In previous research conducted on nonweed species, the efficacy of glyphosate was shown to be greater in unsterile soils compared to sterile soils and soil microorganisms were found to play an important role in glyphosate efficacy. Conducting greenhouse studies in microbe-free soil may therefore produce unreliable data, leading to erroneous conclusions. The objective of this study was to determine the effect of soil microorganisms on the response of glyphosate-resistant and -susceptible biotypes of three problematic weeds of the midwestern United States: giant ragweed, horseweed, and common lambsquarters. A greenhouse dose–response study was conducted on each of the three weed species grown in sterile and unsterile field soil, and the dry weight response of roots and shoots was measured. The three weed species responded differently to glyphosate when grown in the sterile and unsterile soil; that is, in the presence and absence of soil microbes. Soil microbes influenced the response of the susceptible and resistant giant ragweed biotypes and the susceptible common lambsquarters, but not the tolerant common lambsquarters or either horseweed biotype. The different responses of the three species to glyphosate in the presence and absence of soil microbes demonstrates that rhizosphere interactions are fundamental to the mode of action of glyphosate. These findings suggest that the range of tolerance to glyphosate observed in weeds and the evolution of resistance in weed biotypes may also be influenced by rhizosphere interactions. The soil media used in dose–response screenings to identify susceptible and resistant weed biotypes is very important. Unsterile field soil should be incorporated into growth media when conducting dose–response screenings to avoid false-positive results. In addition, researchers performing glyphosate dose–response assays should be aware of these findings.

Root colonization by soil microorganisms has been shown to increase the activity of glyphosate in resistant and susceptible biotypes of giant ragweed and a susceptible common lambsquarters biotype, but not in horseweed biotypes. The objective of this study was to investigate the colonization of roots in glyphosate-resistant and -susceptible giant ragweed and horseweed biotypes, and glyphosate-tolerant and -susceptible biotypes of common lambsquarters after a sublethal glyphosate application. The three weed species were grown separately in sterile and unsterile field soil and treated with glyphosate at two sublethal rates. Soil microbes were isolated from the roots onto sterile media 3 d after the glyphosate treatment. The susceptible biotypes of giant ragweed and horseweed grown in unsterile soil were colonized by more soil microbes at the higher rate of glyphosate, compared to the resistant biotype grown in unsterile soil. Oomycetes were isolated separately on a selective media and they were also more prevalent in the roots of the susceptible biotypes of each weed species grown in the unsterile soil when glyphosate was applied at the highest rate. Therefore, the ability of these three weed species to tolerate a glyphosate application may involve differences in the susceptibility to soil microbial colonization, especially oomycetes.

In a previous study, glyphosate-susceptible and -resistant giant ragweed biotypes grown in sterile field soil survived a higher rate of glyphosate than those grown in unsterile field soil, and the roots of the susceptible biotype were colonized by a larger number of soil microorganisms than those of the resistant biotype when treated with 1.6 kg ae ha 21 glyphosate. Thus, we concluded that soil-borne microbes play a role in glyphosate activity and now hypothesize that the ability of the resistant biotype to tolerate glyphosate may involve microbial interactions in the rhizosphere. The objective of this study was to evaluate differences in the rhizosphere microbial communities of glyphosatesusceptible and -resistant giant ragweed biotypes 3 d after a glyphosate treatment. Giant ragweed biotypes were grown in the greenhouse in unsterile field soil and glyphosate was applied at either 0 or 1.6 kg ha 21 . Rhizosphere soil was sampled 3 d after the glyphosate treatment, and DNA was extracted, purified, and sequenced with the use of Illumina Genome Analyzer next-generation sequencing. The taxonomic distribution of the microbial community, diversity, genera abundance, and community structure within the rhizosphere of the two giant ragweed biotypes in response to a glyphosate application was evaluated by metagenomics analysis. Bacteria comprised approximately 96% of the total microbial community in both biotypes, and differences in the distribution of some microbes at the phyla level were observed. Select soil-borne plant pathogens (Verticillium and Xanthomonas) and plant-growth-promoting rhizobacteria (Burkholderia) present in the rhizosphere were influenced by either biotype or glyphosate application. We did not, however, observe large differences in the diversity or structure of soil microbial communities among our treatments. The results of this study indicate that challenging giant ragweed biotypes with glyphosate causes perturbations in rhizosphere microbial communities and that the perturbations differ between the susceptible and resistant biotypes. However, biological relevance of the rhizosphere microbial community data that we obtained by next-generation sequencing remains unclear. Nomenclature: Glyphosate; giant ragweed, Ambrosia trifida L.