Point Mutations and Cytochrome P450 Can Contribute to Resistance to ACCase-Inhibiting Herbicides in Three Phalaris Species

Species of Phalaris have historically been controlled by acetyl-coenzyme A carboxylase (ACCase)-inhibiting herbicides; however, overreliance on herbicides with this mechanism of action has resulted in the selection of resistant biotypes. The resistance to ACCase-inhibiting herbicides was characterized in Phalaris brachystachys, Phalaris minor, and Phalaris paradoxa samples collected from winter wheat fields in northern Iran. Three resistant (R) biotypes, one of each Phalaris species, presented high cross-resistance levels to diclofop-methyl, cycloxydim, and pinoxaden, which belong to the chemical families of aryloxyphenoxypropionates (FOPs), cyclohexanediones (DIMs), and phenylpyrazolines (DENs), respectively. The metabolism of 14C-diclofop-methyl contributed to the resistance of the P. brachystachys R biotype, while no evidence of herbicide metabolism was found in P. minor or P. paradoxa. ACCase in vitro assays showed that the target sites were very sensitive to FOP, DIM, and DEN herbicides in the S biotypes of the three species, while the R Phalaris spp. biotypes presented different levels of resistance to these herbicides. ACCase gene sequencing confirmed that cross-resistance in Phalaris species was conferred by specific point mutations. Resistance in the P. brachystachys R biotype was due to target site and non-target-site resistance mechanisms, while in P. minor and P. paradoxa, only an altered target site was found.

Detecting ACCase-targeting herbicides effect on ACCase activity utilizing a malachite green colorimetric functional assay

Research was conducted to evaluate acetyl-Coenzyme A carboxylase (ACCase) enzyme activity using a functional malachite green colorimetric assay previously identified as resistant to sethoxydim, and select aryloxyphenoxypropionate (FOPs) herbicides, fenoxaprop, and fluazifop. Two resistant southern crabgrass [Digitaria ciliaris (Retz.) Koeler] biotypes, R1 and R2, containing an Ile-1781-Leu amino acid substitution and previously identified as resistant to sethoxydim, pinoxaden, and fluazifop but not clethodim was utilized as the resistant chloroplastic ACCase source compared to known susceptible (S) ACCase. Dose-response studies with sethoxydim, clethodim, fluazifop-p-butyl, and pinoxaden (0.6 to 40 µM) were conducted to compare the ACCase enzyme-herbicides interaction of R1, R2, and S using the malachite green functional assay. Assay results indicated that R biotypes required more ACCase-targeting herbicides to inhibit ACCase activity compared to S. IC50 values of all four herbicides for R biotypes were consistently an order of magnitude greater than S. No sequencing differences in the carboxyltransferase domain was observed for R1 and R2, however, R2 IC50 values were greater across all herbicides. These results indicate the malachite green functional assay is effective in evaluating ACCase enzyme activity of R and S biotypes in the presence of ACCase-targeting herbicides, which can be used as a replacement for the 14C-based radiometric functional assay.

Evaluation of sequential applications of quizalofop-P-ethyl and florpyrauxifen-benzyl in acetyl CoA carboxylase-resistant rice

Information on performance of sequential treatments of quizalofop-P-ethyl with florpyrauxifen-benzyl on rice is lacking. Field studies were conducted in 2017 and 2018 in Stoneville, MS, to evaluate sequential timings of quizalofop-P-ethyl with florpyrauxifen-benzyl included in preflood treatments of rice. Quizalofop-P-ethyl treatments were no quizalofop-P-ethyl; sequential applications of quizalofop-P-ethyl at 120 g ha−1 followed by (fb) 120 g ai ha−1 applied to rice in the 2- to 3-leaf (EPOST) fb the 4-leaf to 1-tiller (LPOST) growth stages or LPOST fb 10 d after flooding (PTFLD); quizalofop-P-ethyl at 100 g ha−1 fb 139 g ha−1 EPOST fb LPOST or LPOST fb PTFLD; quizalofop-P-ethyl at 139 g ha−1 fb 100 g ha−1 EPOST fb LPOST and LPOST fb PTFLD; and quizalofop-P-ethyl at 85 g ha−1 fb 77 g ha−1 fb 77 g ha−1 EPOST fb LPOST fb PTFLD. Quizalofop-P-ethyl was applied alone and in mixture with florpyrauxifen-benzyl at 29 g ai ha−1 LPOST. Visible rice injury 14 d after PTFLD (DA-PTFLD) was no more than 3%. Visible control of volunteer rice (‘CL151’ and ‘Rex’) 7 DA-PTFLD was similar and at least 95% for each quizalofop-P-ethyl treatment. Barnyardgrass control with quizalofop-P-ethyl at 120 fb 120 g ha−1 LPOST fb PTFLD was greater (88%) in mixture with florpyrauxifen-benzyl. The addition of florpyrauxifen-benzyl to quizalofop-P-ethyl increased rough rice yield when quizalofop-P-ethyl was applied at 100 g ha−1 fb 139 g ha−1 EPOST fb LPOST. Sequential applications of quizalofop-P-ethyl at 120 g ha−1 fb 120 g ha−1 EPOST fb LPOST, 100 g ha−1 fb 139 g ha−1 EPOST fb LPOST, or 139 g ha−1 fb 100 g ha−1 EPOST fb LPOST controlled grass weed species. The addition of florpyrauxifen-benzyl was not beneficial for grass weed control. However, because quizalofop-P-ethyl does not control broadleaf weeds, florpyrauxifen-benzyl could provide broad-spectrum weed control in acetyl coenzyme A carboxylase–resistant rice.

Rate response of select grass weeds to pinoxaden

Pinoxaden is a POST acetyl coenzyme A carboxylase (ACCase) inhibitor in the phenylpyrazolin chemical family and is labelled for turfgrass use at broadcast rates of 35.5 to 71 g ai ha−1 and spot spray rates of 156 to 310 g ai ha−1. A greenhouse rate-response study was conducted to characterize the efficacy of pinoxaden against common grassy weeds. Weed species examined in this study were yellow foxtail, southern sandbur, annual bluegrass, roughstalk bluegrass, large crabgrass, dallisgrass, bahiagrass, goosegrass, and perennial ryegrass. Nonlinear regressions were modelled to determine visible injury rates (the application rate at which 50% of the weed species were injured and the 90% [I90] rate) and weight reduction rates (the application rate at which there was a 50% reduction in fresh weight and 90% reduction [WR90]) for each weed species. Only annual bluegrass, bahiagrass, and goosegrass had visible injury I90 values greater than the maximum labelled spot spray rate of 310 g ai ha−1. Annual bluegrass, bahiagrass, southern sandbur, and goosegrass all had WR90 values greater than the maximum labelled spot spray rate of 310 g ai ha−1. Results from this study indicate that the evaluated weed species can be ranked, according to visible injury I90 values, from most to least susceptible: perennial ryegrass > yellow foxtail > dallisgrass > large crabgrass > southern sandbur > roughstalk bluegrass > bahiagrass > goosegrass > annual bluegrass.

A novel gravity-induced blood flow restriction model augments ACC phosphorylation and PGC-1α mRNA in human skeletal muscle following aerobic exercise: a randomized crossover study

This study tested the hypothesis that a novel, gravity-induced blood flow restricted (BFR) aerobic exercise (AE) model will result in greater activation of the AMPK–PGC-1α pathway compared with work rate-matched non-BFR. Thirteen healthy males (age: 22.4 ± 3.0 years; peak oxygen uptake: 42.4 ± 7.3 mL/(kg·min)) completed two 30-min work rate-matched bouts of cycling performed with their legs below (CTL) and above their heart (BFR) at ∼2 weeks apart. Muscle biopsies were taken before, immediately, and 3 h after exercise. Blood was drawn before and immediately after exercise. Our novel gravity-induced BFR model led to less muscle oxygenation during BFR compared with CTL (O2Hb: p = 0.01; HHb: p < 0.01) and no difference in muscle activation (p = 0.53). Plasma epinephrine increased following both BFR and CTL (p < 0.01); however, only norepinephrine increased more following BFR (p < 0.01). PGC-1α messenger RNA (mRNA) increased more following BFR (∼6-fold) compared with CTL (∼4-fold; p = 0.036). VEGFA mRNA increased (p < 0.01) similarly following BFR and CTL (p = 0.21), and HIF-1α mRNA did not increase following either condition (p = 0.21). Phosphorylated acetyl-coenzyme A carboxylase (ACC) increased more following BFR (p < 0.035) whereas p-PKA substrates, p-p38 MAPK, and acetyl-p53 increased (p < 0.05) similarly following both conditions (p > 0.05). In conclusion, gravity-induced BFR is a viable BFR model that demonstrated an important role of AMPK signalling on augmenting PGC-1α mRNA. Novelty Gravity-induced BFR AE reduced muscle oxygenation without impacting muscle activation, advancing gravity-induced BFR as a simple, inexpensive BFR model. Gravity-induced BFR increased PGC-1α mRNA and ACC phosphorylation more than work rate-matched non-BFR AE. This is the first BFR AE study to concurrently measure blood catecholamines, muscle activation, and muscle oxygenation.

Effect of chronic exposure to monocrotophos on white adipose tissue in rats and its association with metabolic dyshomeostasis

Earlier, we demonstrated that chronic exposure to monocrotophos (MCP) elicits insulin resistance in rats along with increased white adipose tissue (WAT) weights. This study was carried out to delineate the biochemical and molecular changes in adipose tissues of rats subjected to chronic exposure to MCP (0.9 and 1.8 mg/kg bw/d for 180 days). Pesticide-treated rats exhibited increased fasting glucose and hyperinsulinemia as well as dyslipidemia. Tumor necrosis factor-alpha and leptin levels were elevated, while adiponectin level was suppressed in plasma of treated rats. MCP treatment caused discernable increase in the weights of perirenal and epididymal WAT. Acetyl coenzyme A carboxylase, fatty acid synthase, glyceraldehyde-3-phosphate dehydrogenase, lipin-1, and lipolytic activities were elevated in the WAT of MCP-treated rats. Corroborative changes were observed in the expression profile of proteins that are involved in lipogenesis and adipose tissue differentiation. Our results clearly demonstrate that long-term exposure to organophosphorus insecticides (OPIs) such as MCP has far-reaching consequences on metabolic health as evidenced by the association of adipogenic outcomes with insulin resistance, hyperinsulinemia, endocrine dysregulations, and dyslipidemia. Taken together, our results suggest that long-term exposure to OPI may be a risk factor for metabolic dysregulations.