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1.
In this paper we analysed the basis of insecticide resistance in 59 Italian strains of the peach potato aphid Myzus persicae using both molecular and biochemical assays. Our data as a whole clearly indicate that most M. persicae strains (76.3%) have high or extremely high production of an esterase enzyme which sequester and detoxify insecticides with esteric group. Kdr genotypes conferring resistance towards pyrethoids are present in 57.7% of the analysed populations. Moreover, 26.5% of the kdr positive strains possess also the M918T mutation conferring super-kdr phenotype. Strains with modified AChE (MACE) are not so numerous (27.1%), although they can be found almost everywhere in Italy. Considering all the strains analysed, both MACE and kdr phenotypes are associated with high levels of esterase activity. In Central–Southern regions, kdr and MACE resistance mechanisms resulted in linkage disequilibrium. Bioassays performed in order to evaluate the efficacy of a pyrethroid insecticide against a strain possessing a F979S mutation within its para-type sodium channel gene suggests that this amino acid substitution could affect the sodium channel responsivity to pyrethroids.  相似文献   

2.
Genetic linkage maps of Heliothis virescens and Helicoverpa armigera are being used to identify and characterize resistance-conferring genes. The insensitive acetylcholinesterase conferring resistance to organophosphorus insecticides and the insensitive sodium channel conferring resistance to pyrethroids have both been mapped in H. virescens. The linkage mapping approach permits a genetic dissection of resistance, even when the mode of action and lethal target are not precisely known, such as for the insecticidal toxins from the bacterium Bacillus thuringiensis (Bt). We have identified and mapped a major Bt-resistance locus in a strain of H. virescens exhibiting up to 10000-fold resistance to Cry1Ac toxin and are currently developing a linkage map for H. armigera with a set of ‘anchor’ loci to facilitate comparison with H. virescens. Both species are currently experiencing their first significant selective pressure in the field by transgenic cotton expressing Cry1Ac, and timely identification of resistance mechanisms and their underlying genetic basis will be essential in successfully managing the Bt resistance that will eventually appear. ©1997 SCI  相似文献   

3.
The susceptibility of three clones of Aphis gossypii Glover to 15 insecticides was established by bioassay. A high level of resistance towards pirimicarb was confirmed for a clone from Holland (Dutch R) and a clone from Japan (Jap R), while the susceptible clone (S) was killed by very low doses of the insecticide. However, only limited cross-resistance was shown towards other carbamates and organophosphates, and no marked resistance to the pyrethroids tested. The acetylcholinesterase (AChE) of both resistant clones hydrolysed acetyl-choline faster than that of susceptible aphids, with greatest enzyme activity shown by the Dutch R clone. Inter-clone differences in these rates were consistent with differences in catalytic centre activities. Inhibition (I50) of AChE by pirimicarb was approximately 900-fold higher for the resistant clones than for the S clone. First-order kinetics revealed that resistance to pirimicarb in Dutch R and Jap R involved a modified AChE which had a reduced (approximately 350-fold) affinity (Kd) for pirimicarb. The marked change in AChE affinity for pirimicarb was not repeated with the other carbamates tested, ethiofencarb and aldicarb. It was considered that the resistant aphids would not require mechanisms in addition to insensitive AChE in order to show the high level of resistance to pirimicarb shown in the bioassay.  相似文献   

4.
Knockdown resistance (kdr) is a target-site resistance mechanism that confers nerve insensitivity to DDT and pyrethroid insecticides. In the housefly, Musca domestica, molecular cloning of the para-type sodium channel gene has revealed two amino acid mutations that are associated with kdr and super-kdr resistance phenotypes. Both mutations are located in the domain II region of the channel; Leu1014 to Phe in the hydrophobic segment IIS6 and Met918 to Thr in the IIS4-IIS5 linker. To investigate whether these mutations also occur in other insects, we have designed degenerate primers based on conserved sequences in the domain II region of the sodium channel and used these to PCR amplify this region from insecticide-susceptible strains of eight diverse insect species representing four different insect Orders: Helicoverpa armigera, Plutella xylostella, Spodoptera littoralis (Lepidoptera), Blattella germanica (Dictyoptera), Tribolium castaneum (Coleoptera), Myzus persicae, Aphis gossypii and Phorodon humuli (Hemiptera). The primers amplified closely related para-type sodium channel sequences from each insect with a minimum of 85% amino acid identity between species. All of the sequences contained ‘susceptible’ Leu and Met residues at the positions associated with kdr and super-kdr resistance in the housefly. Recent results detailing the presence of a kdr-type Leu to Phe mutation in pyrethroid-resistant strains of two important agricultural pests, P. xylostella and M. persicae, are discussed. ©1997 SCI  相似文献   

5.
In some insect species, knockdown resistance (kdr) to pyrethroids and DDT is linked to point mutations in the sequence of the para-type voltage-dependent sodium channel gene. The effects of pyrethroids were assayed on six Culex pipiens strains: two were susceptible to pyrethroids and the four others displayed various levels of resistance, but, in each case, a kdr-type mechanism was strongly suggested. Degenerate primers were designed on the basis of the corresponding sequences of the para orthologous gene reported from several orders of insects. These primers were used to amplify the region of the sodium channel gene which includes the positions where the kdr and super-kdr mutations have been found in Musca domestica. As expected, the amplified fragment was highly homologous to the para sequences. The super-kdr-like mutation (methionine to threonine at position 918 of the M domestica para sequence) was never detected in any strain. In contrast, the same kdr mutation (leucine to phenylalanine at position 1014) was present in some Culex pyrethroid-resistant samples. An alternative substitution of the same leucine to a serine was detected in one strain slightly resistant to pyrethroids but highly resistant to DDT. These data have allowed us to design a PCR-based diagnostic test on genomic DNA to determine the presence or the absence of the kdr allele in single C pipiens collected in several countries. The validity of this test was checked by comparing the frequency of the resistance allele and the toxicological data. © 1999 Society of Chemical Industry  相似文献   

6.
A range of potential proinsecticides was synthesised and tested against insecticide-susceptible and -resistant clones of Myzus persicae (Sulzer). They were all esters of compounds known to be toxic or pharmacologically active, and were designed to have increased lipophilicity and to be subject to more rapid activation by hydrolysis in resistant than in susceptible aphids due to the increased amount of esterase present in the resistant clones. The most potent toxins were esters of monofluoroacetic acid. When applied topically, the toxicity of these esters to M. persicae was directly proportional to the esterase content of the aphids. Such compounds would not be suitable as commercial insecticides, but the results serve to illustrate the potential benefits of exploiting a resistance mechanism against one class of compounds to render another class more toxic, i.e. to design compounds that show negative cross-resistance. © 1998 SCI  相似文献   

7.
The Colorado beetle, Leptinotarsa decemlineata (Say), is one of the most important pests in many potato‐producing regions. Colorado beetle infestations are normally kept under economic damage thresholds by applying insecticides such as pyrethroids. Pyrethroids are known to act on voltage‐gated sodium channels and have been used for several decades to control L. decemlineata. Their continuous and widespread use has resulted in the development of resistance, which is often linked to a L1014F target‐site mutation in the voltage‐gated sodium channel, known as knockdown resistance (kdr). Since pyrethroids are used in many potato‐growing regions in Germany, more than 140 L. decemlineata samples were collected and analysed for the presence of the kdr allele by pyrosequencing diagnostics. Results showed that kdr is present in many German L. decemlineata populations, but even its homozygous presence does not substantially compromise the efficacy of recommended label rates of pyrethroids, as demonstrated by bioassays and crossing experiments. The implications of these findings for resistance management are briefly discussed.  相似文献   

8.
The effects of pyrethroids were studied upon isolated segmental nerves and neuromuscular junctions in both susceptible (Cooper) and knockdown-resistant (kdr; super-kdr) strains of housefly larvae (Musca domestica L.). Isolated segmental nerves contained neither cell bodies nor synaptic contacts; thus, any effects of pyrethroids were attributed solely to their actions upon voltage-dependent Na+ channels. Threshold concentrations of the type II pyrethroid, deltamethrin, required to elevate the spontaneous firing rate of these nerves were determined. Both resistant strains were about ten times less sensitive to deltamethrin than the susceptible strain, but insensitivity of super-kdr nerves was no greater than in the less resistant kdr strain. At neuromuscular junctions, the minimum concentrations of pyrethroids needed to trigger massive increases in the frequency of miniature excitatory postsynaptic potentials (mEPSPs) were determined for deltamethrin and the type I pyrethroid, fenfluthrin. With fenfluthrin there was no detectable difference between the junctions of kdr and super-kdr strains, which were both about ten-fold less sensitive than Cooper junctions. With deltamethrin, kdr junctions were about 30 times less sensitive than those of Cooper; super-kdr junctions were dramatically insensitive to deltamethrin, being some 10000- and 300-fold less sensitive than those of Cooper and kdr respectively. Thus, in the synaptic assay, super-kdr conferred an extension in resistance over kdr only against the type II pyrethroid, it being ineffective against fenfluthrin. We suggest that kdr resistance comprises at least two site-insensitive areas within the nervous system. One involves insensitivity of the Na+ channel and has similar efficacy in both kdr and super-kdr strains against type I and II pyrethroids; the other is associated with the presynaptic terminal and is particularly effective in super-kdr resistance against type II pyrethroids. The latter could be associated with Ca2+-activated phosphorylation of proteins involved with neurotransmitter release. Such phosphorylation reactions are known to be perturbed by pyrethroids, especially by type II compounds.  相似文献   

9.
BACKGROUND: The accuracy of predicting the survival of insecticide‐resistant aphids following the application of commonly used insecticides from the carbamate, the pyrethroid, a mix of the two or the neonicotinoid chemical classes was evaluated in a potato field in Scotland. Equal proportions of five genotypes of the peach‐potato aphid, Myzus persicae (Sulzer), with none, resistance to dimethyl‐carbamates, resistance to pyrethroids or combinations conferring resistance to both chemical classes were released into potato field plots. The insecticides were sprayed separately onto these plots, the aphid populations were analysed after 6–8 days and the process repeated. RESULTS: For each assessment after the three separate spray events, plots treated with the carbamate had 48, 147 and 28%, those treated with pyrethroid 53, 210 and 89%, those treated with carbamate/pyrethroid 28, 108 and 64% and those treated with neonicotinoid 43, 55 and 11% of the numbers of M. persicae by comparison with untreated controls. Only the proportions of surviving aphids from the genotype containing no insecticide resistance traits and the genotype containing elevated carboxylesterases matched ratios predicted from the selective advantage afforded by the resistance traits alone. Survival of aphids from the other three genotypes that carried 1–3 of the insecticide resistance traits differed from expectations in all cases, possibly owing to physiological differences, including their vulnerability to predators and hymenopterous parasitoids present at the site and/or their carrying unknown insecticide resistance mechanisms. CONCLUSION: Control strategies based on knowledge of the genetically determined insecticide resistance profile of an M. persicae population alone are insufficient. Hence, other important factors contributing to aphid survival under insecticide pressure need to be considered. Copyright © 2012 Society of Chemical Industry  相似文献   

10.
The mechanisms for multiple resistances had been studied with two field resistant strains and the selected susceptible and resistant strains of Spodoptera litura (Fabricius). Bioassay revealed that the two field strains were both with high resistance to pyrethroids (RR: 63-530), low to medium resistance to organophosphates and carbamates, AChE targeted insecticides (RR: 5.7-26), and no resistance to fipronil (RR: 2.0-2.2). Selection with deltamethrin in laboratory could obviously enhance the resistance of this pest to both pyrethroids and AChE targeted insecticides. Synergism test, enzyme analysis and target comparison proved that the pyrethroid resistance in this pest associated only with the enhanced activity of cytochrome P450 monooxygenase (MFO) and esterase. However the resistance to the AChE targeted insecticides depended on the target insensitivity and also the enhanced activity of MFO and esterase. Thus, the cross-resistance between pyrethroids and the AChE targeted insecticides was thought to be resulted from the enhanced activity of MFO and esterase.  相似文献   

11.
Myzus persicae samples were collected from populations present on a range of field crops between 1997 and 2000. A combination of biochemical, DNA-based diagnostics and bioassays was used to assess the presence of three insecticide resistance mechanisms: elevated carboxylesterase (E4 or FE4), insensitive acetylcholinesterase and insensitive sodium channels (knockdown resistance, kdr). For the carboxylesterases, both the levels of enzyme and the type of gene present (E4 or FE4) were determined. The results showed that during the time period studied there was a dramatic reduction in the proportion of aphids with very high levels of E4 and an increase in those with lower levels of FE4. There was also a slightly different E4 gene present in a limited number of samples. The change in esterase genes was accompanied by a virtual loss of the insensitive AChE variant and a maintenance of aphids with kdr. The selection pressures and other factors leading to these changes in field populations of M persicae are discussed.  相似文献   

12.
Insecticide sprays were applied to Myzus persicae (Sulzer) populations carrying various combinations of three insecticide resistance mechanisms (esterase-based metabolic resistance and two target site mechanisms, known as MACE and kdr), supported on host plants growing in field simulator cages. The study showed that MACE confers extreme resistance to pirimicarb and triazamate (carbamate insecticides) but not to deltamethrin + heptenophos (16 + 1) (Decisquick) or dimethoate (an organophosphorus insecticide). Resistance to dimethoate depends solely on levels of esterase-based resistance, while resistance to Decisquick depends on kdr and esterase. None of the four insecticides is effective against aphids carrying MACE combined with extreme esterase-based resistance. This knowledge, in association with current monitoring of the mechanisms, will play an important role in making decisions on insecticide use against M persicae in the UK. © 1999 Society of Chemical Industry  相似文献   

13.
BACKGROUND: The polyphagous cotton‐melon aphid Aphis gossypii Glover is structured into geographically widespread host races comprising a few clones specialised on Cucurbitaceae, cotton, eggplant or pepper. To assess insecticide resistance among and within host races, leaf disc bioassays were conducted on aphid clones collected from Cucurbitaceae (genotypes C4 and C9), cotton (genotypes Burk and Ivo), eggplant (genotype Auber) and pepper (genotype PsP4). Molecular diagnostic (PCR‐RFLP) and enzyme assays were also performed to detect the basic mechanisms underlying insecticide resistance. RESULTS: All six clones were susceptible to acetamiprid (neonicotinoid) or carbosulfan (carbamate). Conversely, all clones were resistant to dimethoate (organophosphate) (RF = 4.1–38.1) and carried mutation S431F in the acetylcholinesterase gene. Auber, PsP4 and Burk also carried mutation A302S in this gene, which possibly conferred moderate resistance (RF = 3.7–6.8) to profenofos and monocrotophos (organophosphates). Auber and Burk were highly resistant (RF = 41.2 and 473 respectively) to cypermethrin (pyrethroid). This resistance was likely associated with point mutation super‐kdr (M918L) in the voltage‐gated sodium channel gene (para gene) or metabolic detoxification mediated by esterase and oxidase enzymes. CONCLUSION: Multiple resistance to a broad range of insecticides and multiple mechanisms of resistance in some clones could explain to some extent the low genetic diversity observed within A. gossypii host races. Copyright © 2009 Society of Chemical Industry  相似文献   

14.
BACKGROUND: Parthenogenetic clones of the green peach aphid, Myzus persicae (Sulzer), and the cotton aphid, Aphis gossypii Glover, were tested with the anthranilic diamide insecticide cyantraniliprole (i.e. DuPont? Cyazypyr?) in systemic‐uptake bioassays to investigate potential for cross‐resistance conferred by mechanisms of insecticide resistance to organophosphates, carbamates and pyrethroids and, in the case of M. persicae, reduced sensitivity to neonicotinoids. These data were compared with the response of field samples of M. persicae and A. gossypii collected from around Europe. RESULTS: Cyantraniliprole was not cross‐resisted by any of the known insecticide resistance mechanisms present in M. persicae or A. gossypii. The compound was equally active against resistant and susceptible aphid strains. The responses of the M. persicae field samples were very consistent with a maximum response ratio of 2.9 compared with a standard laboratory clone. The responses of the A. gossypii field samples were more variable, although a majority of the responses were not statistically different. CONCLUSION: Cyantraniliprole is currently the only anthranilic diamide (IRAC MoA 28) insecticide targeting aphid species such as M. persicae and A. gossypii. There is no evidence to suggest that the performance of this compound is affected by commonly occurring mechanisms that confer resistance to other insecticide chemistries. Cyantraniliprole is therefore a valuable tool for managing insecticide resistance in these globally important pests. Copyright © 2011 Society of Chemical Industry  相似文献   

15.
A point mutation in thepara-homologous sodium channel gene has been shown to be associated with knockdown resistance (kdr) in several insect species including the German cockroach. In this study, we analyzed the genomic organization of the region where thekdrmutation resides and then performed polymerase chain reaction (PCR) and sequencing using genomic DNA as the template to detectkdrmutation in 24 pyrethroid-resistant German cockroach strains, most of which have been collected recently from the field. Thekdrmutation, G to C at nt 2979 resulting in a leucine to phenylalanine amino acid substitution, was detected in 20 strains including 2 strains from overseas (China and Germany). Our results clearly indicate that thekdrmutation is widespread in German cockroach populations. However, the super-kdrmutation detected in super-kdrhouse flies was not found in any of the 4 strains that showed higher levels of knockdown resistance. Little correlation was observed between the presence of thekdrmutation and the level of knockdown resistance, suggesting the existence of multiple resistance mechanisms in many of these strains.  相似文献   

16.
Examination of the enzyme that determines the level of resistance to organophosphorus insecticides and carbamates in Myzus persicae (Sulz.) and bioassays were used to establish the frequency and resistance levels of resistant aphids on outdoor crops in Britain in 1976. The biochemical tests, staining esterase-4 after electrophoresis and total esterase determination, were more sensitive than bioassays. However the dip-test, a simple, rapid and inexpensive bioassay designed to detect resistance and its different levels gave satisfactory results which warrant its use where biochemical detection of resistance is not possible. Carboxylesterase activities of M. persicae collected in 1976 fell into three groups: low, moderate and high, and these were correlated with differences in tolerance to dimethoate, demeton-S-methyl and pirimicarb. Aphids with low esterase activity were susceptible(S). Those with the moderately active enzyme (R1) had five- to seven-fold resistance to the two organophosphorus insecticides and were marginally resistant (about two-fold) to pirimicarb. The insects with the most active esterase (R2) were strongly resistant to dimethoate (resistance factor, RF × 126) and moderately resistant to demeton-S-methyl (RF × 17) and pirimicarb (RF × 8). Some R1, but no S aphids survived the recommended dose of demeton-S-methyl on field crops probably because they were under the lowest leaves and therefore protected from direct contact with the spray. Laboratory tests demonstrated that these R1 aphids tolerated the residual deposit and systemic dose present in the leaves of the treated potato-plants. This enabled their numbers to recover in treated fields much faster than the susceptible insects which could do so by immigration only when the residual dose in the plant was no longer toxic. R1 aphids were common throughout the country particularly in eastern England where susceptible aphids were rare, but in the Shardlow area of Derbyshire susceptible aphids were in the majority throughout the summer. R2 aphids were found only in samples from the west of Scotland and northern England. The implications of the presence of aphids with different levels of resistance for aphid control are discussed.  相似文献   

17.
BACKGROUND: Myzus persicae is a globally important aphid pest that is mainly controlled through the application of chemical insecticides. Recently, a clone of M. persicae exhibiting control‐compromising levels of resistance to neonicotinoid insecticides was described. The resistance of this clone was associated with reduced affinity of imidacloprid for the target site (the nicotinic acetylcholine receptor) as a result of mutation of a key amino acid residue (R81T) in the loop D region of a nAChR β1 subunit. The potent levels of resistance conferred by this mechanism are cause for considerable concern, and the frequency and distribution of the mutation in worldwide populations of M. persicae require careful monitoring. In this study, a high‐throughput assay has been developed that allows detection of the mutation in individual aphids. RESULTS: A real‐time TaqMan assay to detect the R81T substitution was developed that proved to be sensitive and specific in tests of analytical sensitivity and in a blind genotyping trial of DNA extracted from individual aphids comprising the three possible genotypes. The assay was then used to examine the frequency of the R81T mutation in aphids collected and stored in ethanol from peach orchards in southern France. The R81T frequency varied from 33 to 100% in seven populations from the department of Gard, France. CONCLUSIONS: This study describes a rapid and sensitive assay that very effectively detects the R81T mutation in individual aphids. The results also have practical significance for the control of M. persicae in southern France and provide contemporary data to inform current resistance management strategies. Copyright © 2012 Society of Chemical Industry  相似文献   

18.
BACKGROUND: Pediculosis is the most prevalent parasitic infestation of humans. Resistance to pyrethrin‐ and pyrethroid‐based pediculicides is due to knockdown (kdr)‐type point mutations in the voltage‐sensitive sodium channel α‐subunit gene. Early detection of resistance is crucial for the selection of effective management strategies. RESULTS: Kdr allele frequencies of lice from 14 countries were determined using the serial invasive signal amplification reaction. Lice collected from Uruguay, the United Kingdom and Australia had kdr allele frequencies of 100%, while lice from Ecuador, Papua New Guinea, South Korea and Thailand had kdr allele frequencies of 0%. The remaining seven countries investigated, including seven US populations, two Argentinian populations and populations from Brazil, Denmark, Czech Republic, Egypt and Israel, displayed variable kdr allele frequencies, ranging from 11 to 97%. CONCLUSION: The newly developed and validated SISAR method is suitable for accurate monitoring of kdr allele frequencies in head lice. Proactive management is needed where kdr‐type resistance is not yet saturated. Based on sodium channel insensitivity and its occurrence in louse populations resistant to pyrethrin‐ and pyrethroid‐based pediculicides, the T917I mutation appears to be a key marker for resistance. Results from the Egyptian population, however, indicate that phenotypic resistance of lice with single or double mutations (M815I and/or L920F) should also be determined. Copyright © 2010 Society of Chemical Industry  相似文献   

19.
Structure-activity relationships (SARs) for 10 pyrethroids against susceptible, kdr and super-kdr strains of houseflies (Musca domestica L.) were investigated by Principal Components Analysis. In the three strains with kdrLatina' all only slightly to moderately (2.6 to 26-fold) resistant to pyrethroids, no correlation between the structure and Levels of resistance could be discerned. In flies with super-kdr, SARs were influenced by the nature of the alcoholic portion of the ester. Resistance was strongest to esters of a-cyano-3-phenoxybenzyl alcohol (74 to 430-fold) and to permethrin (48 to 55-fold). It was weak (6.2 to 11-fold) to cyclopentenone derivatives, being barely stronger than for flies with kdr (2-6 to 6.3-fold). Two variants of super-kdr (3D and A2) were distinguished on the basis of their differential response to esters of 5-benzyl-3-furylmethanol. It is presumed that kdrLatina, super-kdrA2 and super-kdr3D form an allelic series in which kdrLatina represents ground level insensitivity, and the two super-kdrs the progressive extension of strong resistance to more types of ester. The strong differences in resistance to different pyrethroid esters by super-kdr flies provides scope for improving management of resistance to pyrethroid insecticides and for modifying the SAR of pyrethroids to favour weak resistance.  相似文献   

20.
The susceptibility of Helicoverpa armigera to pyrethroids has been investigated in West Africa by means of laboratory bioassays since 1985, the first year of widespread pyrethroid use. For some years, this survey has shown a tendency for the pest to become more tolerant to pyrethroids. During the 1996 growing season, farmers using calendar‐based spraying programmes reported control failures in various countries. The strong efficacy of cypermethrin on small larvae was confirmed in experimental plots, but the effect decreased quickly in successive instars. Bioassays performed on resistant strains revealed an increase in LD50 that was related to different resistance mechanisms. Metabolic resistance (MFO) appears to be a possible primary mechanism of resistance to pyrethroids. Target modification (kdr) is involved to a small degree and esterases seem to appear only after additional selection pressure. © 2000 Society of Chemical Industry  相似文献   

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