A New V361A Mutation in Amaranthus palmeri PPX2 Associated with PPO-Inhibiting Herbicide Resistance.

Weeds resistant to PPO-inhibiting herbicides threaten the profitability of crop producers relying on this chemistry. In Amaranthus palmeri, mutations at G210 (∆G210) and R128 (R128G/M) of the PPX2 gene were reported to confer PPO-inhibitor resistance. Here, A. palmeri samples from nine states in America, having survived a field application of a PPO-inhibitor, were genotyped to determine the prevalence of these mutations. Less than 5% of the 1828 A. palmeri plants screened contained the ∆G210 mutation. Of the plants lacking ∆G210, a R128 substitution was only found in a single plant. An A. palmeri population from Alabama without mutations at G210 or R128 had a resistance ratio of 3.1 to 3.5 for fomesafen. Of the candidate PPX2 mutations identified in this population, only V361A conferred resistance to lactofen and fomesafen in a transformed bacterial strain. This is the first report of the V361A substitution of PPX2 conferred PPO-inhibiting herbicide resistance in any plant species. Future molecular screens of PPO-inhibitor resistance in A. palmeri and other species should encompass the V361A mutation of PPX2 to avoid false-negative results.

Investigating target-site resistance mechanism to the PPO-inhibiting herbicide fomesafen in waterhemp and interspecific hybridization of Amaranthus species using next generation sequencing.

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.

Differential antioxidant enzyme activity in rapid-response glyphosate-resistant Ambrosia trifida.

BACKGROUND: The giant ragweed (Ambrosia trifida L.) rapid-response (RR) biotype exhibits a sacrificial form of glyphosate resistance whereby an oxidative burst in mature leaves results in foliage loss, while juvenile leaves remain uninjured. This work investigated the safening capacity of antioxidant enzymes in RR juvenile leaves following glyphosate treatment and examined cross tolerance to paraquat. RESULTS: Basal antioxidant enzyme activities were similar between glyphosate-susceptible (GS) and RR biotypes. Lipid peroxidation was first detected in RR mature leaves at 8 h after treatment (HAT) and by 32 HAT was 5.3 and 21.1 times greater than that in RR juvenile leaves and GS leaves, respectively. Preceding lipid peroxidation in the RR biotype at 2 and 4 HAT, the only increase in enzymatic activity was observed in ascorbate-glutathione cycle enzymes in RR juvenile leaves, particularly ascorbate peroxidase, dehydroascorbate reductase, and glutathione reductase. Sensitivity to paraquat was similar between biotypes. CONCLUSION: The RR biotype is not inherently more tolerant to oxidative stress. The difference in tissue damage between RR juvenile and mature leaves following glyphosate treatment is attributable at least partially to the transient increase in antioxidant enzyme expression in juvenile leaves (0-8 HAT), but may also be attributable to lower overall RR induction in juvenile leaves compared with mature leaves. © 2018 Society of Chemical Industry.