HPPD Inhibitors Research Articles

Additive and synergistic interactions of 4-hydroxyphenylpyruvate dioxygenase (HPPD) and photosystem II (PSII) inhibitors for the control of glyphosate-resistant horseweed (Conyza canadensis) in corn

AbstractGlyphosate-resistant (GR) horseweed [Conyza canadensis (L.) Cronquist; syn.: Erigeron canadensis L.] interference can substantially reduce corn (Zea mays L.) yield. The complementary activity of 4-hydroxyphenylpyruvate dioxygenase (HPPD) and photosystem II (PSII) inhibitors has been investigated for the control of several weed species, and in many cases has been synergistic; however, there is little information on the interaction of HPPD- and PSII-inhibiting herbicides for postemergence control of GR C. canadensis in corn. Four field trials were studied over 2 yr (2019, 2020) in Ontario, Canada, in commercial corn fields with natural infestations of GR C. canadensis to evaluate the level of GR C. canadensis control with three HPPD-inhibiting herbicides (mesotrione, tolpyralate, and topramezone) and three PSII-inhibiting herbicides (atrazine, bromoxynil, and bentazon) applied individually and in tank-mix combinations, and to document the interaction of the three HPPD inhibitors tank mixed with the three PSII inhibitors. Mesotrione, tolpyralate, and topramezone controlled GR C. canadensis 83%, 84%, and 72%, respectively, at 8 wk after application (WAA). Bromoxynil and bentazon controlled GR C. canadensis 71% and 79%, respectively, while atrazine provided only 31% control at 8 WAA. The joint application of atrazine, bromoxynil, or bentazon with mesotrione increased GR C. canadensis control from 83% to 100% at 8 WAA. Tolpyralate tank mixed with atrazine, bromoxynil, or bentazon controlled GR C. canadensis 96%, 98%, and 98%, respectively, which was comparable to the mesotrione tank mixes at 8 WAA. Topramezone plus atrazine, bromoxynil, or bentazon controlled GR C. canadensis 91%, 93%, and 95%, respectively, at 8 WAA. Interactions between HPPD and PSII inhibitors were synergistic for all combinations of mesotrione or tolpyralate with atrazine, bromoxynil, or bentazon. The interaction between topramezone and PSII inhibitors was additive. All nine tank mixes controlled GR C. canadensis >90%. This study concludes that bromoxynil or bentazon, instead of atrazine, can be co-applied with mesotrione, tolpyralate, or topramezone without compromising GR C. canadensis control in corn.

Molecular Docking and QSAR Studies for Modeling the Inhibitory Activity of Pyrazole-benzimidazolone Hybrids as Novel Inhibitors of Human 4-hydroxyphenylpyruvate dioxygenase Against Type I Tyrosinemia Disease

Pyrazole-benzimidazolone is a novel inhibitor of human 4-Hydroxyphenylpyruvate dioxygenase of the HPPD receptor. To analyze this inhibitory activity, a QSAR study was performed using 33 compounds. The MLR multiple linear regression method and the NN neural network were used to establish the predicted model. This model was validated by the external validation method. The molecular electronic descriptors were calculated by the DFT method. This study shows that the activity of the compounds is strongly correlated with the five descriptors selected by the MLR method. The correlation coefficients calculated by MLR and then by NN are R =0.878 and R =0.978; they allow us to evaluate the proposed quantitative model. The latter allowed us to predict the activity of new HPPD receptor inhibitors and to show the good predictive competence of the established QSAR model. Based on these results, we performed docking in the active site of HPPD protein with the most active compound (12), and we deduced that this compound specifically interacts with are GLN 307, ASN 423, and PHE 392 at the HPPD protein binding pocket; thus, it showed additional interactions with VAL 228, PRO 280, SER 267, LEU 265, LYS 421, GLN 379, GLY 420, and LEU 368, This interaction is similar to that of NTBC with the active site.

DISCOVERY OF TRIKETONE-QUINOXALINE HYBRIDS AS POTENT HPPD INHIBITORS USING STRUCTURE-BASED DRUG DESIGN

<List> <ListItem><ItemContent> • HPPD is one of the most promising targets for new herbicides. </ItemContent></ListItem> <ListItem><ItemContent> • A family of novel HPPD inhibitors based on the triketone-quinoxaline scaffold was designed and synthesized. </ItemContent></ListItem> <ListItem><ItemContent> • One particular product (7d) gave the highest inhibition of HPPD of the newly synthesized derivatives. </ItemContent></ListItem> <ListItem><ItemContent> • Triketone-quinoxaline derivatives provide a useful molecular scaffold for the discovery of novel HPPD-inhibiting herbicides. </ItemContent></ListItem></List> <i>p</i>-Hydroxyphenylpyruvate dioxygenase (EC 1.13.11.27, HPPD) belongs to the family of Fe(II)-dependent non-heme oxygenases that occur in the majority of aerobic organisms. HPPD has proved to be a promising target in herbicide research and development. A battery of novel triketone-quinoxaline compounds has been designed using a structure-based drug design strategy and then prepared. Enzyme inhibition assays show that these synthesized derivatives possess favorable inhibition capability against <i>Arabidopsis thaliana</i> HPPD with IC₅₀ values ranging from 0.317 to 0.891 μmol·L⁻¹. Subsequently, the molecular docking results indicate that two adjacent carbonyls of the triketone moiety of the representative compound 2-(2,3-dimethyl-8-(o-tolyl)quinoxaline-6-carbonyl)-3-hydroxycyclohex-2-en-1-one (7d) engage in chelation with the ferrous ion of <i>A. thaliana</i> HPPD in a bidentate pose, and its quinoxaline scaffold forms two sets of parallel π-stacking interaction between two phenylalanine residues (Phe424 and Phe381). In addition, the extended phenyl group also interacts with Phe392 in a π-π stacking way. This study indicates that triketone-quinoxaline is a promising scaffold for discovering HPPD inhibitors with substantially increased potency, providing insight into the molecular design of new herbicides.

Characterization and inheritance of dicamba resistance in a multiple-resistant waterhemp (Amaranthus tuberculatus) population from Illinois

AbstractWaterhemp [Amaranthus tuberculatus (Moq.) Sauer] is one of the most troublesome agronomic weeds in the midwestern United States. The rapid evolution and selection of herbicide-resistance traits in A. tuberculatus is a major challenge in managing this species. An A. tuberculatus population, designated CHR, was identified in 2012 in Champaign County, IL, and previously characterized as resistant to herbicides from six site-of-action groups: 2,4-D (Group 4), acetolactate synthase inhibitors (Group 2), protoporphyrinogen oxidase inhibitors (Group 14), 4-hydroxyphenylpyruvate dioxygenase inhibitors (Group 27), photosystem II inhibitors (Group 5), and very-long-chain fatty-acid synthesis inhibitors (Group 15). Recently, ineffective control of CHR was observed in the field after dicamba application. Therefore, this research was initiated to confirm dicamba resistance, quantify the resistance level, and investigate its inheritance in CHR. Multiple field trials were conducted at the CHR location to confirm poor control with dicamba and compare dicamba treatments with other herbicides. Greenhouse trials were conducted to quantify the resistance level in CHR and confirm genetic inheritance of the resistance. In field trials, dicamba did not provide more than 65% control, while glyphosate and glufosinate provided at least 90% control. Multiple accessions were generated from controlled crosses and evaluated in greenhouse trials. Greenhouse dicamba dose–response experiments indicated a resistance level of 5- to 10-fold relative to a sensitive parental line. Dose–response experiments using F1 lines indicated that dicamba resistance was an incompletely dominant trait. Segregation analysis with F2 and backcross populations indicated that dicamba resistance had moderate heritability and was potentially a multigenic trait. Although dicamba resistance was predominantly inherited as a nuclear trait, minor maternal inheritance was not completely ruled out. To our knowledge, CHR is one of the first cases of dicamba resistance in A. tuberculatus. Further studies will focus on elucidating the genes involved in dicamba resistance.

Design, synthesis and biological activity of novel triketone-containing quinoxaline as HPPD inhibitor.

4-Hydroxyphenyl pyruvate dioxygenase (EC 1.13.11.27, HPPD) is one of the important target enzymes used to address the issue of weed control. HPPD-inhibiting herbicides can reduce the carotenoid content in plants and hinder photosynthesis, eventually causing albinism and death. Exploring novel HPPD-inhibiting herbicides is a significant direction in pesticide research. In the process of exploring new high-efficiency HPPD inhibitors, a series of novel quinoxaline derivatives were designed and synthesized using an active fragment splicing strategy. The title compounds were unambiguously characterized by infrared, 1 H NMR, 13 C NMR, and high-resolution mass spectroscopy. The results of the in vitro tests indicated that the majority of the title compounds showed potent inhibition of Arabidopsis thaliana HPPD (AtHPPD). Preliminary bioevaluation results revealed that a number of novel compounds displayed better or excellent herbicidal activity against broadleaf and monocotyledonous weeds. Compound III-5 showed herbicidal effects comparable to those of mesotrione at a rate of 150 g of active ingredient (ai)/ha for post-emergence application. The results of molecular dynamics verified that compound III-5 had a more stable protein-binding ability. Molecular docking results showed that compound III-5 and mesotrione shared homologous interplay with the surrounding residues. In addition, the enlarged aromatic ring system adds more force, and the hydrogen bond formed can enhance the synergy with π-π stacking. The present work indicates that compound III-5 may be a potential lead structure for the development of new HPPD inhibitors.

On the use of inhibitors of 4-hydroxyphenylpyruvate dioxygenase as a vector-selective insecticide in the control of mosquitoes.

Blood-sucking insects incorporate many times their body weight of blood in a single meal. Because proteins are the major component of vertebrate blood, its digestion in the gut generates extremely high concentrations of free amino acids. Previous reports showed that the tyrosine degradation pathway plays an essential role in adapting these animals to blood feeding. Inhibition of 4-hydroxyphenylpyruvate dioxygenase (HPPD), the rate-limiting step of tyrosine degradation, results in the death of insects after a blood meal. Therefore, it has been suggested that compounds that block the catabolism of tyrosine could act selectively on blood-feeding insects. Here, we evaluated the toxicity against mosquitoes of three HPPD inhibitors currently used as herbicides and in human health. Of the compounds tested, nitisinone (NTBC) proved to be more potent than mesotrione (MES) and isoxaflutole (IFT) in Aedes aegypti. NTBC was lethal to Ae. aegypti in artificial feeding assays [median lethal dose (LD50 ): 4.53 μm] and in topical application (LD50 : 0.012 nmol/mosquito). NTBC was also lethal to Ae. aegypti populations that were resistant to neurotoxic insecticides, and to other mosquito species (Anopheles and Culex). HPPD inhibitors, particularly NTBC, represent promising new drugs for mosquito control. Because they affect only blood-feeding organisms, they represent a safer and more environmentally friendly alternative to conventional neurotoxic insecticides. © 2021 Society of Chemical Industry.

Design, Synthesis, Structure-Activity Relationship, Molecular Docking, and Herbicidal Evaluation of 2-Cinnamoyl-3-Hydroxycyclohex-2-en-1-one Derivatives as Novel 4-Hydroxyphenylpyruvate Dioxygenase Inhibitors.

Cinnamic acid, isolated from cinnamon bark, is a natural product with excellent bioactivity, and it effectively binds with cyclohexanedione to form novel 4-hydroxyphenylpyruvate dioxygenase (HPPD) inhibitors. According to the active sub-structure combination principle, a series of novel 3-hydroxy-2-cinnamoyl-2-en-1-one derivatives were designed and synthesized. The title compounds were characterized by infrared, 1H NMR, 13C NMR, and HRMS. The in vitro inhibitory activity of AtHPPD verified that compound II-13 showed the most activity with a half-maximal inhibitory concentration (IC50) value of 0.180 μM, which was superior to that of mesotrione (0.206 μM) in vitro. The preliminary herbicidal activity tests demonstrated that some compounds had good herbicidal activity especially compound II-13 at a concentration of 150 g ai/ha. The binding mode of AtHPPD through molecular docking indicated that two oxygens of compounds II-13 formed bidentate interactions with metal ions, and the benzene ring formed π-π accumulation effects with Phe-381 and Phe-424. The results of molecular dynamics simulations showed that compound II-13 exhibited a more stable binding ability with AtHPPD than mesotrione. This study provided insights into the development of natural and efficient herbicides in the future.

Quinoxaline derivatives as herbicide safeners by improving Zea mays tolerance

Isoxaflutole (IXF), a 4-hydroxyphenylpyruvate dioxygenase (HPPD) inhibitor, causes injury to crops leading to reductions in grain yield. In order to solve the phytotoxicity caused by IXF, the present work evaluated the protective response of the substituted quinoxaline derivatives as potential safeners on Zea mays. The bioassay results showed that all of the test compounds displayed protection against IXF. In particular, safener I-6 exhibited excellent safener activity against IXF injury via enhancing glutathione (GSH) content, glutathione S transferases (GSTs) and cytochrome P450 monooxygenases (CYP450) activity. The tested compounds induced the activity of CYP450 and GSTs in Z. mays. The physicochemical properties and ADMET properties of safener I-6, benoxacor and diketonitrile (DKN, IXF metabolite) were compared to predict pharmaceutical behavior. The present work demonstrates that the safener I-6 could be considered as a potential candidate for developing novel safeners in the future.

Mechanism of action and selectivity of a novel herbicide, fenquinotrione.

Fenquinotrione is a novel herbicide that can control a wide range of broadleaf and sedge weeds with excellent rice selectivity. We revealed that fenquinotrione potently inhibited the 4-hydroxyphenylpyruvate dioxygenase (HPPD) activity in Arabidopsis thaliana with an IC50 of 44.7 nM. The docking study suggested that the 1,3-diketone moiety of fenquinotrione formed a bidentate interaction with Fe(II) at the active site. Furthermore, π–π stacking interactions occurred between the oxoquinoxaline ring and the conserved Phe409 and Phe452 rings, indicating that fenquinotrione competes with the substrate, similar to existing HPPD inhibitors. A more than 16-fold difference in the herbicidal activity of fenquinotrione in rice and the sedge, Schoenoplectus juncoides, was observed. However, fenquinotrione showed high inhibitory activity against rice HPPD. Comparative metabolism study suggested that the potent demethylating metabolism followed by glucose conjugation in rice was responsible for the selectivity of fenquinotrione.

Genetic architecture underlying HPPD-inhibitor resistance in a Nebraska Amaranthus tuberculatus population.

Amaranthus tuberculatus is a problematic weed species in Midwest USA agricultural systems. Inhibitors of 4-hydroxyphenylpyruvate dioxygenase (HPPD) are an important chemistry for weed management in numerous cropping systems. Here, we characterize the genetic architecture underlying the HPPD-inhibitor resistance trait in an A. tuberculatus population (NEB). Dose-response studies of an F1 generation identified HPPD-inhibitor resistance as a dominant trait with a resistance factor of 15.0-21.1 based on dose required for 50% growth reduction. Segregation analysis in a pseudo-F2 generation determined the trait is moderately heritable (H2 = 0.556) and complex. Bulk segregant analysis and validation with molecular markers identified two quantitative trait loci (QTL), one on each of Scaffold 4 and 12. Resistance to HPPD inhibitors is a complex, largely dominant trait within the NEB population. Two large-effect QTL were identified controlling HPPD-inhibitor resistance in A. tuberculatus. This is the first QTL mapping study to characterize herbicide resistance in a weedy species.

Assessment of genetically modified cotton GHB811 for food and feed uses, under Regulation (EC) No 1829/2003 (application EFSA-GMO-ES-2018-154).

Cotton GHB811 was developed to confer tolerance to glyphosate and HPPD inhibitor herbicides. The molecular characterisation data and bioinformatic analyses do not identify issues requiring food/feed safety assessment. None of the identified differences in the agronomic/phenotypic and compositional characteristics tested between cotton GHB811 and its conventional counterpart needs further assessment, except for % lint, lint length and dihydrosterculic acid, which do not raise nutritional and safety concerns. The GMO Panel does not identify safety concerns regarding the toxicity and allergenicity of the 2mEPSPS and HPPD W336 proteins as expressed in cotton GHB811 and finds no evidence that the genetic modification would change the overall allergenicity of cotton GHB811. In the context of this application, the consumption of food and feed from cotton GHB811 does not represent a nutritional concern in humans and animals. The GMO Panel concludes that cotton GHB811 is as safe as the conventional counterpart and non‐GM cotton reference varieties tested, and no post‐market monitoring of food/feed is considered necessary. In the case of accidental release of viable cotton GHB811 seeds into the environment, this would not raise environmental safety concerns. The post‐market environmental monitoring plan and reporting intervals are in line with the intended uses of cotton GHB811. The GMO Panel concludes that cotton GHB811 is as safe as its conventional counterpart and the tested non‐GM cotton reference varieties with respect to potential effects on human and animal health and the environment.

Recurrent Selection with Sub-Lethal Doses of Mesotrione Reduces Sensitivity in Amaranthus palmeri.

Amaranthus palmeri, ranked as the most prolific and troublesome weed in North America, has evolved resistance to several herbicide sites of action. Repeated use of any one herbicide, especially at lower than recommended doses, can lead to evolution of weed resistance, and, therefore, a better understanding of the process of resistance evolution is essential for the management of A. palmeri and other difficult-to-control weed species. Amaranthus palmeri rapidly developed resistance to 4-hydroxyphenylpyruvate dioxygenase (HPPD) inhibitors such as mesotrione. The objective of this study was to test the potential for low-dose applications of mesotrione to select for reduced susceptibility over multiple generations in an A. palmeri population collected from an agricultural field in 2001. F0 plants from the population were initially treated with sub-lethal mesotrione rates and evaluated for survival three weeks after treatment. All F0 plants were controlled at the 1× rate (x = 105 g ai ha−1). However, 2.5% of the F0 plants survived the 0.5× treatment. The recurrent selection process using plants surviving various mesotrione rates was continued until the F4 generation was reached. Based on the GR50 values, the sensitivity index was determined to be 1.7 for the F4 generation. Compared to F0, HPPD gene expression level in the F3 population increased. Results indicate that after several rounds of recurrent selection, the successive generations of A. palmeri became less responsive to mesotrione, which may explain the reduced sensitivity of this weed to HPPD-inhibiting herbicides. The results have significance in light of the recently released soybean and soon to be released cotton varieties with resistance to HPPD inhibitors.

Unravelling Phytotoxicity and Mode of Action of Tripyrasulfone, a Novel Herbicide.

Tripyrasulfone is a novel herbicide post-emergence applied in paddy fields. In this study, tripyrasulfone phytotoxicity and its mode of action were investigated. Within 3-7 days after treatment (DAT), tripyrasulfone caused strong bleaching symptoms on newly developed leaves of Echinochloa crus-galli followed by necrosis prior to death within 14 DAT. By investigating pigment composition, photosynthetic activity and energy dissipation of E. crus-galli treated with tripyrasulfone, the accumulation of phytoene and significant decreases in total carotenoids were observed; the photosystem II complex (PSII) reaction center and PSII-PSI electron transport chain were damaged; and the non-photochemical energy quenching and reactive oxygen species were significantly increased. Based on the reversion of bleaching symptoms in treated Spirodela polyrrhiza by the addition of homogentisic acid, it was hypothesized that tripyrasulfone blocks the biosynthesis of HGA, possibly by the inhibition of 4-hydroxyphenylpyruvate dioxygenase (HPPD). However, based on its chemical structure, tripyrasulfone may tend to be hydrolyzed in plants. Indeed, the hydrolyzed tripyrasulfone (HDT) inhibited the activity of HPPD from Arabidopsis thaliana produced by Escherichia coli, which was approximately 6 times less effective than mesotrione. Molecular docking showed that the HDT formed a stable bidentate interaction with the active center Fe2+ chelation of A. thaliana HPPD.

Synthesis and Herbicidal Activity of Triketone-Aminopyridines as Potent p-Hydroxyphenylpyruvate Dioxygenase Inhibitors.

Exploring novel p-hydroxyphenylpyruvate dioxygenase (EC 1.13.11.27, HPPD) inhibitors has become one of the most promising research directions in herbicide innovation. On the basis of our tremendous interest in exploiting more powerful HPPD inhibitors, we designed a family of benzyl-containing triketone-aminopyridines via a structure-based drug design (SBDD) strategy and then synthesized them. Among these prepared derivatives, the best active 3-hydroxy-2-(3,5,6-trichloro-4-((4-isopropylbenzyl)amino)picolinoyl)cyclohex-2-en-1-one (23, IC50 = 0.047 μM) exhibited a 5.8-fold enhancement in inhibiting Arabidopsis thaliana (At) HPPD activity over that of commercial mesotrione (IC50 = 0.273 μM). The predicted docking models and calculated energy contributions of the key residues for small molecules suggested that an additional π-π stacking interaction with Phe-392 and hydrophobic contacts with Met-335 and Pro-384 were detected in AtHPPD upon the binding of the best active compound 23 compared with that of the reference mesotrione. Such a molecular mechanism and the resulting binding affinities coincide with the proposed design scheme and experimental values. It is noteworthy that inhibitors 16 (3-hydroxy-2-(3,5,6-trichloro-4-((4-chlorobenzyl)amino)picolinoyl)cyclohex-2-en-1-one), 22 (3-hydroxy-2-(3,5,6-trichloro-4-((4-methylbenzyl)amino)picolinoyl)cyclohex-2-en-1-one), and 23 displayed excellent greenhouse herbicidal effects at 150 g of active ingredient (ai)/ha after postemergence treatment. Furthermore, compound 16 showed superior weed-controlling efficacy against Setaria viridis (S. viridis) versus that of the positive control mesotrione at multiple test dosages (120, 60, and 30 g ai/ha). These findings imply that compound 16, as a novel lead of HPPD inhibitors, possesses great potential for application in specifically combating the malignant weed S. viridis.

Interaction of 4-hydroxyphenylpyruvate dioxygenase (HPPD) and atrazine alternative photosystem II (PS II) inhibitors for control of multiple herbicide–resistant waterhemp (Amaranthus tuberculatus) in corn

Abstract The complementary activity of 4-hydroxphenylpyruvate dioxygenase (HPPD) inhibitors and atrazine is well documented, but the use of atrazine is restricted in some geographic areas, including the province of Quebec in Canada, necessitating the evaluation of atrazine alternatives and their interactions with HPPD inhibitors. The objectives of this study were to determine whether mixing HPPD inhibitors with atrazine alternative photosystem II (PS II) inhibitors, such as metribuzin and linuron applied PRE or bromoxynil and bentazon applied POST, results in similar control of multiple herbicide–resistant (MHR) waterhemp [Amaranthus tuberculatus (Moq.) Sauer] in corn (Zea mays L.). Ten field trials, five with herbicides applied PRE and five with herbicides applied POST, were conducted in Ontario, Canada, in fields infested with MHR A. tuberculatus. Isoxaflutole, applied PRE, controlled MHR A. tuberculatus 58% to 76%; control increased 17% to 34% with the addition of atrazine, metribuzin, or linuron at three of five sites across 2, 4, 8, and 12 wk after application (WAA). The interaction between isoxaflutole and PS II inhibitors, applied PRE, was additive for MHR A. tuberculatus control and biomass and density reduction. Mesotrione, tolpyralate, and topramezone, applied POST, controlled MHR A. tuberculatus 54% to 59%, 61%, and 44% to 45%, respectively, at two of five sites across 4, 8, and 12 WAA. The addition of atrazine, bromoxynil, or bentazon to mesotrione improved MHR A. tuberculatus control 29%, 34%, and 22%; to tolpyralate, improved control 2%, 20%, and 10%; and to topramezone, improved control 3%, 14%, and 8%, respectively. Interactions between HPPD and PS II inhibitors were mostly additive; however, synergistic responses were observed with mesotrione + bromoxynil or bentazon, and tolpyralate + bromoxynil. Mixing atrazine alternatives metribuzin or linuron with isoxaflutole, applied PRE, and bromoxynil or bentazon with mesotrione or tolpyralate, applied POST, resulted in similar or better control of MHR A. tuberculatus in corn.

Assessment of genetically modified soybean GMB151 for food and feed uses, under Regulation (EC) No 1829/2003 (application EFSA-GMO-NL-2018-153).

Soybean GMB151 was developed to confer tolerance to 4‐hydroxyphenylpyruvate dioxygenase (HPPD) inhibitor herbicides and resistance to nematodes. The molecular characterisation data and bioinformatic analyses do not identify issues requiring food/feed safety assessment. None of the identified differences in the agronomic/phenotypic and compositional characteristics tested between soybean GMB151 and its conventional counterpart needs further assessment, except for palmitic acid and heptadecenoic acid in seeds and carbohydrate and crude protein in forage, which does not raise nutritional and safety concerns. The GMO Panel does not identify safety concerns regarding the toxicity and allergenicity of the HPPD‐4 and Cry14Ab‐1 proteins as expressed in soybean GMB151, and finds no evidence that the genetic modification would change the overall allergenicity of soybean GMB151. In the context of this application, the consumption of food and feed from soybean GMB151 does not represent a nutritional concern in humans and animals. The GMO Panel concludes that soybean GMB151 is as safe as the conventional counterpart and non‐GM soybean reference varieties tested, and no post‐market monitoring of food/feed is considered necessary. In the case of accidental release of viable soybean GMB151 seeds into the environment, this would not raise environmental safety concerns. The post‐market environmental monitoring plan and reporting intervals are in line with the intended uses of soybean GMB151. The GMO Panel concludes that soybean GMB151 is as safe as its conventional counterpart and the tested non‐GM soybean reference varieties with respect to potential effects on human and animal health and the environment.

Design, synthesis, herbicidal activity and CoMFA of aryl-formyl piperidinone HPPD inhibitors

Isoxazole, nicotinic acid and benzoic acid are important components in many natural products and useful synthons to build macrostructures having valuable biological activities. In continuation of our effort to discover 4-hydroxyphenylpyruvate dioxygenase (HPPD, EC 1.13.11.27) inhibitors and search for active fragments from natural products, a series of substituted aryl-formyl piperidinone derivatives with natural product fragments was rationally designed, synthesized and tested for their herbicidal activity. Compound I-9 was considered the most effective candidate with an IC50 value of 0.260 μM. The molecular docking results showed that the triketone group of compound I-9 forms a bidentate complex with a metal ion, and the benzene ring interacted with Phe424 and Phe381 via π-π stacking, which was similar to the mechanisms of mesotrione. The present work indicates that compound I-9 may serve as a potential lead compound for further development of green HPPD inhibitors.

The structure of 4-hydroxylphenylpyruvate dioxygenase complexed with 4-hydroxylphenylpyruvic acid reveals an unexpected inhibition mechanism

4-Hydroxyphenylpyruvate dioxygenase (HPPD) is an important target for both drug and pesticide discovery. As a typical Fe(II)-dependent dioxygenase, HPPD catalyzes the complicated transformation of 4-hydroxyphenylpyruvic acid (HPPA) to homogentisic acid (HGA). The binding mode of HPPA in the catalytic pocket of HPPD is a focus of research interests. Recently, we reported the crystal structure of Arabidopsis thaliana HPPD (AtHPPD) complexed with HPPA and a cobalt ion, which was supposed to mimic the pre-reactive structure of AtHPPD-HPPA-Fe(II). Unexpectedly, the present study shows that the restored AtHPPD-HPPA-Fe(II) complex is still nonreactive toward the bound dioxygen. QM/MM and QM calculations reveal that the HPPA resists the electrophilic attacking of the bound dioxygen by the trim of its phenyl ring, and the residue Phe381 plays a key role in orienting the phenyl ring. Kinetic study on the F381A mutant reveals that the HPPD-HPPA complex observed in the crystal structure should be an intermediate of the substrate transportation instead of the pre-reactive complex. More importantly, the binding mode of the HPPA in this complex is shared with several well-known HPPD inhibitors, suggesting that these inhibitors resist the association of dioxygen (and exert their inhibitory roles) in the same way as the HPPA. The present study provides insights into the inhibition mechanism of HPPD inhibitors.

Herbicide strategies for managing glyphosate-resistant and -susceptible kochia (Bassia scoparia) in spring wheat

Kochia [Bassia scoparia (L.) A.J. Scott] is a summer annual tumbleweed that is tolerant of heat, drought, and salinity and capable of causing large yield losses in spring wheat (Triticum aestivum L). Increased incidence of glyphosate- and acetolactate synthase (ALS) inhibitor-resistant kochia in western Canada warrants investigation of alternative herbicides to manage these biotypes. Herbicides applied pre- or post-emergence in spring wheat were evaluated based on crop tolerance and control of ALS inhibitor-resistant kochia accessions with and without the glyphosate resistance trait in five environments near Lethbridge and Coalhurst, Alberta, from 2013 to 2015. The most effective and consistent treatments for kochia management included sulfentrazone applied pre-emergence and fluroxypyr/bromoxynil/2,4-D or pyrasulfotole/bromoxynil applied post-emergence. All of these treatments resulted in ≥90% visible control in all environments and ≥90% kochia biomass reduction compared with the untreated control in Lethbridge 2014 and 2015. MCPA/dichlorprop-p/mecoprop-p, dicamba/2,4-D/mecoprop-p, and dicamba/fluroxypyr resulted in acceptable control among environments (≥80% visible control in all environments and ≥80% kochia biomass reduction in Lethbridge 2014 and 2015); however, the latter two options caused unacceptable (&gt;10%) wheat visible injury in Coalhurst 2014. Recent confirmations of auxinic herbicide-resistant kochia in western Canada—due, in part, to use of synthetic auxins to manage glyphosate-resistant kochia in small-grain cereals—will limit kochia management options. When implemented with non-chemical tools as part of an integrated weed management program, alternative herbicide modes of action like protoporphyrinogen oxidase inhibitors before and photosystem II or 4-hydroxyphenylpyruvate dioxygenase inhibitor(s) within spring wheat could mitigate selection for multiple herbicide-resistant kochia.

Structure-Guided Discovery of Silicon-Containing Subnanomolar Inhibitor of Hydroxyphenylpyruvate Dioxygenase as a Potential Herbicide.

4-Hydroxyphenylpyruvate dioxygenase (HPPD, EC 1.13.11.27) has been recognized as one of the most promising targets in the field of herbicide innovation considering the severity of weed resistance currently. In a persistent effort to develop effective HPPD-inhibiting herbicides, a structure-guided strategy was carried out to perform the structural optimization for triketone-quinazoline-2,4-diones, a novel HPPD inhibitor scaffold first discovered in our lab. Herein, starting from the crystal structure of Arabidopsis thaliana (At)HPPD complexed with 6-(2-hydroxy-6-oxocyclohex-1-ene-1-carbonyl)-1,5-dimethyl-3-(o-tolyl)quinazoline-2,4(1H,3H)-dione (MBQ), three subseries of quinazoline-2,4-dione derivatives were designed and prepared by optimizing the hydrophobic interactions between the side chain of the core structure at the R1 position and the hydrophobic pocket at the active site entrance of AtHPPD. 6-(2-Hydroxy-6-oxocyclohex-1-ene-1-carbonyl)-1,5-dimethyl-3-(3-(trimethylsilyl)prop-2-yn-1-yl)quinazoline-2,4(1H,3H)-dione (60) with the best inhibitory activity against AtHPPD was identified to be the first subnanomolar-range AtHPPD inhibitor (Ki = 0.86 nM), which significantly outperformed that of the lead compound MBQ (Ki = 8.2 nM). Further determination of the crystal structure of AtHPPD in complex with compound 60 (1.85 Å) and the binding energy calculation provided a molecular basis for the understanding of its high efficiency. Additionally, the greenhouse assay indicated that 6-(2-hydroxy-6-oxocyclohex-1-ene-1-carbonyl)-1,5-dimethyl-3-propylquinazoline-2,4(1H,3H)-dione (28) and compound 60 showed acceptable crop safety against peanut and good herbicidal activity with a broad spectrum. Moreover, compound 28 also showed superior selectivity for wheat at the dosage of 120 g ai/ha and favorable herbicidal efficacy toward the gramineous weeds at the dosage of as low as 30 g ai/ha. We believe that compounds 28 and 60 have promising prospects as new herbicide candidates for wheat and peanut fields.