Field experiments were conducted over 3 yr at three locations in Illinois to evaluate the efficacy of glyphosate in glyphosate-resistant soybean planted in rows spaced 19, 38, and 76 cm. Minimal soybean injury (less than 10%) was observed from any glyphosate treatment. Glyphosate treatments controlled 82 to 99% of giant foxtail. Common waterhemp control was increased as soybean row spacing was decreased. Applying sequential glyphosate applications or increasing the glyphosate rate from 420 g ae/ha to 840 g/ha frequently increased common waterhemp control in 76-cm rows. Velvetleaf control with glyphosate was variable, ranging from 48 to 99%. Decreasing soybean row spacing, utilizing sequential glyphosate applications, or increasing the glyphosate rate improved velvetleaf control in at least four of eight site-years. Glyphosate treatments generally resulted in weed control and soybean yield equal to or greater than the standard herbicide treatments. However, glyphosate treatments yielded less than the hand-weeded control in four of eight site-years, suggesting that weed control from glyphosate treatments was sometimes inadequate.

A field study was conducted in 2007 and 2008 near Murphysboro, IL to determine the effect of plant height and addition of glyphosate on control of glyphosate-resistant horseweed with saflufenacil. Saflufenacil was applied at rates ranging from 25 to 125 g ai ha−1alone and in combination with glyphosate at 840 g ae ha−1, and the efficacy compared to paraquat at 840 g ai ha−1. Control of horseweed with glyphosate applied alone was less than 30%, confirming the presence of glyphosate-resistant plants. At 14 d after application, all treatments with saflufenacil or paraquat provided at least 90% control. Saflufenacil applied alone at the lowest rate of 25 g ha−1provided less control (92%) than all other treatments that included saflufenacil, and efficacy was reduced as horseweed height at application increased. Horseweed control from saflufenacil at 50 g ha−1was reduced as plant height increased in 2007 but not in 2008. However, saflufenacil applied at 50 g ha−1or greater resulted in at least 98% control, regardless of horseweed height at application or tank mixture with glyphosate. Combining glyphosate with saflufenacil at 25 g ha−1increased horseweed control compared with saflufenacil applied alone and resulted in control similar to saflufenacil applied at 50 g ha−1. Control of horseweed from paraquat declined over time as the growth continued from the apical meristem. The extent of horseweed regrowth from applications of saflufenacil alone was less than that observed from paraquat. The addition of glyphosate to saflufenacil further reduced the frequency of horseweed regrowth compared with saflufenacil applied alone.

Waterhemp resistance to foliar applications of protoporphyrinogen oxidase (PPO)–inhibiting herbicides has become increasingly disconcerting given the widespread distribution of glyphosate resistance. Fortunately, soil-residual PPO-inhibiting herbicides remain efficacious in waterhemp populations resistant to PPO-inhibiting herbicides; however, these herbicides should theoretically select for the resistant biotype as herbicide concentrations diminish in the soil. Accordingly, the objectives of this research were twofold: (1) evaluate the efficacy of three PPO-inhibiting herbicides, foliar- and soil-applied, on PPO-resistant (PPO-R) and PPO-susceptible (PPO-S) waterhemp, and (2) investigate the differential effects of PPO-inhibiting herbicides on an R biotype and an S biotype during several discrete developmental events relevant to soil–residual herbicide activity (i.e., radicle protrusion, radicle elongation, and waterhemp emergence). Greenhouse and growth chamber experiments indicated that the R biotype was least sensitive to the diphenylether herbicide fomesafen, followed by sulfentrazone and flumioxazin; however, fomesafen pluss-metolachlor improved soil-residual efficacy over fomesafen alone. Growth stage considerably influenced the R : S ratio, decreasing from 38× to 3.4×, when comparing ratios generated from foliar applications and soil-residual applications measuring radicle protrusion, respectively. Overall, this research supports the use of full soil-residual herbicide rates, reinforcing the importance of best management practices to manage the spread of herbicide resistance.

BACKGROUND Waterhemp (Amaranthus tuberculatus (Moq.) J. D. Sauer) is one of the most pernicious weeds in cropping systems of the USA due to its evolved resistance against several herbicide sites‐of‐action, including protoporphyrinogen oxidase inhibitors (PPO‐R). Currently, the only source of PPO‐R documented in waterhemp is ΔG210 of PPX2. Gene flow may not only lead to a transfer of herbicide‐resistant alleles, but also produce a hybrid genotype more competitively fit than one or both parents. However, investigating gene flow of Amaranthus species has been of interest in the past two decades with limited evidence. RESULTS Here, a high‐throughput MiSeq amplicon sequencing method was used to investigate alterations of the PPX2 gene in 146 PPO‐R waterhemp populations across five Midwest states of the USA. Five R128 codons of PPX2, novel to waterhemp, were found including AGG (R), GGA (G), GGG (G), AAA (K) and ATA (I). R128G, R128I, and R128K were found in 11, 3, and 2 populations, respectively. R128G and R128I, but not R128K, conferred fomesafen resistance in a bacterial system. Sequence alignment of the R128 region of PPX2 identified a tumble pigweed (Amaranthus albus)‐type and Palmer amaranth (Amaranthus palmeri)‐type PPX2 allele to be present and widespread in the surveyed waterhemp populations, thus providing strong evidence of gene flow between Amaranthus species. CONCLUSION Using a next‐generation sequencing method, we identified two PPO target‐site mutations R128G/I novel to waterhemp and provided evidence of gene flow of Amaranthus species in a large group of screened waterhemp populations from five Midwest states of the USA. © 2019 Society of Chemical Industry

Molecular assays are often implemented by weed scientists for detection of herbicide-resistant individuals; however, the utility of these assays can be limited if multiple mechanisms of evolved resistance exist. Waterhemp resistant to protoporphyrinogen oxidase (PPO)– inhibiting herbicides is conferred by a target-site mutation in PPX2L (a gene coding for PPO), resulting in the loss of a glycine at position 210 (ΔG210). This ΔG210 mutation of PPX2L is the only known mechanism responsible for PPO-inhibitor resistance (PPO-R) in waterhemp from five states (Illinois, Indiana, Iowa, Kansas, and Missouri); however, a limited number of populations have been tested, especially in Illinois. To verify the ubiquity of the ΔG210 in PPO-R waterhemp populations in Illinois, a previously published allele-specific PCR (asPCR) was used for the detection of the ΔG210 mutation to associate this mutation with phenotypic resistance in 94 Illinois waterhemp populations. The ΔG210 mutation was detected in all populations displaying phenotypic resistance to lactofen (220 g ai ha−1), indicating the deletion is likely the only mechanism of resistance. With evidence that the ΔG210 mutation dominates PPO-R waterhemp biotypes, molecular detection techniques have considerable utility. Unfortunately, the previously published asPCR is time consuming, very sensitive to PCR conditions, and requires additional steps to eliminate the possibility of false negatives. To overcome these limitations, a streamlined molecular method using the TaqMan® technique was developed, utilizing allele-specific, fluorescent probes for high-throughput, robust discrimination of each allele (resistant and susceptible) at the 210th amino acid position of PPX2L.