Protoporphyrinogen oxidase (PPO)-inhibiting herbicides remain an important and useful chemistry 60 yr after their first introduction. In this review, based on topics introduced at the Weed Science Society of America 2021 symposium titled “A History, Overview, and Plan of Action on PPO Inhibiting Herbicides,” we discuss the current state of PPO-inhibiting herbicides. Renewed interest in the PPO-inhibiting herbicides in recent years, due to increased use and increased cases of resistance, has led to refinements in knowledge regarding the mechanism of action of PPO inhibitors. Herein we discuss the importance of the two isoforms of PPO in plants, compile a current knowledge of target-site resistance mechanisms, examine non–target site resistance cases, and review crop selectivity mechanisms. Consistent and reproducible greenhouse screening and target-site mutation assays are necessary to effectively study and compare PPO-inhibitor resistance cases. To this end, we cover best practices in screening to accurately identify resistance ratios and properly interpret common screens for point mutations. The future of effective and sustainable PPO-inhibitor use relies on development of new chemistries that maintain activity on resistant biotypes and the promotion of responsible stewardship of PPO inhibitors both new and old. We present the biorational design of the new PPO inhibitor trifludimoxazin to highlight the future of PPO-inhibitor development and discuss the elements of sustainable weed control programs using PPO inhibitors, as well as how responsible stewardship can be incentivized. The sustained use of PPO inhibitors in future agriculture relies on the effective and timely communication from mode of action and resistance research to agronomists, Extension workers, and farmers.

Protoporphyrinogen oxidase (PPO) is an important target for discovering new herbicides that interfere with the synthesis of porphyrin. To discover new PPO inhibitors with improved biological activity, a series of new diphenyl ethers containing tetrahydrophthalimide were designed and synthesized. Among them, J6.1 (IC50 = 4.7 nM) and J6.3 (IC50 = 30.0 nM) show higher maize (Zea mays L.) PPO inhibitory activity than the commercial herbicides oxyfluorfen (IC50 = 117.9 nM) and flumioxazin (IC50 = 157.1 nM). The greenhouse herbicidal activity of J6.3 is comparable to that of oxyfluorfen, and it is greater than that of flumioxazin. Even at a dose of 300 g ai ha−1, cotton (Gossypium hirsutum L.) and peanut (Arachis hypogaea L.) show greater tolerance to J6.3, suggesting that J6.3 could be used for further development of new herbicide candidates in those fields. In addition, molecular docking has been used to further study the mechanism of action of J6.3. The results show that the introduction of a nitro group and tetrahydrophthalimide into the diphenyl ether structure is beneficial to biological activity.

A two-year herbicide residue study was conducted at a commercial nursery to determine the presence of herbicide residues in pond water and sediment following normal production cycles. The runoff water from irrigation and rainfall was contained on the nursery and reused for irrigation. The nursery site was approximately 60 ha with two interconnected ponds, one for containment of runoff water and one serving as a source of irrigation water. The most commonly used herbicides were oxyfluorfen plus pendimethalin (granular formulation-Ornamental Herbicide 2); approximately twice as much herbicide was applied during the second year of the study. Herbicide levels found in pond water and sediment were approximately two-fold greater during the second year corresponding to the increase in active ingredients applied. The highest concentration of oxyfluorfen found in water and sediment was 0.04 μg/ml and 4.0 μg/g, respectively. The highest concentration of pendimethalin found in water and sediment was 0.008 μg/ml and 14.3 μg/g, respectively. In irrigation water samples, the highest concentration of oxyfluorfen and pendimethalin detected was 0.005 μg/ml and 0.002 μg/ml, respectively. The herbicides did not accumulate in water or sediment over the two-year period. No landscape plant phytotoxicity problems are likely at the herbicide levels detected in irrigation water samples.

Protein damage, as a primary phytotoxic consequence of in vivo lipid peroxidation, induced by the diphenyl ether herbicide oxyfluorfen [2-chloro-1-(3-ethoxy-4-nitrophenoxy)-4-(trifluoromethyl)benzene] at a concentration of 10 μM, was measured with the green alga Scenedesmus acutus. In the light, water-soluble proteins are destroyed by a herbicide-induced peroxidation process that can be detected by production of fluorescent products and loss of specific amino acid residues of proteins. The water-soluble cytochrome c-553 and the membrane-bound cytochrome f-553, components of the photosynthetic electron transport, were specifically used as sensitive markers for protein damage, measured as decrease of redox reactions of the cytochromes. Under peroxidizing conditions, destruction of the algal cytochrome c is significantly higher than destruction of membrane-bound components, such as cytochrome f and chlorophyll. Protection against protein loss is achieved by the nonbiological antioxidant ethoxyquin (1,2-dihydro-6-ethoxy-2,2,4-trimethylquinoline) or the photosynthesis inhibitor diuron [N′-(3,4-dichlorophenyl)-N,N-dimethylurea].

Oat (Avena sativa L. ‘Elgin′) and soybean (Glycine max L. ‘Evans′) were treated with 14C-diclofop-methyl {methyl ester of 2-[4-(2,4-dichlorophenoxy)phenoxy] propionic acid]} or 14C-diclofop alone or in combination with 2,4-D [(2,4-dichlorophenoxy)acetic acid] or bentazon [3-isopropyl-1H-2,1,3-benzothiadiazin-4(3H)-one 2,2-dioxide] and 14C-2,4-D alone or in combination with diclofop-methyl or diclofop. More radioactivity was recovered in the treatment zone after 14C-diclofop-methyl applications, alone or in combination, than after similar 14C-diclofop treatments in oat and soybean. Basipetal movement of radioactivity was 4 and 1% and acropetal movement was 1 and 4% in oat and soybean, respectively, regardless of the diphenyl ether treatment or time. Addition of 2,4-D or bentazon did not reduce translocation of radioactivity from 14C-diclofop-methyl treatments in either plant species. Basipetal movement of radioactivity from 14C-diclofop-methyl was greater than from 14C-diclofop in both oat and soybean. The addition of diclofop-methyl or diclofop did not affect the pattern or amount of 14C-2,4-D radioactivity translocated. In oats, radioactivity appeared to accumulate within the intercalary meristematic region with 14C-2,4-D and 14C-diphenyl-ether applications. Diclofop-methyl at 1 kg/ha applied either to an entire oat plant at the three-leaf stage or the apical meristemic region resulted in plant mortality. The extent to which transport contributes to diclofop-methyl efficacy is questioned.

A collection of common purslane (Portulaca oleracea L.) ecotypes from various locations around the world was assembled at the University of Illinois. Applications of nitrofen (2,4-dichlorophenyl-p-nitrophenyl ether) to the various ecotypes showed that 3 of the 26 ecotypes [India, Mexico, and Florida (Sanford)] exhibited some degree of tolerance. This marginal tolerance rapidly disappeared as the rate of nitrofen increased beyond 0.56 kg/ha. All of the ecotypes tested were susceptible to applications of oxyfluorfen [2-chloro-2-(3-ethoxy-4-nitrophenoxy)-4-(trifluoromethyl)benzene]. Oxyfluorfen was toxic to common purslane plants at rates as low as 5 g/ha. Treatment of ‘Hybelle’ (tolerant) and ‘Rio Verde’ (susceptible) cabbage (Brassica olerace var. capitata L.) cultivars with 3.36 kg/ha nitrofen reduced plant fresh weight to 82% and 62%, respectively, of control. Oxyfluorfen at 0.15 kg/ha reduced fresh weight to 60% and 27% for Hybelle and Rio Verde, respectively. Applications of nitrofen at 3.36 kg/ha reduced plant fresh weight in Hybelle the same amount as 100 g/ha oxyfluorfen and 70 g/ha in Rio Verde.

Reducing herbicide rates can reduce production costs and the potential for carryover injury in subsequent crops, but may also compromise residual weed control. When used at the highest recommended rates (0.35 kg ai/ha), fomesafen may cause carryover injury on sweet corn. One of the most valuable attributes of fomesafen use in snap and dry beans is control of common ragweed. Field experiments were conducted in 1999, 2000, and 2002 to investigate both the residual ragweed control from fomesafen at 0.09, 0.18, and 0.36 kg ai/ha and the potential for a second application to extend the duration of residual control, while reducing the total amount applied per season. Results were significantly affected by weather conditions. In a very wet year (2000), with the greatest ragweed pressure, a single application of fomesafen at 0.20 kg/ha was predicted to provide at least 90% control of ragweed throughout snap and dry bean seasons. In the driest year (1999), 0.09 kg ai/ha provided greater than 90% control for the same period. Regardless of whether a single or double application was made, the total amount of fomesafen required per season was less than the manufacturer's recommended rate. Compared with making a single application, two low-rate applications did not reduce the total fomesafen rate required per season. To help insure at least 90% control of ragweed throughout the snap and dry bean seasons a grower should apply 0.20 kg ai/ha early in the season. Lower rates may be used, but in wet seasons, a second application may be needed to prevent yield loss or interference of ragweed with mechanical harvesting.

Studies were conducted to determine acifluorfen and lactofen absorption, translocation, and metabolism in protox-inhibiting herbicide-susceptible and -resistant common waterhemp. Acifluorfen and lactofen absorption was similar in both biotypes. Herbicide absorption was 12% in both susceptible and resistant common waterhemp 6 h after treatment (HAT). Absorption increased to 32 and 42% in susceptible and resistant plants, respectively, at 72 HAT. Translocation was similar in both biotypes for both herbicides. Herbicide translocation out of the treated leaf ranged between 5 and 15%. In a separate study, resistant common waterhemp plants were treated with acifluorfen or lactofen, alone or with tridiphane. Acifluorfen or lactofen injury to resistant common waterhemp was not altered with the addition of tridiphane. Treatments of 14C-acifluorfen or -lactofen on susceptible and resistant common waterhemp resulted in similar lactofen metabolism in both biotypes, but acifluorfen was not metabolized in either biotype within 24 HAT. This data indicate that differences in herbicide absorption, translocation, or metabolism are not the mechanism of common waterhemp resistance to protox-inhibiting herbicides.

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.