Age-dependent response to insecticides and enzymatic variation in susceptible and resistant codling moth larvae

Insecticide resistance in the codling moth, Cydia pomonella, partly results from increased metabolic detoxification. The aim of this study was to follow the age variations in larval susceptibility to deltamethrin and teflubenzuron in one susceptible (S) strain, and two resistant (Rv and Rt) ones selected for resistance to deltamethrin and diflubenzuron, respectively. The age variation of the activities of cytochrome P450-dependent monooxygenase (MFO), glutathione S-transferases (GST), and esterases in S and both resistant strains were simultaneously investigated. The highest levels of insecticide resistance were recorded in late instars in both resistant strains, although Rv neonates exhibited enhanced resistance to deltamethrin. The involvement of an additional deltamethrin-specific mechanism of resistance, which could be mainly expressed in early instars, was supported by previous demonstration of a kdr point mutation in the Rv strain. The cross-resistance between deltamethrin and teflubenzuron indicated the involvement of non-specific metabolic pathways in resistance to teflubenzuron, rather than target site modification. A positive correlation between enhanced GST activities and deltamethrin resistance suggested that this mechanism might take place into the adaptive response of C. pomonella to pyrethroids treatments. Enhanced MFO activity was recorded in each instar of the two resistant strains compared to the susceptible one. But these activities were not correlated to the responses to deltamethrin nor to teflubenzuron. In the light of these findings, studying age-dependence of responses to selection is central to the implementation of monitoring tests of resistances, especially if the target instars are difficult to collect in the field.     

Resistance of Spanish codling moth (Cydia pomonella) populations to insecticides and activity of detoxifying enzymatic systems

Resistance of codling moth, Cydia pomonella (L.) (Lepidoptera: Tortricidae), to insecticides has become a major problem in many apple and pear production areas. Our aim was to determine the level of insecticide resistance in Spanish field populations. Seven field populations collected from apple, Malus domestica Borkhausen (Rosaceae), orchards, and three laboratory susceptible strains of codling moth were studied. Damage at harvest in all the conventional orchards from which codling moth populations were collected was higher than the economic threshold. The efficacy of eight insecticides, with five modes of action, was evaluated by topical application of the diagnostic concentrations on post‐diapausing larvae. The enzymatic activity of mixed‐function oxidases (MFOs), glutathione transferases (GSTs), and esterases (ESTs) was evaluated for each population. The susceptibility to insecticides and the biochemical activity of the three laboratory strains and one organic orchard population were not significantly different. Field populations were less susceptible to the tested insecticides than the susceptible strains, especially for azinphos‐methyl, diflubenzuron, fenoxycarb, and phosalone. The efficacy of all insecticides was significantly dependent on the activity of MFOs. Only the toxicity of the three insecticides most used in Spain when the populations were collected (azinphos‐methyl, fenoxycarb, and phosalone) was also dependent on the activity of ESTs and GSTs activity. We conclude that the control failures were because of the existence of populations resistant to the main insecticides used.     

Sex linkage of CpGV resistance in a heterogeneous field strain of the codling moth Cydia pomonella (L.)

The occurrence of codling moth populations in European apple orchards that were not controlled by Cydia pomonella granulovirus (CpGV) is the first reported case of field resistance against a baculovirus control agent. A monogenic dominant sex-linked mode of inheritance was previously demonstrated in single-pair crosses between a homogeneous resistant (CpRR1) and a susceptible (CpS) laboratory strain of codling moth. However, resistant field populations (CpR) are more heterogeneous in their levels of resistance, and the possibility that they could harbor different resistance genes to CpRR1 had not been directly addressed. Here we report single pair crossing experiments using a resistant codling moth strain collected from an apple orchard in the southwest of Germany. Single-pair crosses within the field strain revealed a genetic basis to the heterogeneity of CpR concerning CpGV resistance. Hybrid crosses to a susceptible laboratory strain and backcrosses of the F1 generation to the resistant CpR strain confirmed that the homogeneous CpRR1 and the heterogeneous field strain CpR share the same mode of inheritance. Thus the variable levels of CpGV resistance in field populations is likely due to frequency differences of the same resistance-conferring gene, rather than different genes, which will facilitate future efforts to monitor and manage resistance.     

Releases of Trichogramma platneri (Hymenoptera: Trichogrammatidae) in Apple Orchards under a Sterile Codling Moth Release Program

Based on the premise that augmented host numbers may help multiply and support parasitoid populations, the egg parasitoid Trichogramma platneri Nargarkatti was released in apple orchards which were participating in a sterile codling moth, Cydia pomonella (L.), release program. Nonviable eggs resulting from matings involving at least one sterile codling moth partner can be successfully parasitized by T. platneri. Grain moth-reared, as well as codling moth-reared, T. platneri were released either in the spring or in the spring and summer oviposition period(s) of wild codling moth. Sentinel codling moth eggs were hung weekly, for 3-day periods, from May until September to determine fluctuations in T. platneri populations both during and between releases. Low numbers of wild or nonviable codling moth eggs or other susceptible host eggs resulted in sufficient eggs to maintain low spring- or summer-introduced T. platneri populations. T. platneri reduced codling moth damage in trees in which the Trichogramma were released.     

Effect of radiation on fecundity and fertility of codling moth Cydia pomonella (Linnaeus) (Lepidoptera: Tortricidae) from South Africa

Codling moth Cydia pomonella (Linnaeus) (Lepidoptera: Tortricidae) is the key pest of pome fruit in South Africa, and it’s control in apple and pear orchards relies on the application of insecticides and pheromone‐mediated mating disruption. Development of resistance to insecticides and placement of restrictions on the use of certain insecticides has made control of codling moth in South Africa increasingly problematic. The use of the sterile insect technique (SIT) as a control tactic for codling moth is under investigation as a potential addition to the current control strategy. We investigated the radiosensitivity of a laboratory strain of codling moth that was established from moths collected from commercial and organic orchards in the Western Cape, South Africa. Fecundity and fertility of this strain following radiation were consistent with values for the codling moth strain in the Canadian rearing facility in British Columbia. For both strains, the female codling moth was considerably more radiosensitive than the male. At a radiation dose of 100 Gy or higher, treated females were 100% sterile. The fertility of the South African strain was higher (86.3%) than for the Canadian strain (71.9%). This difference in fertility between the two strains was maintained when the dose of radiation was 100 Gy. However, the level of fertility was very similar between the two strains for doses ≥150 Gy. Therefore, based upon previously published work and the data from this study, an operational dose of 150 Gy is recommended for future codling moth SIT programmes in South Africa.     

Costs of insecticide resistance in Cydia pomonella (Lepidoptera: Tortricidae)

The obvious benefits associated with insecticide resistance for pest species may come at a cost to other life-history traits. In this study, we compared the larval and pupal developmental times, pupal mass wing surface area and wing fluctuating asymmetry (FA) in insecticide resistant and control strains of codling moth, Cydia pomonella (L.) (Lepidoptera: Tortricidae), collected from apple (Malus spp.) orchards in central Canada. Resistant strains had significantly longer larval developmental times and lower pupal masses compared with the susceptible strain. Although the forewings of resistant moths were smaller in resistant than control strain, no difference in wing FA was detected. Longer developmental times could increase exposure time to natural enemies, and reduced adult size could affect longevity and total reproductive output.     

Enzymatic diagnosis of resistance to deltamethrin in diapausing larvae of the codling moth,Cydia pomonella (L.)

The resistance to deltamethrin was evaluated in diapausing larvae of 14 field populations of the codling moth (Cydia pomonella L.) From the main French orchard areas using biological assays. Resistance to deltamethrin was compared to mixed-function-oxidase (mfo) activity measured at the individual level through ethoxycoumarin-O-deethylase (ECOD) activity using a fluorescence microplate reader. The larvae were collected in corrugated paper band traps in the autumn of 1995. Analysis was previously performed on two laboratory strains, one susceptible and one resistant to deltamethrin, in order to characterize the changes in resistance during diapause development. Resistance to deltamethrin as well as the ECOD activity were stable during the diapause, and ECOD activity was always significantly lower in the susceptible strain than in the resistant one. The ECOD activity was significantly correlated to the frequency of resistant moths in the field populations. This strongly suggested the involvement of the mfo system in the resistance to deltamethrin in these populations. The intra-strain variabilities in ECOD activity of both laboratory and field resistant insects indicated that other resistance mechanisms might also be involved. Further investigations on these mechanisms are needed in order to develop complete diagnostic methods and to define suitable control strategies against each resistant population. Arch. Insect Biochem. Physiol. 39:55–64, 1998. © 1998 Wiley-Liss, Inc.     

Genetic analysis of Cydia pomonella (Lepidoptera: Tortricidae) populations with different levels of sensitivity towards the Cydia pomonella granulovirus (CpGV)

Microsatellite (simple sequence repeats, SSR) and mitochondrial DNA markers were used to assess the structure of European codling moth populations showing different levels of susceptibility towards one of the most important biocontrol agents used in apple production, the Cydia pomonella granulovirus CpGV-M. In 638 C. pomonella individuals from 33 different populations a total of 92 different alleles were scored using six SSR loci. The global estimate of genetic differentiation for all 33 populations was not significantly different from zero, thus indicating a lack of genetic differentiation. AMOVA analysis revealed a very weak but significant variance among C. pomonella populations from different geographic regions, however, no significant variation was evident between CpGV-M resistant or susceptible C. pomonella populations. Sequence analysis of a fragment of the cytochrome oxidase subunit 1 in eight C. pomonella populations resulted in 27 haplotypes, which were grouped in two distinct clusters. Again, no genetic differentiation between CpGV-M resistant and susceptible codling moth populations was detectable. In addition, Structure analysis using microsatellites and association tests with mtDNA haplotypes found neither population-level nor individual correlations associated with CpGV-M resistance. Accordingly, this lack of population structure does not allow discriminating between one or several, separate origins of CpGV-M resistance.     

Variability in diapause propensity within populations of a temperate insect species: interactions between insecticide resistance genes and photoperiodism

Temperate insects generally use day length as a reliable cue for long-term seasonal changes in their environment. Significant variation in photoperiodism between and within populations is thought to be associated with genetic variation resulting from local adaptation. In this study, we investigated whether genetic variation associated with selection for insecticide resistance may be a source of divergence in the photoperiodic timing of diapause through pleiotropic interactions. Critical photoperiods for diapause induction were estimated in one susceptible and two insecticide-resistant homozygous strains of the codling moth Cydia pomonella , as well as in their reciprocal F₁ progeny. Diapause responses to naturally decreasing day length were subsequently followed in the laboratory strains and in two field populations of C. pomonella in south-eastern France. We found higher critical photoperiods for diapause induction in homozygous resistant individuals than in both homozygous susceptible and heterozygous ones. This partly explained the significantly earlier timings of diapause found in homozygous resistant individuals of both laboratory and field populations of C. pomonella under natural photoperiods, relative to those found in both homozygous susceptible and heterozygous ones. We assume that adaptive genetic changes associated with selection for insecticide resistance may generate substantial variation in the seasonal timing of diapause, an essential ecological feature of the fitness of insecticide resistance genes in this species.  © 2004 The Linnean Society of London, Biological Journal of the Linnean Society , 2004, 83 , 341–351.     

Mating behaviour and reproductive output in insecticide‐resistant and ‐susceptible strains of the maize weevil (Sitophilus zeamais)

Insecticide resistance is a broadly recognised and well‐studied management problem resulting from intensive insecticide use, which also provides useful evolutionary models of newly adapted phenotypes to changing environments. Two common assumptions in such models are the existence of fitness costs associated with insecticide resistance, which will place the resistant individuals at a disadvantage in insecticide‐free environments, and the prevalence of random mating among insecticide‐resistant and ‐susceptible individuals. However, cases of insecticide resistance lacking apparent fitness disadvantages do exist impacting the evolution and management of insecticide resistance. Assortative mating, although rarely considered, may also favour the evolution and spread of insecticide resistance. Thus, the possible existence of both conditions in the maize weevil (Sitophilus zeamais), a key pest of stored cereals, led to the assessment of the mating behaviour and reproductive fitness of insecticide‐resistant and ‐susceptible weevil strains and their reciprocal crosses. The patterns of female and male mating choice also were assessed. Although mating behaviour within and between weevil strains was similar without mate choice, mating within the resistant strain led to higher reproductive output than within the susceptible strain; inter‐strain matings led to even higher fertility. Thus, no apparent fitness cost associated with resistance seems to exist in these weevils, favouring the evolution of this phenotype that is further aided by the higher fertility of inter‐strain matings. Mate choice reduced latency to mate and no inter‐strain preference was detected, but female weevils were consistent in their mate selection between 1st and 2nd matings indicating existence of female mating preference among maize weevils. Therefore, if female mate selection comes to favour trait(s) associated with insecticide resistance, higher reproductive fitness will be the outcome of such matings favouring the evolution and spread of insecticide resistance among maize weevil populations reverting into a management concern.     

Improvement of the sterile insect technique for codling moth Cydia pomonella (Linnaeus) (Lepidoptera Tortricidae) to facilitate expansion of field application

The codling moth Cydia pomonella (L.) (Lepidoptera Tortricidae) is a key pest of pome fruit (apple, pear and quince) and walnut orchards in most temperate regions of the world. Efforts to control the codling moth in the past mostly relied on the use of broad spectrum insecticide sprays, which has resulted in the development of insecticide resistance, and the disruption of the control of secondary pests. In addition, the frequent reliance and use of these insecticides are a constant threat to the environment and human health. Consequently, there have been increased demands from the growers for the development of codling moth control tactics that are not only effective but also friendly to the environment. In that respect, the sterile insect technique (SIT) and its derivative, inherited sterility (IS), are, together with mating disruption and granulosis virus, among the options that offer great potential as cost‐effective additions to available control tactics for integration in area‐wide integrated pest‐management approaches. In support of the further development of the SIT/IS for codling moth control, the Joint FAO/IAEA Programme of Nuclear Techniques in Food and Agriculture implemented a 5‐year Coordinated Research Project (CRP) entitled ‘Improvement of codling moth SIT to facilitate expansion of field application’. Research focussed on sterile codling moth quality and management (e.g. mobility and life‐history traits in relation to rearing strategy, dispersal, flight ability, radiosensitivity and mating compatibility) and a better understanding of the basic genetics of codling moth to assist the development of genetic sexing strains (e.g. cytogenetics, the development of dominant conditional lethal mutations, molecular characterization of the sex chromosomes, sex identification in embryos and cytogenetic markers). The results of the CRP are presented in this special issue.     

Fitness costs and morphological change of laboratory‐selected thiamethoxam resistance in the B‐type Bemisia tabaci (Hemiptera: Aleyrodidae)

Thiamethoxam has been used as a key insecticide to control the whitefly, B‐type Bemisia tabaci, for several years in China with no known cases of resistance in field populations. To evaluate the risk of resistance, a field population was collected and resistant strains were developed by exposure to thiamethoxam in the laboratory. After selection for 36 generation, a strain with 60‐fold resistance was successfully identified. Fitness analysis by constructing life tables, demonstrated that resistant B‐type whiteflies had obvious fitness disadvantages in their development and reproduction. The fitness of resistant B‐type whiteflies decreased dramatically, to only one‐half that of the susceptible strain. Some changes in the morphological characteristics of the resistant strain were observed. The lengths of first, second and third instars of the resistant strain were significantly smaller than those of the susceptible strain, and the width of the first and the fourth instars were also significantly smaller than in the susceptible strain. Our results suggest that the B‐type B. tabaci has the potential to develop high resistance to thiamethoxam, and that the resistance changed the morphology of the insects. The slow development of resistance and the lower fitness of resistant B. tabaci strains may result in a quick recovery of sensitivity when the population is no longer in contact with thiamethoxam in the field.     

Trichogramma platneri (Hym.: Trichogrammatidae): Host choices between viable and nonviable codling moth,Cydia pomonella,and three-lined leafroller,Pandemis limitata (Lep.: Tortricidae) eggs

In laboratory host-preference studies,Trichogramma platneri Nagarkatti (Hym.: Trichogrammatidae) females, reared on viable codling moth,Cydia pomonella (L.) (Lep.: Tortricidae) eggs, parasitized significantly more viable than nonviable codling moth eggs produced by cobalt 60 irradiated adults. This ovipositional preference was maintained when competition increased per host egg. Similarly, one, three and fiveT. platneri females consistently oviposited more eggs into viable versus nonviable host eggs. A single femaleT. platneri parasitized a similar number of viableC. pomonella and three-lined leafroller,Pandemis limitata (Robinson) (Lep.: Tortricidae) eggs. However, as the number of females per host eggs increased, a significantly larger number of codling moth versus leafroller eggs were parasitized. One, three and fiveTrichogramma females parasitized significantly moreP. limitata versus nonviableC. pomonella eggs. NonviableC. pomonella eggs deflated to less than 25% of their original volume in four and ten days when held under dry and humid conditions respectively. The blackened prepupal stage of the parasitoid generally occurred in the nonviableC. pomonella eggs four days post-parasitization whereas the parasitized viable eggs blackened zero to one day earlier. Incubation under dry or humid conditions did not appear to influence the number of parasitized eggs that were able to blacken and show evidence of parasitism. Summerland Research Centre Contribution No. 938.     

Bidirectional selection for body mass and correlated response of pyrethroid resistance and fitness in Sitophilus zeamais

Responses to artificial selection on body mass in the maize weevil Sitophilus zeamais (Coleoptera: Curculionidae) were investigated to determine whether changes in body mass are associated with insecticide susceptibility, rate of population growth, and metabolic rate. Two strains of the maize weevil differing in susceptibility to pyrethroid insecticides were subjected to bidirectional selection on body mass. The susceptible strain responded to selection resulting in individuals with lower or higher body mass, but the resistant strain responded significantly only to selection for lower body mass. The resistant strain selected for low body mass increased its level of deltamethrin resistance in 44 × . In contrast, selection for low body mass in the susceptible parental strain led to increased deltamethrin susceptibility (50 × ) and selection for high body mass increased deltamethrin resistance (4 × ). Thus, the correlated response of insecticide resistance to selection for body mass differed between strains, a likely consequence of their distinct genetic background. Regardless, body mass was positively correlated with fitness (reproductive output) (r = 0.79; P < 0.001), while such correlation with respiration rate was significant only at P = 0.07 (r = 0.44). Therefore, the association between body mass and deltamethrin resistance is population‐dependent in the maize weevil, and the confluence of deltamethrin resistance and high body mass in a given strain will likely favour its energy metabolism and lead to the mitigation of fitness costs usually associated with insecticide resistance. The genetic background and selection history of insecticide resistant populations should not be neglected since they may favour the confluence of insecticide resistance with mitigation mechanisms of its associated fitness costs limiting the tactics available to their management.     

不同品系小菜蛾成虫脑突触体 蛋白质磷酸化的研究

对小菜蛾Plutella xylostella(L.)敏感品系、抗溴氰菊酯品系、抗杀虫双品系和抗杀螟丹品系的成虫脑突触体蛋白质磷酸化进行了研究比较。结果表明：蛋白质磷酸化在各个品系中的表现是不一样的。cAMP和钙加钙调蛋白对不同品系小菜蛾脑蛋白质磷酸化都有不同程度的刺激作用;3种杀虫剂均对各品系小菜蛾的磷酸化反应有影响,杀虫双、杀螟丹表现为抑制,溴氰菊酯表现为加强。这种影响在敏感品系中表现得比抗性品系中要强烈。     

Overcoming the resistance of codling moth against conventional Cydia pomonella granulovirus (CpGV-M) by a new isolate CpGV-I12

Recently, codling moth (CM, Cydia pomonella L.) populations with a significantly reduced susceptibility to C. pomonella granulovirus (CpGV) products have been observed in Germany. A novel CpGV isolate, designated CpGV-I12, is able to overcome the CpGV resistance. CpGV-I12 originated from Iran and showed superior efficacy in laboratory bioassays against a resistant CM strain (CpR), which has a 100-fold reduced susceptibility to commercially used isolate CpGV-M. Determination of the median lethal concentration (LC₅₀) indicated that CpGV-I12 is nearly as efficient in resistant CpR as CpGV-M in a susceptible CM strain (CpS). Beyond, CpGV-I12 caused superior mortality in CpS. Infection experiments showed that the resistance breaking effect can be observed in all instars of CpR. CpGV-I12 is a promising alternative control agent of CM in orchards where conventional CpGV products fail. In addition, we demonstrate in bioassays with recombinant expressed Cry1Ab that cross-resistance to CpGV and Bacillus thuringiensis products is not likely.     

Factors influencing codling moth larval response to α-farnesene

Behavioral responses of newly-emerged codling moth (Cydia pomonella L.) larvae to α-farnesene were compared for a laboratory-reared strain (‘lab’, 160 generations inbred) and a recently collected strain (‘wild’, 3 generations). Video recordings of single larvae placed 20 mm from Bond papers releasing a range of seven α-farnesene concentrations were scored for head turning, head lifting, head direction, and movement across a Petri dish. The laboratory strain was significantly less successful at finding the treated papers than the wild strain. The lab strain was also significantly less active, shown by more random orientation, slower walking speed, less head turning and less head lifting. Both strains showed lower response to α-farnesene concentrations less than 10⁻⁶ (w/v) α-farnesene. Orientation, walking speed, and head turning rate were significantly higher in the presence of α-farnesene concentrations >10⁻⁷ w/v, for both strains. Head lifting did not show any effect from α-farnesene. Head turning was associated with both head lifting and walking speed and this association improved with the presence of α-farnesene. Increased attraction to odour source was correlated with improved direction finding at concentrations above 10⁻⁷ w/v. Activity factors such as head turning and walking speed influenced time to locate odour source more than orientation factors. Odour-guided orientation to α-farnesene in codling moth larvae was composed of both locomotory, and to a lesser extent, orientation responses, which increased in a directed fashion to the stimulus.     

白纹伊蚊抗高效氯氰菊酯和抗敌百虫品系的生物学特性及种群参数

白纹伊蚊Aedes albopictus Skuse抗高效氯氰菊酯和抗敌百虫品系的某些种群参数及生物学特性与敏感品系的比较有显著差异,两个抗药性品系之间差异也较明显。抗高效氯氰菊酯品系(Rc)的繁殖力强、平均世代历期短,雌性个体比率高。抗敌百虫品系(Rt)有适合度缺陷、平均世代历期长,雌性个体比率低。Rc和Rt的孵化率均低于敏感品系。成虫死亡率由高到低依次为Rt、Rc、S。综合分析得知,高效氯氰菊酯刺激白纹伊蚊生殖、种群增长快。抗敌百虫白纹伊蚊明显地表现出生殖不利性,种群加倍较慢。     

Pheromone pre-exposure and mating modulate codling moth (Lepidoptera: Tortricidae) response to host plant volatiles

Abstract 1 Two codling moth Cydia pomonella kairomonal attractants, ethyl (E,Z)‐2,4‐decadienoate (pear ester) and (E)‐β‐farnesene, were tested in an insecticide‐sprayed apple orchard and an orchard treated for mating disruption with synthetic pheromone (E,E)‐8,10‐dodecadienol (codlemone). Male captures with pear ester were higher in the pheromone‐treated than in the insecticide‐treated orchard, whereas captures with (E)‐β‐farnesene were not different. Subsequent wind tunnel experiments confirmed that pre‐exposure to sex pheromone codlemone increased the behavioural response of codling moth males to pear ester. This supports the idea that male attraction to the plant volatile pear ester and sex pheromone codlemone is mediated through the same sensory channels. 2 Pear ester is a bisexual codling moth attractant and even captures of female moths were significantly increased in the pheromone‐treated orchard. In the laboratory wind tunnel, pheromone pre‐exposure had no effect on female response to pear ester, but significantly more mated than unmated codling moth females flew upwind towards a pear ester source. Differences in mating status in insecticide‐treated vs. pheromone‐treated orchards may thus account for the differences in female trap captures with pear ester. 3 These findings are important with respect to monitoring of codling moth with pear ester in mating disruption orchards. They also emphasize the importance of host plant volatiles in pheromone‐mediated mating disruption, which has been neglected to date.     

Fitness costs of resistance to Bacillus thuringiensis in the Indianmeal moth, Plodia interpunctella

Genetic changes in insects that result in insecticide resistance can also affect their fitness. Here, we report measurements of development time and survival of the Indianmeal moth, Plodia interpunctella (Hübner), to compare the relative fitness of Bacillus thuringiensis (Bt)-susceptible and -resistant colonies. Measurements of larval development time and survival indicated that a fitness cost was associated with resistance to Bt in some Bt-resistant colonies but not others. Comparisons of geographically different populations revealed inherent differences in development time and survival. In most cases, Bt-resistant moths suffered no disadvantage when feeding on a Bt-treated diet. In many cases, the development of Bt-resistant moths on Bt-treated diet was slower than the unselected moths on untreated diet, but it is unclear whether these differences would affect the successful mating of susceptible and resistant moths.