Weedy species of the genus Amaranthus, commonly referred to as pigweeds, have increased in frequency and severity over the past few years. Identification of these weeds is difficult because of similar morphological characteristics among species and variation within species. Studies were initiated to develop a molecular marker identification system utilizing restriction enzyme analysis of amplified ribosomal DNA (rDNA). A set of polymerase chain reaction (PCR) markers was developed to distinguish 10 weedy species of pigweeds. Restriction-site variation, utilizing five endonucleases, within the internal transcribed spacers (ITS) of the rDNA allowed for the positive identification of eight species and one pair of species. These markers will be useful for biological and ecological studies on the genus.

A waterhemp population (MCR) previously characterized as resistant to 4-hydroxyphenylpyruvate dioxygenase and photosystem II inhibitors demonstrated both moderate and high levels of resistance to acetolactate synthase (ALS) inhibitors. Plants from the MCR population exhibiting high resistance to ALS inhibitors contained the commonly found Trp574Leu ALS amino acid substitution, whereas plants with only moderate resistance did not have this substitution. A subpopulation (JG11) was derived from the MCR population in which the moderate-resistance trait was isolated from the Trp574Leu mutation. Results from DNA sequencing and ALS enzyme assays demonstrated that resistance to ALS inhibitors in the JG11 population was not due to an altered site of action. This nontarget-site ALS-inhibitor resistance was characterized with whole-plant dose–response experiments using herbicides from each of the five commercialized families of ALS-inhibiting herbicides. Resistance ratios ranging from 3 to 90 were obtained from the seven herbicides evaluated. Nontarget-site resistance to ALS has been rarely documented in eudicot weeds, and adds to the growing list of resistance traits evolved in waterhemp.

A survey of soybean fields containing waterhemp infestations was conducted just prior to harvest in 2008 and 2009 to determine the frequency and distribution of glyphosate-resistant waterhemp in Missouri, and to determine if there are any in-field parameters that may serve as indicators of glyphosate resistance in this species in future crop production systems. Glyphosate resistance was confirmed in 99 out of 144, or 69%, of the total waterhemp populations sampled, which occurred in 41 counties of Missouri. Populations of glyphosate-resistant waterhemp were more likely to occur in fields with no other weed species present at the end of the season, continuous cropping of soybean, exclusive use of glyphosate for several consecutive seasons, and waterhemp plants showing obvious signs of surviving herbicide treatment compared to fields characterized with glyphosate-susceptible waterhemp. Therefore, it is suggested that these four site parameters, and certain combinations of these parameters, serve as predictors of glyphosate resistance in future waterhemp populations.

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 plus s-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.

The extensive use of foliar-applied protoporphyrinogen oxidase (PPO)-inhibiting herbicides indisputably contributes to the continued selection of waterhemp resistant to PPO-inhibiting herbicides (PPO-R). However, the role of soil-residual applications of PPO-inhibiting herbicides in the selection of PPO-R has been clouded by the efficacy that these herbicides have on PPO-R waterhemp. The aim of the present study was to understand if soil-residual PPO-inhibiting herbicides have the potential to influence the proportion of resistant waterhemp in emerging plants as herbicide concentrations diminish in the soil. Greenhouse and field experiments were conducted in a PRE dose-response experiment testing fomesafen or fomesafen plus s-metolachlor in the presence of mixed seed populations of PPO-R and -susceptible (S) waterhemp. The first 20 (greenhouse) or 10 (field) waterhemp plants that survived the residual herbicide treatment were sampled for genotypic analysis to detect the presence of the allele responsible for PPO-R in waterhemp (ΔG210). Relative to the nontreated control, the highest rate of fomesafen increased the frequency of resistance (FOR) by 70% in the greenhouse experiments and 20% in the field experiments. The addition of s-metolachlor did not reduce the fomesafen-induced increase in the FOR in the surviving plants from field or greenhouse experiments. However, the additional herbicide from an alternate site of action (s-metolachlor) substantially improved soil-residual herbicide efficacy over fomesafen alone, which limited the number of waterhemp plants surviving the herbicide application. Thus, the selection for resistance can be delayed with the addition of a herbicide from an alternative site of action by postponing and/or reducing waterhemp emergence. These data strongly suggest that soil-residual PPO-inhibiting herbicides can influence the FOR in a field population, placing even greater importance on the implementation of best management practices such as full herbicide use rates and using herbicides from multiple sites of action to mitigate the risk of selecting for herbicide-resistant weed biotypes.