Clements J, Sanchez-Sedillo B, Bradfield CA, Groves RL. PLoS ONE. 2018;13(10):e0205881. doi: 10.1371/journal.pone.0205881.
The Colorado potato beetle, Leptinotarsa decemlineata (Say), is an agricultural pest of commercial potatoes in parts of North America, Europe, and Asia. Plant protection strategies within this geographic range employ a variety of pesticides to combat not only the insect, but also plant pathogens. Previous research has shown that field populations of Leptinotarsa decemlineata have a chronological history of resistance development to a suite of insecticides, including the Group 4A neonicotinoids. The aim of this study is to contextualize the transcriptomic response of Leptinotarsa decemlineata when exposed to the neonicotinoid insecticide imidacloprid, or the fungicides boscalid or chlorothalonil, in order to determine whether these compounds induce similar detoxification mechanisms. We found that chlorothalonil and imidacloprid induced similar patterns of transcript expression, including the up-regulation of a cytochrome p450 and a UDP-glucuronosyltransferase transcript, which belong to protein families associated with xenobiotic metabolism. Further, transcriptomic responses varied among individuals within the same treatment group, suggesting individual insects' responses vary within a population and may cope with chemical stressors in a variety of manners.
Ruiz-Arroyo V, García-Robles I, Ochoa-Campuzano C, et al. Insect Mol Biol. 2017;26(2):204-214. doi: 10.1111/imb.12285.
Bacillus thuringiensis parasporal crystal proteins (Cry proteins) are insecticidal pore-forming toxins that bind to specific receptor molecules on the brush border membrane of susceptible insect midgut cells to exert their toxic action. In the Colorado potato beetle (CPB), a coleopteran pest, we previously proposed that interaction of Cry3Aa toxin with a CPB ADAM10 metalloprotease is an essential part of the mode of action of this toxin. Here, we annotated the gene sequence encoding an ADAM10 metalloprotease protein (CPB-ADAM10) in the CPB genome sequencing project, and using RNA interference gene silencing we demonstrated that CPB-ADAM10 is a Cry3Aa toxin functional receptor in CPB. Cry3Aa toxicity was significantly lower in CPB-ADAM10 silenced larvae and in vitro toxin pore-forming ability was greatly diminished in lipid planar bilayers fused with CPB brush border membrane vesicles (BBMVs) prepared from CPB-ADAM10 silenced larvae. In accordance with our previous data that indicated this toxin was a substrate of ADAM10 in CPB, Cry3Aa toxin membrane-associated proteolysis was altered when CPB BBMVs lacked ADAM10. The functional validation of CPB-ADAM10 as a Cry3Aa toxin receptor in CPB expands the already recognized role of ADAM10 as a pathogenicity determinant of pore-forming toxins in humans to an invertebrate species.
Gaddelapati SC, Kalsi M, Roy A, Palli SR. Insect Biochem Mol Biol. 2018;99:54-62. doi: 10.1016/j.ibmb.2018.05.006.
The Colorado potato beetle (CPB), Leptinotarsa decemlineata developed resistance to imidacloprid after exposure to this insecticide for multiple generations. Our previous studies showed that xenobiotic transcription factor, cap 'n' collar isoform C (CncC) regulates the expression of multiple cytochrome P450 genes, which play essential roles in resistance to plant allelochemicals and insecticides. In this study, we sought to obtain a comprehensive picture of the genes regulated by CncC in imidacloprid-resistant CPB. We performed sequencing of RNA isolated from imidacloprid-resistant CPB treated with dsRNA targeting CncC or gene coding for green fluorescent protein (control). Comparative transcriptome analysis showed that CncC regulated the expression of 1798 genes, out of which 1499 genes were downregulated in CncC knockdown beetles. Interestingly, expression of 79% of imidacloprid induced P450 genes requires CncC. We performed quantitative real-time PCR to verify the reduction in the expression of 20 genes including those coding for detoxification enzymes (P450s, glutathione S-transferases, and esterases) and ABC transporters. The genes coding for ABC transporters are induced in insecticide resistant CPB and require CncC for their expression. Knockdown of genes coding for ABC transporters simultaneously or individually caused an increase in imidacloprid-induced mortality in resistant beetles confirming their contribution to insecticide resistance. These studies identified CncC as a transcription factor involved in regulation of genes responsible for imidacloprid resistance. Small molecule inhibitors of CncC or suppression of CncC by RNAi could provide effective synergists for pest control or management of insecticide resistance.
Clements J, Schoville S, Clements N, Chapman S, Groves RL. Pest Manag Sci. 2017;73(3):641-650. doi: 10.1002/ps.4480.
BACKGROUND: The 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. RESULTS: We 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. CONCLUSION: Temporal 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.
Arain MS, Wan P, Shakeel M, et al. Phytoparasitica. 2017;45(1):103-111. doi: 10.1007/s12600-016-0560-z.
The speed of toxic action of an insecticide is an indicator for control efficacy and has considerable practical importance. For agricultural pest control, fast-acting is an important feature for an insecticide to consistently reduce the amount of feeding damage. Butene-fipronil is a novel compound obtained via the structural modification of fipronil. However, information about the toxicity and speed of toxic action is still limited. In the present paper, we compared the toxic feature of butene-fipronil with seven other insecticides, of which imidacloprid and abamectin are slow-acting insecticides, and acephate, endosulfan, methomyl, α-cypermethrin and spinosad are fast-acting insecticides. We found that the contact and stomach toxicities of butene-fipronil were among the highest ever estimated to Leptinotarsa decemlineata and Drosophila melanogaster. The speed of toxic action of butene-fipronil was determined using median lethal time (LT50) at a dose (concentration) equivalent to LD80 values. For L. decemlineata, the values for butene-fipronil, imidacloprid, abamectin, acephate, endosulfan, methomyl, cypermethrin and spinosad were calculated to be 39.9, 36.5, 37.5, 20.2, 22.4, 23.8, 16.4 and 23.1 h, respectively. Those for D. melanogaster were 29.8, 31.5, 29.4, 14.0, 20.3, 18.1, 13.5, and 20.1 h, respectively. ANOVA analysis showed that butene-fipronil, imidacloprid, abamectin had similar LT50 values, whereas acephate, endosulfan, methomyl, spinosad and cypermethrin had comparable LT50 values. Thus, butene-fipronil belongs to slow-acting insecticides. Our results provide more empirical information for butene-fipronil potential application.
Maliszewska J, Tęgowska E. Int J Pest Manage. 2017;63(4):331-340.
The effectiveness of insecticides differs with changes in temperature, but insecticide toxicities are determined at constant temperatures. Constant thermal conditions do not occur in the field, where insects can change their behaviors to achieve a preferred temperature. The aim of this study was to assess whether the choice of ambient temperature affects the mortality rate of intoxicated firebugs and Colorado potato beetles. The insects' mortality following insecticide exposure was monitored at constant temperatures (15, 25, and 35°C) as well as in a thermal gradient system, where the insects could freely select their preferred ambient temperature. Firebugs treated with oxadiazine showed 58% higher mortality when held at a constant temperature post-treatment compared to mortality levels seen when able to choose a preferred temperature in a thermal gradient. Similar results were seen in Colorado potato beetles treated with oxadiazine (15%-33% higher mortality in constant vs. preferred temperature) or organophosphate (36% higher mortality in constant vs. preferred temperature). The insects' ability to mitigate the impacts of pesticide exposure by selecting more beneficial thermal conditions is an important consideration for pest management. Therefore, the application rates of insecticides used under field conditions should be additionally analyzed to take this factor into account.
Xu Q, Meng Q, Shi J, Deng P, Guo W, Li G. Pestic Biochem Physiol. 2017;143:173-180. doi: 10.1016/j.pestbp.2017.07.010.
To accomplish consistent, long-term, integrated management (IPM) of the Colorado potato beetle, Leptinotarsa decemlineata (Say), research assessing the potential of novel, IPM-compatible insecticides is essential. Novaluron is a potent benzoylurea insecticide. In the present paper, we found that novaluron ingestion by the fourth-instar larvae inhibited foliage consumption, reduced larval fresh weight, and delayed development period, in a dose dependent manner. Most of the resulting larvae fail to pupate, and died at prepupae stage, with larvicidal activity comparable with those of cyhalothrin and spinosad but lower than those of fipronil and abamectin. Moreover, many surviving pupae that fed novaluron failed to emerge as adults, in a dose dependent pattern. Furthermore, feeding of novaluron significantly decreased chitin contents in body carcass (without midgut) and integument specimen, whereas the chitin concentration in the midgut peritrophic matrix was not affected. Furthermore, uridine diphosphate-N-acetylglucosamine-pyrophosphorylase gene (LdUAP1) and chitin synthase Aa (LdChSAa), which were mainly responsible for chitin biosynthesis in ectodermally-derived tissues, were suppressed and activated respectively after novaluron ingestion. Therefore, novaluron is an effective benzoylurea insecticide to L. decemlineata fourth-instar larvae. It inhibited chitin biosynthesis in ectodermally-derived tissues, disrupted ecdysis, impaired pupation and adult emergence, and led to death in juvenile life stages.
Crossley MS, Chen YH, Groves RL, Schoville SD. Mol Ecol. 2017;26(22):6284-6300. doi: 10.1111/mec.14339.
The ability of insect pests to rapidly and repeatedly adapt to insecticides has long challenged entomologists and evolutionary biologists. Since Crow's seminal paper on insecticide resistance in 1957, new data and insights continue to emerge that are relevant to the old questions about how insecticide resistance evolves: such as whether it is predominantly mono- or polygenic, and evolving from standing vs. de novo genetic variation. Many studies support the monogenic hypothesis, and current management recommendations assume single- or two-locus models. But inferences could be improved by integrating data from a broader sample of pest populations and genomes. Here, we generate evidence relevant to these questions by applying a landscape genomics framework to the study of insecticide resistance in a major agricultural pest, Colorado potato beetle, Leptinotarsa decemlineata (Say). Genome-environment association tests using genomic variation from 16 populations spanning gradients of landscape variables associated with insecticide exposure over time revealed 42 strong candidate insecticide resistance genes, with potentially overlapping roles in multiple resistance mechanisms. Measurements of resistance to a widely used insecticide, imidacloprid, among 47 L. decemlineata populations revealed heterogeneity at a small (2 km) scale and no spatial signature of origin or spread throughout the landscape. Analysis of nucleotide diversity suggested candidate resistance loci have undergone varying degrees of selective sweeps, often maintaining similar levels of nucleotide diversity to neutral loci. This study suggests that many genes are involved in insecticide resistance in L. decemlineata and that resistance likely evolves from both de novo and standing genetic variation.
Tryjanowski P, Sparks TH, Blecharczyk A, Małecka-Jankowiak I, Switek S, Sawinska Z. American Journal of Potato Research. 2018;95(1):26-32. doi: 10.1007/s12230-017-9611-3.
Potato Solanum tuberosum is one of the world's four most important crops. Its cultivation is steadily increasing in response to the need to feed a growing world population. The yield of potato is influenced inter alia by both climate and pests. The main defoliator pest of potato is Colorado potato beetle Leptinotarsa decemlineata. Using data from a long-term experiment (1958-2013) in western Poland, we show that increasing temperature has affected the trophic relationship between potato and Colorado potato beetle. The planting, leafing, flowering and harvest dates for potato were advanced, after controlling for different cultivars, by 2.00 days, 3.04 days, 3.80 days and 3.42 days respectively for every 1°C increase in temperature. In contrast, first treatment against Colorado potato beetle advanced by 4.66 days for every 1°C increase in temperature, and, furthermore, the number of treatments against the beetle increased by 0.204 per 1°C increase in temperature. This suggests that the beetle responds faster to increasing temperature than the plant does, but both parts of the system are probably greatly modified by farming practices.
Kitaev, K. A., I. S. Mardanshin, E. V. Surina, T. L. Leontieva, M. B. Udalov, and G. V. Benkovskaya. Russ J Genet Appl Res (2017) 7: 36. doi:10.1134/S2079059717010063
The main approach to pest control consists in the application of chemical insecticides. The efficacy of insecticides is reduced due to the development of resistance by pest populations. This is an especially important problem with the Colorado potato beetle. There are different strategies for the use of insecticides to slow the formation of resistance. Based on the results of long-term studies, we propose a hypothesis on delaying the development of resistance by applying insecticides at low doses. To test this hypothesis, we built predictive discrete genetic models of resistance in the Colorado potato beetle populations. The model based on the classical equations of population genetics has been supplemented by various factors. Calculations of the survival rates of individuals of the Colorado potato beetle were carried out taking into account the statistical regularities of the distribution of toxic substance after treatment by insecticides. We calculated the survival rates for different genotypes using a lognormal distribution after at least doubling the insecticide dose. The factor of differential mortality during winter was additionally introduced into the model. The use of phenetic markers of nonspecific resistance to environmental factors allowed us to compute the model with mediated intergenic interactions. Various hypotheses on the strategies of overcoming resistance have been tested using this model. The calculations demonstrated that use of insecticides at the minimum effective dose (low dose) leads to a slower increase in the proportion of resistant individuals in the populations of the Colorado potato beetle for two seasons. Resistance develops much more slowly following alternate treatment with insecticides from different chemical classes. The best strategy is through off-season treatment with lower doses of insecticides of different chemical classes.