Quantitative genetics of adult behavioral response and larval physiological tolerance to permethrin in diamondback moth (Lepidoptera: Plutellidae)

We investigated the genetic basis of adult behavioral response and larval physiological tolerance to permethrin within two diamondback moth, Plutella xylostella (L.) (Lepidoptera: Plutellidae), populations from Wooster and Celeryville, OH, with different average levels of larval tolerance. The adult behavioral response was measured as oviposition site preference and was investigated using full-sib design and parent-offspring regression. Additive genetic variance (0.134 +/- 0.02) and the heritability (h2 = 0.31 +/- 0.08) for the behavioral response was significant for the Celeryville population, suggesting that in this population, a high proportion of phenotypic variation for adult behavioral response to permethrin was heritable genetic variation. The larval physiological response was measured with a topical application bioassay and was investigated using a half-sib design. Significant additive genetic variances and heritabilities for physiological tolerance to permethrin were detected in both populations. The genetic correlation between adult behavioral response and larval physiological tolerance to permethrin were negative, but significant only in the Celeryville population; indicating that adults from this population that are more behaviorally responsive produced offspring that are more susceptible to permethrin. Our findings have implications for the evolution and management of insecticide resistance in the diamondback moth. The adult behavioral response can lower the exposure of larvae to the insecticide, lowering selection pressure for physiological resistance in larvae. Furthermore, to the extent that the adult behavioral response increases fitness, it can indirectly select for larval susceptibility because of the negative correlation between the two traits.     

Inheritance of Resistance to the Bacillus thuringiensis Toxin Cry1C in the Diamondback Moth

Laboratory selection increased resistance to the Bacillus thuringiensis toxin Cry1C in a strain of diamondback moth (Plutella xylostella). The selected strain was derived from a field population that had evolved high levels of resistance to Bacillus thuringiensis subsp. kurstaki and moderate resistance to Cry1C. Relative to the responses of a susceptible strain of diamondback moth, the resistance to Cry1C of the selected strain increased to 62-fold after six generations of selection. The realized heritability of resistance was 0.10. Analysis of F(inf1) hybrid progeny from reciprocal crosses between the selected strain and a susceptible strain showed that resistance to Cry1C was autosomally inherited. The dominance of resistance to Cry1C depended on the concentration; inheritance was increasingly dominant as the concentration decreased. Responses of progeny from single-pair families showed that resistance to Cry1C and resistance to Cry1Ab were inherited independently, which enhances opportunities for managing resistance. However, compared with projections based on previously reported recessive inheritance of resistance to Cry1A toxins, the potentially dominant inheritance of resistance to Cry1C observed here could accelerate evolution of resistance.     

Geographical variation in larval susceptibility of the diamondback moth, Plutella xylostella (Lepidoptera: Plutellidae) to Bacillus thuringiensis spore-crystal mixtures and purified crystal proteins and associated resistance development in India

The susceptibility of larvae of the diamondback moth, Plutella xylostella Linnaeus to purified crystal proteins and spore-crystal preparations of Bacillus thuringiensis was investigated for 13 populations from seven states in India. The LC50 (microg ml(-1), 48 h) values of Cry proteins for different populations of P. xylostella ranged from 0.14-3.74 (Cry1Aa), 0.007-1.25 (Cry1Ab), 0.18-2.47 (Cry1Ac) and 0.12-3.0 (Cry1C). The LC50 (mg (ai) l(-1), 48 h) of spore-crystal preparations ranged from 0.02-0.98 (HD-1) and 0.06-2.14 (HD-73). Significantly higher LC50 values for all tested toxins and strains were obtained with populations collected from Iruttupallam and Ottanchathiram in the southern state of Tamil Nadu, whereas some of the populations collected from the northern part of India were more susceptible than the susceptible IARI 17-65 population. The high levels of resistance in the Iruttupallam and Ottanchathiram populations to Cry1Ab suggested selection pressure by Cry1Ab, which is the predominant toxin in B. thuringiensis formulations used in India. Cry1Ab was found to be more toxic than the other toxins. The population from Iruttupallam showed increased resistance following selection with Cry1Ab in the laboratory (LC50 from 1.25 to 4.31 microg ml(-1) over two generations) and also showed cross resistance to CrylAa and CrylAc. The resistance to Biobit in the field population from Iruttupallam declined slowly; requiring c. 33 generations for an overall 10-fold decline in LC50 when the insects were reared in the laboratory without exposure to B. thuringensis.     

Phenotypic variation in adult behavioral response and offspring fitness in Plutella xylostella (Lepidoptera: Plutellidae) in response to permethrin

Phenotypic variation in behavioral response and physiological tolerance to permethrin was measured and compared between two populations of diamondback moth, Plutella xylostella (L.). A field population from Celeryville, OH, and a laboratory population originally collected from Wooster, OH, were compared. In laboratory choice bioassays, females from both populations were less likely to oviposit on cabbage leaf disks and seedlings treated with permethrin. The oviposition-deterrent effect was positively related to permethrin concentration. The laboratory population was significantly more behaviorally responsive to the insecticide and showed a significantly greater avoidance than the field population of the highest concentration of permethrin tested (1.50 g [AI] /liter). The physiological response of each population was measured by feeding bioassays, and the laboratory population was physiologically more susceptible to the permethrin. Larvae from the laboratory population that were fed permethrin-treated leaves had significantly lower growth rate, higher mortality, and lower adult fecundity compared with larvae from the field population. These data suggest that female moths that are more behaviorally responsive to permethrin produce offspring that tend to be more susceptible to the same insecticide, at least for the populations tested. The relationship between adult behavioral response and larval physiological tolerance to permethrin is discussed.     

High Genetic Variability for Resistance to Bacillus thuringiensis Toxins in a Single Population of Diamondback Moth

The long-term benefit of insecticidal products based on Cry toxins, either in sprays or as transgenic crops, is threatened by the development of resistance by target pests. The models used to predict evolution of resistance to Cry toxins most often are monogenic models in which two alleles are used. Moreover, the high-dose/refuge strategy recommended for implementation with transgenic crops relies on the assumption that the resistance allele is recessive. Using selection experiments, we demonstrated the occurrence in a laboratory colony of diamondback moth of two different genes (either allelic or nonallelic) that confer resistance to Cry1Ab. At the concentration tested, resistance was dominant in one selection line and partially recessive in the other. Resistant insects from the two selection lines also differed in their cross-resistance patterns. The diamondback moth colony was derived from a field population from the Philippines, which originally showed a different resistance phenotype. This is the first time that an insect population has been directly shown to carry more than one gene conferring resistance to the same Cry toxin.     

High genetic variability for resistance to Bacillus thuringiensis toxins in a single population of diamondback moth.

The long-term benefit of insecticidal products based on Cry toxins, either in sprays or as transgenic crops, is threatened by the development of resistance by target pests. The models used to predict evolution of resistance to Cry toxins most often are monogenic models in which two alleles are used. Moreover, the high-dose/refuge strategy recommended for implementation with transgenic crops relies on the assumption that the resistance allele is recessive. Using selection experiments, we demonstrated the occurrence in a laboratory colony of diamondback moth of two different genes (either allelic or nonallelic) that confer resistance to Cry1Ab. At the concentration tested, resistance was dominant in one selection line and partially recessive in the other. Resistant insects from the two selection lines also differed in their cross-resistance patterns. The diamondback moth colony was derived from a field population from the Philippines, which originally showed a different resistance phenotype. This is the first time that an insect population has been directly shown to carry more than one gene conferring resistance to the same Cry toxin.     

Cross-resistance and stability of resistance to Bacillus thuringiensis toxin Cry1C in diamondback moth.

We tested toxins of Bacillus thuringiensis against larvae from susceptible, Cry1C-resistant, and Cry1A-resistant strains of diamondback moth (Plutella xylostella). The Cry1C-resistant strain, which was derived from a field population that had evolved resistance to B. thuringiensis subsp. kurstaki and B. thuringiensis subsp. aizawai, was selected repeatedly with Cry1C in the laboratory. The Cry1C-resistant strain had strong cross-resistance to Cry1Ab, Cry1Ac, and Cry1F, low to moderate cross-resistance to Cry1Aa and Cry9Ca, and no cross-resistance to Cry1Bb, Cry1Ja, and Cry2A. Resistance to Cry1C declined when selection was relaxed. Together with previously reported data, the new data on the cross-resistance of a Cry1C-resistant strain reported here suggest that resistance to Cry1A and Cry1C toxins confers little or no cross-resistance to Cry1Bb, Cry2Aa, or Cry9Ca. Therefore, these toxins might be useful in rotations or combinations with Cry1A and Cry1C toxins. Cry9Ca was much more potent than Cry1Bb or Cry2Aa and thus might be especially useful against diamondback moth.     

Assessing permethrin resistance in the stable fly (Diptera: Muscidae) in Florida by using laboratory selections and field evaluations

Insecticide resistance in the stable fly, Stomoxys calcitrans (L.) (Diptera: Muscidae),has been demonstrated previously, but mostly with insecticides that are no longer used, such as the organochlorines. Resistance to commonly used pyrethroids has been evaluated twice, but only in the midwestern United States. Stable fly susceptibility to a commonly used pyrethroid, permethrin, was determined in Florida to assess the possibility of resistance development. Diagnostic concentration evaluations of three stable fly field strains demonstrated a maximum of 57 and 21% survival to permethrin residues of 3x and 10x the LC99 of a susceptible strain, respectively. Stable flies from an equine facility with no reported insecticide use demonstrated approximately 20% survival with a 3x diagnostic concentration. Despite a distance of 91-km between field collection sites, survival profiles of field-collected stable fly strains were similar. Although an established stable fly colony collected from a local dairy previously expressed low level resistance to permethrin residues, five generations of laboratory permethrin selection increased resistance 15-fold.     

Transgenic plants expressing two Bacillus thuringiensis toxins delay insect resistance evolution

Preventing insect pests from developing resistance to Bacillus thuringiensis (Bt) toxins produced by transgenic crops is a major challenge for agriculture. Theoretical models suggest that plants containing two dissimilar Bt toxin genes ('pyramided' plants) have the potential to delay resistance more effectively than single-toxin plants used sequentially or in mosaics. To test these predictions, we developed a unique model system consisting of Bt transgenic broccoli plants and the diamondback moth, Plutella xylostella. We conducted a greenhouse study using an artificial population of diamondback moths carrying genes for resistance to the Bt toxins Cry1Ac and Cry1C at frequencies of about 0.10 and 0.20, respectively. After 24 generations of selection, resistance to pyramided two-gene plants was significantly delayed as compared with resistance to single-gene plants deployed in mosaics, and to Cry1Ac toxin when it was the first used in a sequence. These results have important implications for the development and regulation of transgenic insecticidal plants.     

Cross-Resistance and Stability of Resistance to Bacillus thuringiensis Toxin Cry1C in Diamondback Moth

We tested toxins of Bacillus thuringiensis against larvae from susceptible, Cry1C-resistant, and Cry1A-resistant strains of diamondback moth (Plutella xylostella). The Cry1C-resistant strain, which was derived from a field population that had evolved resistance to B. thuringiensis subsp. kurstaki and B. thuringiensis subsp. aizawai, was selected repeatedly with Cry1C in the laboratory. The Cry1C-resistant strain had strong cross-resistance to Cry1Ab, Cry1Ac, and Cry1F, low to moderate cross-resistance to Cry1Aa and Cry9Ca, and no cross-resistance to Cry1Bb, Cry1Ja, and Cry2A. Resistance to Cry1C declined when selection was relaxed. Together with previously reported data, the new data on the cross-resistance of a Cry1C-resistant strain reported here suggest that resistance to Cry1A and Cry1C toxins confers little or no cross-resistance to Cry1Bb, Cry2Aa, or Cry9Ca. Therefore, these toxins might be useful in rotations or combinations with Cry1A and Cry1C toxins. Cry9Ca was much more potent than Cry1Bb or Cry2Aa and thus might be especially useful against diamondback moth.     

南京地区小菜蛾的抗药性检测及初步分析

利用浸叶法对南京郊区小菜蛾Plutella xylostella的抗药性进行了监测,发现其对拟除虫菊酯类药剂的抗性较高,而对氟虫腈、辛硫磷、毒死蜱、多杀菌素和虫酰肼依然处于敏感阶段。利用抗性小菜蛾测试发现,氧化胡椒基丁醚（增效醚,简称PBO）对拟除虫菊酯具有显著的增效作用。室内敏感性恢复实验表明,在不接触药剂的条件下,小菜蛾对拟除虫菊酯的抗性迅速下降,繁殖10代以后,抗性维持在低抗水平。因此, 小菜蛾的抗药性治理,应充分利用不同药剂的轮换使用,避免单一使用某一品种,以延缓抗性的发展,同时可以利用PBO增强拟除虫菊酯的防治效果,保证这类药剂在小菜蛾防治中的作用。     

阿维菌素对小菜蛾的抗性选育及其对解毒酶活性的影响

用阿维菌素对小菜蛾Plutella xylostella (L.)进行了抗性选育,并对选育过程中小菜蛾解毒酶的活性进行了研究。选育从F₀至F₂₁代,抗性缓慢波动上升,达到选育前的122.91倍;F₂₁至F₂₇代,抗性迅速增长,达到选育前的812.73倍,抗性发展趋势呈现S型曲线。随着选育代数的增加,对乙酰胆碱酯酶(AChE)没有明显的影响;羧酸酯酶(CarE)活性,F₂₇是F₀的1.5倍,从F₂₂开始,活性在较高水平上波动;谷胱甘肽转移酶(GST)活性F₂₇是F₀的2.2倍,且从F₁₈开始,活性在较高水平上波动。选育的抗性品系,增效醚对阿维菌素增效6.34倍。     

Effect of insecticides on the diamondback moth (Lepidoptera: Plutellidae) and its parasitoid Diadegma insulare (Hymenoptera: Ichneumonidae)

Studies were conducted to evaluate the toxicity of insecticides to adult Diadegma insulare (Cresson) and its host the diamondback moth, Plutella xylostella (L.). Leaf-dip and direct-dip bioassays for diamondback moth larvae and residual bioassays for adults of diamondback moth and D. insulare were used to assess mortalities. Larval mortalities at field rates were significantly higher with carbaryl, permethrin, spinosad, and tebufenozide when compared with Bacillus thuringiensis, or imidacloprid in the larval-dip bioassay 72 h after treatment. In the leaf-dip and residual bioassays, both permethrin and spinosad caused 100% mortalities to diamondback moth larvae and adults, respectively, 72 h after treatment. Of all the materials tested, only B. thuringiensis and tebufenozide were not toxic to D. insulare 24 h after treatment. Spinosad was not toxic to D. insulare 30 min after treatment. However, 100% mortality was observed 8 h after treatment.     

Selection for starvation resistance in Drosophila melanogaster : physiological correlates,enzyme activities and multiple stress responses

Correlated responses to artificial selection for stress tolerance can provide insight into underlying genetic variation and the physiological basis of stress resistance . Lines of Drosophila melanogaster held in the absence of food or with an unsuitable resource, specifically decomposing lemon, responded to selection by becoming starvation resistant. The lemon-selected lines also adapted by evolving a resource-based induction response. Compared to control lines, the selected lines tended to store more lipid, develop slower and have a larger body size. Additional responses included resistance to desiccation and acetone fumes, suggesting multiple stress resistance is a correlated result of selection for starvation resistance. The specific metabolic rate was lower in the starvation selected lines and enzyme activities changed in response to selection. In particular, enzyme activities indirectly associated with lipid biogenesis increased in both types of selected lines. The correlated responses to the two selection regimes were sufficiently consistent to indicate a common basis for starvation resistance. Specific responses to starvation selection appeared to oppose the short-term phenotypic responses to starvation. Thus, a common response to stress selection may be to ameliorate the immediate physiological impact of the stress factor.     

Genetic Divergence under Uniform Selection. II. Different Responses to Selection for Knockdown Resistance to Ethanol among DROSOPHILA MELANOGASTER Populations and Their Replicate Lines

We have tested the hypothesis that genetic differences among conspecific populations may result in diverse responses to selection, using natural populations of Drosophila melanogaster. Selection for ethanol tolerance in a tube measuring knockdown resistance was imposed on five West Coast populations. In 24 generations the selected lines increased their mean knockdown times, on average, by a factor of 2.7. An initially weak latitudinal cline was steepened by selection. The two southernmost populations showed the same increases in the selected character, but differed consistently in their correlated responses in characters related to ethanol tolerance. This result indicates that the populations responded to selection by different genetic changes. Selection decreased female body weight and increased resistance to acetone, suggesting components of the response unrelated to ethanol metabolism. The Adhs allele was favored by selection in all populations at the onset, but increased in frequency only in the selected lines of the southernmost population. There was a correlation between latitude and Adh frequency changes, suggesting that fitnesses of the Adh alleles were dependent on the genetic background. Genetic background also had a large effect on the loss of fitness due to selection. Genetic drift between replicate lines caused more variation in selection response than initial genetic differences between populations. This result demonstrates the importance of genetic drift in divergence among natural populations undergoing uniform selection, since the effective population sizes approached those of small natural populations. Drift caused greater divergence between selected replicates than control replicates. Implications of this result for the genetic model of selection response are discussed.     

Variation in susceptibility of Diadegma insulare (Hymenoptera: Ichneumonidae) to permethrin

Six field populations of the parasitoid Diadegma insulare (Cresson) were collected from Mexico and the United States and tested for their susceptibility to permethrin by using a glass-vial residue bioassay. Significant differences existed in susceptibility, with one population significantly more susceptible than the other five. Susceptibility of D. insulare increased when they were reared for several generations in the greenhouse in the absence of selection. D. insulare was less suceptible to permethrin than the adult stage of its host, Plutella xylostella (L.), from a laboratory colony, but more susceptible than those hosts collected from a commercial field. Our data suggest that a field population of D. insulare appeared to increase its tolerance to permethrin much more slowly than P. xylostella.     

High stability and no fitness costs of the resistance of codling moth to Cydia pomonella granulovirus (CpGV-M)

Resistance against the biocontrol agent Cydia pomonella granulovirus (CpGV-M) was previously observed in field populations of codling moth (CM, C. pomonella) in South-West Germany. Incidental observations in a laboratory reared field colony (CpR) indicated that this resistance is rather stable, even in genetically heterogeneous CM colonies consisting of both susceptible and resistant individuals. To test this hypothesis, the resistance level of CpR that was 1000times less susceptible to CpGV-M was followed for more than 60 generations of rearing. Even without virus selection pressure, the high level of resistance, expressed as median lethal concentration, remained stable for more than 30 generations and declined only by a factor of 10 after 60 generations. When cohorts of the F32 and F56 generations of the same colony were selected to CpGV-M for five and two generations, respectively, the resistance level increased to factor of >1,000,000 compared to a susceptible control colony. Laboratory reared colonies of CpR, did not exhibit any measurable fitness costs under laboratory conditions in terms of fecundity and fertility. Resistance testing of seven selected codling moth field populations collected between 2003 and 2008 in commercial orchards in Germany that were repeatedly sprayed with CpGV products gave evidence of different levels of resistance and a more than 20-fold increase of the resistance in 1-3years when selection by CpGV-M was continued. A maximum 1,000,000-fold level of resistance to CpGV-M that could be induced in the laboratory under virus pressure had been also observed in one field population. The high stability of resistance observed in the genetically heterogenous colony CpR indicates that resistance to CpGV-M is not very costly.     

Induced plant defense responses against chewing insects. Ethylene signaling reduces resistance of Arabidopsis against Egyptian cotton worm but not diamondback moth

The induction of plant defenses by insect feeding is regulated via multiple signaling cascades. One of them, ethylene signaling, increases susceptibility of Arabidopsis to the generalist herbivore Egyptian cotton worm (Spodoptera littoralis; Lepidoptera: Noctuidae). The hookless1 mutation, which affects a downstream component of ethylene signaling, conferred resistance to Egyptian cotton worm as compared with wild-type plants. Likewise, ein2, a mutant in a central component of the ethylene signaling pathway, caused enhanced resistance to Egyptian cotton worm that was similar in magnitude to hookless1. Moreover, pretreatment of plants with ethephon (2-chloroethanephosphonic acid), a chemical that releases ethylene, elevated plant susceptibility to Egyptian cotton worm. By contrast, these mutations in the ethylene-signaling pathway had no detectable effects on diamondback moth (Plutella xylostella) feeding. It is surprising that this is not due to nonactivation of defense signaling, because diamondback moth does induce genes that relate to wound-response pathways. Of these wound-related genes, jasmonic acid regulates a novel beta-glucosidase 1 (BGL1), whereas ethylene controls a putative calcium-binding elongation factor hand protein. These results suggest that a specialist insect herbivore triggers general wound-response pathways in Arabidopsis but, unlike a generalist herbivore, does not react to ethylene-mediated physiological changes.     

Assessment of insecticide resistance after the outbreak of diamondback moth (Lepidoptera: Plutellidae) in California in 1997

During an outbreak of the diamondback moth, Plutella xylostella (L.), in California in 1997, nine populations were collected from the major broccoli areas throughout the state. Populations were assayed for their susceptibility to currently used materials (Bacillus thuringiensis subsp. kurstaki, permethrin, and methomyl) and to newer materials that had not yet been commercially used in California (spinosad, emamectin benzoate, and chlorfenapyr). For the currently used insecticides, elevated levels of resistance were seen only with permethrin and seven of the nine populations had tolerance ratios (TR) of > 100. With the newer chemistries, TR values were all < 15. To compare potential cross-tolerance, TR values of the currently used insecticides were compared with TR values of the newer insecticides. There were significant relationships found between: methomyl and emamectin benzoate, methomyl and spinosad, and permethrin and spinosad. Further biochemical studies are needed to confirm the actual mechanisms that lead to these relationships and field tests are needed to determine what impact, if any, such TR levels would have on control in the field. These data indicate that resistance to at least one of the commonly used insecticides (permethrin) may have played a role in the outbreak during 1997. However, other factors may have been at least equally important. The winter of 1996-1997 was warmer than normal, and during the period from February through August of 1997 the amount of rainfall was < 50% of normal. Hot and dry conditions are known to be conducive to outbreaks of P. xylostella. These data add to an overall knowledge about the geographic variation of resistance in P. xylostella populations within the United States. They also serve as a baseline for monitoring changes in susceptibility to these newer insecticides and can also help explain the occurrence of outbreaks caused by factors other than insecticide resistance.     

Insect resistance to Bacillus thuringiensis: uniform or diverse

Resistance to the insecticidal proteins produced by the soil bacterium Bacillus thuringiensis (Bt) has been documented in more than a dozen species of insect. Nearly all of these cases have been produced primarily by selection in the laboratory, but one pest, the diamondback moth (Plutella xylostella), has evolved resistance in open-field populations. Insect resistance to Bt has immediate and widespread significance because of increasing reliance on Bt toxins in genetically engineered crops and conventional sprays. Furthermore, intense interest in Bt provides an opportunity to examine the extent to which evolutionary pathways to resistance vary among and within species of insect. One mode of resistance to Bt is characterized by more than 500-fold resistance to at least one Cry1A toxin, recessive inheritance, little or no cross-resistance to Cry1C, and reduced binding of at least one Cry1A toxin. Analysis of resistance to Bt in the diamondback moth and two other species of moth suggests that although this particular mode of resistance may be the most common, it is not the only means by which insects can attain resistance to Bt.