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BACKGROUND: A population of Echinochloa colona infesting agricultural fields in the northern region of Western Australia evolved glyphosate resistance after 10 years of glyphosate selection. This study identified two phenotypic (susceptible S versus resistant R) lines from within a segregating glyphosate-resistant population. Estimation of survival, growth and reproductive rates of the phenotypes in response to glyphosate selection helped to characterise the level of resistance, fitness and the selection intensity for glyphosate in this species. RESULTS: Estimations of LD50 (lethal dose) and GR50 (growth rate) showed an eightfold glyphosate resistance in this population. The resistant index based on the estimation of seed number (SYn50) showed a 13-fold resistance. As a result of linear combination of plant survival and fecundity rates, plant fitness values of 0.2 and 0.8 were estimated for the S and R phenotypes when exposed to the low dose of 270 g glyphosate ha−1. At the recommended dose of 540 g glyphosate ha−1, fitness significantly decreased (fivefold) in S plants but remained markedly similar (0.7) in plants of the R phenotype. Thus, the calculated selection intensity (SI) at 540 g glyphosate ha−1 was much greater (SI = 17) than at 270 g glyphosate ha−1 (SI = 4). CONCLUSIONS: The assessment of plant survival and fecundity in response to glyphosate selection in the S and R phenotypes allowed a greater accuracy in the estimation of population fitness of both phenotypes and thus of glyphosate selection intensity in E. colona. The estimation of seed number or mass of phenotypes under herbicide selection is a true ecological measure of resistance with implications for herbicide resistance evolution. Fil: Sheng Goh, Sou. University of Western Australia; Australia Fil: Vila Aiub, Martin Miguel. University of Western Australia; Australia. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Parque Centenario. Instituto de Investigaciones Fisiológicas y Ecológicas Vinculadas a la Agricultura. Universidad de Buenos Aires. Facultad de Agronomía. Instituto de Investigaciones Fisiológicas y Ecológicas Vinculadas a la Agricultura; Argentina Fil: Busi, Roberto. University of Western Australia; Australia Fil: Powles, Stephen B.. University of Western Australia; Australia

The rate of herbicide resistance evolution in plants depends on fitness traits endowed by alleles in both the presence and absence (resistance cost) of herbicide selection. The effect of two Lolium rigidum spontaneous homozygous target-site resistance-endowing mutations (Ile-1781-Leu, Asp-2078-Gly) on both ACCase activity and various plant growth traits have been investigated here. Relative growth rate (RGR) and components (net assimilation rate, leaf area ratio), resource allocation to different organs, and growth responses in competition with a wheat crop were assessed. Unlike plants carrying the Ile-1781-Leu resistance mutation, plants homozygous for the Asp-2078-Gly mutation exhibited a significantly lower RGR (30%), which translated into lower allocation of biomass to roots, shoots, and leaves,and poor responses to plant competition. Both the negligible and significant growth reductions associated, respectively, with the Ile-1781-Leu and Asp-2078-Gly resistance mutations correlated with their impact on ACCase activity. Whereas the Ile-1781-Leu mutation showed no pleiotropic effects on ACCase kinetics, the Asp-2078-Gly mutation ledto a significant reduction in ACCase activity. The impaired growth traits are discussed in the context of resistance costs and the effects of each resistance allele on ACCase activity. Similar effects of these two particular ACCase mutations on the ACCase activity of Alopecurus myosuroides were also confirmed. Fil: Vila Aiub, Martin Miguel. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Parque Centenario. Instituto de Investigaciones Fisiológicas y Ecológicas Vinculadas a la Agricultura; Argentina. University of Western Australia. School of Plant Biology. Australian Herbicide Resistance Initiative; Australia Fil: Yu, Quin. University of Western Australia. School of Plant Biology. Australian Herbicide Resistance Initiative; Australia Fil: Han, Heping. University of Western Australia. School of Plant Biology. Australian Herbicide Resistance Initiative; Australia Fil: Powles, Stephen B.. University of Western Australia. School of Plant Biology. Australian Herbicide Resistance Initiative; Australia

Background Gene mutations that endow herbicide resistance may cause pleiotropic effects on plant ecology and physiology. This paper reports on the effect of a number of known and novel target-site resistance mutations of the ALS gene (Ala-122-Tyr, Pro-197-Ser, Asp-376-Glu or Trp-574-Leu) on vegetative growth traits of the weed Raphanus raphanistrum. Results The results from a series of experiments have indicated that none of these ALS resistance mutations imposes negative pleiotropic effects on relative growth rate (RGR), photosynthesis and resource-competitive ability in R. raphanistrum plants. The absence of pleiotropic effects on plant growth occurs in spite of increased (Ala-122-Tyr, Pro-197-Ser, Asp-376-Glu) and decreased (Trp-574-Leu) extractable ALS activity. Conclusion The absence of detrimental pleiotropic effects on plant growth associated with the ALS target-site resistance mutations reported here is a contributing factor in resistance alleles being at relatively high frequencies in ALS-herbicide-unselected R. raphanistrum populations. Fil: Li, Mei. University of Western Australia. School of Plant Biology. Australian Herbicide Resistance Initiative; Australia. Shandong Academy of Agricultural Sciences. Institute of Plant Protection; China Fil: Yu, Qin. University of Western Australia. School of Plant Biology. Australian Herbicide Resistance Initiative; Australia Fil: Han, Heping. University of Western Australia. School of Plant Biology. Australian Herbicide Resistance Initiative; Australia Fil: Vila Aiub, Martin Miguel. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Parque Centenario. Instituto de Investigaciones Fisiológicas y Ecológicas Vinculadas a la Agricultura; Argentina. University of Western Australia. School of Plant Biology. Australian Herbicide Resistance Initiative; Australia Fil: Powles, Stephen B.. University of Western Australia. School of Plant Biology. Australian Herbicide Resistance Initiative; Australia

International audience; Modern herbicides greatly contribute to world agricultural production but their sustainability is threatened by the widespread evolution of herbicide resistant weedy plant populations. Despite the commercial and scientific importance of resistance, there has not been an experimental model system for pro-actively evaluating the potential for herbicide resistance evolution. Here, utilizing the rapidly growing, unicellular photosynthetic microalgae Chlamydomonas reinhardtii (Dangeard), a ratchet protocol has been developed that solves the problem of maintaining both large populations and strong herbicide selection. The ratchet protocol is a progressive set of cycles, each cycle commencing with a population of approximately one million individuals apportioned amongst three herbicide doses for 14 days. Whenever the evolving population demonstrates growth across the three herbicide selection intensities, then the population ratchets to the next cycle of higher herbicide dose. Therefore, by always maintaining large populations under selection pressure, this system offers the opportunity for beneficial mutations to arise and be enriched. Using the well-characterized atrazine herbicide, the ratchet protocol resulted in rapid evolution of populations with different levels of resistance. This robust laboratory based Chlamydomonas system is proposed for application in establishing the respective propensity for resistance evolution to herbicides or other selecting agents.

Unlike in the pesticide and antibiotic resistance literature, potential social costs and externalities associated with herbicide resistance have not generally been considered by economists. The economics of managing herbicide resistance in weeds has focused on cost-effective responses by growers to the development of resistance at the individual farm and field level. Economic analyses of optimal herbicide use have focused on optimising farmer returns in the long run. Weeds have been considered less mobile, compared to insects and diseases, suggesting that externalities resulting from resistance spread will be minimal and any consequent social costs low. Glyphosate is the world's most widely used broad-spectrum non-selective herbicide. Declining glyphosate prices, the adoption of no-till and minimum-till systems and the adoption of glyphosate-tolerant crops, have combined to cause a rapid increase in the use of glyphosate, and resistance is now appearing. In this paper we argue that the increasing possibility of widespread glyphosate resistance, exacerbated in some situations by spread through resistance mobility, presents a case where social costs associated with glyphosate resistance need to be considered when assessing optimal use of this herbicide resource at the farm level. Possible social costs associated with the loss of glyphosate efficacy include potential failure of herbicide-resistant crop systems, reduced use of conservation tillage techniques, and potentially more reliance on herbicides with greater environmental and health risks.

In grasses, residues homologous to residues Ile-1,781 and Ile-2,041 in the carboxyl-transferase (CT) domain of the chloroplastic acetyl-coenzyme A (CoA) carboxylase (ACCase) from the grass weed black-grass (Alopecurus myosuroides [Huds.]) are critical determinants for sensitivity to two classes of ACCase inhibitors, aryloxyphenoxypropionates (APPs) and cyclohexanediones. Using natural mutants of black-grass, we demonstrated through a molecular, biological, and biochemical approach that residues Trp-2,027, Asp-2,078, and Gly-2,096 are also involved in sensitivity to ACCase inhibitors. In addition, residues Trp-2,027 and Asp-2,078 are very likely involved in CT activity. Using three-dimensional modeling, we found that the side chains of the five residues are adjacent, located at the surface of the inside of the cavity of the CT active site, in the vicinity of the binding site for APPs. Residues 1,781 and 2,078 are involved in sensitivity to both APPs and cyclohexanediones, whereas residues 2,027, 2,041, and 2,096 are involved in sensitivity to APPs only. This suggests that the binding sites for these two classes of compounds are overlapping, although distinct. Comparison of three-dimensional models for black-grass wild-type and mutant CTs and for CTs from organisms with contrasted sensitivity to ACCase inhibitors suggested that inhibitors fitting into the cavity of the CT active site of the chloroplastic ACCase from grasses to reach their active sites may be tight. The three-dimensional shape of this cavity is thus likely of high importance for the efficacy of ACCase inhibitors.