Resistance and cross-resistance to imidacloprid and thiamethoxam in the Colorado potato beetle Leptinotarsa decemlineata

One of the major challenges in managing the Colorado potato beetle, Leptinotarsa decemlineata (Say) is its remarkable ability to develop resistance to virtually every insecticide that has ever been used against it. Resistance is particularly common throughout northeastern USA as far north as Maine. The first instances of resistance to imidacloprid have already been reported from several locations in New York, Delaware and southern Maine. Rotating insecticides with different modes of action may delay insecticide resistance, but successful implementation of this technique depends on a good understanding of resistance and cross-resistance patterns in populations of target pests. LC(50) values were measured for imidacloprid and thiamethoxam in Colorado potato beetle populations from a variety of locations in the USA and Canada using diet incorporation bioassays. The field performance of imidacloprid, thiamethoxam and clothianidin against imidacloprid-resistant beetles on a commercial potato farm in southern Maine was also evaluated. Correlation between LC(50) values for imidacloprid and thiamethoxam was highly significant, even when populations previously exposed to thiamethoxam were excluded from the analysis. There was no statistically detectable difference in the LC(50) values between populations exposed to both insecticides and to imidacloprid alone. Applications of neonicotinoid insecticides at planting delayed build-up of imidacloprid-resistant beetle populations on field plots by 1-2 weeks but failed to provide adequate crop protection. Consistently with bioassay results, there was also substantial cross-resistance among the three tested neonicotinoid insecticides. Results of the present study support the recommendation to avoid rotating imidacloprid with thiamethoxam as a part of a resistance management plan.     

Neural actions of imidacloprid and their involvement in resistance in the Colorado potato beetle, Leptinotarsa decemlineata (Say)

BACKGROUND: The development of resistance to imidacloprid in eastern US populations of the Colorado potato beetle (CPB), Leptinotarsa decemlineata (Say), threatens this critical use for neonicotinoid insecticides. Previous pharmacokinetic studies with resistant adult CPBs provided no explanation for the high resistance level (over 200-fold) to topically applied imidacloprid. The authors assessed the neural activity of imidacloprid by recording spontaneous activity from a motor nerve leaving the isolated central nervous system to compare the sensitivity of the latter to imidacloprid between susceptible and resistant CPBs. RESULTS: On the isolated central nervous system, imidacloprid was initially neuroexcitatory, and neuroinhibitory at higher concentrations. The neuroexcitatory action of imidacloprid was blocked by coapplication of a specific nAChR antagonist, methyllycaconitine, indicating that it is a result of action on nAChRs. The sensitivity to the neuroexcitatory and inhibitory activities of imidacloprid varied independently among individuals in each population. The sensitivity of the central nervous system of resistant CPBs to excitation by imidacloprid did not differ from that of susceptible insects, but the sensitivity to inhibition by imidacloprid was reduced 52- to 58-fold, indicating a possible change in the sensitivity of at least one subgroup of nAChRs. CONCLUSION: This study provides evidence that reduced nerve sensitivity to the blocking action of imidacloprid is associated with imidacloprid resistance in the CPB.     

Ingested RNA interference for managing the populations of the Colorado potato beetle, Leptinotarsa decemlineata

BACKGROUND: RNA interference (RNAi) is a breakthrough technology for conducting functional genomics studies and also as a potential tool for crop protection against insect pests. The major challenge for efficient pest control using RNAi in the field is the development of efficient and reliable methods for production and delivery of double‐stranded RNA (dsRNA). In this paper, the potential of feeding dsRNA expressed in bacteria or synthesized in vitro to manage populations of Colorado potato beetle, Leptinotarsa decemlineata (Say) (CPB), was investigated. RESULTS: Feeding RNAi successfully triggered the silencing of all five target genes tested and caused significant mortality and reduced body weight gain in the treated beetles. This study provides the first example of an effective RNAi response in insects after feeding dsRNA produced in bacteria. CONCLUSION: These results suggest that the efficient induction of RNAi using bacteria to deliver dsRNA is a possible method for management of CPB. This could be also a promising bioassay approach for genome‐wide screens to identify effective target genes for use as novel RNAi‐based insecticides. Copyright © 2010 Society of Chemical Industry     

Effect of strip-intercropping potato and annual alfalfa on populations of Leptinotarsa decemlineata Say and its predators

Intercropping systems are practiced to reducing pest density, enhancing predator's diversity and stabling crop yield. We evaluated the effect of strip-intercropping potato and annual alfalfa on populations of Leptinotarsa decemlineata Say, predator's biodiversity and potato yield over two seasons (2014 and 2015). Our results revealed that the densities of egg masses, eggs, larvae and adults of Colorado potato beetle (CPB) were significantly lower in intercrops than in monoculture. The main CPB predators recorded on potato plants (ladybirds and the green lacewing) showed a significant increase in the density at each of the three intercrops than in monoculture. The percentage of dry tubers weight loss was 40.9%–42.6% in monoculture, 16.3%–18.7% in 6P: 2A and <10% in 2P: 2A and 4P: 2A in two studied years. Our results suggest that strip-intercropping potato with annual alfalfa, particularly in 2P: 2A and 4P: 2A patterns may be an effective way in integrated management of CPB for reducing pest density, enhancing the presence of predators in potato fields and improving potato yield.     

The Red Queen in a potato field: integrated pest management versus chemical dependency in Colorado potato beetle control

Originally designed to reconcile insecticide applications with biological control, the concept of integrated pest management (IPM) developed into the systems‐based judicious and coordinated use of multiple control techniques aimed at reducing pest damage to economically tolerable levels. Chemical control, with scheduled treatments, was the starting point for most management systems in the 1950s. Although chemical control is philosophically compatible with IPM practices as a whole, reduction in pesticide use has been historically one of the main goals of IPM practitioners. In the absence of IPM, excessive reliance on pesticides has led to repeated control failures due to the evolution of resistance by pest populations. This creates the need for constant replacement of failed chemicals with new compounds, known as the ‘insecticide treadmill’. In evolutionary biology, a similar phenomenon is known as the Red Queen principle – continuing change is needed for a population to persevere because its competitors undergo constant evolutionary adaptation. The Colorado potato beetle, Leptinotarsa decemlineata (Say), is an insect defoliator of potatoes that is notorious for its ability to develop insecticide resistance. In the present article, a review is given of four case studies from across the United States to demonstrate the importance of using IPM for sustainable management of a highly adaptable insect pest. Excessive reliance on often indiscriminate insecticide applications and inadequate use of alternative control methods, such as crop rotation, appear to expedite evolution of insecticide resistance in its populations. Resistance to IPM would involve synchronized adaptations to multiple unfavorable factors, requiring statistically unlikely genetic changes. Therefore, integrating different techniques is likely to reduce the need for constant replacement of failed chemicals with new ones. © 2014 Society of Chemical Industry     

Acute and sublethal toxicity of novaluron, a novel chitin synthesis inhibitor, to Leptinotarsa decemlineata (Coleoptera: Chrysomelidae)

The acute and sublethal toxicities of novaluron, a novel chitin synthesis inhibitor, to a laboratory-reared insecticide-susceptible strain of Colorado potato beetle, Leptinotarsa decemlineata (Say), were determined. Novaluron exhibited excellent residual (120 h LC(50) = 0.42 mg litre(-1)) and good direct contact (120 h LC(50) = 27 mg litre(-1)) activity against second-instar larvae (L2). Hatch of eggs exposed by direct contact to novaluron solutions > or =100 mg litre(-1) was significantly reduced, as was the ability of emerged first-instar larvae to moult. L2 from eggs exposed to > or =100 mg litre(-1) novaluron weighed significantly less (P < 0.0001) than those from untreated eggs. However, L2 from eggs treated with 1 mg litre(-1) novaluron weighed significantly more (P < or = 0.05) than those from untreated eggs, suggesting novaluron can have a hormetic effect on L decemlineata larval development. Leptinotarsa decemlineata mating pairs fed foliage treated with novaluron at 25 or 75 g AI ha(-1) produced approximately 25% fewer egg masses and eggs per mass. Hatch of eggs on treated foliage was almost completely suppressed, and longevity of male beetles was reduced by approximately 50% when fed foliage treated with novaluron at 75 g AI ha(-1).     

Evaluation of the resistance of transgenic potato plants expressing various levels of Cry3A against the Colorado potato beetle (Leptinotarsa decemlineata Say) in the laboratory and field

BACKGROUND: The Colorado potato beetle (CPB), Leptinotarsa decemlineata Say, is a destructive pest. The CPB is a quarantine pest in China, but has now invaded the Xinjiang Uygur Autonomous Region and is continuing to spread eastwards. To control the damage and overspreading, transgenic potato plants expressing Cry3A toxin were developed, and their resistance to CPB was evaluated by bioassays in the laboratory and field in 2009, 2010 and 2011. RESULTS: The insect resistance of the high‐dose (HD) transgenic lines was significantly greater than the middle‐dose (MD) and low‐dose (LD) transgenic lines regarding leaf consumption, biomass accumulation and mortality. The HD and MD transgenic lines showed 100% mortality when inoculated with first‐ and second‐instar larvae; however, the LD transgenic lines showed about 50% mortality. The HD transgenic lines exhibited a significantly higher yield than the MD and LD transgenic lines owing to their high CPB resistance. CONCLUSION: Commercially available transgenic potato plants with above 0.1% Cry3A of total soluble protein and NT control refugia could control damage, delay adaptation and halt dispersion eastwards. The two HD transgenic lines developed in this study, PAH1 and PAH2, are ideal for use as cultivars or germplasm to breed new cultivars. Copyright © 2012 Society of Chemical Industry     

Detection of organophosphate and pyrethroid resistance alleles in Czech Leptinotarsa decemlineata (Coleoptera: Chrysomelidae) populations by molecular methods

BACKGROUND: Pyrethroids and organophosphates are the most frequently used insecticides for Colorado potato beetle, Leptinotarsa decemlineata (Say), control in the Czech Republic. Based on molecular methods, organophosphate and pyrethroid resistance alleles have been detected in samples from three sites. The accuracy of restriction fragment length polymorphism (RFLP) and bidirectional polymerase chain reaction amplification of specific alleles (Bi‐PASA) for detection of resistance alleles is compared. RESULTS: Leptinotarsa decemlineata from three sites showed higher frequencies of resistance alleles to organophosphates than to pyrethroids. The rates of occurrence of individuals homozygous resistant (RR) to pyrethroids ranged from 20.0 to 22.9%, while the rates of occurrence of individuals RR to organophosphates ranged from 52.9 to 66.7%. The incidences of individuals with resistance alleles to both organophosphates and pyrethroids ranged from 8.6 to 13.6%. No relationship was found between incidence of the pyrethroid resistance allele and site, while incidence of the organophosphate resistance allele differed significantly according to site. CONCLUSION: Both RFLP and Bi‐PASA were suitable for detecting resistance alleles to pyrethroids, and in most cases also for detecting resistance alleles to organophosphates. In contrast to Bi‐PASA, RFLP was also suitable for samples with lower DNA quality when testing for the resistance allele to pyrethroids. On the other hand, RFLP was not as accurate as Bi‐PASA in detection of the organophosphate resistance allele. Copyright © 2010 Society of Chemical Industry     

Selection for fungicide resistance throughout a growing season in populations of Plasmopara viticola

A method for evaluating the potential threat of selection for resistance to organically-based fungicides in populations of P. viticola is needed to screen a large panel of products alternative to copper in organic viticulture. Populations from an unexposed plot were compared throughout one season with a population sprayed with azoxystrobin (Quadris), reported as engendering selection pressure and resistance, and a population sprayed with an organically-based fungicide (Mycosan). The evolution of the three populations was followed with neutral specific SSR markers and with the specific marker for strobilurin resistance, as control of selection for resistant mutants. A reduction in genetic diversity of the P. viticola population was observed in the population sprayed with azoxystrobin, consistent with directional selection toward higher resistance, confirmed by an enhanced frequency of resistant mutants with respect to the unexposed population. In contrast, a higher diversity and a reduced frequency of resistant mutants were observed in the population sprayed with the organically-based fungicide. Assessing a reduction of genotypic diversity allows the detection of selection for resistance and constitutes a valid instrument for screening a large panel of products with non-specific, different and possibly indirect modes of action.     

Functional analysis of mutations in expressed acetylcholinesterase that result in azinphosmethyl and carbofuran resistance in Colorado potato beetle

The functional attributes of specific point mutations, R30K, S291G, and I392T, associated with full-length acetylcholinesterase (AChE) cDNAs of organophosphate (OP)- and carbamate-resistant Colorado potato beetles (CPB), were determined using site-directed mutagenesis and baculovirus expression. Enzymatic and inhibitory properties of altered recombinant acetylcholinesterases (rAChEs) were examined. S291G increased the hydrolysis of substrates with larger substituted alkyl groups (e.g., BTC vs ATC) and increased the inhibitory action of inhibitors with larger alkyl groups (e.g., paraoxon, DFP, and N-propyl carbofuran vs. azinphosmethyl-oxon and N-methyl carbofuran). R30K in conjunction with S291G enhanced the hydrolysis activity of larger substrates and the inhibitory action of larger inhibitors. I392T attenuated the effects of S291G in that the altered rAChE with both S291G and I392T elicited substrate specificity and inhibitory properties more similar to the susceptible form of AChE without mutations.     

Synergism of resistance to phosalone and comparison of kinetic properties of acetylcholinesterase from four field populations and a susceptible strain of Colorado potato beetle

The susceptibility to phosalone and biochemical characteristics of acetylcholinesterase (AChE) were compared between susceptible (SS) strain and four field populations of Colorado potato beetle (CPB) collected from commercial potato fields of Hamedan Province in west of Iran. Bioassays involving topical application of phosalone to fourth instars revealed up to 252 fold resistance in field populations compared with the SS strain. Synergism studies showed that although esterase and/or glutathione S-transferase metabolic pathways were present and active against phosalone, they were not selected for and did not have a major role in resistance. It is likely that piperonyl butoxide (PBO) reduced phosalone toxicity by inhibiting bio-activation of phosalone. The affinity (K_m) and hydrolyzing efficiency (V_max) of AChE to selected substrates, namely, acetylthiocholine iodide (ATC), propionylthiocholine iodide (PTC), and butyrylthiocholine iodide (BTC) were examined. AChE inhibition by higher substrate concentration was evident only in the SS strain. In resistant field populations, Aliabad (Aa), Bahar (B) and Dehpiaz (Dp), substrate inhibition at higher concentrations was not seen. There was no definite optimal concentration found for any of the substrates examined. When ATC, PTC, and BTC were used as substrate, the reaction rates of AChE from Yengijeh (Yg) population increased as the concentration of all three substrates were increased, but were almost constant at concentration of ATC ? 3.98, PTC ? 2.8, and BTC ? 5 mM. The susceptible form of AChE had the most efficient ATC hydrolysis but very low BTC hydrolysis activity. In contrast, AChEs from field populations elicited relatively reduced ATC hydrolysis, but relatively increased BTC hydrolysis. The in vitro inhibition potency of some organophosphates (OPs), on AChEs of the field populations and SS strain was determined. The rank order from the most potent inhibitor to the least as determined by their bimolecular reaction constants (K_i) was ethyl paraoxon > diazoxon > methyl paraoxon for AChE from Aa, B, Dp, and Yg populations, respectively, whereas the rank order for the susceptible strain was methyl paraoxon > ethyl paraoxon > diazoxon.     

中国水稻主产区褐飞虱对3种杀虫剂的抗性监测

2006-2009年,用稻茎浸渍法连续监测了广西南宁市、广东阳江市、湖南东安县、福建福清市、江西上高县、湖北孝感市、浙江金华市、江苏通州市和安徽和县共9个地区褐飞虱种群对吡虫啉、噻嗪酮和氟虫腈的抗性变化。结果表明:褐飞虱种群对吡虫啉的抗性仍处于高水平至极高水平抗性阶段(105.5~459.7倍),但2009年监测到东安、孝感、上高种群对吡虫啉的抗性已有下降趋势;褐飞虱种群对氟虫腈的抗性有增长趋势,2006到2009年褐飞虱种群对氟虫腈由敏感至低水平抗性(<6.9倍)发展到了中水平至高水平抗性(13.5~43.3倍);由于2005年吡虫啉在高抗地区的禁用,褐飞虱种群对噻嗪酮的抗性上升速度加快,2009年已处于低水平至中水平抗性阶段(7.0~14.4倍)。这表明在吡虫啉、氟虫腈被禁用后,大面积单一使用噻嗪酮进行防治,褐飞虱对噻嗪酮的抗性有可能加速发展。     

Monitoring for imidacloprid resistance in the tobacco‐adapted form of the green peach aphid,Myzus persicae (Sulzer) (Hemiptera: Aphididae), in the eastern United States

BACKGROUND: Imidacloprid is the primary insecticide for controlling the tobacco‐adapted form of the green peach aphid (TGPA), Myzus persicae (Sulzer), a major pest of tobacco worldwide. This study used leaf‐dip bioassays to assess TGPA resistance to imidacloprid in the eastern United States from 2004 through 2007. RESULTS: When combined over the 4 year study, 18, 14 and 3% of the TGPA had imidacloprid resistance ratios (RRs) of 10–20‐fold, 20–30‐fold and 30–90‐fold, respectively, compared with the most susceptible colony tested. This indicates that some colonies have developed moderate levels of resistance to imidacloprid. A colony collected near Clayton, North Carolina, had the highest RR of 91 (LC₅₀ value = 31 mg L^?1). This resistance declined for six tests over a 3 year period in the laboratory culture from >130‐fold RR (LC₅₀ = 48 mg L^?1) to 40‐fold RR (LC₅₀ = 15 mg L^?1). Over the same period, the most susceptible colony and a standard colony not exposed to imidacloprid for over 7 years had consistently low LC₅₀ values. CONCLUSION: Moderate levels of resistance to imidacloprid are noticed among TGPA colonies from the eastern United States. The variation in resistance indicates that the factors responsible are present in the populations at low frequencies and are just not enough to cause field failures yet. Copyright © 2010 Society of Chemical Industry     

Selection for imidacloprid resistance in silverleaf whiteflies from the Imperial Valley and development of a hydroponic bioassay for resistance monitoring

A field-collected population of the silverleaf whitefly, Bemisia argentifolii, was selected with the nicotinyl compound, imidacloprid, over 32 generations to determine if resistance would develop when maintained under continuous selection pressure in a greenhouse. Resistance was slow to increase at first with low to moderate levels of resistance (RR from 6- to 17-fold) in the first 15 generations of selection. Further selection steadily led to higher levels of resistance, with the greatest resistance ratio at 82-fold, the gradual rise suggesting the involvement of a polygenic system. At the end of the selection, slopes of probit regressions were substantially steeper than earlier, indicating increased homogeneity of imidacloprid resistance in this strain. A hydroponic bioassay featuring systemic uptake of imidacloprid through roots was developed to monitor the changes in resistance to imidacloprid in the selected whitefly strain and in seven field-collected strains from Imperial Valley, California. Six out of seven field-collected strains exhibited low LC₅₀ values (0·002 to 0·512 mg ml^-1) compared to the selected resistant strain, with one exception where the LC₅₀ was 0·926 mg ml^-1 (RR=15·0). Variation in responses to imidacloprid in the field strains suggest that this technique is sufficiently sensitive to detect differences in susceptibilities of whitefly populations. The imidacloprid-resistant strain showed no cross-resistance to endosulfan, chlorpyrifos or methomyl (RR ranging from 0·4- to 1·5-fold). A low level of cross-resistance was observed to bifenthrin in the IM-R strain at 7-fold. The success of selection for resistance to imidacloprid has serious implications for whitefly control programs that rely heavily on imidacloprid. ©1997 SCI     

Monitoring of insecticide resistance in damson hop aphid,Phorodon humuli Schrank (Hemiptera: Aphididae) from German hop gardens

The damson hop aphid, Phorodon humuli (Schrank) (Hemiptera: Aphididae), is one of the most important sucking pests of many hop-growing areas world-wide. In this study we determined the efficacy of several insecticides against strains collected throughout the year 2001. All strains were collected in different hop gardens in the Hallertau (Bavaria), Germany, the largest hop-growing area of the world. First of all we established a leaf dip bioassay, carried out using six-well tissue culture plates and appropriate for monitoring susceptibility against imidacloprid, oxydemeton-methyl, cyfluthrin, amitraz, pymetrozine and pirimicarb. Four of these compounds, imidacloprid, cyfluthrin, pymetrozine and amitraz, are currently registered for the control of sucking pests in German hop gardens and are useful against P. humuli. The leaf-dip bioassay system turned out to be very reliable and robust. Ten P. humuli strains were collected in May 2001 and maintained in the laboratory to assess their resistance to the different insecticides in comparison with two laboratory reference strains (H2 and H5). Using diagnostic concentrations, resistance monitoring for imidacloprid and cyfluthrin was investigated during July and August 2001 on 53 populations from 30 sites around the Hallertau, an area of ca 2500 km2. Resistance to diagnostic concentrations (LC95 for reference strains) of imidacloprid, amitraz and pymetrozine was not detected in any strain received in 2001, but late-season (August) populations seemed to respond more heterogeneously than those collected mid-season (July). Overall composite mean mortalities to diagnostic concentrations of imidacloprid (13 mg litre(-1)) in collections from May, July and August were 95 (+/-2.5), 98 (+/-2.3) and 87 (+/-5.9)%, respectively. Moderate resistance to pyrethroids was observed in all strains collected in May and August using a diagnostic concentrations of 10 mg litre(-1) cyfluthrin (LC95 of the susceptible reference strain H5). Slight to moderate resistance to diagnostic concentrations of oxydemeton-methyl and pirimicarb was observed in some, but not all, strains collected early season. The results are discussed in terms of the implemention of hop aphid resistance management strategies in German hop-cultivation areas.     

Application of DNA-based genotyping techniques for the detection of kdr-like pyrethroid resistance in field populations of Colorado potato beetle

Permethrin resistance in 16 field populations of Colorado potato beetle was established by bioassay. Permethrin-resistant genotypes were detected using two genotyping techniques, bidirectional polymerase chain reaction amplification of specific alleles (bi-PASA) and single stranded conformational polymorphism (SSCP). The bi-PASA technique was 86% and SSCP was 95% reliable. In terms of accuracy, bi-PASA determined the correct genotype compared to the actual sequencing data 98% and SSCP 93% of the time. There was marked correlation between increasing survivorship (increasing LD₅₀values) and increasing proportions of resistant genotypes (increasing percentage of resistant allele). These techniques allowed the detection of a resistant allele 98% of the time when sampling only 384 beetles from an overwintering adult population (4995 beetles/ha) that possessed a 1% resistant allele frequency at a cost of $500/ha, substantiating the cost effectiveness of the genotyping approaches to monitoring.     

Biochemical study of resistance to imidacloprid in B biotype Bemisia tabaci from Guatemala

Systemic uptake bioassays using excised cotton leaves confirmed resistance to imidacloprid in a Guatemalan population of the tobacco whitefly Bemisia tabaci Gennadius. Polyacrylamide gel electrophoresis of naphthyl esterases identified the insects as B-types. Upon collection from the field, resistance was determined to be 58-fold relative to a susceptible strain originating in the Imperial Valley of California. Resistance levels increased to 126-fold in this population during its continuous exposure to systemically treated cotton. In biochemical investigations, there was no detectable NADPH-dependent mixed function oxidase metabolism of 14C-imidacloprid at any time during the selection process. In contrast, microsomal preparations from housefly abdomens readily produced significant amounts of the mono-hydroxy and olefin derivatives of the parent compound. Detoxification of imidacloprid by housefly MFOs may account for reports of lower toxicity of the insecticide towards this insect compared with whiteflies, despite similar binding properties between imidacloprid and the nicotinic acetylcholine receptors in both species.