly higher in Res strain than in Sus strain. CYP6AY1 was found to be the most different expressed P450 gene and its mRNA level in Res strain was 17.9 times of that in Sus strain. By expressing in E. coli cells, CYP6AY1 was found to metabolize imidacloprid efficiently with initial velocity calculated of 0.851 ± 0.073 pmol/min/pmol P450. When CYP6AY1 mRNA levels in Res strain was reduced by RNA interference, imidacloprid susceptibility was recovered. In four field populations with different resistance levels, high levels of CYP6AY1 transcript were also found. In vitro and in vivo studies provided evidences that the over-expression of CYP6AY1 was one of the key factors contributing to imidacloprid resistance in the laboratory selected strain Res, which might also be the important mechanism for imidacloprid resistance in field populations, when the target site mutation was not prevalent at present.

Migration analysis of Nilaparvata lugens (Hemiptera: Delphacidae) from the Philippines to Taiwan under typhoon-induced windy conditions

The population of Nilaparvata lugens (Stal) in Taiwan is believed to belong to the East Asian population of the species, which has low susceptibility to the insecticide imidacloprid. The population in the Philippines belongs to the Southeast Asian population, which has high susceptibility to imidacloprid. In this study, long-winged adults of N. lugens collected in paddy fields in Taitung, southeastern Taiwan, just after a typhoon, were tested for imidacloprid susceptibility after rearing for 4–17 generations. The migration source was also estimated by backward trajectory analysis. Our findings were, first, that its curved dosage–mortality plots for imidacloprid suggested that the Taitung population is heterogeneous in its susceptibility to the insecticide. Source estimation found areas on Luzon Island of the Philippines, mainly, and southern China, partially, as possible sources. Before the typhoon approached, however, light trap monitoring data indicated some local populations might have occurred in the survey area. Therefore, with other pieces of circumstantial evidence, this study suggested that migration of N. lugens to Taiwan from the Philippines, and, simultaneously, possibly southern China occurred because of the typhoon, with the insects partly crossing the boundary of the Asian populations, and that a mixture of individuals with different susceptibility to the insecticide happened to form the Taitung population at the time of collection.

Imidacloprid Susceptibility Survey and Selection Risk Assessment in Field Populations of Nilaparvata lugens (Homoptera: Delphacidae)

Imidacloprid has been used for many years to control planthopper Nilaparvata lugens (Stål) (Homoptera: Delphacidae) in China. To provide resistance assessment for the national insecticide resistance management program, we collected a total of 42 samples of the planthoppers from 27 locations covering eight provinces to monitor their dose responses and susceptibility changes to imidacloprid over an 11-yr period (1996–2006). Results showed that most field populations maintained susceptibility from 1996 to 2003 except for a population from Guilin, Guangxi, in 1997, which showed a low level of resistance to imidacloprid. However, surveys conducted in 2005 indicated that 16 populations from six provinces quickly developed resistance with resistance ratios ranging from 79 to 811. The data collected in 2006 revealed that the resistance levels in 12 populations collected from seven different provinces decreased slightly (RR = 107–316), except the Tongzhou population (Jiangsu Province), which developed 625-fold resistance. Dominant and intensive use of imidacloprid in a wide range of rice, Oryza savita L., growing areas might be a driving force for the resistance development. Migration of the insect also significantly boosted the resistance levels due to extensive and intensive use of imidacloprid in emigrating areas and continuous postmigration sprays of the chemical. In addition, laboratory resistance selection using imidacloprid showed that resistance ratio increased to 14-fold after 27 generations, suggesting that quick resistance development might be associated with more frequent applications of the insecticide in recent years.