Fenvalerate Resistance Research Articles

Identification of insect cuticular protein genes LCP17 and SgAbd5 from Helicoverpa armigera and evaluation their roles in fenvalerate resistance

Insect cuticular protein (ICP) plays an important role in insect growth and development. However, research on the role of ICP in insecticide resistance is very limited. In this study, insect cuticular protein genes LCP17 and SgAbd5 were cloned and characterized in Helicoverpa armigera based on previous transcriptome data. The functions of LCP17 and SgAbd5 genes in fenvalerate resistance were assessed by RNA interference (RNAi), and their response to fenvalerate was further detected. The results showed that LCP17 and SgAbd5 were overexpressed in the fenvalerate-resistant strain comparing with a susceptible strain. The open reading frames of LCP17 and SgAbd5 genes were 423 bp and 369 bp, encoding 141 and 123 amino acids, respectively. LCP17 and SgAbd5 genes were highly expressed in the larval stage, but less expressed in the adult and pupal stages. The expression level of LCP17 and SgAbd5 genes increased significantly after fenvalerate treatment at 24 h. When the cotton bollworms larvae were exposed to fenvalerate at LD50 level, RNAi-mediated silencing of LCP17 and SgAbd5 genes increased the mortality from 50.68% to 68.67% and 63.89%, respectively; the mortality increased to even higher level, which was 73.61%, when these two genes were co-silenced. Moreover, silencing of these two genes caused the cuticle lamellar structure to become loose, which led to increased penetration of fenvalerate into the larvae. The results suggested that LCP17 and SgAbd5 may be involved in the resistance of cotton bollworm to fenvalerate, and LCP17 and SgAbd5 could serve as potential targets for H. armigera control.

Correlation between pyrethroid knockdown resistance and mutation frequency of voltage-gated sodium channel and its application in Aedes aegypti management

Aedes aegypti, the primary vector responsible for transmitting dengue fever in southern Taiwan, has developed a relatively high resistance to synthetic pyrethroids. It has evolved four amino acid substitutions in the voltage-gated sodium channel (VGSC), namely S996P, V1023G, F1565C, and D1794Y. To unveil the distribution and correlation of VGSC mutations and pyrethroid resistance among different field populations, Ae. aegypti collected from various districts in Kaohsiung and Tainan Cities underwent tests for resistance development against different pyrethroids and frequency of S996P, V1023G, F1565C, and D1794Y substitutions. The adult knockdown assay revealed a relatively high knockdown resistance in the Ae. aegypti populations from Kaohsiung and Tainan against permethrin, cypermethrin, and fenvalerate (averaging >50-fold). Conversely, less resistance was observed against α-cypermethrin, deltamethrin, λ-cyhalothrin, cyfluthrin, and etofenprox (averaging <35-fold). Using Polymerase Chain Reaction/restriction fragment length polymorphism analysis, four mutant haplotypes were identified in these field populations. Notably, the SIAVFD and SIBVFD wild haplotypes were absent. Analysis utilizing IBM SPSS Statistics 20.0 and Spearman's rank correlation coefficient indicated that Haplotype C (PIAGFD), especially P allele, frequency displayed a significant positive correlation with five Type II pyrethroid resistance, while 1023G and 1023G/G exhibited a significant association with permethrin and fevalerate resistance. Conversely, Haplotype E (SIBVCD) negatively correlated with pyrethroid resistance, particularly fenvalerate resistance (−0.776). Haplotype C and E were the most prevalent and widely distributed among the investigated field populations. This prevalence of haplotype C is likely tied to the extensive and excessive use of Type II pyrethroids for dengue control over the past three decades. Given the significant positive correlation, the best-fit lines and R2 values were established to facilitate the swift prediction of knockdown resistance levels to various pyrethroids based on VGSC mutation frequency. This predictive approach aims to guide insecticide usage and the management of pyrethroid resistance in the field populations of Ae. aegypti in Taiwan.

Biochemical mechanisms and molecular analysis of fenvalerate resistant population of Spodoptera litura (Fabricius)

Spodoptera litura (Fabricius) is an important polyphagous insect pest of several host plants worldwide and has developed resistance to many insecticides. The present study aimed at analysing the effect of selection with fenvalerate on a resistant field population from Sangrur, Punjab, India (SGR) through synergist and biochemical assays. Following selection with fenvalerate for 14 generations, the selected population (SGR-Sel) was analysed through bioassays to assess changes in levels of resistance, synergism and detoxification enzyme activities. Resistance ratio increased from 12.61 to 52.81 fold when compared to the laboratory susceptible strain (H-Sus). Synergism of piperonyl butoxide (6.19 fold) and triphenyl phosphate (5.88) was observed with SGR-Sel, which suggested the involvement of mainly monooxygenases (mixed function oxidases or MFOs) and esterases in resistance to fenvalerate. A strong positive correlation (r = 0.98) was observed between MFO activity and fenvalerate resistance in SGR-Sel, the activity was 4.26- and 1.84- fold that of H-Sus and SGR. Similarly, esterase activity was 3.70- and 1.73- fold that of H-Sus and SGR populations, with a strong positive correlation between esterases and fenvalerate resistance (r = 0.97). Alignment between COI sequences of SGR-Sel and SGR identified the existence of a single base/nucleotide polymorphism at position 421 represented by replacement of C in SGR with T in SGR-Sel. In the translated protein sequence, serine in SGR was replaced by phenylalanine in SGR-Sel population at position 139.

Point mutations in the voltage-gated sodium channel gene associated with pyrethroid resistance in Iranian populations of the European red mite Panonychus ulmi

The European red mite Panonychus ulmi (Koch) is a major pest of apple trees worldwide and causes significant damage to apple orchards in Iran. Pyrethroid insecticides/acaricides, such as fenpropathrin and fenvalerate, are widely used to control P. ulmi, but their long-term use may lead to low efficacy. Earlier studies investigating pyrethroid resistance in closely related mites such as Tetranychus urticae revealed that pyrethroid resistance was associated with point mutations in the voltage-gated sodium channel gene (vgsc). The aim of this study was to investigate the biochemical and molecular mechanisms of fenpropathrin and fenvalerate resistance in Iranian populations of P. ulmi. Pyrethroid toxicity bioassays were carried out on different P. ulmi field populations. Marand (resistance ratio, RR = 149), Maraqeh (RR = 90) and Mianeh2 (RR = 71) populations exhibited high levels of resistance to fenpropathrin, compared to a susceptible field population (Shahin Dej). Resistance was also observed for fenvalerate with resistance ratio's ranging from 2- to 20-fold. Synergism experiments and enzyme activity assays predicted a minor role for classical detoxification enzymes. In contrast, two amino acid substitutions in the VGSC, L1024V and F1538I, that were previously shown to confer pyrethroid resistance, were detected in all three resistant P. ulmi populations and point towards target-site insensitivity as the most likely resistance mechanism. Furthermore, sequencing after cloning of vgsc fragments from single haploid males revealed the presence of multiple copies of vgsc in a highly resistant strain. The link between resistance mutations and vgsc copy number variation should be the subject of future study, as this might be used to develop molecular markers for monitoring pyrethroid resistance of P. ulmi in the field.

Biochemical Resistance Mechanisms to Fenvalerate in Plutella xylostella (Lepidoptera: Plutellidae).

The diamondback moth, Plutella xylostella (Linnaeus) (Lepidoptera: Plutellidae), is one of the most important pests of cruciferous crops in Iran and is controlled mostly by fenvalerate. The susceptibility to fenvalerate was investigated for four populations of P. xylostella. Bioassay results indicated significant differences among the populations tested. The highest level of resistance to fenvalerate was obtained for the Khuzestan (Khz) population (resistance ratio = 9.5). Survival was suppressed by diethyl maleate, piperonylbutoxide, and triphenyl phosphate, which confirmed that resistance to fenvalerate is caused by glutathione S-transferases (GSTs), mixed function oxidases, and esterases, respectively. Up to 8.6-, 2-, 2.7-, and 1.75-fold increases in GSTs, esterase (α-naphthylacerate and β-naphthyl acetate as substrate), and cytochrome P450 monooxygenases activities in resistant strains when compared with the susceptible one, were observed, respectively. The expression of six GST genes of P. xylostella including GSTs1, GSTe2, GSTe4, GSTo4, GSTd4, and GSTd5 were analyzed. The quantitative PCR analysis showed that three of the PxGSTs had the highest expression levels in the Khz population. Two of the GSTs (GSTd4 and PxGSTe2) exhibited highest expression level in both Khz and Alborz (Alb) populations. Therefore, PxGST genes were involved in fenvalerate resistance in P. xylostella. Overall, the mechanisms of insecticide resistance in diamondback moth populations in four regions of Iran were related to GST, esterase, and cytochrome P450 monooxygenase activities.

Pyrethroid and carbamate resistance in Australian Helicoverpa armigera (Lepidoptera: Noctuidae) from 2008 to 2015: what has changed since the introduction of Bt cotton?

Pyrethroid and carbamate resistance was evaluated in Helicoverpa armigera from 2008 to 2015. Insects were collected as eggs primarily from cultivated hosts in the major cropping areas of New South Wales and Queensland, Australia. Larvae reared from eggs were tested for resistance to fenvalerate, bifenthrin or methomyl in the F0 generation using a topical application of a discriminating dose of insecticide. In 2008-2009, resistance to fenvalerate was 71% and no resistance to bifenthrin was recorded. In the following two seasons, resistance to pyrethroids was relatively stable with fenvalerate resistance ranging from 63% to 67% and bifenthrin resistance ranging from 5.6% and 6.4% in 2009-2010 and 2010-2011, respectively. However, in 2011-2012, pyrethroid resistance had increased to 91% and 36% for fenvalerate and bifenthrin, respectively. Resistance remained above 90% for fenvalerate and above 35% for bifenthrin in the following three seasons from 2012 to 2015. In 2008-2009, methomyl resistance was 33% and declined to 22% and 15% in 2009-2010 and 2010-2011, respectively. Methomyl resistance remained at moderate levels from 2011-12 to 2014-15, ranging from 21% to 40%. Factors that influenced selection pressure of pyrethroid and carbamate insecticides and impacted resistance frequency in H. armigera may have been associated with changes in the composition of the cropping landscape. The rapid expansion of the pulse industry and the commensurate increased use of insecticide may have played a role in reselection of high-level pyrethroid resistance, and highlights the need for an urgent and strategic response to insecticide resistance management in the Australian grains industry.

Heterologous expression of Helicoverpa armigera cytochrome P450 CYP6B7 in Pichia pastoris and interactions of CYP6B7 with insecticides.

Previous studies indicated that constitutive over-expression of cytochrome P450 CYP6B7 was involved in fenvalerate resistance in Helicoverpa armigera. In this study, the CYP6B7 gene from H. armigera (namely HaCYP6B7), was heterologously expressed in Pichia pastoris GS115. A vector pPICZA-HaCYP6B7 was constructed and transformed into P. pastoris GS115, the transformant of pPICZA-HaCYP6B7-GS115 was then cultured and induced by 1% (v/v) methanol and the heterologous expression of HaCYP6B7 protein in P. pastoris was confirmed by SDS-PAGE and western blot. Microsomes containing the expressed HaCYP6B7 showed activities against model substrate p-nitroanisole and 7-ethoxycoumarin, with p-nitroanisole O-demethylation (PNOD) and 7-ethoxycoumarin O-deethylation (ECOD) activities of 15.66- and 4.75-fold of the control, respectively. Moreover, it showed degradation activities against the insecticides bifenthrin, fenvalerate and chlorpyrifos, with clearance activities of 6.88-, 1.49- and 2.27-fold of the control, respectively. The interactions of HaCYP6B7 with insecticides were further confirmed by molecular docking in silico with binding scores of 5.450, 5.295 and 2.197 between putative HaCYP6B7 protein and bifenthrin, fenvalerate and chlorpyrifos, respectively. The results of present study provided more direct and important evidence on the role of HaCYP6B7 conferring pyrethroid resistance in H. armigera. © 2017 Society of Chemical Industry.

Over-expression of multiple cytochrome P450 genes in fenvalerate-resistant field strains of Helicoverpa armigera from north of China

Pyrethroid resistance was one of the main reasons for control failure of cotton bollworm Helicoverpa armigera (Hübner) in China. The promotion of Bt crops decreased the application of chemical insecticides in controlling H.armigera. However, the cotton bollworm still kept high levels of resistance to fenvalerate. In this study, the resistance levels of 8 field-collected strains of H. armigera from north of China to 4 insecticides, as well as the expression levels of related P450 genes were investigated. The results of bioassay indicated that the resistance levels to fenvalerate in the field strains varied from 5.4- to 114.7-fold, while the resistance levels to lambda-cyhalothrin, phoxim and methomyl were low, which were ranged from 1.5- to 5.2-, 0.2- to 1.6-, and 2.9- to 8.3- fold, respectively, compared to a susceptible strain. Synergistic experiment showed that PBO was the most effective synergist in increasing the sensitivity of H. armigera to fenvalerate, suggesting that P450 enzymes were involved in the pyrethroid resistance in the field strains. The results of quantitative RT-PCR indicated that eight P450 genes (CYP332A1, CYP4L11, CYP4L5, CYP4M6, CYP4M7, CYP6B7, CYP9A12, CYP9A14) were all significantly overexpressed in Hejian1 and Xiajin1 strains of H. armigera collected in 2013, and CYP4L5 was significantly overexpressed in all the 6 field strains collected in 2014. CYP332A1, CYP6B7 and CYP9A12 had very high overexpression levels in all the field strains, indicating their important roles in fenvalerate resistance. The results suggested that multiple P450 genes were involved in the high-level fenvalerate-resistance in different field strains of H. armigera collected from north of China.

Variation in P450-mediated fenvalerate resistance levels is not correlated with CYP337B3 genotype in Chinese populations of Helicoverpa armigera

Metabolic resistance to synthetic pyrethroids in Helicoverpa armigera has recently been associated with the chimeric cytochrome P450 enyzme CYP337B3. One variant of the latter, CYP337B3v1, accounts for 40–50 fold resistance to fenvalerate in an Australian population while a second variant, CYP337B3v2, has been associated with ~7 fold resistance to cypermethrin in a Pakistani population. Here we show that ~250–1200 fold resistance to fenvalerate in populations of the species from northern and northwestern China is largely due to P450-based metabolism, and that CYP337B3v2 is also at high frequency in these populations but absent in a susceptible control strain. However we find little correlation between the level of resistance and CYP337B3v2 frequency, either across the resistant populations studied, or over time within them. While there is variation between populations in the levels of CYP337B3v2 expression, this is not correlated with the level of resistance either. These data suggest that much of the variation in the level of fenvalerate resistance in China is explained by P450s other than CYP337B3. We also find that both the level of resistance and CYP337B3v2 frequency decline in field populations transferred to the laboratory and remained there without fenvalerate exposure, suggesting a fitness cost to both characters in the absence of the pesticide pressure. However the declines in the two characters are not well correlated across populations, again consistent with a large contribution to the variation in resistance levels from factors other than CYP337B3.

An independent occurrence of the chimeric P450 enzyme CYP337B3 of Helicoverpa armigera confers cypermethrin resistance in Pakistan

The increasing resistance level of insect pest species is a major concern to agriculture worldwide. The cotton bollworm, Helicoverpa armigera, is one of the most important pest species due to being highly polyphagous, geographically widespread, and resistant towards many chemical classes of insecticides. We previously described the mechanism of fenvalerate resistance in Australian populations conferred by the chimeric cytochrome P450 monooxygenase CYP337B3, which arose by unequal crossing-over between CYP337B1 and CYP337B2. Here, we show that this mechanism is also present in the cypermethrin-resistant FSD strain from Pakistan. The Pakistani and the Australian CYP337B3 alleles differ by 18 synonymous and three nonsynonymous SNPs and additionally in the length and sequence of the intron. Nevertheless, the activity of both CYP337B3 proteins is comparable. We demonstrate that CYP337B3 is capable of metabolizing cypermethrin (trans- and especially cis-isomers) to the main metabolite 4'-hydroxycypermethrin, which exhibits no intrinsic toxicity towards susceptible larvae. In a bioassay, CYP337B3 confers a 7-fold resistance towards cypermethrin in FSD larvae compared to susceptible larvae from the Australian TWB strain lacking CYP337B3. Linkage analysis shows that presence of CYP337B3 accounts for most of the cypermethrin resistance in the FSD strain; up-regulation of other P450s in FSD plays no detectable role in resistance. The presence or absence of CYP337B3 can be easily detected by a simple PCR screen, providing a powerful tool to rapidly distinguish resistant from susceptible individuals in the field and to determine the geographical distribution of this resistance gene. Our results suggest that CYP337B3 evolved twice independently by unequal crossing-over between CYP337B2 and two different CYP337B1 alleles.

Current Status of Insecticide Resistance in &lt;I&gt;Helicoverpa armigera&lt;/I&gt; After 15 Years of Bt Cotton Planting in China

Insecticide resistance was an important factor responsible for outbreaks of Helicoverpa armigera (Hübner) in China in the early 1990s. Bt cotton has been adopted in China since 1997, and has resulted in a reduction of insecticide use for H. armigera control. After 15 yr of Bt cotton planting, in 2011 we surveyed resistance to fenvalerate, phoxim, and emamectin benzoate in 16 field populations of H. armigera collected from major cotton production areas of China. Fourteen populations from northern China showed very strong resistance to fenvalerate (from 43- to 830-fold) and low levels of resistance to phoxim (3.0- to 8.9-fold) when compared with the susceptible SCD strain of H. armigera, whereas two populations from northwestern China showed low levels of resistance to fenvalerate (3.0- and 10-fold) and no resistance to phoxim (0.7- and 0.9-fold). Synergist bioassays demonstrated that oxidase-based detoxification was involved in fenvalerate resistance and esterase-based detoxification in phoxim resistance in the resistant field populations. In comparison with the resistance in field populations before Bt cotton adoption, we observed a maintenance of high levels of fenvalerate resistance, but a reversion of phoxim resistance from high levels to low levels in the field populations of H. armigera from northern China. All 16 field populations from both northern China and northwestern China were susceptible to emamectin benzoate (with about two-fold variations in LD50s among populations), but the SCD strain has an inherent tolerance of 11-fold compared with the most susceptible field population (Xiajin-1). Emamectin benzoate is not cross resistant to fenvalerate and may provide an alternative option for H. armigera control in China, if the efficacy of Bt cotton is compromised by Bt resistance in the field.

Knockdown of several components of cytochrome P450 enzyme systems by RNA interference enhances the susceptibility of Helicoverpa armigera to fenvalerate

The function of cytochrome P450 proteins (P450s) in the metabolism of a variety of compounds by oxidation and reduction is well elucidated, but its interactions with other electron transfer components in the pyrethroid resistance of insect pests have been a mystery for a long time. In previous studies the authors cloned and characterised CYP6B7 and cytochrome b(5) (Cyt-b(5)) in the fenvalerate-resistant HDFR strain of cotton bollworm (Helicoverpa armigera Hübner) and showed that CYP6B7 mRNA was overexpressed and important for resistance to fenvalerate. In the present study, the functional interactions of CYP6B7, NADPH-dependent cytochrome P450 reductase (CPR) and Cyt-b(5) were assessed using RNA interference (RNAi) strategies and monitoring for fenvalerate resistance levels. RT-qPCR analyses indicated that the expression levels of CYP6B7, CPR and Cyt-b(5) mRNA were decreased drastically in the midgut of fourth-instar larvae of the H. armigera HDFR strain after corresponding double-stranded RNA (dsRNA) injection, compared with that of the control. The knockdown of CYP6B7, CPR and Cyt-b(5) transcripts was time course dependent during a 12-48 h period after dsRNA injection. At the earlier time points analysed, significant suppression of CYP6B7 mRNA levels was observed in larvae injected with dsCYP6B7-313 as compared with controls, and further suppression was observed in larvae injected with dsCYP6B7-313, dsCPR-403 and dsCyt-b(5) . The injection of dsCYP6B7-313 together with dsCPR-403 and dsCyt-b(5) increased larval susceptibility of the HDFR strain to fenvalerate. The results demonstrated that silencing of CYP6B7 alone or CYP6B7 together with CPR and/or Cyt-b(5) increased the susceptibility of H. armigera to fenvalerate, suggesting that CYP6B7, CPR and Cyt-b(5) collaboratively participated in enhanced metabolism of fenvalerate and played an important role in the resistance of H. armigera to fenvalerate.

Effect of fenvalerate on the reproduction and fitness costs of the brown planthopper, Nilaparvata lugens and its resistance mechanism

The brown planthopper (BPH), Nilaparvata lugens Stål, is a primary insect pest of cultivated rice, and its effective control is essential for crop production. However, in recent years, outbreaks of the brown planthopper have occurred more frequently in China. In order to determine the causes and mechanisms of insecticide-induced BPH resurgence and perform population management, we conducted the following studies. By the topical application method, our results showed that, fenvalerate acted as stimulus of fecundity from 3.50 × 10 −3 to 2.02 × 10 −2 μg/female in the BPH. Apart from 7.00 × 10 −3 μg/female, the number of hatched nymphs was increased gradually with an increase in application dose from 3.50 × 10 −3 to 1.74 × 10 −2 μg/female. After continuous selection with fenvalerate for 11 generations by the rice-stem dipping method, a resistant strain was achieved with medium resistance to fenvalerate (RR 39.22). Life table study indicated that the resistant strain (G4 and G8) showed reproductive advantages, including increased female ratio, copulation rate and fecundity. But the hatchability of resistant strain was lower. The survival rate and emergence rate were significantly lower in G4 and G8 resistant strain. Resistant strains in G4 and G8 showed a fitness advantage (1.04 and 1.11), and the number of offspring in G8 generation was higher than that in G4 generation. The significant difference detected between resistant insects (G4, G5, G8 and G9) and S-strain contains not only the effect of resistant selection but also the effect of continuous rearing itself. Hence it was concluded that the BPH had the potential to develop high resistance against fenvalerate and the induction of the nymphs by sublethal doses of fenvalerate was of importance in the BPH population management, particularly in the predicting. Further studies demonstrated that triphenyl phosphate (TPP) and diethyl maleate (DEM) had no synergism on fenvalerate. However, piperonyl butoxide (PBO) displayed significant synergism in susceptible strain (1.97) and resistant strain (2.73). We concluded that esterase and glutathione S-transferase play little role in fenvalerate detoxification. The increase of the P450-monooxygenases detoxification is an important mechanism for fenvalerate resistance. Because their resistant populations had a fitness advantage, we should pay close attention to the occurrence of BPH and use other functionally different insecticides to control the BPH.

Cloning and expression of cytochrome P450 CYP6B7 in fenvalerate‐resistant and susceptible Helicoverpa armigera (Hübner) from China

AbstractOur previous studies have found that the levels of cytochrome P450 and oxidative activities of cytochrome P450 monooxygenases were enhanced in a fenvalerate‐resistant strain (HDFR) of Helicoverpa armigera (Hübner) in China, comparing with those of a susceptible strain (HDS). Here, we report the molecular cloning and gene expression of a specific P450 CYP6B7 from HDFR and HDS strain of H. armigera. Comparison of the deduced amino acid sequence showed that there were three substitutions of amino acids in the coding region of CYP6B7 cDNA of HDFR strain, comparing with that of HDS strain. Meanwhile, there were three mutative nucleotides in the intron of CYP6B7 genomic DNA in HDFR strain. Northern blotting analysis showed that the expression of CYP6B7 mRNA in HDFR strain of H. armigera was more than 5‐fold higher than that in HDS strain. Further studies indicated that the expression of CYP6B7 mRNA increased as the larvae developed; however, it was rarely detected in the pupae and adult stages. The results suggested that cytochrome P450 CYP6B7 plays an important role in the pyrethroid insecticide fenvalerate resistance of HDFR strain of H. armigera; over‐expression of CYP6B7 combined with relevant substitution of amino acids might be responsible for fenvalerate‐resistance in H. armigera from China.

Identification of candidate genes for fenvalerate resistance in Helicoverpa armigera using cDNA‐AFLP

A cDNA-amplified fragment length polymorphisms approach was undertaken to screen for candidate genes associated with fenvalerate resistance in the AN02 strain of Helicoverpa armigera. Larvae and adults of this strain manifest approximately 50-fold resistance, which is suppressible by piperonyl butoxide and controlled by the semidominant gene RFen1 previously mapped to AFLP Linkage Group 13. Two cytochrome P450s (CYP337B1 and CYP4S1), one carboxylesterase-like protein and one glutathione transferase were found to be constitutively upregulated in resistant insects. Mapping of these potential detoxification genes showed that one of them, the novel P450 CYP337B1, was tightly linked to the resistance locus. This suggests that the RFen1(R) allele has a cis-acting effect on CYP337B1 expression, and possible trans-acting effects on expression of other genes.

Genetic relationships between development of insecticide resistance and reduction of egg size as a negative effect on the fitness of the diamondback moth Plutella xylostella (Lepidoptera: Yponomeutidae)

Reciprocal crosses between susceptible (SS) and resistant (RR) lines to fenvalerate were conducted to determine whether or not there are any genetic relationships between insecticide resistance and reduction in egg size, as well to whether or not they affect the offspring fitness of the diamondback moth, Plutella xylostella. The dose-mortality regression lines of the F1 hybrid of susceptible females and resistant males (SR) and resistant females and susceptible males (RS) were roughly intermediate between those of parental lines, i.e., SS and RR. This result indicates that the mode of inheritance of fenvalerate resistance was neither complete dominance nor recessive, with no sex-linkage. The egg size of RR lines was significantly smaller than that of the SS lines. The RS and SR lines have an egg size close to that of their own matrilineage. We considered that the mode of inheritance of resistance and that of egg size were different from each other, although the selection experiment showed that there is a negatively correlated response between these parameters. The survival rate of the RR lines was significantly lower than that of the SS lines. However, the survival rate of the RS lines did not differ from that of the SR lines. Reproductive fitness is discussed with regard to different egg sizes among parental lines and reciprocal crosses.

Correlation between Fenvalerate Resistance and Cytochrome P450-mediated &lt;I&gt;O&lt;/I&gt;-Demethylation Activity in &lt;I&gt;Helicoverpa armigera&lt;/I&gt; (Lepidoptera: Noctuidae)

Cytochrome P450 monooxygenases are a major metabolic mechanism responsible for pyrethroid resistance in Helicoverpa armigera (Hübner) from Asia. Cytochrome P450-mediated O-demethylation activity toward p-nitroanisole (PNOD) of individual fourth instars was determined in five strains of H. armigera by using a microplate reader. The four resistant strains of YS, HD, YGF, and YG59 had 6-, 71-, 2540-, and 11,800-fold resistance, respectively, to fenvalerate in comparison with the susceptible BK77 strain. Their mean PNOD activity was 4-, 10-, 24-, and 60-fold, respectively, compared with the BK77 strain. A strong positive correlation (correlation coefficient r = 0.98) between PNOD activity and fenvalerate resistance was found. Of 48 larvae from each strain, only 4% larvae of the susceptible BK77 strain had detectable PNOD activity, whereas 25, 33, 79, and 96% of larvae from the resistant strains YS, HD, YGF, and YG59 exhibited PNOD activity, respectively. There was a clear discrimination of patterns of PNOD frequency distribution between H. armigera strains and their magnitudes of fenvalerate resistance. The PNOD activity can be used as a biochemical marker for monooxygenase-mediated pyrethroid resistance in field populations of H. armigera.

Host resistance to an insecticide favors selection of resistance in the parasitoid, Cotesia plutellae (Hymenoptera: Braconidae)

Abstract Laboratory selection was conducted for fenvalerate resistance in two colonies of the parasitoid, Cotesia plutellae Kurdjumov, each reared on a different colony of its host, Plutella xylostella (L.), with different level of resistance to the insecticide. In the selection regime the insecticide was applied to the host larvae that harbored the parasitoid larvae. Compared with the unselected parasitoid colony, the parasitoid colony selected with the susceptible hosts acquired 4.5-fold resistance after 14 selection cycles, while the colony selected with the more resistant hosts acquired 13.6-fold resistance after 13 selection cycles. These results demonstrate that parasitoid larvae could be exposed to insecticide selection via the hosts, selection with more resistant hosts could accelerate development of resistance in the parasitoid, and resistance genes selected during larval development could be expressed at the adult stage. This information can help improve selection procedures for insecticide resistance in parasitoids.

Mechanisms of resistance to DDT and pyrethroids in Patagonian populations of Simulium blackflies.

Mixed populations of the pest blackflies Simulium bonaerense Coscarón & Wygodzinsky, S. wolffhuegeli (Enderlein) and S. nigristrigatum Wygodzinsky & Coscarón (Diptera: Simuliidae) are highly resistant to DDT and pyrethroids in the Neuquén Valley, a fruit-growing area of northern Patagonia, Argentina. As these insecticides have not been used for blackfly control, resistance is attributed to exposure to agricultural insecticides. Pre-treatment with the synergist piperonyl butoxide (PBO) reduced both DDT and fenvalerate resistance, indicating that resistance was partly due to monooxygenase inhibition. Pre-treatment with the synergist tribufos to inhibit esterases slightly increased fenvalerate toxicity in the resistant population. Even so, biochemical studies indicated almost three-fold higher esterase activity in the resistant population, compared to the susceptible. Starch gel electrophoresis confirmed higher frequency and staining intensity of esterase electromorphs in the resistant population. Incomplete synergism against metabolic resistance indicates additional involvement of a non-metabolic resistance mechanism, such as target site insensitivity, assumed to be kdr-like in this case. Glutathione S-transferase activities were low and inconsistent, indicating no role in Simulium resistance. Knowing these spectra of insecticide activity and resistance mechanisms facilitates the choice of more effective products for Simulium control and permits better coordination with agrochemical operations.

Mechanisms of Fenvalerate Resistance in the German Cockroach,Blattella germanica(L.)

The Munsyana strain of German cockroach, originally collected in Muncie, Indana, was found to have high-level resistance to fenvalerate, displaying 825-fold levels of resistance by topical application compared to a susceptible laboratory strain (Johnson Wax). Pretreatment with the cytochrome P450 monooxygenase inhibitors piperonyl butoxide (PBO) or MGK-264, or the esterase inhibitorS,S,S-tributyl phosphorotrithioate (DEF), reduced fenvalerate LD50resistance ratios by 12-fold, 55-fold, and 10-fold, respectively. Detoxication enzyme assays revealed elevated activity of microsomal oxidases (N-demethylation), esterases (PNPA hydrolysis), and glutathione-S-transferases (CDNB conjugation).In vivopenetration studies employing [14C]fenvalerate indicated reduced accumulation in the resistant strain relative to the susceptible strain.In vitrometabolism of [14C]fenvalerate by 105,000-g supernatant fractions with and without DEF inhibition suggested that hydrolysis was occurring. In the microsomal fraction, PBO also reduced the quantity of metabolitesin vitro. Overall, results suggest that monooxygenase, hydrolase, glutathione-S-transferase and decreased cuticular penetration are involved in fenvalerate resistance in the Munsyana strain. However, resistance to fenvalerate was not completely eliminated by either PBO, MGK-246, or DEF, suggesting that additional mechanisms, possibly including sodium channel insensitivity, are involved in this resistance.