Large-Scale Monitoring of the Frequency of Ryanodine Receptor Target-Site Mutations Conferring Diamide Resistance in Brazilian Field Populations of Fall Armyworm, Spodoptera frugiperda (Lepidoptera: Noctuidae).

Fall armyworm (FAW), Spodoptera frugiperda, is an important lepidopteran pest in the Americas, and recently invaded the Eastern Hemisphere. In Brazil, FAW is considered the most destructive pest of corn and cotton. FAW has evolved resistance to many insecticides and Bacillus thuringiensis (Bt) proteins. Here, a large-scale monitoring was performed between 2019 and 2021 to assess diamide insecticide susceptibility in more than 65 FAW populations sampled in corn and cotton. We did not detect a significant shift in FAW susceptibility to flubendiamide, but a few populations were less affected by a discriminating rate. F2 screen results of 31 selected FAW populations across regions confirmed that the frequency of diamide resistance alleles remained rather stable. Two laboratory-selected strains exhibited high resistance ratios against flubendiamide, and cross-resistance to anthranilic diamides. Reciprocal crosses indicated that resistance is autosomal and (incompletely) recessive in both strains. F1 backcrosses suggested monogenic resistance, supported by the identification of an I4734M/K target-site mutation in the ryanodine receptor (RyR). Subsequent genotyping of field-collected samples employing a TaqMan-based allelic discrimination assay, revealed a low frequency of RyR I4790M/K mutations significantly correlated with phenotypic diamide resistance. Our findings will help to sustainably employ diamides in FAW resistance management strategies across crops.

Sublethal Effects of Diamide Insecticides on Development and Flight Performance of Chloridea virescens (Lepidoptera: Noctuidae): Implications for Bt Soybean Refuge Area Management.

High-dose and refuge are the most important strategies for delaying resistance evolution in Bt crops. Insecticide sprays in refuge areas could be necessary and may limit refuge effectiveness. Here, we evaluated the sublethal effects of two diamide insecticides (chlorantraniliprole and flubendiamide) on Chloridea virescens life history traits and flight performance. Sublethal concentrations of chlorantraniliprole and flubendiamide increased larval and pre-pupal development times and decreased larval weight; flubendiamide increased pupal development times. Chlorantraniliprole increased adult male longevity and reduced female fertility, while flubendiamide reduced fecundity. Overall life table parameters were negatively impacted by both treatments. Males exposed to either insecticide showed significant reductions in flight duration and distance for unsustained flights (<30 min). The duration and distance of the first flights were reduced when exposed to chlorantraniliprole. Sustained flights (>30 min) were generally unaffected by insecticide exposure and both sexes flew >6400 m in a single flight. The sublethal effects of flubendiamide and chlorantraniliprole on C. virescens' population dynamics could lead to generation asynchrony and provide insufficient susceptible moths when sprayed on refuge crops. However, the distance and duration of flight may still be sufficient to ensure mixing of potentially resistant and susceptible populations from refuge plots.

Selection and characterization of the inheritance of resistance of Spodoptera frugiperda (Lepidoptera: Noctuidae) to chlorantraniliprole and cross-resistance to other diamide insecticides.

BACKGROUND: Understanding the genetic basis of insect resistance to insecticides can help to implement insecticide resistance management (IRM) strategies. In this study, we selected a strain of Spodoptera frugiperda (J. E. Smith) (Lepidoptera: Noctuidae) resistant to chlorantraniliprole using the F2 screen method, characterized the inheritance of resistance and evaluated patterns of cross-resistance to other diamide insecticides. RESULTS: The chlorantraniliprole-resistant strain (Chlorant-R) was selected from a field-collected population with an estimated allele frequency of 0.1316. The estimated median lethal concentration (LC50 ) values were 0.011 and 2.610 µg a.i. cm-2 for the susceptible (Sus) and Chlorant-R strains, respectively, resulting in a resistance ratio (RR) of 237-fold. The LC50 values of the reciprocal crosses were 0.155 and 0.164 µg a.i. cm-2 , indicating that resistance is autosomally inherited. Resistance was characterized as incompletely recessive and monogenic at concentrations close to the recommended field rates of chlorantraniliprole. Survival of the resistant strain and heterozygous larvae in maize plants treated at the field rate was ∼ 60 (similar to the susceptible strain on untreated maize) and 15%, respectively. The Chlorant-R strain showed low cross-resistance to cyantraniliprole (RR ∼ 27-fold) and very high cross-resistance to flubendiamide (RR > 42 000-fold). CONCLUSIONS: The frequency of chlorantraniliprole resistance allele was moderately high in a field-collected population of S. frugiperda. The inheritance of chlorantraniliprole resistance was characterized as autosomal, incompletely recessive and monogenic. S. frugiperda showed cross-resistance to other diamide insecticides. These results provide essential information for the implementation of IRM programs to preserve the useful life of diamide insecticides for controlling S. frugiperda in Brazil. © 2019 Society of Chemical Industry.

Field-Evolved Resistance and Cross-Resistance of Brazilian Tuta absoluta (Lepidoptera: Gelechiidae) Populations to Diamide Insecticides.

Tuta absoluta (Meyrick), one of the most important tomato pests worldwide, is heavily controlled by the application of insecticides. Diamide insecticides represent a new class of products recently registered to control T. absoluta After 6 yr of use, control failures have been reported in populations of this pest, suggesting a hypothetical resistance development. Therefore, a resistance survey was performed using nine populations of T. absoluta that were collected in open fields, including from areas with reports of a reduced efficacy of diamides in the Northeast and Central regions of Brazil. Initial surveys with diagnostic and label doses proved the reduced efficacy of diamides against most populations. The LC50 values of chlorantraniliprole varied from 0.0044 (Brasília) to 1,263 (América Dourada) mg AI liter-1 (the resistance ratios [RR50] ranged from 1.0- to 288,995-fold), whereas the LC50 values for cyantraniliprole and flubendiamide, respectively, varied from 0.015 (Brasília) to 281 (América Dourada) mg AI liter-1 and from 0.038 (Guaraciaba do Norte) to 3,018 (Gameleira 1) mg AI liter-1 The resistance ratios (RR50) ranged from 1.0- to 18,423-fold for cyantraniliprole and from 1.0- to 80,413-fold for flubendiamide. The log LC50 values of pairwise diamides were strongly and significantly correlated, which denoted cross-resistance among them. Very high resistance to diamides in T. absoluta was observed in this study, suggesting that strategies to mitigate resistance and thereby control the pest must not include only insecticides. Other control tactics must be carefully implemented over time to increase the life span of diamides, including rotational practices with other molecules.

Susceptibility of Tuta absoluta (Lepidoptera: Gelechiidae) Brazilian populations to ryanodine receptor modulators.

BACKGROUND: Phthalic and anthranilic diamides comprise a new insecticide class recently registered in Brazil to control Lepidoptera such as Tuta absoluta (Meyrick). Therefore, the baseline of susceptibility was determined for eight representative field populations of this species to establish a resistance monitoring programme. The potential for cross-resistance as well as detoxification metabolism was assessed in order to fine-tune the resistance management programme. RESULTS: Brazilian populations were very susceptible to chlorantraniliprole (LC50 values varied from 3.17 to 29.64 µg AI L(-1) ), cyantraniliprole (LC50 values varied from 8.61 to 28.95 µg AI L(-1) ) and flubendiamide (LC50 values varied from 94 to 230 µg AI L(-1) ), with respective resistance ratios of 9.33-, 3.36- and 2.45-fold between most susceptible and tolerant populations. Anthranilic diamides showed significant correlations between log LC50 values among themselves, suggesting a high risk of cross-resistance. However, the log LC50 values of T. absoluta to phthalic diamide did not show any correlation with anthranilic diamides. Cytochrome- P450-dependent monooxygenase activity showed a weak correlation with log LC50 values of T. absoluta populations to anthranilic diamides, which suggests a potential route for evolving resistance to anthranilic diamides. CONCLUSION: The diamides were highly effective against T. absoluta, with populations showing a homogeneous response to them. Cross-resistance is very likely between anthranilic diamides in T. absoluta. Populations of this pest may evolve resistance by increasing cytochrome- P450-dependent monooxygenases.