Protoporphyrinogen Oxidase-inhibiting Herbicide Research Articles

Glyphosate-resistant and susceptible downy brome (Bromus tectorum) management with soil-applied residual herbicides

Abstract Downy brome is a cleistogamous facultative winter-annual grass weed that invades cropland, pastureland, and ruderal areas in western North America. Glyphosate-resistant downy brome, the first known glyphosate-resistant grass weed in Canada, was confirmed in a glyphosate-resistant canola field in southern Alberta in 2021. A controlled-environment study was conducted to determine the impact of preemergence soil-applied residual herbicides on glyphosate-resistant and susceptible downy brome in two field soils. Flumioxazin/pyroxasulfone (70/89 g ai ha−1), carfentrazone/pyroxasulfone (18/150 g ai ha−1), sulfentrazone/pyroxasulfone (100/100 or 150/150 g ai ha−1), and saflufenacil/pyroxasulfone (36/120 g ai ha−1) resulted in excellent (≥90%) visible control and downy brome biomass reduction 8 wk after treatment (WAT). The low rate of carfentrazone/pyroxasulfone (12/100 g ai ha−1) resulted in good (≥80%) visible control and biomass reduction 8 WAT, while the low and medium rates of saflufenacil/pyroxasulfone (18/60 or 25/84 g ai ha−1) resulted in ≥80% biomass reduction but suppression only (66% to 75%) based on visible control. Flumioxazin alone (105 g ai ha−1) resulted in good visible control (81%) 8 WAT in a sandy loam soil, but poor (13%) control in a clay loam soil. Soil type affected plant growth as evidenced by reduced growth in the untreated sandy loam soil compared to clay loam soil. The glyphosate-resistant population emerged and grew more vigorously than the glyphosate-susceptible population resulting in greater plant densities in the untreated control and some less-effective herbicide treatments. These results suggest that mixtures of a protoporphyrinogen oxidase-inhibiting herbicide with the very-long-chain fatty acid elongase inhibitor pyroxasulfone applied preemergence at ≥89 g ai ha−1 could be effective components of an herbicide layering strategy targeting glyphosate-resistant and glyphosate-susceptible downy brome.

Vertical mobility and cotton tolerance to trifludimoxazin, a new protoporphyrinogen oxidase-inhibiting herbicide, in three West Texas soils

AbstractTrifludimoxazin, a new protoporphyrinogen oxidase–inhibiting herbicide, is being evaluated for possible use as a soil-residual active herbicide treatment in cotton for control of small-seeded annual broadleaf weeds. Laboratory and greenhouse studies were conducted to compare vertical mobility and cotton tolerance of trifludimoxazin to flumioxazin and saflufenacil, which are two currently registered protoporphyrinogen oxidase–inhibiting herbicides for use in cotton, in three West Texas soils. Vertical soil mobility of trifludimoxazin was similar to flumioxazin in Acuff loam and Olton loam soils, but was more mobile than flumioxazin in the Amarillo loamy sand soil. The depth of trifludimoxazin movement after a 2.5-cm irrigation event ranged from 2.5 to 5.0 cm in all soils, which would not allow for crop selectivity based on herbicide placement, because ideal cotton seeding depth is from 0.6 to 2.54 cm deep. Greenhouse studies indicated that PRE treatments were more injurious than the 14 d preplant treatment when summarized across soils for the three herbicides (43% and 14% injury, respectively). No differences in visual cotton response or dry weight was observed after trifludimoxazin preplant as compared with the nontreated control within each of the three West Texas soils and was similar to the flumioxazin preplant across soils. On the basis of these results, a use pattern for trifludimoxazin in cotton may be established with the use of a more than 14-d preplant restriction before cotton planting.

Characterization of multiple herbicide–resistant waterhemp (Amaranthus tuberculatus) populations from Illinois to VLCFA-inhibiting herbicides

AbstractField experiments were conducted in 2016 and 2017 in Champaign County, IL, to study a waterhemp [Amaranthus tuberculatus(Moq.) J. D. Sauer] population (CHR) resistant to 2,4-D and 4-hydroxyphenylpyruvate dioxygenase (HPPD)-, photosystem II–, acetolactate synthase (ALS)-, and protoporphyrinogen oxidase–inhibiting herbicides. Two field experiments were designed to investigate the efficacy of very-long-chain fatty-acid (VLCFA)-inhibiting herbicides, including a comparison of active ingredients at labeled use rates and a rate titration experiment.Amaranthus tuberculatusdensity and control were evaluated at 28 and 42 d after treatment (DAT). Nonencapsulated acetochlor, alachlor, and pyroxasulfone provided the greatest PRE control of CHR (56% to 75%) at 28 DAT, while metolachlor,S-metolachlor, dimethenamid-P, and encapsulated acetochlor provided less than 27% control. In the rate titration study, nonencapsulated acetochlor controlled CHR more than equivalent field use rates ofS-metolachlor. Subsequent dose–response experiments with acetochlor,S-metolachlor, dimethenamid-P, and pyroxasulfone in the greenhouse included three multiple herbicide–resistant (MHR)A. tuberculatuspopulations: CHR-M6 (progeny generated from CHR), MCR-NH40 (progeny generated from Mclean County, IL), and ACR (Adams County, IL), in comparison with a sensitive population (WUS). Both CHR-M6 and MCR-NH40 are MHR to atrazine and HPPD, and ALS inhibitors and demonstrated higher survival rates (LD50) toS-metolachlor, acetochlor, dimethenamid-P, or pyroxasulfone than ACR (atrazine resistant but HPPD-inhibitor sensitive) and WUS. Based on biomass reduction (GR50), resistant to sensitive (R:S) ratios between CHR-M6 and WUS were 7.5, 6.1, 5.5, and 2.9 forS-metolachlor, acetochlor, dimethenamid-P, and pyroxasulfone, respectively. Values were greater for MCR-NH40 than CHR-M6, and ACR was the most sensitive to all VLCFA inhibitors tested. Complete control of all populations was achieved at or below a field use rate of acetochlor. In summary, field studies demonstrated CHR is not controlled by several VLCFA-inhibiting herbicides. Greenhouse dose–response experiments corroborated field results and generated R:S ratios (LD50) ranging from 4.5 to 64 for CHR-M6 and MCR-NH40 among the four VLCFA-inhibiting herbicides evaluated.

Effect of Sequential Applications of Protoporphyrinogen Oxidase-Inhibiting Herbicides on Palmer Amaranth (Amaranthus palmeri) Control and Peanut Response

Two experiments were conducted in 2013 and 2014 in Florida to evaluate the effects of protoporphyrinogen oxidase (PPO)-inhibiting herbicides and single versus sequential applications on Palmer amaranth control and peanut injury. Protoporphyrinogen oxidase-inhibiting herbicides are among the last available herbicides for the POST control of acetolactate synthase (ALS)-resistant Palmer amaranth in peanut. Lactofen (219 g ai ha–1) applied 5 d after the initial application provided the highest level of Palmer amaranth control 7 and 14 d after initial application (DAIT). Delaying sequential applications of lactofen to 15 d resulted in the highest level of Palmer amaranth control 21 and 28 DAIT. Similar to Palmer amaranth control, foliar injury to peanut was often highest from lactofen applications, and by 28 DAIT lactofen treatments were the only treatments that caused foliar injury. Although no statistical difference was observed between yields of plots treated with acifluorfen (280 g ai ha–1), bentazon (560 g ai ha–1), 2,4-DB (280 g ae ha–1) alone or in combination with each other, plots treated with sequential applications of lactofen 5 or 15 DAIT produced the lowest yields. Sequential applications of lactofen applied 15 DAIT controlled Palmer amaranth more effectively than any other treatment but also caused the highest level of peanut injury. The use of sequential applications of lactofen was the most effective method for control of Palmer amaranth in this study, but did reduce peanut yield.

Constitutive active/androstane receptor, peroxisome proliferator-activated receptor α, and cytotoxicity are involved in oxadiazon-induced liver tumor development in mice

Oxadiazon (OX) is a protoporphyrinogen oxidase-inhibiting herbicide that induces porphyria and liver tumors in rodents. Although porphyria is generally considered to be a risk factor for liver tumor development, the mechanisms through which OX mediates tumor development are unclear. Therefore, in this study, we investigated the mechanisms of tumor development by focusing on constitutive active/androstane receptor (CAR), which is essential for the development of tumors in response to several chemicals. After 1, 4, or 13 weeks of dietary treatment with 1000 ppm OX, hepatic Cyp2b10 expression was induced in wild-type (WT) mice. However, this effect was blocked in CAR-knockout (CARKO) mice. Hepatic Cyp4a10 expression, indicative of peroxisome proliferator-activated receptor α (PPARα) activation, and cytotoxic changes in hepatocytes were also observed in both groups of mice. After initiation by diethylnitrosamine, 26-week treatment with OX resulted in an increase in proliferative lesions, including foci and adenomas, in both genotypes, and the incidence and multiplicity of proliferative lesions in CARKO mice were higher than those in control mice but lower than those in WT mice. These results suggested that CAR, PPARα activation, and cytotoxicity were involved in the development of liver tumors. Moreover, porphyrin was not apparently involved in OX-induced tumor development.

Understanding resistance mechanism of protoporphyrinogen oxidase-inhibiting herbicides: insights from computational mutation scanning and site-directed mutagenesis.

The potential of protoporphyrinogen oxidase (PPO) to develop resistance against five PPO-inhibiting herbicides has been studied using computational mutation scanning (CMS) protocol, leading to valuable insights into the resistance mechanisms and structure-resistance relationship of the PPO inhibitors. The calculated shifts in the binding free energies caused by the mutations correlated very well with those derived from the corresponding experimental data obtained from site-directed mutagenesis of PPO, leading to valuable insights into the resistance mechanisms of PPO inhibitors. The calculated entropy change was related to the conformational flexibility of the inhibitor, which demonstrated that inhibitors with appropriate conformational flexibility may inhibit both the wild type and mutants simultaneously. The reasonable correlation between the computational and experimental data further validate that CMS protocol is valuable for predicting resistance associated with amino acid mutations on target proteins.

Synthesis, in vitro protoporphyrinogen oxidase inhibition, and herbicidal activity of N-(benzothiazol-5-yl)hexahydro-1H-isoindole-1,3-diones and N-(benzothiazol-5-yl)hexahydro-1H-isoindol-1-ones.

Protoporphyrinogen oxidase (EC 1.3.3.4) is one of the most significant targets for a large family of herbicides. As part of our continuous efforts to search for novel protoporphyrinogen oxidase-inhibiting herbicides, N-(benzothiazol-5-yl)tetrahydroisoindole-1,3-dione was selected as a lead compound for structural optimization, leading to the syntheses of a series of novel N-(benzothiazol-5-yl)hexahydro-1H-isoindole-1,3-diones (1a-o) and N-(benzothiazol-5-yl)hexahydro-1H-isoindol-1-ones (2a-i). These newly prepared compounds were characterized by elemental analyses, (1) H NMR, and ESI-MS, and the structures of 1h and 2h were further confirmed by X-ray diffraction analyses. The bioassays indicated that some compounds displayed comparable or higher protoporphyrinogen oxidase inhibition activities in comparison with the commercial control. Very promising, compound 2a, ethyl 2-((6-fluoro-5-(4,5,6,7-tetrahydro-1-oxo-1H-isoindol-2(3H)-yl)benzo[d]thiazol-2-yl)-sulfanyl)acetate, was recognized as the most potent candidate with K(i) value of 0.0091 μm. Further greenhouse screening results demonstrated that some compounds exhibited good herbicidal activity against Chenopodium album at the dosage of 150 g/ha.

Influence of Flumioxazin Application Timing and Rate on Cotton Emergence and Yield

Palmer amaranth is one of the most troublesome weeds in the southeast. Significant reductions in cotton yield because of Palmer amaranth competition warrant intense control efforts consisting of both PRE and POST herbicides. Flumioxazin is a soil-active, protoporphyrinogen oxidase-inhibiting herbicide that is labeled for use in cotton 14 to 21 d before planting; however, shorter preplant application intervals could increase the duration of control from this herbicide. Flumioxazin was applied at 3 rates (0.03 and 0.06 kg ai ha−1in 2009 and an additional rate of 0.09 kg ai ha−1in 2010 and 2011) and 6 application timings (30, 20, 15, 10, 5, and 0 d before planting cotton). Cotton emergence, height, and yield were documented. In 2009, at the Jay and Citra, FL, sites, cotton emergence, plant height, and yield were not affected by any herbicide rate or timing. At Dawson, GA, in the same year, significant reduction in cotton stand counts were observed with application timings < 10 d before planting. Cotton height was reduced similarly at Dawson, GA, but recovered to levels equal to the control by 45 d after planting (DAP). It is believed that rainfall during cotton emergence resulted in this significant level of injury at Dawson, GA. In 2010 and 2011, at Citra, FL, cotton emergence was only reduced when 0.06 and 0.09 kg ha−1were applied at planting. Cotton height showed a similar pattern with additional reductions in height at 0.03 kg ha−1applied at planting and 0.09 kg ha−1applied 5 d before planting. In 2010 and 2011, at Citra, FL, yield was reduced when 0.09 kg ha−1flumioxazin was applied 5 d before planting and when 0.06 and 0.09 kg ha−1were applied at planting. These results indicate that flumioxazin application intervals can be shortened with little crop impact likely to be seen at lower use rates. However, rainfall at crop emergence has the potential to significantly injure cotton and reduce yield.

Experimental hepatic uroporphyria induced by the diphenyl-ether herbicide fomesafen in male DBA/2 mice

Hepatic uroporphyria can be readily induced by a variety of treatments in mice of the C57BL strains, whereas DBA/2 mice are almost completely resistant. However, feeding of the protoporphyrinogen oxidase-inhibiting herbicide fomesafen (0.25% in the diet for 18 weeks) induced hepatic uroporphyria in male DBA/2N mice (liver porphyrin content up to 150 nmol/g, control animals 1 nmol/g), whereas fomesafen-treated male C57BL/6N mice displayed only a slight elevation of liver porphyrins (∼5 nmol/g). The profile of accumulated hepatic porphyrins in fomesafen-treated DBA/2N mice resembled the well-characterised uroporphyria induced by polyhalogenated aromatic hydrocarbons, while histological examination confirmed the presence of uroporphyria-specific cytoplasmic inclusions in the hepatocytes. Uroporphyrinogen decarboxylase activity decreased to about 30% of control values in fomesafen-treated DBA/2N mice; microsomal methoxyresorufin O-dealkylase activity was slightly reduced. The amount of CYP1A1 and CYP1A2 mRNA, as determined by real-time PCR, was not significantly changed; mRNA encoding the housekeeping 5-aminolevulinic acid synthase was elevated 10-fold. Total liver iron was slightly increased. A similar uroporphyria was induced by the herbicide formulation Blazer, containing a structurally related herbicide acifluorfen, when fed to DBA/2N mice at a dose corresponding to 0.25% of acifluorfen in the diet. Since DBA/2 mice are almost completely resistant to all well-characterised porphyrogenic chemicals, the results suggest the possible existence of a yet unknown mechanism of uroporphyria induction, to which the DBA/2 mouse strain is more sensitive than the C57BL strain.

Effects of Protoporphyrinogen Oxidase Inhibitors on Soybean (Glycine max L.) Response, Sclerotinia sclerotiorum Disease Development, and Phytoalexin Production by Soybean1

Sclerotinia stem rot is an important soybean disease. An increase in phytoalexin production with herbicide treatments may reduce the incidence of this disease in soybean. Research was conducted to determine soybean response, Sclerotinia sclerotiorum lesion development, and phytoalexin production in glyphosate-resistant and -susceptible soybean cultivars treated with protoporphyrinogen oxidase–inhibiting herbicides. Necrosis of soybean leaves 7 d after postemergence application of oxyfluorfen at 17.5 g ai/ha, carfentrazone at 1.8 g ai/ha, sulfentrazone at 9.0 g ai/ha, fomesafen at 280 g ai/ha, acifluorfen at 425 g ai/ha, flumiclorac at 30 g ai/ha, CGA-248757 at 4 g ai/ha, and oxadiazon at 280 g ai/ha was equal to or less than lactofen at 70 g ai/ha. In a detached leaf bioassay, S. sclerotiorum lesion diameter was reduced by oxyfluorfen, carfentrazone, sulfentrazone, lactofen, fomesafen, flumiclorac, and oxadiazon compared with the untreated control. Furthermore, lesion diameter on untreated leaves of soybean treated with oxyfluorfen, carfentrazone, sulfentrazone, lactofen, fomesafen, acifluorfen, flumiclorac, CGA-248757, and oxadiazon was reduced compared with the untreated control. Lactofen and sulfentrazone increased leaf phytoalexin production similarly, but neither herbicide affected stem phytoalexin production compared with the untreated control. Glyphosate-resistant and near-isogenic–susceptible cultivars responded similarly when inoculated with S. sclerotiorum in the detached leaf bioassay. Glyphosate-resistant S20-B9 and P93B01 produced more phytoalexins than glyphosate-susceptible S 19-90 and P9281. Herbicide treatments may increase phytoalexin production in leaves of treated plants, but levels in the stem do not explain protection from Sclerotinia stem rot.Nomenclature: Acifluorfen; carfentrazone; CGA-248757, [[2-chloro-4-fluoro-5-[(tetrahydro-3-oxo-1H,3H-[1,3,4]thiadiazolo[3,4-a]pyridazin-1-yliden)amino]phenyl]thio]acetate (proposed common name, fluthiacet-methyl); flumiclorac; fomesafen; lactofen; oxadiazon; oxyfluorfen; sulfentrazone; soybean, Glycine max (L.) Merr. ‘Novartis S 19-90’ (‘S 19-90’), ‘Novartis S20-B9’ (‘S20-B9’), ‘Great Lakes 2415’ (‘GL2415’), ‘Great Lakes 2600’ (‘GL2600’), ‘Pioneer 9281’ (‘P9281’), ‘Pioneer 93B01’ (‘P93B01’); Sclerotinia stem rot, Sclerotinia sclerotiorum (Lib.) de Bary.Additional index words: Aryl triazinone, cultivar, cyclic imide, diphenyl ether, herbicide resistant, isolines, oxadiazole, phytotoxicity, postemergence, protoporphyrinogen inhibitor, Sclerotinia stem rot.Abbreviations: DAT, days after treatment; NIS, nonionic surfactant; GR, glyphosate resistant; protox, protoporphyrinogen oxidase; TLC, thin-layer chromatography; UAN, 28% urea ammonium nitrate.

Synthesis, herbicidal activity, and mode of action of IR 5790.

IR 5790, an arylthiadiazolone herbicide structurally related to oxadiargyl and oxadiazon, was synthesized. The herbicidal activity and mode of action of IR 5790 were investigated. This herbicide has broad-spectrum pre-emergence activity against both dicotyledonous and monocotyledonous weeds. The phenotypic responses of susceptible plants, such as interruption of growth and light-dependent development of necrotic areas on the foliage, are consistent with those observed with protoporphyrinogen oxidase-inhibiting herbicides. Tissues exposed to IR 5790 in darkness accumulated protoporphyrin IX, which led to a photodynamic loss of membrane integrity upon exposure to light. Consistent with these physiological symptoms, IR 5790 strongly inhibited protoporphyrinogen oxidase, with an I(50) value of 3 nM. The presence of a sulfur atom did not significantly alter the molecular properties of the thiadiazolone ring, relative to the oxadiazolone ring of oxadiargyl, which explains why IR 5790 has the same mode of action as this herbicide.

Determination of Porphyrin-Induced Peroxidation of Plant Lipids by Oxygen Electrode.

Protoporphyrinogen oxidase-inhibiting herbicides cause massive accumulation of protoporphyrin IX (Proto IX) in treated plant cells. Reactive oxygen species (ROS) induced by the accumulated Proto IX is thought to cause peroxidative damage, however, the process of peroxidation is still unclear. To investigate the mechanism of the peroxidation of lipids by Proto IX-induced ROS, we used an oxygen electrode. By measuring dissolved oxygen consumption in the reaction mixture, the rate of peroxidation was evaluated in vitro. Production of lipid hydroperoxide was confirmed by measuring thiobarbituric acid reacting substances (TBARS). The linear decrease of dissolved oxygen and linear increase of lipid hydroperoxides occurred immediately after light irradiation, suggesting that the method used in this study is applicable as a model system to investigate lipid peroxidation by Proto IX-induced ROS. Determination of the antioxidants effect on the reduction of dissolved oxygen indicated that both superoxide and hydrogen peroxide scavenging enzymes as well as singlet oxygen scavengers remarkably suppressed the peroxidative reaction. These results suggest that in addition to singlet oxygen, other ROS are involved in the Proto IX-induced peroxidation processes in plant cells. Furthermore, comparison of porphyrin intermediates derived from chlorophyll synthesis showed that the peroxidizing ability of Proto IX was not higher than other porphyries. This suggests that greater accumulation of Proto IX is necessary to initiate lipid peroxidation in plant cells.

Protoporphyrinogen Destruction by Plant Extracts and Correlation with Tolerance to Protoporphyrinogen Oxidase-Inhibiting Herbicides

Herbicidal damage by photobleaching diphenylether herbicides is the indirect result of inhibition of an enzyme in chlorophyll biosynthesis. The substrate of the inhibited enzyme, protoporphyrinogen, accumulates and is subsequently converted to protoporphyrin, a potent photoactive compound which causes light-dependent membrane damage. In the present study, we report characteristics of a factor in the soluble fraction of leaves which can decompose protoporphyrinogen to nonporphyrin products. This process may be important in protecting plants from herbicide damage, since it would interfere with accumulation of the phototoxic porphyrin, protoporphyrin. We found that this protoporphyrinogen destruction is associated with the protein fraction of the soluble leaf homogenate, suggesting its enzymatic nature. Protoporphyrinogen destruction is stable to mild heat, but is eliminated by boiling. Protoporphyrinogen destruction is present in the soluble leaf homogenate but is not localized within the stromal fraction of the chloroplast. The reductants dithiothreitol and β-mercaptoethanol, but not glutathione, inhibit protoporphyrinogen destruction at high concentrations. In contrast, ascorbic acid markedly inhibits destruction even at low concentrations, suggesting a role for cellular ascorbic acid in protecting protoporphyrinogen from destruction, thereby enhancing herbicide action. Protoporphyrinogen destruction was least active in young cucumber leaves, a plant highly susceptible to herbicides. Higher levels of protoporphyrinogen destruction were found in leaves of broadleaf mustard and radish, two plants exhibiting herbicide tolerance. For cucumber, the extent of destruction increased with the age of the plant. These findings suggest a correlation between increased protoporphyrinogen destruction and herbicide tolerance in some plant species.

The effect of protoporphyrinogen oxidase inhibitors on microsomal and mitochondrial cytochromes.

Neurological dysfunction is a characteristic feature of acute porphyrias, unexplained until now. One of the possible explanations is a deficiency of heme in the central nervous system, caused by a block in porphyrin biosynthesis. To test this possibility, the content of brain mitochondrial cytochrome a3 was determined after intracerebroventricular administration of the protoporphyrinogen oxidase-inhibiting herbicide fomesafen. It was established in in vitro experiments that fomesafen is a potent inhibitor of brain mitochondrial protoporphyrinogen oxidase (PROTOOX). Addition of 10(-6) M fomesafen to incubation mixture decreased PROTOOX from 1.02 nmol/mg/h (controls) to 0.42 nmol/mg/h. 10(-5)M of fomesafen decreased this activity to 0.27 nmol/mg/h. In in vivo experiments, 5 microleters of fomesafen solution containing 0.2 M fomesafen/l was administered to male rats by intracerebroventricular injection. The content of brain mitochondrial cytochrome a3 was determined 72 hours later. A slight decrease of the a3 content was observed (control rats 0.25 nmol/mg protein, treated rats 0.22 nmol/mg). Brain cytochrome P450 activities were below detection limits in both control and treated groups. In a separate experiment, male ICR mice were fed 1000 ppm of the protoporphyrinogen oxidaseinhibiting herbicide oxadiazon in the diet for 10 days. Liver microsomal cytochrome P450 content was decreased and liver porphyrins increased. An increase of porphyrin content was also observed in the testes of oxadiazon-fed mice, but testicular cytochrome a3 content was unchanged. The results indicate that, contrary to liver microsomal cytochromes P450, the mitochondrial cytochromes are not susceptible to changes in heme biosynthesis.

Effect of the protoporphyrinogen oxidase-inhibiting herbicide fomesafen on liver uroporphyrin and heptacarboxylic porphyrin in two mouse strains

The effect of the protoporphyrinogen oxidase-inhibiting herbicide fomesafen on liver porphyrin accumulation was studied in long-term high-dose experiments. Fomesafen caused liver accumulation of uroporphyrin and heptacarboxylic porphyrin when fed at 0.25% in the diet to male ICR mice for 5 months (fomesafen-treated mice: 52 nmol uroporphyrin, 21 nmol heptacarboxylic porphyrin/g liver; control mice: traces of uroporphyrin, heptacarboxylic porphyrin not detected). Uroporphyrinogen decarboxylase activity was depressed to about 25% of control values. Iron treatment accelerated the development of this porphyria cutanea tarda-like experimental porphyria both in ICR and C57B1/6J mice. In contrast to other uroporphyrinogen decarboxylase inhibitors, fomesafen treatment did not increase the cytochrome P450IA-related activities and the amount of P450IA2 protein was shown to be significantly decreased by Western immunoblotting. Thus, unique chemical that inhibits both the oxidation of protoporphyrinogen as well as the conversion of uroporphyrinogen to coproporphyrinogen. However, the accumulation of highly carboxylated porphyrins is evident only after prolonged treatment with high doses of the herbicide.

Physiological Basis for Differential Sensitivities of Plant Species to Protoporphyrinogen Oxidase-Inhibiting Herbicides

With a leaf disc assay, 11 species were tested for effects of the herbicide acifluorfen on porphyrin accumulation in darkness and subsequent electrolyte leakage and photobleaching of chlorophyll after exposure to light. Protoporphyrin IX (Proto IX) was the only porphyrin that was substantially increased by the herbicide in any of the species. However, there was a wide range in the amount of Proto IX accumulation caused by 0.1 millimolar acifluorfen between species. Within species, there was a reduced effect of the herbicide in older tissues. Therefore, direct quantitative comparisons between species are difficult. Nevertheless, when data from different species and from tissues of different age within a species were plotted, there was a curvilinear relationship between the amount of Proto IX caused to accumulate during 20 hours of darkness and the amount of electrolyte leakage or chlorophyll photobleaching caused after 6 and 24 hours of light, respectively, following the dark period. Herbicidal damage plateaued at about 10 nanomoles of Proto IX per gram of fresh weight. Little difference was found between in vitro acifluorfen inhibition of protoporphyrinogen oxidase (Protox) of plastid preparations of mustard, cucumber, and morning glory, three species with large differences in their susceptibility at the tissue level. Mustard, a highly tolerant species, produced little Proto IX in response to the herbicide, despite having a highly susceptible Protox. Acifluorfen blocked carbon flow from delta-aminolevulinic acid to protochlorophyllide in mustard, indicating that it inhibits Protox in vivo. Increasing delta-aminolevulinic acid concentrations (33-333 micromolar) supplied to mustard with 0.1 millimolar acifluorfen increased Proto IX accumulation and herbicidal activity, demonstrating that mustard sensitivity to Proto IX was similar to other species. Differential susceptibility to acifluorfen of the species examined in this study appears to be due in large part to differences in Proto IX accumulation in response to the herbicide. In some cases, differences in Proto IX accumulation appear to be due to differences in activity of the porphyrin pathway.