Levels Of Herbicide Resistance Research Articles

Herbicide resistance in Bromus and Hordeum spp. in the Western Australian grain belt

Random surveys conducted in the Western Australian (WA) grain belt have shown that herbicide-resistant Lolium rigidum and Raphanus raphanistrum are a widespread problem across the cropping region. In 2010, a random survey was conducted to establish the levels of herbicide resistance for common weed species in crop fields, including the minor but emerging weeds Bromus and Hordeum spp. This is the first random survey in WA to establish the frequency of herbicide resistance in these species. For the annual grass weed Bromus, 91 populations were collected, indicating that this species was present in >20% of fields. Nearly all populations were susceptible to the commonly used herbicides tested in this study; however, a small number of populations (13%) displayed resistance to the acetolactate synthase-inhibiting sulfonylurea herbicides. Only one population displayed resistance to the acetyl-coenzyme A carboxylase-inhibiting herbicides. Forty-seven Hordeum populations were collected from 10% of fields, with most populations being susceptible to all herbicides tested. Of the Hordeum populations, 8% were resistant to the sulfonylurea herbicide sulfosulfuron, some with cross-resistance to the imidazolinone herbicides. No resistance was found to glyphosate or paraquat, although resistance to these herbicides has been documented elsewhere in Australia for Hordeum spp. (Victoria) and Bromus spp. (Victoria, South Australia and WA).

Intensive cropping systems select for greater seed dormancy and increased herbicide resistance levels in Lolium rigidum (annual ryegrass).

Lolium rigidum (annual ryegrass) is a widespread annual crop weed that has evolved high levels of resistance to selective herbicides. Anecdotal evidence suggests that intensive cropping also leads to higher seed dormancy in L. rigidum. This was quantified by measuring dormancy levels in L. rigidum populations collected from paired sites (one with nil to low cropping intensity, the other intensively cropped) located throughout the Western Australian grain belt. Populations from non-cropped fields or those with low cropping intensity showed higher and faster germination than populations from fields with a medium- or high-intensity cropping regime. Resistance to selective herbicides was also higher in the medium- and high-intensity cropping fields than in the low-intensity cropping fields. High-intensity cropping systems are likely to impose greater selection pressures for seed dormancy and selective herbicide resistance, because late-emerging seedlings avoid preplanting weed control practices (tillage and non-selective herbicide application) but are exposed to selective in-crop herbicides.

Bioassay and Characterization of Several Palmer Amaranth (<i>Amaranthus palmeri</i>) Biotypes with Varying Tolerances to Glyphosate

The wide distribution of Palmer amaranth (Amaranthus palmeri) in the southern US became a serious weed control problem prior to the extensive use of glyphosate-resistant crops. Currently glyphosate-resistant populations of Palmer amaranth occur in many areas of this geographic region creating an even more serious threat to crop production. Investigations were undertaken using four biotypes (one glyphosate-sensitive, one resistant from Georgia and two of unknown tolerance from Mississippi) of Palmer amaranth to assess bioassay techniques for the rapid detection and level of resistance in populations of this weed. These plants were characterized with respect to chlorophyll, betalain, and protein levels and immunological responses to an antibody of 5-enolpyruvylshikimate-3-phosphate synthase (EPSPS) the target site of glyphosate. Only slight differences were found in four biotypes grown under greenhouse conditions regarding extractable soluble protein and chlorophyll content, but one biotype was found to be devoid of the red pigment, betalain. Measurement of early growth (seedling shoot elongation) of seedlings was a useful detection tool to determine glyphosate resistance. A leaf disc bioassay (using visual ratings and/or chlorophyll analysis) and an assay for shikimate accumulation were effective methods for determining herbicide resistance levels. The two unknown biotypes were found to be resistant to this herbicide. Some differences were found in the protein profiles of the biotypes, and western blots demonstrated a weak labeling of antibody in the glyphosate-sensitive biotype, whereas strong labeling occurred in the resistant plants. This latter point supports research by others, that increased copy number of the EPSPS gene (and increased EPSPS protein levels) is the resistance mechanism in this species. Results indicate the utility of certain bioassays for the determination of resistance and provide useful comparative information on the levels of inherent constituents among closely related plants.

Evolved polygenic herbicide resistance in Lolium rigidum by low‐dose herbicide selection within standing genetic variation

The interaction between environment and genetic traits under selection is the basis of evolution. In this study, we have investigated the genetic basis of herbicide resistance in a highly characterized initially herbicide-susceptible Lolium rigidum population recurrently selected with low (below recommended label) doses of the herbicide diclofop-methyl. We report the variability in herbicide resistance levels observed in F1 families and the segregation of resistance observed in F2 and back-cross (BC) families. The selected herbicide resistance phenotypic trait(s) appear to be under complex polygenic control. The estimation of the effective minimum number of genes (NE), depending on the herbicide dose used, reveals at least three resistance genes had been enriched. A joint scaling test indicates that an additive-dominance model best explains gene interactions in parental, F1, F2 and BC families. The Mendelian study of six F2 and two BC segregating families confirmed involvement of more than one resistance gene. Cross-pollinated L. rigidum under selection at low herbicide dose can rapidly evolve polygenic broad-spectrum herbicide resistance by quantitative accumulation of additive genes of small effect. This can be minimized by using herbicides at the recommended dose which causes high mortality acting outside the normal range of phenotypic variation for herbicide susceptibility.

Comparability of imazapyr-resistant Arabidopsis created by transgenesis and mutagenesis

The Arabidopsis CSR1 gene codes for the enzyme acetohydroxyacid synthase (AHAS, EC 2.2.1.6), also known as acetolactate synthase, which catalyzes the first step in branched-chain amino acid biosynthesis. It is inhibited by several classes of herbicides, including the imidazolinone herbicides, such as imazapyr; however, a substitution mutation in csr1-2 (Ser-653-Asn) confers selective resistance to the imidazolinones. The transcriptome of csr1-2 seedlings grown in the presence of imazapyr has been shown in a previous study (Manabe in Plant Cell Physiol 48:1340-1358, 2007) to be identical to that of wild-type seedlings indicating that AHAS is the sole target of imazapyr and that the mutation is not associated with pleiotropic effects detectable by transcriptome analysis. In this study, a lethal null mutant, csr1-7, created by a T-DNA insertion into the CSR1 gene was complemented with a randomly-inserted 35S/CSR1-2/NOS transgene in a subsequent genetic transformation event. A comparison of the csr1-2 substitution mutant with the transgenic lines revealed that all were resistant to imazapyr; however, the transgenic lines yielded significantly higher levels of resistance and greater biomass accumulation in the presence of imazapyr. Microarray analysis revealed few differences in their transcriptomes. The most notable was a sevenfold to tenfold elevation in the CSR1-2 transcript level. The data indicate that transgenesis did not create significant unintended pleiotropic effects on gene expression and that the mutant and transgenic lines were highly similar, except for the level of herbicide resistance.

Genetic Resistance to Acetyl‐Coenzyme A Carboxylase‐Inhibiting Herbicides in Grain Sorghum

ABSTRACTWeed management is one of the most important considerations impacting sorghum [Sorghum bicolor (L.) Moench] production in the United States today. The objectives of this study were to evaluate the level of resistance, type of inheritance, and causal mutation of wild sorghum accessions that are resistant to acetyl‐coenzyme A carboxylase (ACCase)‐inhibiting herbicides compared to susceptible grain sorghum seed parents. Acetyl‐coenzyme A carboxylase‐inhibiting herbicides used in this study were fluazifop‐P and quizalofop‐P from the aryloxyphenoxypropionate (APP) family and clethodim and sethoxydim from the cyclohexanedione (CHD) family. Dose–response studies indicated the level of resistance was very high for APP herbicides but low to nonexistent for CHD herbicides. Estimates of the resistance factors to APP herbicides based on herbicide rate required for 50% growth reduction (GR50) were 54 to 64 for homozygous plants and 9 to 19 for heterozygous plants. Resistance to CHD herbicides was very low with resistance factors ranging from one to about five. Genetic segregation studies indicated a single gene is the cause of resistance to APP herbicides. Polymerase chain reaction (PCR) amplicons from the carboxyl transferase (CT) domain of the chloroplast directed ACCase gene were sequenced and a single point mutation that results in a cysteine replacing tryptophan (conversion from tryptophan at amino acid position 2027 to cysteine [Trp‐2027‐Cys]) was discovered. Given the high level of herbicide resistance and simple inheritance, these sources of APP resistance may provide useful herbicide resistance traits for use in sorghum.

Towards large‐scale prediction of Lolium rigidum emergence. II. Correlation between dormancy and herbicide resistance levels suggests an impact of cropping systems

Owen MJ, Michael PJ, Renton M, Steadman KJ & Powles SB (2011). Towards large‐scale prediction of Lolium rigidum emergence. II. Correlation between dormancy and herbicide resistance levels suggests an impact of cropping systems. Weed Research51, 133–141.SummaryThis study investigated a possible link between seed dormancy and herbicide resistance status of Lolium rigidum (annual or rigid ryegrass). Mature seeds were collected from 406 populations across the 14‐million hectare grain belt of southern Western Australia. For each population, initial dormancy and change in dormancy over a 6‐month period were measured, and resistance status of seedlings to four herbicides (diclofop‐methyl, sethoxydim, clethodim and sulfometuron‐methyl) was assessed. Greater seed dormancy correlated with higher levels of herbicide resistance for all four herbicides tested. The herbicides represented two modes of action (acetyl CoA carboxylase‐ and acetolactate synthase inhibitors) and a contrast of generalist (metabolic) and target‐site mutation mechanisms. The coexistence of dormancy and herbicide resistance is suggested to be an adaptation to decades of intense cropping; the plants that are most likely to successfully reproduce are those that exhibit delayed germination (avoiding pre‐seeding weed control strategies) and possess herbicide resistance (surviving subsequent in‐crop herbicide application). We propose that herbicide resistance status may have a role as a predictive tool in modelling dormancy in L. rigidum at a large spatial scale.

Herbicide-Resistant Weed Seeds Contaminate Grain Sown in the Western Australian Grainbelt

Preventing the introduction of weeds into the farming system through sowing of clean seeds is an essential component of weed management. The weed seed contamination of cleaned grain and herbicide resistance levels of the recovered weed seeds were examined in a study conducted across 74 farms in the Western Australian grainbelt. Most farmers grew and conserved their own crop seed. The majority of cleaned samples had some level of seed contamination from 11 foreign weed and volunteer crop species, with an average of 62 seeds 10 kg−1grain, substantially higher than the 28 seeds 10 kg−1grain expected by farmers. The most common weed contaminants across all samples were rigid ryegrass, wild radish, brome, and wild oat. When categorized by crop type, rigid ryegrass was the most frequent contaminant of cereal crops (barley and wheat), however wild radish was the most frequent contaminant of lupin crops. Uncleaned crop seed samples had almost 25 times more contamination than cleaned crop seed. Herbicide resistance was highly prevalent within rigid ryegrass populations recovered from cleaned grain except for glyphosate, which controlled all populations tested. Some resistance was also found in wild radish and wild oat populations; however, brome was susceptible to fluazifop. This study has shown that farmers are unknowingly introducing weed seeds into their farming systems during crop seeding, many of which have herbicide resistance.

The origin of herbicide‐resistant Chenopodium album: analysis of genetic variation and population structure

Aper J, De Riek J, Mechant E, De Cauwer B, Bulcke R & Reheul D (2010). The origin of herbicide‐resistant Chenopodium album: analysis of genetic variation and population structure. Weed Research50, 235–244.Summary Chenopodium album is a predominantly self‐fertilising weed species common in temperate regions of the world that has developed resistance to photosystem II inhibitors. The genetic variation and population structure of eight populations from North West Europe were investigated using 416 amplified fragment length polymorphism (AFLP) markers. The populations were checked for their level of herbicide resistance by pot experiments and the investigated plants were genotyped for the Ser264 to Gly mutation by cleaved amplified polymorphic sequence. Five populations were sampled in sugar beet, one population from a maize monoculture served as a triazine‐resistant reference and the two other populations were susceptible reference populations. Nei’s gene diversity within populations for the whole dataset was low (0.073), whereas, the differentiation among populations was significant (ΦST = 0.26, amova, P < 0.001). There was no correlation between geographical and genetic distance of population pairs. The allelic richness (estimated by ‘band richness’ and percentage of polymorphic loci) was lowest in the two populations with a complete set of herbicide‐resistant plants and in one population with a mixture of herbicide‐resistant and ‐susceptible plants, giving evidence for a historical bottleneck. However, the loss in genetic variation was not that large, indicating that herbicide‐resistant populations may maintain their adaptive capacity. Furthermore, this study shows that different techniques can be used to extract information from AFLP markers for the investigation of the genetic background of weed populations. Knowledge about the origin and spread of herbicide‐resistant weed populations may give opportunities to manage the resistance problem, with its associated ecological and economic consequences.

Tactical management of pasture fallows in Western Australian cropping systems

Agricultural systems in the Western Australian wheatbelt are increasingly moving towards specialist crop production due to elevated cereal prices, depressed markets for livestock products, and ongoing labour scarcity. However, the profitability of crop-only farms is threatened by increasing levels of herbicide resistance and declining soil fertility. This study determines the value of ungrazed pasture fallows grown tactically between crop phases to address these agronomic constraints. A novel metaheuristic optimisation technique, compressed annealing, is used to determine profitable flexible land-use sequences in a complex simulation model. Tactical use of single-year, ungrazed pasture phases is found to be more valuable than the use of break crops in crop-only systems. In contrast to previous analysis, it is identified that rotation of short periods of crop and pasture is more profitable than extended phases of both. Effective weed control without dependence on selective herbicides is the key agronomic characteristic determining the value of intermittent pasture phases. Accordingly, the number of single-year pasture phases employed in a sequence should increase with the severity of herbicide resistance. Compressed annealing is shown to be a practical method of identifying profitable land-use sequences that respond to information that unfolds dynamically.

Economic contribution of French serradella (Ornithopus sativus Brot.) pasture to integrated weed management in Western Australian mixed‐farming systems: an application of compressed annealing*

Sowing phases of French serradella (Ornithopus sativus Brot.) pasture between extended cropping sequences in the Western Australian wheatbelt can sustain grain production through restoring soil fertility and reducing selective herbicide use. The objective of this article is to investigate the profitability of rotations involving this pasture under a variety of weed management scenarios to obtain greater insight into its value for mixed farming systems in this region. A stochastic search procedure, compressed annealing, is used to identify profitable sets of weed management strategies in a simulation model representing a large number of potential combinations of chemical and non‐chemical forms of weed control. In contrast to a continuous‐cropping sequence, the inclusion of a serradella phase in a rotation is profitable at high weed densities and with increasing levels of herbicide resistance. A single year of pasture in the rotation is optimal if resistance to Group A selective herbicides is present at the beginning of the planning horizon, but a three‐year phase is required if resistance to multiple herbicide groups is observed. Sowing a serradella pasture twice over a two‐year phase is also shown to be economically attractive given benefits of successive high weed kills.

Stability of Herbicide Resistance over 8 Years of Coppice in Field-Grown, Genetically Engineered Poplars

Abstract Herbicide resistance may be useful for reducing costs and environmental impacts, and improving yields, during weed control in poplar plantations. However, genetically engineered traits can sometimes show instability, which would compromise their commercial value. To study the stability of herbicide resistance, we analyzed resistance to the contact herbicide glufosinate in 384 transgenic plants originating from 32 gene-insertion events created within two Populus hybrids (P. tremula × P. alba and P. tremula × P. tremuloides). Resistance was measured at the start and end of an 8-year period during which trees were cut and allowed to resprout in the field several times. The transgenic events had widely varying levels of resistance, ranging from complete tolerance to nearly complete sensitivity. When analyzed as three classes (tolerant, intermediate, and sensitive), the levels of resistance remained stable, and there were no cases of resistance breakdown. The level of theresistance-conferring PAT protein, based on enzyme-linked immunosorbent assays, was strongly correlated with resistance class; thus, simple protein assays should enable early screening for acceptable resistance levels. Our data suggest that commercial levels of herbicide resistance and stability can be introduced into elite clones of hybrid poplar with as little as 2–3 years of transformation and field testing.

Diversity of Acetyl-Coenzyme A Carboxylase Mutations in Resistant Lolium Populations: Evaluation Using Clethodim

The acetyl-coenzyme A carboxylase (ACCase)-inhibiting cyclohexanedione herbicide clethodim is used to control grass weeds infesting dicot crops. In Australia clethodim is widely used to control the weed Lolium rigidum. However, clethodim-resistant Lolium populations have appeared over the last 5 years and now are present in many populations across the western Australian wheat (Triticum aestivum) belt. An aspartate-2078-glycine (Gly) mutation in the plastidic ACCase enzyme has been identified as the only known mutation endowing clethodim resistance. Here, with 14 clethodim-resistant Lolium populations we revealed diversity and complexity in the molecular basis of resistance to ACCase-inhibiting herbicides (clethodim in particular). Several known ACCase mutations (isoleucine-1781-leucine [Leu], tryptophan-2027-cysteine [Cys], isoleucine-2041-asparagine, and aspartate-2078-Gly) and in particular, a new mutation of Cys to arginine at position 2088, were identified in plants surviving the Australian clethodim field rate (60 g ha(-1)). Twelve combination patterns of mutant alleles were revealed in relation to clethodim resistance. Through a molecular, biochemical, and biological approach, we established that the mutation 2078-Gly or 2088-arginine endows sufficient level of resistance to clethodim at the field rate, and in addition, combinations of two mutant 1781-Leu alleles, or two different mutant alleles (i.e. 1781-Leu/2027-Cys, 1781-Leu/2041-asparagine), also confer clethodim resistance. Plants homozygous for the mutant 1781, 2078, or 2088 alleles were found to be clethodim resistant and cross resistant to a number of other ACCase inhibitor herbicides including clodinafop, diclofop, fluazifop, haloxyfop, butroxydim, sethoxydim, tralkoxydim, and pinoxaden. We established that the specific mutation, the homo/heterozygous status of a plant for a specific mutation, and combinations of different resistant alleles plus herbicide rates all are important in contributing to the overall level of herbicide resistance in genetically diverse, cross-pollinated Lolium species.

Genetic variation and population structure in black‐grass (Alopecurus myosuroides Huds.), a successful, herbicide‐resistant, annual grass weed of winter cereal fields

Black-grass (Alopecurus myosuroides) is an allogamous grass weed common in cereal fields of northern Europe, which developed resistance to a widely used family of herbicides, the ACCase-inhibiting herbicides. Resistance is caused by mutations at the ACCase gene and other, metabolism-based, mechanisms. We investigated the genetic structure of 36 populations of black-grass collected in one region of France (Côte d'Or), using 116 amplified fragment length polymorphism (AFLP) loci and sequence data at the ACCase gene. The samples were characterized for their level of herbicide resistance and genotyped for seven known ACCase mutations conferring resistance. All samples contained herbicide-resistant plants, and 19 contained ACCase mutations. The genetic diversity at AFLP loci was high (H(T) = 0.246), while differentiation among samples was low (F(ST) = 0.023) and no isolation by distance was detected. Genetic diversity within samples did not vary with the frequency of herbicide resistance. A Bayesian algorithm was used to infer population structure. The two genetic clusters inferred were not associated with any geographical structure or with herbicide resistance. A high haplotype diversity (H(d) = 0.873) and low differentiation (G(ST) = 0.056) were observed at ACCase. However, haplotype diversity within samples decreased with the frequency of ACCase-based resistance. We suggest that the genetic structure of black-grass is affected by its recent expansion as a weed. Our data demonstrate that the strong selection imposed by herbicides did not modify the genome-wide genetic structure of an allogamous weed that probably has large effective population sizes. Our study gives keys to a better understanding of the evolution of successful, noxious weeds in modern agriculture.

Use of Myxococcus xanthus protoporphyrinogen oxidase as a selectable marker for transformation of rice

Protoporphyrinogen oxidase (PPO) is the target enzyme of peroxidizing herbicides. The overexpression of Myxococcus xanthus PPO ( Mx PPO) confers a high level of herbicide resistance in rice. Among the peroxidizing herbicides, butafenacil has an efficiency ∼1000-fold that of oxadiazon, as judged by calli susceptibility tests upon herbicide treatment. Butafenacil (0.1 μM) was used to select transgenic rice plants expressing Mx PPO under the control of the constitutive maize ubiquitin promoter. The ectopic expression of the Mx PPO transgene was investigated in the T 0 generation by Northern blot and Western blot analysis. The T 0 transgenic plants expressing the Mx PPO gene were resistant to butafenacil based on in vitro leaf disk and in vivo foliar spray tests.

P‐Hydroxyphenylpyruvate dioxygenase inhibitor‐resistant plants

The enzyme p-hydroxyphenylpyruvate dioxygenase (HPPD) catalyzes the formation of homogentisic acid, the aromatic precursor of plastoquinone and vitamin E. HPPD is the specific target of several herbicide families: isoxazoles, triketones and pyroxazoles. Its inhibition results in the depletion of the plant plastoquinone and vitamin E pools, leading to bleaching symptoms. These herbicides are very potent for the selective pre- and in some cases post-emergence control of a wide range of broadleaf and grass weeds in maize and rice. Their herbicidal potential raised interest in the development of highly resistant transgenic crops. This goal was first achieved by over-expression of a bacterial HPPD in crop plants, and an increased level of resistance was obtained by using a mutant enzyme. A second strategy based on bypassing HPPD in the production of homogentisate was then developed. Recently, a third strategy of resistance based on the increase of p-hydroxyphenylpyruvate substrate flux has been developed. This was achieved by the introduction of the yeast prephenate dehydrogenase gene (PDH) into transgenic plants already overexpressing HPPD. In addition to a high level of herbicide resistance, a massive accumulation of vitamin E, mainly tocotrienols, was observed in leaves of the transgenic HPPD-PDH plants.

The glutamate dehydrogenase gene gdhA increased the resistance of tobacco to glufosinate

SummaryThe gene gdhA from Escherichia coli, that encodes a NADPH‐dependent glutamate dehydrogenase (GDH), directs a novel pathway in transgenic plants that allows an increase in ammonium assimilation. Glufosinate leads to plant death by the irreversible inhibition of glutamate synthetase (GS) leading to a disruption of subsequent GS‐related processes resulting in elevated ammonium and disruption of photorespiration. Therefore, it was speculated that the gdhA‐transformed plants may exhibit a novel mechanism of resistance to glufosinate by altered activity of the GDH‐directed pathway(s) and subsequently related processes. Studies were conducted in the greenhouse to evaluate the resistance of tobacco plants containing the gdhA gene to glufosinate. Five tobacco genotype lines were investigated including a non‐transformed control line, a positive control line and three transformed lines with levels of increasing GDH activity directed by the gdhA gene. Plants transformed with the gdhA gene expressed up to six times increased level of resistance (GR50) to glufosinate compared with the non‐transformed control, which is 100 times less resistant than plants transformed with the bar gene. The GDH activity among lines was highly correlated (r2 = 0.9903) with the level of herbicide resistance. Thus, the use of the E. coli gdhA gene in plant transformations can provide an additional mechanism for resistance to glufosinate.

The economic value of haying and green manuring in the integrated management of annual ryegrass and wild radish in a Western Australian farming system

Most cropping farms in Western Australia must deal with the management of herbicide-resistant populations of weeds such as annual ryegrass (Lolium rigidum) and wild radish (Raphanus raphanistrum). Farmers are approaching the problem of herbicide resistance by adopting integrated weed management systems, which allow weed control with a range of different techniques. These systems include non-herbicide methods ranging from delayed seeding and high crop seeding rates to the use of non-cropping phases in the rotation. In this paper, the Multi-species RIM (resistance and integrated management) model was used to investigate the value of including non-cropping phases in the crop rotation. Non-crop options investigated here were haying and green manuring. Despite them providing excellent weed control, it was found that inclusion of these non-cropping phases did not increase returns, except in cases of extreme weed numbers and high levels of herbicide resistance.

Evolution of resistance to diclofop-methyl in ryegrass (Lolium multiflorum): investigation of the role of introgression with related species

Populations of Lolium multiflorum (ryegrass) exposed to increasing levels of herbicide selection have variability in the rate of evolution of herbicide resistance. We tested the hypothesis that herbicide dose and gene flow with related species are important factors regulating the rate of evolution of herbicide resistance in L. multiflorum. Seeds of the latter were planted in pure stands and in mixture with Festuca rubra and subjected to four herbicide rates. The level of herbicide resistance attained by the offspring after two years of selection was evaluated. Evolution of resistance observed in the field was compared to that calculated by Gressel and Segel's rotational model. ©1997 SCI

Evolution of resistance to diclofop‐methyl in ryegrass (Lolium multiflorum): investigation of the role of introgression with related species

Populations of Lolium multiflorum (ryegrass) exposed to increasing levels of herbicide selection have variability in the rate of evolution of herbicide resistance. We tested the hypothesis that herbicide dose and gene flow with related species are important factors regulating the rate of evolution of herbicide resistance in L. multiflorum. Seeds of the latter were planted in pure stands and in mixture with Festuca rubra and subjected to four herbicide rates. The level of herbicide resistance attained by the offspring after two years of selection was evaluated. Evolution of resistance observed in the field was compared to that calculated by Gressel and Segel's rotational model. ©1997 SCI