Assessing fitness costs of the resistance of Spodoptera frugiperda (Lepidoptera: Noctuidae) to pyramided Cry1 and Cry2 insecticidal proteins on different host plants.

Fall armyworm (FAW), Spodoptera frugiperda (Smith), is one of the major pests targeted by transgenic crops expressing insecticidal proteins from Bacillus thuringiensis (Bt) Berliner. However, FAW presents a high capacity to develop resistance to Bt protein-expressing crop lines, as reported in Brazil, Argentina, Puerto Rico and the southeastern U.S. Here, FAW genotypes resistant to pyramided maize events expressing Cry1F/Cry1A.105/Cry2Ab2 (P-R genotype) and Cry1A.105/Cry2Ab2 (Y-R genotype) from Brazil were used to investigate the interactions between non-Bt hosts (non-Bt maize, non-Bt cotton, millet and sorghum) and fitness costs. We also tested a FAW genotype susceptible to Bt maize and F1 hybrids of the resistant and susceptible genotypes (heterozygotes). Recessive fitness costs (i.e., costs affecting the resistant insects) were observed for pupal and neonate to adult survival of the P-R genotype on non-Bt cotton; larval developmental time of the P-R genotype on millet and sorghum; larval and neonate-to-adult developmental time of the Y-R genotype on non-Bt cotton and sorghum; the fecundity of the Y-R genotype on non-Bt cotton; and mean generation time of both resistant genotypes. However, on non-Bt cotton and non-Bt maize, the P-R genotype had a higher fitness (i.e., fitness benefits), displaying greater fecundity and rates of population increases than the Sus genotype. Non-recessive fitness costs (i.e., costs affecting heterozygotes) were found for fecundity and population increases on millet and sorghum. These findings suggest that, regardless of the disadvantages of the resistant genotypes in some hosts, the resistance of FAW to Cry1 and Cry2 Bt proteins is not linked with substantial fitness costs, and may persist in field conditions once present.

Inheritance patterns, cross-resistance and synergism in Spodoptera frugiperda (Lepidoptera: Noctuidae) resistant to emamectin benzoate.

BACKGROUND: Fall armyworm, Spodoptera frugiperda (J. E. Smith), is a relevant global pest due to severe damage caused on agricultural crops and its capacity to evolve resistance to insecticides. Here, we selected a strain of S. frugiperda resistant to emamectin benzoate under laboratory conditions to understand the inheritance patterns, cross-resistance and synergism involved in the resistance. RESULTS: The emamectin benzoate-resistant (Ben-R) strain was isolated by using F2 screen in a field population collected in Lucas do Rio Verde, Mato Grasso state, Brazil. After ten generations of selection pressure with emamectin benzoate, the estimated LC50 of the Ben-R strain was 678.38 µg a.i. mL-1 whereas that of the susceptible (Sus) strain was 0.29 µg a.i.mL-1 , resulting in a resistance ratio (RR) of ~ 2340-fold. The LC50 values of the offspring from reciprocal crosses of Sus and Ben-R strains were 93.37 and 105.32 µg a.i. mL-1 , suggesting that resistance is an autosomal incompletely dominant trait. The high survival of heterozygous and Ben-R strains (>92%) on non-Bt maize sprayed with the field rate of emamectin benzoate confirmed that resistance is functionally dominant. The minimum number of segregations influencing resistance was 3.55, suggesting a polygenic effect. Low cross-resistance was detected between emamectin benzoate and the insecticides methomyl, chlorpyrifos, lambda-cyhalothrin, spinetoram, indoxacarb and chlorantraniliprole (RR <5.75-fold). There was no effect of synergists piperonyl butoxide, diethyl maleate and S, S, S-tributyl phosphorotrithiotate on the Ben-R strain, suggesting a minor role of metabolic resistance. CONCLUSIONS: Our results showed a high risk of resistance evolution of S. frugiperda to emamectin benzoate, based on incompletely dominant inheritance. Rotation of insecticides with different modes of action can be one of the resistance management strategies to be implemented to delay the evolution of resistance of S. frugiperda to emamectin benzoate in Brazil.

Performance of Seed Treatments Applied on Bt and Non-Bt Maize Against Fall Armyworm (Lepidoptera: Noctuidae).

Fall armyworm (FAW), Spodoptera frugiperda (J. E. Smith), is the main pest of maize in Brazil, attacking plants from emergence to reproductive stages. Here, we conducted studies to evaluate the efficacy of two seed treatments (chlorantraniliprole alone and imidacloprid combined with thiodicarb) on Bt and non-Bt maize in laboratory bioassays with distinct FAW strains that are susceptible, selected for resistance to Bt-maize single (Cry1F) or pyramided (Cry1A.105 + Cry2Ab2) events and F1 hybrids of the selected and susceptible strains (heterozygotes), and in the field against a natural infestation. In the laboratory, leaf-discs from seed treated Bt-maize plants at 7 d after emergence (DAE) increased the mortality of FAW resistant, heterozygote, and susceptible strains up to 24.8%, when compared with the respective maize grown without a seed treatment. In the field against natural infestations of FAW, Bt maize with a seed treatment had ~30% less FAW damage than non-Bt maize with the same seed treatment at 7 and 14 DAE. No differences in FAW damage was observed between Bt and non-Bt maize grown with and without a seed treatment at 21 DAE. Maize seeds treated with chlorantraniliprole alone or imidacloprid and thiodicarb combined presented limited protection against early infestations of FAW strains under laboratory and field studies.

Field-evolved resistance to chitin synthesis inhibitor insecticides by soybean looper, Chrysodeixis includens (Lepidoptera: Noctuidae), in Brazil.

Soybean looper (SBL), Chrysodeixis includens (Walker), is an economically important soybean and cotton pest in Brazil. Here, we selected an SBL strain resistant to teflubenzuron using F2 screening, estimated the resistance allele frequency, characterized the inheritance of resistance, investigated fitness costs, evaluated patterns of cross-resistance, and determined the magnitude of resistance. The teflubenzuron-resistant strain (Teflu-R) was selected from field-collected populations with an estimated allele frequency of 0.1700. Estimated LC50 values were 0.010 and 363.61 µg a.i. cm-2 for the susceptible (Sus) and Teflu-R strains, respectively, representing a 36,361-fold resistance ratio (RR). The LC50 values of reciprocal crosses were 1.02 and 0.59 µg a.i. cm-2, suggesting that resistance is autosomally inherited. The low survival of reciprocal crosses (16 and 20%) on teflubenzuron-sprayed leaves indicates incomplete recessive resistance. The number of segregations influencing resistance was 2.72, suggesting a polygenic effect. The Teflu-R strain showed longer development periods as well as lower survival and population growth than the Sus strain, revealing fitness costs. The Teflu-R strain also showed high cross-resistancesto other chitin inhibitor insecticides, such as novaluron (RR = 6147-fold) and lufenuron (RR = 953-fold), but low cross-resistance to methoxyfenozide, flubendiamide, and indoxacarb (RR < 3.45-fold). On discriminatory concentrations of teflubenzuron and novaluron, populations of SBL showed survival rates from 15 to 52%, indicating field resistance to these insecticides. Our findings indicated that resistance to teflubenzuron in SBL is autosomal, recessive, polygenic, and associated with fitness cost. We also found a high cross-resistance to other benzoylphenylureas and a high frequency of resistance to this mode-of-action in SBL in Brazil.

Soybean aphid (Hemiptera: Aphididae) response to lambda-cyhalothrin varies with its virulence status to aphid-resistant soybean.

BACKGROUND: Soybean aphid, Aphis glycines, is an invasive insect in North America, considered one of the most important pests of soybean. Their management relies heavily on foliar insecticides, but there is growing effort to expand these tools to include aphid-resistant varieties. We explored if the LC50 and LC25 of lambda-cyhalothrin varied between virulent (resistant to Aphis glycines (Rag) soybeans) and avirulent (susceptible to Rag-genes soybeans) populations of soybean aphid with a leaf-dip bioassay. We also investigated the response to the LC25 of lambda-cyhalothrin on adults (F0) and their progeny (F1) for both avirulent and virulent soybean aphid. RESULTS: The LC50 of the virulent aphid population was significantly higher compared with the LC50 of the avirulent population. The LC25 significantly reduced fecundity of the F0 generation of avirulent soybean aphid, but no significant effect was observed for virulent aphids. In addition, the LC25 significantly shortened the adult pre-oviposition period (APOP) and lengthened the total pre-oviposition period (TPOP) of avirulent aphids, while the mean generation time (T) was significantly increased. For the virulent aphid, sublethal exposure significantly lengthened development time of first and third instars, TPOP, and adult longevity. In addition, all demographic parameters of virulent soybean aphid were significantly affected when they were exposed to the LC25 of lambda-cyhalothrin. CONCLUSION: Our results demonstrate lambda-cyhalothrin is less toxic to virulent aphids and exposure to the LC25 can trigger hormesis, which may have implications for the long-term management of this pest with this insecticide as well as with aphid-resistant varieties of soybean. © 2019 Society of Chemical Industry.

No cross-resistance between ChinNPV and chemical insecticides in Chrysodeixis includens (Lepidoptera: Noctuidae).

Chrysodeixis includens nucleopolyhedrovirus (ChinNPV: Baculoviridae: Alphabaculovirus) is an active ingredient of a biological-based insecticide (Chrysogen®) recommended against soybean looper (SBL), Chrysodeixis includens (Walker, [1858]), in soybean in Brazil. We investigated if SBL strains resistant to chemical insecticides are cross-resistant to the baculovirus ChinNPV. In droplet feeding bioassays, SBL strains resistant to lambda-cyhalothrin and teflubenzuron showed equivalent susceptibility to ChinNPV as heterozygous and susceptible strains, indicating no cross-resistance between ChinNPV and chemical insecticides in SBL. Therefore, the ChinNPV is a valuable new "mode-of-action" tool for SBL resistance management in Brazil.

Susceptibility of Brazilian Populations of Chrysodeixis includens (Lepidoptera: Noctuidae) to Selected Insecticides.

Soybean looper (SBL), Chrysodeixis includens (Walker) (Lepidoptera: Noctuidae), is an important pest of soybean and cotton in Brazil. The use of insecticides is one of the main control tactics against this pest. To support Integrated Pest Management (IPM) and Insect Resistance Management (IRM) programs, we characterized the susceptibility of Brazilian populations of SBL to insecticides. Field populations were collected from soybean fields during the 2016-2017 and 2017-2018 crop seasons. In the laboratory, late L2/early L3 larvae were exposed to insecticides in diet-overlay or topical bioassays. Field populations of SBL showed high susceptibility to spinetoram (LC50 = 0.074-0.25 µg a.i. per cm2), indoxacarb (LC50 = 0.46-0.94 µg a.i. per cm2), thiodicarb (LC50 = 9.14-36.61 µg a.i. per cm2), chlorantraniliprole (LC50 = 0.15-0.57 µg a.i. per cm2), flubendiamide (LC50 = 0.45-2.01 µg a.i. per cm2), and chlorfenapyr (LC50 = 0.15-0.25 µg a.i. per cm2); the resistance ratios were less than 16-fold. In contrast, SBL populations have reduced susceptibility to lambda-cyhalothrin (LC50 = 3.71-9.54 µg a.i./cm2), methoxyfenozide (LC50 = 0.67-4.23 µg a.i. per cm2), novaluron (LC50 = 27.52-77.63 µg a.i. per cm2), and teflubenzuron (LC50 = 13.41-73.02 µg a.i. per cm2). The resistance ratios relative to a Lab population (susceptible of reference) was up to 38-, 63-, 1,553-, and 5,215-fold, respectively. These high resistance ratios can be associated with resistance evolution. Implications of these findings to IPM and IRM programs are discussed.

Laboratory and field survival of Spodoptera frugiperda (Lepidoptera: Noctuidae) on Bt and non-Bt maize and its susceptibility to insecticides.

BACKGROUND: Field-evolved resistance of fall armyworm (FAW), Spodoptera frugiperda (Smith), has been reported to Bt maize technologies in Brazil. The control failures of FAW by Bt maize increased the use of insecticides for their control. However, no information is available on the interaction between resistant FAW and their response to insecticides. Here, we evaluated the survival of FAW strains on Bt and non-Bt maize in laboratory and field conditions, and their susceptibility to insecticides. RESULTS: In the laboratory, resistant FAW larvae reared on Bt and non-Bt maize showed a similar susceptibility to spinetoram (LC50 = 0.16-0.18 µg a.i. cm-2 ) and chlorfenapyr (LC50 = 0.17-0.20 µg a.i. cm-2 ). However, their susceptibility was lower than that of the susceptible strain reared on non-Bt maize: LC50 = 0.05 (spinetoram) and 0.08 (chlorfenapyr) µg a.i. cm-2 . In contrast, heterozygous strains had similar susceptibility to the susceptible strain. In field trials, no differences in FAW survival were detected between strains when the commercial dose of two insecticides was applied in Bt and non-Bt maize. CONCLUSION: FAW strains surviving on Bt and non-Bt maize, at the same development stage, have similar susceptibility to insecticides. The integrated pest management practices and insect resistance management importance of these results are discussed. © 2019 Society of Chemical Industry.

Baseline Susceptibility of Brazilian Populations of Chrysodeixis includens (Lepidoptera: Noctuidae) to C. includens Nucleopolyhedrovirus and Diagnostic Concentration for Resistance Monitoring.

The Chrysodeixis includens nucleopolyhedrovirus (ChinNPV: Baculoviridae: Alphabaculovirus) is a registered insecticide for the management of soybean looper, Chrysodeixis includens (Walker, [1858]) in Brazil. We conducted studies of baseline susceptibility of Brazilian populations of C. includens to the ChinNPV (Chrysogen, AgBiTech, Fort Worth, TX) as valuable knowledge in support of Integrated Pest Management and Insect Resistance Management programs. In bioassays, neonates were infected with different concentrations of ChinNPV using the droplet feeding bioassay method. Larvae were then transferred to artificial diet and mortality was assessed at 7 d. Results confirm that neonates from Brazilian populations of C. includens are susceptible to ChinNPV. Concentrations from 1.0 × 103 to 1.0 × 108 occlusion bodies (OBs) per ml caused mortality from 1.5 to 99%, respectively. The LC50 ranged from 1.4 × 105 to 7.7 × 105 OBs per ml for populations of C. includens (5.5-fold variation). Similar variation was detected for the LC90 which ranged from 1.6 × 107 to 7.7 × 107 OBs per ml (4.8-fold variation). Importantly, the field-collected populations showed equivalent susceptibility to the reference susceptible population. This indicates a low interpopulation variation in susceptibility of Brazilian populations of C. includens to ChinNPV, representing natural geographic variation and not variation caused by previous selection pressure. The candidate diagnostic concentration of 2.9 × 108 OBs per ml was estimated based on the pooled data and caused mortality ranging from 98.6 to 100%. This concentration will be used in proactive resistance monitoring programs. The Chrysogen will be a valuable tool as a new mode of action in C. includens resistance management in Brazil.