Greenhouse studies were conducted in 2003 at the Stine–Haskell Research Center to determine whether herbicide inhibitors of six specific sites in the carotenoid biosynthesis pathway would elicit synergistic responses when applied postemergence (POST) in combination with the photosystem II (PSII) inhibitor atrazine. Based on data analysis with the Isobole method, synergistic responses were observed on red morningglory, common cocklebur, and giant foxtail when atrazine was applied in mixtures with the deoxy-D-xylulose-5-phosphate reductoisomerase (DOXP reductoisomerase) inhibitor fosmidomycin, thep-hydroxyphenylpyruvate dioxygenase (HPPD) inhibitor mesotrione, and the DuPont proprietary zeta-carotene desaturase (ZDS) inhibitor DFPC. Clomazone (its metabolite ketoclomazone is the actual enzyme inhibitor), an inhibitor of 1-deoxy-D-xylulose-5-phosphate synthatase (DOXP synthase), provided synergistic responses on red morningglory, but antagonistic responses on both common cocklebur and giant foxtail when applied in mixtures with atrazine. Combinations of the lycopene cyclase (LC) inhibitor, CPTA, with atrazine produced synergistic responses on both common cocklebur and giant foxtail but were antagonistic on red morningglory. Norflurazon, a phytoene desaturase (PDS) inhibitor, applied in mixtures with atrazine provided synergistic responses on red morningglory, antagonistic responses on giant foxtail, and independent responses on common cocklebur. Because carotenoids have been determined to play a key role in quenching singlet oxygen species in the chloroplast and also assist in the maintenance of the D1 protein in PSII, this might help explain the synergistic responses with atrazine observed in our studies.

In pharmaceutical drug discovery and agricultural crop product discovery, in vivo bioassay experiments are used to identify promising compounds for further research. The reproducibility and accuracy of the bioassay is crucial to be able to correctly distinguish between active and inactive compounds. In the case of agricultural product discovery, a replicated dose-response of commercial crop protection products is assayed and used to monitor test quality. The activity of these compounds on the test organisms, the weeds, insects, or fungi, is characterized by a dose-response curve measured from the bioassay. These curves are used to monitor the quality of the bioassays. If undesirable conditions in the bioassay arise, such as equipment failure or problems with the test organisms, then a bioassay monitoring procedure is needed to quickly detect such issues. In this paper we illustrate a proposed nonlinear profile monitoring method to monitor the variability of multiple assays, the adequacy of the dose-response model chosen, and the estimated dose-response curves for aberrant cases in the presence of heteroscedasticity. We illustrate these methods with in vivo bioassay data collected over one year from DuPont Crop Protection.

BACKGROUNDSulfonylurea (SU) herbicides are effective because they inhibit acetolactate synthase (ALS), a key enzyme in branched-chain amino acid synthesis required for plant growth. A soybean line known as W4-4 was developed through rounds of seed mutagenesis and was demonstrated to have a high degree of ALS-based resistance to both post-emergence and pre-emergence applications of a variety of SU herbicides. This report describes the molecular and phenotypic characterization of the Als1 and Als2 mutations that confer herbicide resistance to SUs and other ALS inhibitors.RESULTSThe mutations are shown to occur in two different ALS genes that reside on different chromosomes: Als1 (P178S) on chromosome 4 and Als2 (W560L) on chromosome 6 (P197S and W574L in Arabidopsis thaliana).CONCLUSIONAlthough the Als1 and Als2 genes are unlinked, the combination of these two mutations is synergistic for improved tolerance of soybeans to ALS-inhibiting herbicides. © 2014 DuPont Pioneer. Pest Management Science published by JohnWiley & Sons Ltd on behalf of Society of Chemical Industry.

Researchers, product registration personnel, and growers desire the ability to chemically detect residual amounts of herbicides in soil at concentrations below those necessary to cause phytotoxicity to sensitive nontarget or rotational crop plants. Alfalfa, cotton, soybean, and sunflower, crops sensitive to low concentrations of aminocyclopyrachlor in soil, were planted at field test sites approximately 1 yr after aminocyclopyrachlor methyl was applied. Soil samples were collected when rotational crops were planted and were analyzed for aminocyclopyrachlor by a method based on high performance liquid chromatography tandem mass spectrometry (HPLC/MS/MS), with a limit of detection (LOD) of 0.1 part per billion (ppb) (soil oven-dry weight basis). Loglogistic dose–response analysis correlated visual phytotoxic plant responses to residual concentrations of aminocyclopyrachlor in the soil. Concentrations of aminocyclopyrachlor estimated to cause 25% phytotoxicity to alfalfa, cotton, soybean, and sunflower were 5.4, 3.2, 2.0, and 6.2 ppb, respectively, 20 to 60 times greater than the LOD of the analytical method available for soil analysis. Results from these studies suggest this HPLC/MS/MS method of analysis can be used to indicate potential risk and severity of plant response for alfalfa, cotton, soybean, and sunflower, and for other plant species once dose–response curves for these additional species are established. This chemical assay may be particularly important if researchers desire to study the concentration, movement, and dissipation of aminocyclopyrachlor in soil or as part of a forensic investigation to better understand the cause of an unanticipated or undesirable plant response.

Vapor movement of synthetic auxin herbicides can injure desirable plants outside the treatment zone. Vapor movement of the synthetic auxin herbicides aminocyclopyrachlor and aminocyclopyrachlor methyl was compared with that of the relatively volatile herbicide dicamba and the low volatile herbicide aminopyralid with a soybean bioassay under greenhouse and field conditions. Soybean is very sensitive to these active ingredients. Under greenhouse conditions, 82 (61 to 104) mg ae ha−1of aminocyclopyrachlor, 26 (18 to 33) mg ae ha−1of aminocyclopyrachlor methyl, 82 (69 to 95) mg ae ha−1of aminopyralid, and 61 (47 to 75) mg ae ha−1of dicamba produced an estimated 25% visual soybean phytotoxicity response when soybean was treated POST at the V3 growth stage (GR25[95% confidence interval]). In field studies, aminocyclopyrachlor, aminocyclopyrachlor methyl, and aminopyralid were applied at 70 g ae ha−1and dicamba was applied at 560 g ae ha−1(labeled application rates) to soybean at the V3 growth stage. All herbicides were applied within an enclosed chamber (3 m by 3 m by 1 m) to mitigate movement of spray droplets. The enclosures were removed shortly after spray application and soybean response immediately surrounding the treated area was recorded in each of eight directions approximately 10 d after treatment. On the basis of bioassay responses, relative amount of vapor movement was dicamba > aminocyclopyrachlor methyl > aminopyralid ≈ aminocyclopyrachlor. Vapor movement of aminocyclopyrachlor was very low indicating that the risk of phytotoxic response of sensitive plants due to volatility of aminocyclopyrachlor is negligible.