Twenty-two amino acid substitutions at seven conserved amino acid residues in the acetohydroxyacid synthase (AHAS) gene have been identified to date that confer target-site resistance to AHAS-inhibiting herbicides in biotypes of field-evolved resistant weed species. However, the effect of resistance mutations on AHAS functionality and plant growth has been investigated for only a very few mutations. This research investigates the effect of various AHAS resistance mutations in Lolium rigidum on AHAS functionality and plant growth. The enzyme kinetics of AHAS from five purified L. rigidum populations, each homozygous for the resistance mutations Pro-197-Ala, Pro-197-Arg, Pro-197-Gln, Pro-197-Ser or Trp-574-Leu, were characterized and the pleiotropic effect of three mutations on plant growth was assessed via relative growth rate analysis. All these resistance mutations endowed a herbicide-resistant AHAS and most resulted in higher extractable AHAS activity, with no-to-minor changes in AHAS kinetics. The Pro-197-Arg mutation slightly (but significantly) increased the K_m for pyruvate and remarkably increased sensitivity to feedback inhibition by branched chain amino acids. Whereas the Pro-197-Ser and Trp-574-Leu mutations exhibited no significant effects on plant growth, the Pro-197-Arg mutation resulted in lower growth rates. It is clear that, at least in L. rigidum, these five AHAS resistance mutations have no major impact on AHAS functionality and hence probably no plant resistance costs. These results, in part, explain why so many Pro-197 AHAS resistance mutations in AHAS have evolved and why the Pro-197-Ser and the Trp-574-Leu AHAS resistance mutations are frequently found in many weed species.

Costs of resistance are predicted to reduce plant productivity in herbicide-resistant weeds. Lolium rigidum herbicide-susceptible individuals (S), individuals possessing cytochrome P450-based herbicide metabolism (P450) and multiple resistant individuals possessing a resistant ACCase and enhanced cytochrome P450 metabolism (ACCase/P450) were grown in the absence of mutual plant interaction to estimate plant growth traits. Both P450 and ACCase/P450 resistant phenotypes produced less above-ground biomass than the S phenotype during the vegetative stage. Reduced biomass production in the resistant phenotypes corresponded to a reduced relative growth rate and a lower net assimilation rate and rate of carbon fixation. There were no significant differences between the two resistant phenotypes, suggesting that costs of resistance are associated with P450 metabolism-based resistance. There were no differences in reproductive output among the three phenotypes, indicating that the cost of P450 resistance during vegetative growth is compensated during the production of reproductive structures. The P450-based herbicide metabolism is shown to be associated with physiological resistance costs, which may be manipulated by agronomic management to reduce the evolution of herbicide resistance.

Summary: Quantification of fitness differences between herbicide‐resistant and herbicide‐susceptible weeds permits better prediction of herbicide resistance, and the design of weed management strategies to exploit those traits that result in reduced ecological performance. Reported here is the first attempt to compare the germination and seedling emergence characteristics of one herbicide‐susceptible and two herbicide‐resistant phenotypes from a single weed population. A series of experiments was carried out in controlled conditions to study seed germination and emergence in Lolium rigidum phenotypes possessing target‐site and non‐target‐site mechanisms of herbicide resistance. Lines composed of herbicide‐susceptible (S), metabolism‐based (P450) and target site‐based (ACCase) resistant individuals, isolated from a single multiple‐resistant population (SLR31), were used in the comparative experiments. No major differences in seedling emergence were found among the phenotypes when exposed to an alternating 25/15 °C cycle with a 12‐hourly photoperiod. However, the absence of light associated with soil burial (1–8 cm) markedly inhibited total germination and seedling emergence in the ACCase phenotype compared with the S and P450 phenotypes. Germination at constant temperatures was also inhibited in seeds of the ACCase phenotype, which showed the highest base temperature (T_b) for germination and required more time to reach 50% emergence (t_E50) than the S and P450 phenotypes. Seedling emergence from deep burial (8 cm) in soil promoted significantly higher fatal germination in the S and P450 phenotypes compared with the ACCase phenotype. Despite fatal germination, the S phenotype produced greater emergence from deep burial than both herbicide‐resistant phenotypes. Synthesis and applications. This study demonstrates differential germination and emergence responses to light and thermal environments between herbicide‐susceptible phenotypes and phenotypes possessing target‐site (ACCase) and non‐target site (P450) herbicide resistance within a single L. rigidum population. Shallow seed burial (1 cm) by any cultivation tool will potentially inhibit seedling recruitment of the ACCase phenotype in contrast with the S and P450 phenotypes, which require less restricted conditions to germinate and emerge.