Assessing the Efficacy of Bacillus thuringiensis (Bt) Pyramided Proteins Cry1F, Cry1A.105, Cry2Ab2, and Vip3Aa20 Expressed in Bt Maize Against Lepidopteran Pests in Brazil.

Field studies across four states in maize-producing areas of Brazil were conducted to characterize the efficacy of a new pyramided Bacillus thuringiensis (Bt) Berliner technology in maize, Zea mays L., and compare it to existing single and pyramided commercial Bt technologies, to control Helicoverpa zea Boddie (Lepidoptera: Noctuidae), Elasmopalpus lignosellus Zeller (Lepidoptera: Pyralidae), Agrotis ipsilon (Hufnagel) (Lepidoptera: Noctuidae), and Diatraea saccharalis F. (Lepidoptera: Crambidae). Bt maize expressing Vip3Aa20 protein and pyramided Bt maize expressing proteins Cry1F + Cry1A.105 + Cry2Ab2 + Vip3Aa20 provided excellent protection against kernel feeding by H. zea compared to Bt technologies expressing only Cry1F or Cry1A.105 + Cry2Ab2. Bt maize expressing Cry1F, Cry1A.105 + Cry2Ab2, Cry1F + Cry1A.105 + Cry2Ab2, and Cry1F + Cry1A.105 + Cry2Ab2 + Vip3Aa20 resulted in less than 5% of plants injured by E. lignosellus, significantly less than Bt maize expressing only Vip3Aa20 and a non-Bt maize iso-hybrid with and without a thiamethoxam seed treatment. The highest protection against plant cutting injury caused by A. ipsilon was observed in the pyramid Bt maize technology expressing Cry1F + Cry1A.105 + Cry2Ab2 + Vip3Aa20. However, it did not differ statistically from the Bt maize expressing Vip3Aa20, Cry1F, or Cry1F + Cry1A.105 + Cry2Ab2. All Bt maize hybrids evaluated in our study were highly effective in reducing tunneling injury caused by D. saccharalis. These results show that a new maize technology expressing pyramided Bt proteins Cry1F + Cry1A.105 + Cry2Ab2 + Vip3Aa20 offers a higher level of protection from feeding by the above lepidopteran pest complex compared to maize with a single Bt protein or double pyramided Bt proteins.

Impact of transgenic soybean expressing Cry1Ac and Cry1F proteins on the non-target arthropod community associated with soybean in Brazil.

Field-scale studies that examine the potential for adverse effects of Bt crop technology on non-target arthropods may supplement data from laboratory studies to support an environmental risk assessment. A three year field study was conducted in Brazil to evaluate potential for adverse effects of cultivating soybean event DAS-81419-2 that produces the Cry1Ac and Cry1F proteins. To do so, we examined the diversity and abundance of non-target arthropods (NTAs) in Bt soybean in comparison with its non-Bt near isoline, with and without conventional insecticide applications, in three Brazilian soybean producing regions. Non-target arthropod abundance was surveyed using Moericke traps (yellow pan) and pitfall trapping. Total abundance (N), richness (S), Shannon-Wiener (H'), Simpson's (D) and Pielou's evenness (J) values for arthropod samples were calculated for each treatment and sampling period (soybean growth stages). A faunistic analysis was used to select the most representative NTAs which were used to describe the NTA community structure associated with soybean, and to test for effects due to the treatments effects via application of the Principal Response Curve (PRC) method. Across all years and sites, a total of 254,054 individuals from 190 taxa were collected by Moericke traps, while 29,813 individuals from 100 taxa were collected using pitfall traps. Across sites and sampling dates, the abundance and diversity measurements of representative NTAs were not significantly affected by Bt soybean as compared with non-sprayed non-Bt soybean. Similarly, community analyses and repeated measures ANOVA, when applicable, indicated that neither Bt soybean nor insecticide sprays altered the structure of the NTA communities under study. These results support the conclusion that transgenic soybean event DAS-81419-2 producing Cry1Ac and Cry1F toxins does not adversely affect the NTA community associated with soybean.

Synthesis and biological activity of a new class of insecticides: the N-(5-aryl-1,3,4-thiadiazol-2-yl)amides.

BACKGROUND: Optimization studies on a high-throughput screening (HTS) hit led to the discovery of a series of N-(6-arylpyridazin-3-yl)amides with insecticidal activity. It was hypothesized that the isosteric replacement of the pyridazine ring with a 1,3,4-thiadiazole ring could lead to more potent biological activity and/or a broader sap-feeding pest spectrum. The resulting N-(5-aryl-1,3,4-thiadiazol-2-yl)amides were explored as a new class of insecticides. RESULTS: Several methods for 2-amino-1,3,4-thiadiazole synthesis were used for the preparation of key synthetic intermediates. Subsequent coupling to variously substituted carboxylic acid building blocks furnished the final targets, which were tested for insecticidal activity against susceptible strains of Aphis gossypii (Glover) (cotton aphid), Myzus persicae (Sulzer) (green peach aphid) and Bemisia tabaci (Gennadius) (sweetpotato whitefly). CONCLUSION: Structure-activity relationship (SAR) studies on both the amide tail and the aryl A-ring of novel N-(5-aryl-1,3,4-thiadiazol-2-yl)amides led to a new class of insecticidal molecules active against sap-feeding insect pests. © 2016 Society of Chemical Industry.

Cross-resistance relationships of the sulfoximine insecticide sulfoxaflor with neonicotinoids and other insecticides in the whiteflies Bemisia tabaci and Trialeurodes vaporariorum.

BACKGROUND: Insecticides are important tools for managing damaging insect pests. Compounds that are effective against pests such as the whiteflies Bemisia tabaci and Trialeurodes vaporariorum, which show resistance to a range of insecticidal modes of action (MOA), have particular value as components of resistance management programmes. The sulfoximine insecticides are chemically unique as the first to incorporate a sulfoximine functional group. Sulfoxaflor is the first sulfoximine compound under commercial development for the control of sap-feeding insects. Its cross-resistance relationships were investigated by comparing the responses of field-collected strains with those of insecticide-susceptible laboratory strains of B. tabaci and T. vaporariorum. RESULTS: Sulfoxaflor exhibited very low (less than threefold) resistance ratios (RR) when tested against strains of B. tabaci that produced RR of up to 1000-fold to imidacloprid and cross-resistance to other neonicotinoid insecticides. Similarly, sulfoxaflor was not cross-resistant in a strain of B. tabaci exhibiting resistance to a pyrethroid (deltamethrin) and an organophosphate (profenophos). No cross-resistance was observed between sulfoxaflor and imidacloprid in T. vaporariorum. One population of the three field strains tested showed slightly reduced susceptibility to sufloxaflor with an RR of 4.17. By comparison, this same population exhibited an RR of more than 23.8-fold for imidacloprid relative to the susceptible population. CONCLUSION: In spite of sharing a target site with neonicotinoids (the nicotinic acetylcholine receptor), sulfoxaflor was largely unaffected by existing cases of neonicotinoid resistance in B. tabaci and T. vaporariorum. Neonicotinoid resistance mechanisms in these whitefly species are known to be primarily based on enhanced detoxification of insecticide. This lack of cross-resistance indicates that sulfoxaflor is a valuable new tool for the management of sap-feeding pests already resistant to established insecticide groups.

Novel nicotinic action of the sulfoximine insecticide sulfoxaflor.

The novel sulfoximine insecticide sulfoxaflor is as potent or more effective than the neonicotinoids for toxicity to green peach aphids (GPA, Myzus persicae). The action of sulfoxaflor was characterized at insect nicotinic acetylcholine receptors (nAChRs) using electrophysiological and radioligand binding techniques. When tested for agonist properties on Drosophila melanogaster Dα2 nAChR subunit co-expressed in Xenopus laevis oocytes with the chicken ß2 subunit, sulfoxaflor elicited very high amplitude (efficacy) currents. Sulfoximine analogs of sulfoxaflor were also agonists on Dα2/ß2 nAChRs, but none produced maximal currents equivalent to sulfoxaflor nor were any as toxic to GPAs. Additionally, except for clothianidin, none of the neonicotinoids produced maximal currents as large as those produced by sulfoxaflor. These data suggest that the potent insecticidal activity of sulfoxaflor may be due to its very high efficacy at nAChRs. In contrast, sulfoxaflor displaced [(3)H]imidacloprid (IMI) from GPA nAChR membrane preparations with weak affinity compared to most of the neonicotinoids examined. The nature of the interaction of sulfoxaflor with nAChRs apparently differs from that of IMI and other neonicotinoids, and when coupled with other known characteristics (novel chemical structure, lack of cross-resistance, and metabolic stability), indicate that sulfoxaflor represents a significant new insecticide option for the control of sap-feeding insects.

Biological characterization of sulfoxaflor, a novel insecticide.

BACKGROUND: The commercialization of new insecticides is important for ensuring that multiple effective product choices are available. In particular, new insecticides that exhibit high potency and lack insecticidal cross-resistance are particularly useful in insecticide resistance management (IRM) programs. Sulfoxaflor possesses these characteristics and is the first compound under development from the novel sulfoxamine class of insecticides. RESULTS: In the laboratory, sulfoxaflor demonstrated high levels of insecticidal potency against a broad range of sap-feeding insect species. The potency of sulfoxaflor was comparable with that of commercial products, including neonicotinoids, for the control of a wide range of aphids, whiteflies (Homoptera) and true bugs (Heteroptera). Sulfoxaflor performed equally well in the laboratory against both insecticide-susceptible and insecticide-resistant populations of sweetpotato whitefly, Bemisia tabaci Gennadius, and brown planthopper, Nilaparvata lugens (Stål), including populations resistant to the neonicotinoid insecticide imidacloprid. These laboratory efficacy trends were confirmed in field trials from multiple geographies and crops, and in populations of insects with histories of repeated exposure to insecticides. In particular, a sulfoxaflor use rate of 25 g ha(-1) against cotton aphid (Aphis gossypii Glover) outperformed acetamiprid (25 g ha(-1) ) and dicrotophos (560 g ha(-1) ). Sulfoxaflor (50 g ha(-1) ) provided a control of sweetpotato whitefly equivalent to that of acetamiprid (75 g ha(-1) ) and imidacloprid (50 g ha(-1) ) and better than that of thiamethoxam (50 g ha(-1) ). CONCLUSION: The novel chemistry of sulfoxaflor, its unique biological spectrum of activity and its lack of cross-resistance highlight the potential of sulfoxaflor as an important new tool for the control of sap-feeding insect pests.

Discovery and characterization of sulfoxaflor, a novel insecticide targeting sap-feeding pests.

The discovery of sulfoxaflor [N-[methyloxido[1-[6-(trifluoromethyl)-3-pyridinyl]ethyl]-λ(4)-sulfanylidene] cyanamide] resulted from an investigation of the sulfoximine functional group as a novel bioactive scaffold for insecticidal activity and a subsequent extensive structure-activity relationship study. Sulfoxaflor, the first product from this new class (the sulfoximines) of insect control agents, exhibits broad-spectrum efficacy against many sap-feeding insect pests, including aphids, whiteflies, hoppers, and Lygus, with levels of activity that are comparable to those of other classes of insecticides targeting sap-feeding insects, including the neonicotinoids. However, no cross-resistance has been observed between sulfoxaflor and neonicotinoids such as imidacloprid, apparently the result of differences in susceptibility to oxidative metabolism. Available data are consistent with sulfoxaflor acting via the insect nicotinic receptor in a complex manner. These observations reflect the unique structure of the sulfoximines compared with neonicotinoids.

Discovery and characterization of field resistance to Bt maize: Spodoptera frugiperda (Lepidoptera: Noctuidae) in Puerto Rico.

Transgenic maize, Zea mays L., event TC1507 produces the Cry1F protein to provide protection from feeding by several important lepidopteran pests, including Spodoptera frugiperda (J.E. Smith) (Lepidoptera: Noctuidae). Reports of reduced field performance against this species in Puerto Rico were investigated, and laboratory bioassays showed that S. frugiperda collected from the affected area exhibited lower sensitivity to the Cry1F protein compared with typical colonies from other regions. The resistance was shown to be autosomally inherited and highly recessive. The Puerto Rico colony was shown to be moderately less sensitive than susceptible laboratory strains to Cry1Ab and Cry1Ac, but the differences in sensitivity were dramatically smaller than for Cry1F. Potential contributory factors to the emergence of resistance to Cry1F in Puerto Rico populations of S. frugiperda include the tropical island geography, unusually large population sizes in 2006, and drought conditions reducing the availability of alternative hosts. In response to this resistance incident, the technology providers have stopped commercial sales of TC1507 maize in Puerto Rico pending potential reversion to susceptibility.

Field measures of western corn rootworm (Coleoptera: Chrysomelidae) mortality caused by Cry34/35Ab1 proteins expressed in maize event 59122 and implications for trait durability.

Maize, Zea mays L., has been transformed to express the Cry34Ab1 and Cry35Ab1 proteins from Bacillus thuringiensis strain PS149B1. These two proteins act together as a binary insecticidal protein that is effective against corn rootworm (Coleoptera: Chrysomelidae) species. The design of the resistance management plan to preserve the long-term durability of this trait largely depends on the level of rootworm mortality induced by Cry34/35Ab1 corn rootworm-protected maize (frequently referred to as "dose" in this context). Here, we report on studies that showed Cry34/35Ab1-expressing maize event 59122 caused 99.1 to 99.98% mortality of western corn rootworm, Diabrotica virgifera virgifera LeConte, larvae, after adjusting adult emergence numbers for density-dependent mortality. In two of three studies, there was a short delay in time to 50% adult emergence from 59122 maize plots compared with control plots, although emergence was completed at approximately the same time from both types of maize. These data support an expectation that alleles conferring resistance to the Cry34/35Ab1 proteins in western corn rootworm will be functionally nearly completely to completely recessive on 59122 maize and that there is unlikely to be assortative mating of Cry34/35Ab1-resistant and susceptible rootworms. When incorporated into simulation models of rootworm adaptation to transgenic maize, these findings suggest that a 20% refuge is likely to be highly effective at prolonging the durability of 59122 maize.