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Straw from no-till cropping systems, in addition to increasing the soil organic matter content, may also impede the movement of applied herbicides into the soil and, thus, alter the behavior and fate of these compounds in the environment. Rain or irrigation before or after an herbicide treatment can either help or hinder its movement through the straw, influencing weed control. Our objective was to develop a system for herbicide application and rain simulation, enabling the evaluation of the movement of various herbicides either in dry or wet straw under different rainfall volumes (25, 50, 75, and 100 mm). The amount of the applied herbicides that moved through the straw were collected and measured using a liquid chromatograph with a tandem mass spectrometry system (LC-MS/MS). Measurements obtained with the developed system showed a high herbicide treatment uniformity across all replications. The movement of the active ingredients through the straw showed variability that was a function of the applied herbicide, ranging from 17% to 99%. In wet straw, the collected herbicide remained constant from 50 to 100 mm of simulated rainfall. For the wet straw, the decreasing percentages of the herbicide movement through straw to the soil were sulfentrazone (99%), atrazine and diuron (91% each), hexazinone (84%), fomesafen (80.4%), indaziflam (79%), glyphosate (63%), haloxyfop-p-methyl (45%), and S-metolachlor (27%). On the dry straw, the decreasing percentages of the herbicide movement were fomesafen (88%), sulfentrazone (74%), atrazine (69.4%), hexazinone (69%), diuron (68.4%), glyphosate (48%), indaziflam (34.4%), S-metolachlor (22%), and haloxyfop-p-methyl (18%). Overall, herbicide movement was higher in wet straw (with a previous 25 mm simulated rainfall layer) than in dry straw. Some herbicides, like haloxyfop-p-methyl and indaziflam, exhibited over 50% higher movement in wet straw than dry straw after 100 mm of simulated rain. The developed system can be adapted for various uses, serving as a valuable tool to evaluate the behavior of hazardous substances in different agricultural and environmental scenarios.

Brazil, the world’s largest soybean producer, owes its success to the cultivation of glyphosate-resistant (GR) cultivars. However, the soybean yields lag behind those obtained in areas managed for high productivity. Glyphosate-induced hormesis holds promise for increasing crop yields, but the potential evolution of resistance in certain weed species poses a challenge to foliar applications under field conditions. This study assessed the effects of a hormesis-inducing glyphosate dose [90 g acid equivalent (ae) ha−1] on the agronomic characteristics and yield of four GR soybean cultivars. The evaluation was conducted in field settings across various Brazilian locations, considering foliar, seed, and seed + foliar treatments. The results showed variations in dry mass, root nodules, nutrient composition, plant height, pods, and yield, primarily influenced by environmental conditions, soil quality, and, ultimately, the interaction between GR cultivars and treatments. Total dry mass consistently increased with glyphosate, with seed and seed + foliar treatments showing the most substantial increases (7–21%). All three treatments increased nodulation by up to 36% across locations and cultivars, with seed + foliar treatment causing notable increases in nodule dry mass (up to 56%), followed by seed treatment (41%). Nutrient composition, especially for N, P, Br, and Fe, displayed location-dependent variations. Plant height varied among locations and cultivars, with minimal differences between treatments. Glyphosate treatments increased pod numbers (10 to 35%) and yields (11 to 42%) of soybean in seed and seed + foliar treatments. The findings highlight the potential of glyphosate hormesis as a viable tool for improving yields of GR soybean cultivars at the field level. However, the extent of benefits depends on the agronomic conditions of location, choice of cultivars, and herbicide application method.

Glyphosate is the most widely used herbicide for weed control in citrus orchards in Brazil; therefore, it is likely that several species have gained resistance to this herbicide and that more than one resistant species can be found in the same orchard. The objective was to identify weeds resistant to glyphosate in citrus orchards from different regions of the São Paulo State (SP) and determine how many resistant species are present within the same orchard. Seeds of Amaranthus deflexus, A. hybridus, Bidens pilosa, Chloris elata, Conyza bonariensis, Digitaria insularis, Solanum Americanum, and Tridax procumbens, which, as reported by growers, are suspected to be resistant to glyphosate, were collected from plants that survived the last application of this herbicide (>720 g of acid equivalent [ae] ha–1) in sweet orange and Tahiti acid lime orchards. Based on dose–response and shikimic acid accumulation assays, all populations of A. deflexus, A. hybridus, B. pilosa, and T. procumbens were sensitive to glyphosate. However, populations of B. pilosa from the Olimpia region (R-NS, R-PT and R-OdA) showed signs of resistance based on plant mortality rates by 50% within a population (LD50 = 355–460 g ae ha−1). All populations of C. bonariensis, C. elata, and D. insularis were resistant to glyphosate, presenting resistance ratios from 1.9 to 27.6 and low shikimate accumulation rates. Solanum americanum also showed resistance, with resistance ratios ranging from 4.3 to 25.4. Most of the citrus orchards sampled presented the occurrence of more than one species resistant to glyphosate: Nossa Senhora—one species; Olhos D’agua and Passatempo—two species; Araras—four species; and Cordeiropolis and Mogi-Mirim—up to five species. The results reported in this paper provide evidence of multiple species in citrus orchards from São Paulo that have exhibited resistance to glyphosate. This underscores the difficulties in managing glyphosate-resistant weeds which are prevalent throughout the country, such as C. bonariensis and D. insularis. The presence of these resistant species further complicates the control of susceptible species that may also develop resistance. In addition, the glyphosate resistance of S. americanum was identified for the first time.

The presence of weeds in the sunflower crop is one of the main factors linked to the low increase in productivity of this crop, and to determine the most appropriate management of weeds, it is essential to carry out a diagnosis through the phytosociological survey. The objective of this study was to assess the influence of chemical control on the phytosociological community of weeds in three areas cultivated with sunflower in the Parecis region (Brazil). The areas were treated with 2,4-D + glyphosate for desiccation; S-metolachlor was used for pre-emergence control in the three areas; meanwhile, sulfentrazone and flumioxazin were applied only in one area; and, finally, clethodim was applied for post-emergence weed management. Sampling was carried out at two different times, in the initial and pre-harvest stages (at 35 and 100 days after the emergence of the crop, respectively), using a quadrate, in which weeds were identified and quantified to determine the frequency, relative frequency, density, relative density, abundance, relative abundance, importance index, and similarity index between areas and times. Seventeen weed species were found in the sunflower crop (70.6% dicot and 29.4% monocotyledonous) in the two seasons, grouped into nine botanical families, with Poaceae being the most diverse family. The dicots Tridax procumbens and Acanthospermum hispidium were present in low frequency only in the initial stages of development of the sunflower crop. The weeds with the highest importance index values in the initial and pre-harvest stages were Euphorbia hirta (104 and 91%) and Bidens pilosa (45 and 66%, respectively), both belonging to the dicots group. These two species were present in the two evaluated periods and in the three experimental areas, demonstrating that there was a similarity index between them with values above 93%. These results of the phytosociological study may contribute to determining more efficient management strategies for weed chemical control in the sunflower crop.

The characterization of the mechanisms conferring resistance to herbicides in weeds is essential for developing effective management programs. This study was focused on characterizing the resistance level and the main mechanisms that confer resistance to glyphosate in a resistant (R) Steinchisma laxum population collected in a Colombian rice field in 2020. The R population exhibited 11.2 times higher resistance compared to a susceptible (S) population. Non-target site resistance (NTSR) mechanisms that reduced absorption and impaired translocation and glyphosate metabolism were not involved in the resistance to glyphosate in the R population. Evaluating the target site resistance mechanisms by means of enzymatic activity assays and EPSPS (5-enolpyruvylshikimate-3-phosphate synthase) gene sequencing, the mutation Pro106Ser was found in R plants of S. laxum. These findings are crucial for managing the spread of S. laxum resistance in Colombia. To effectively control S. laxum in the future, it is imperative that farmers use herbicides with different mechanisms of action in addition to glyphosate and adopt Integrate Management Programs to control weeds in rice fields of the central valleys of Colombia.

Herbicide-resistant weeds have been identified and recorded on every continent where croplands are available. Despite the diversity of weed communities, it is of interest how selection has led to the same consequences in distant regions. Brassica rapa is a widespread naturalized weed that is found throughout temperate North and South America, and it is a frequent weed among winter cereal crops in Argentina and in Mexico. Broadleaf weed control is based on glyphosate that is used prior to sowing and sulfonylureas or mimic auxin herbicides that are used once the weeds have already emerged. This study was aimed at determining whether a convergent phenotypic adaptation to multiple herbicides had occurred in B. rapa populations from Mexico and Argentina by comparing the herbicide sensitivity to inhibitors of the acetolactate synthase (ALS), 5-enolpyruvylshikimate-3-phosphate (EPSPS), and auxin mimics. Five B. rapa populations were analyzed from seeds collected in wheat fields in Argentina (Ar1 and Ar2) and barley fields in Mexico (Mx1, Mx2 and MxS). Mx1, Mx2, and Ar1 populations presented multiple resistance to ALS- and EPSPS-inhibitors and to auxin mimics (2,4-D, MCPA, and fluroxypyr), while the Ar2 population showed resistance only to ALS-inhibitors and glyphosate. Resistance factors ranged from 947 to 4069 for tribenuron-methyl, from 1.5 to 9.4 for 2,4-D, and from 2.7 to 42 for glyphosate. These were consistent with ALS activity, ethylene production, and shikimate accumulation analyses in response to tribenuron-methyl, 2,4-D, and glyphosate, respectively. These results fully support the evolution of the multiple- and cross-herbicide resistance to glyphosate, ALS-inhibitors, and auxinic herbicides in B. rapa populations from Mexico and Argentina.

Weeds that usually grow in non-agricultural areas have become increasingly common invading perennial crops. Species of the genus Filago, in addition to invading Spanish olive groves, have developed certain levels of natural tolerance to the acetolactate synthase (ALS) inhibiting herbicide flazasulfuron. The objective of this study was to determine the level and the mechanism involved in the tolerance to flazasulfuron in Filago pyramidata L., which occurs in olive groves of southern Spain, as well as to identify possible cross- or multiple-tolerances by evaluating alternative herbicides for its control. A population resistant (R) to flazasulfuron and a susceptible (S) one of Conyza canadensis were used as references. The accessions of F. pyramidata presented LD50 values (from 72 to 81 g active ingredient (ai) ha−1) higher than the field dose of flazasulfuron (50 g ai ha−1), being 11–12.5 times more tolerant than the S population of C. canadensis, but less than half the R population (170 g ai ha−1). Enzymatically, F. pyramidata was as sensitive to flazasulfuron (I50 = 17.3 μM) as the S population of C. canadensis. Filago pyramidata plants treated with flazasulfuron, combined with 4-chloro-7-nitro-2,1,3-benzoxadiazole, had a growth reduction of up to 85%, revealing the participation of glutathione-S-transferases in herbicide metabolism. Filago pyramidata presented cross-tolerance to the different chemical groups of ALS inhibitors, except triazolinones (florasulam). Synthetic auxins (2,4-D and fluroxypyr) presented good control, but some individuals survived (low multiple resistance). Cellulose synthesis, 5-enolpyruvylshikimate-3-phosphate, 4-hydroxyphenylpyruvate dioxygenase, protoporphyrinogen oxidase, photosystem I, and photosystem II inhibitor herbicides, applied in PRE or POST-emergence, presented excellent levels of control of F. pyramidata. These results confirmed the natural tolerance of F. pyramidata to flazasulfuron and cross-tolerance to most ALS-inhibiting herbicides. The mechanism involved was enhanced metabolism mediated by glutathione-S-transferases, which also conferred low multiple tolerance to synthetic auxins. Even so, herbicides with other mechanisms of action still offer excellent levels of control of F. pyramidata.

Abstract The effect of the phosphorus (P) and glyphosate interactions on the growth and nutrition of Arabica coffee cultivars (Coffea arabica), as well as on the control of Ipomoea grandifolia and Urochloa decumbens, was evaluated. Catuaí-Amarelo/IAC-62 and Catuaí-Vermelho/IAC-144 cultivars did not show glyphosate poisoning, regardless of the soil P content. However, glyphosate reduced the growth of Catuaí-Vermelho/IAC-144. In addition, the soil P content influenced the height, leaf area and dry matter of Catuaí-Amarelo/IAC-62, and the absorption of P and Ca in both cultivars. On the other hand, glyphosate efficiently controlled U. decumbens but not I. grandifolia. Glyphosate effectiveness on I. grandifolia decreased as the soil P content increased. In addition, the soil P content and the glyphosate influenced the P content in I. grandifolia and U. decumbens plants. The soil P content influenced the growth and absorption of other nutrients by coffee plants as well as glyphosate effectiveness on weed control.

In recent decades, cereal agriculture across main producing areas in Spain has progressively adopted direct sowing, associated with an increased use of herbicides in pre-sowing. Weedy species from genus Avena have been observed after the application of glyphosate in wheat. Here, Avena fatua (two accessions), Avena byzantina and Avena sterilis subsp. sterilis, one accession each, were taxonomically characterized by a biometric study. Dose-response trials confirmed that one A. fatua accession evolved to resistance, because it was four times more resistant (R) than the others, ascribed as susceptible (S). In addition, based on LD50, A. byzantina and A. sterilis had low susceptibility to glyphosate, with 604 and 458 g ae ha−1. Shikimic acid accumulation was able to discriminate between high susceptible (A. fatua (S)), low susceptible (A. byzantina and A. sterilis), and low resistant (A. fatua (R)) populations. On the other hand, the study revealed that A. fatua (R), A. byzantina and A. sterilis had low foliar uptake and decreased movement of glyphosate. In addition, the metabolism study showed less metabolite accumulation in A. byzantina and A. fatua (S). However, at 96 h after glyphosate application, A. fatua (R) and A. sterilis were able to convert more than 30% of glyphosate to aminomethylphosphonic acid (AMPA) and sarcosine. Enzyme basal activity and I50 values predicted high affinity between the herbicide and the target enzyme for all accessions, thus indicating that a target-site (TS) mechanism is probably not involved in the differences in glyphosate susceptibility. In closing, non-target site (NTS) mechanisms could participate both in A. fatua R to glyphosate, and low susceptibility in A. byzantina and A. sterilis from Spain.

Introducción. El uso continuo de herbicidas, frecuentemente con múltiples aplicaciones durante el mismo ciclo agrícola, provoca el surgimiento de biotipos/poblaciones de malezas resistentes. La resistencia es la capacidad hereditaria de algunos individuos para crecer y reproducirse sexualmente después del tratamiento de un herbicida a dosis de campo letal para los individuos susceptibles de la población (WSSA, 1998). Objetivos. Entender qué es la resistencia a herbicidas, cuáles son los tipos mecanismos que la confieren y la importancia de su caracterización sirven para implementar estrategias de manejo asertivas tanto de prevención como de manejo de la resistencia. Métodos. Los mecanismos que confieren resistencia pueden ser relacionados con el sitio de acción (mutaciones o sobreexpresión del gen que codifica la enzima-objetivo del herbicida) o no relacionados con el sitio de acción (reducida absorción y traslocación, metabolismo, exudación radicular, exclusión celular del herbicida, muerte celular rápida) (Gaines et al., 2020), los cuales son caracterizados por técnicas de biología molecular, pruebas in vivo usando plantas completas, ensayos enzimáticos in vitro, radiológicas ( 14 C), plataformas analíticas (HPLC, LC-MS/MS, GC-MS) entre otras (Dayan et al., 2015; Déyle et al., 2015; Nandula y Vencill, 2015). Resultados y discusión. La identificación de biotipos de malezas resistentes en campo no debe estar relacionado a un mal uso de herbicidas y sí a su falta de eficacia en especies que previamente eran controladas. Conocer el tipo de mecanismos que confieren resistencia a herbicidas en una población de malezas es fundamental para desarrollar estrategias de manejo adecuadas (Beckie and Harker, 2017). Los mecanismos del sitio de acción son gobernados por un solo gen (resistencia monogénica); por lo tanto, una rotación de herbicidas con modo de acción diferente podría ser suficiente para controlar la resistencia. Sin embargo, los mecanismos no relacionados con el sitio de acción son gobernados por múltiples genes (resistencia poligénica) y cada gen contribuye con cierto nivel de resistencia. Además, estos genes también pueden conferir resistencia múltiple a herbicidas con diferentes mecanismos de acción que no fueron previamente utilizados en el área; por lo tanto, además de rotación de herbicidas, se requieren de estrategias de manejo de la resistencia no químicas. Conclusión. Distinguir poblaciones resistentes (identificar el problema) oportunamente a través de prospecciones de campo, pero principalmente, caracterizar los mecanismos de resistencia (diagnosticar la causa) es esencial para elaborar y ejecutar programas de manejo de resistencia eficientes para evitar el aumento de los niveles de resistencia, así como de la selección de resistencia cruzada o múltiple.

Herbicides have become one of the most widespread weed-control tools in the world since their advent in the mid-20th century [...]

Short-spike canarygrass (Phalaris brachystachys (Link.)) from Iranian wheat fields has developed resistance to acetyl-coenzyme A carboxylase (ACCase) inhibitors due to a target-site mutation (Ile-1781-Thr). Target-site resistance mutations may confer pleiotropic effects in weeds. In this paper, the possible effect of the Ile-1781-Thr mutation on the ecological fitness during life cycles in P. brachystachys plants was investigated. ACCase genes of P. brachystachys populations resistant (R) and susceptible (S) to ACCase inhibitors were sequenced and the vegetative growth and reproductive characteristics of the plants were assessed. In the final growth stage (217 days after planting—DAP), R sub-population plants were 30 cm taller than the S plants. Additionally, the R sub-population produced up to 12 leaves and 2.8 tillers more per plant, and accumulated double the dry weight (2850 g m−2) compared to the S sub-population. The leaf area index (LAI) of the R sub-population was 1.1 times higher than that of the S sub-population. In addition, the net assimilation rate (NAR) and plant growth rate (PGR) between 114 and 182 DAP of the R sub-population were 0.11 and 13 g m−2 d−1 higher than the S sub-population, but the relative growth rate (RGR) was similar between R and S sub-populations. The number of spikes (6 vs. 3.8), the spike length (8.4 vs. 5.5), and number of seeds per plant (1276 vs. 751 seed plant−1) of the R sub-population were higher than the S ones, but the weight (3.25 g) and size (11.6 mm2) of 1000 seeds were similar between populations. The R sub-population of P. brachystachys exhibited higher plant growth and reproductive parameters than the S one, which may increase the frequency of resistance in the population in the absence of adequate weed-control methods.

Atta and Acromyrmex are the main genera of leaf-cutting ants present in North and South America, causing extensive damage to agroforestry. Control of the ants requires high handling costs with few effective methods available to decrease the losses. The symbiosis between the leaf-cutting ants and the fungus Leucoagaricus gongylophorus is essential for ant nest survival. Therefore, L. gongylophorus may be a key target in controlling leaf-cutting ants, since its reduction may cause an imbalance in the symbiosis necessary to maintain the nest. Among the options for natural fungal control, plant species are considered important sources of compounds belonging to several classes of natural products that show potential as antifungal agents. This review also presents studies that establish that the antagonist fungi from the Escovopsis and Trichoderma genera effectively reduce the development of L. gongylophorus. The development of nanostructured delivery systems, which have shown advantages over conventional formulations, is suggested for ant control; no commercial nanotechnology-based product has yet been developed, and this appears to be a new approach for future studies.

The levels of resistance to glyphosate of 13 barnyard grass (Echinochloa crus-galli) populations harvested across different agriculture areas in the Southern Iberian Peninsula were determined in greenhouse and laboratory experiments. Shikimate accumulation fast screening separated the populations regarding resistance to glyphosate: susceptible (S) E2, E3, E4, and E6 and resistant (R) E1, E5, E7, E8, E9, E10, E11, E12, and E13. However, resistance factor (GR50 E1–E13/GR50 E6) values separated these populations into three groups: (S) E2, E3, E4, and E6, (R) E1, E5, E7, E8, and E9, and very resistant (VR) E10, E11, E12, and E13. 14C-glyphosate assays performed on two S populations (E2 and E6) showed greater absorption and translocation than those found for R (E7 and E9) and VR (E10 and E12) populations. No previous population metabolized glyphosate to amino methyl phosphonic acid (AMPA) and glyoxylate, except for the E10 population that metabolized 51% to non-toxic products. The VR populations showed two times more 5-enolpyruvylshikimate-3-phosphate synthase (EPSPS) activity without herbicide than the rest, while the inhibition of the EPSPS activity by 50% (I50) required much higher glyphosate in R and VR populations than in S populations. These results indicated that different target-site and non-target-site resistance mechanisms were implicated in the resistance to glyphosate in E. crus-galli. Our results conclude that resistance is independent of climate, type of crop, and geographic region and that the level of glyphosate resistance was mainly due to the selection pressure made by the herbicide on the different populations of E. crus-galli studied.

Different Lolium species, common weeds in cereal fields and fruit orchards in Chile, were reported showing isolated resistance to the acetyl CoA carboxylase (ACCase), acetolactate synthase (ALS) and 5-enolpyruvylshikimate-3-phosphate synthase (EPSPS) inhibiting herbicides in the late 1990s. The first case of multiple resistance to these herbicides was Lolium multiflorum found in spring barley in 2007. We hypothesized that other Lolium species may have evolved multiple resistance. In this study, we characterized the multiple resistance to glyphosate, diclofop-methyl and iodosulfuron-methyl-sodium in Lolium rigidum, Lolium perenne and Lolium multiflorum resistant (R) populations from Chile collected in cereal fields. Lolium spp. populations were confirmed by AFLP analysis to be L. rigidum, L. perenne and L. multiflorum. Dose-response assays confirmed multiple resistance to glyphosate, diclofop-methyl and iodosulfuron methyl-sodium in the three species. Enzyme activity assays (ACCase, ALS and EPSPS) suggested that the multiple resistance of the three Lolium spp. was caused by target site mechanisms, except the resistance to iodosulfuron in the R L. perenne population. The target site genes sequencing revealed that the R L. multiflorum population presented the Pro-106-Ser/Ala (EPSPS), Ile-2041-Asn++Asp-2078-Gly (ACCase), and Trp-574-Leu (ALS) mutations; and the R L. rigidum population had the Pro-106-Ser (EPSPS), Ile-1781-Leu+Asp-2078-Gly (ACCase) and Pro-197-Ser/Gln+Trp-574-Leu (ALS) mutations. Alternatively, the R L. perenne population showed only the Asp-2078-Gly (ACCase) mutation, while glyphosate resistance could be due to EPSPS gene amplification (no mutations but high basal enzyme activity), whereas iodosulfuron resistance presumably could involve non-target site resistance (NTSR) mechanisms. These results support that the accumulation of target site mutations confers multiple resistance to the ACCase, ALS and EPSPS inhibitors in L. multiflorum and L. rigidum from Chile, while in L. perenne, both target and NTSR could be present. Multiple resistance to three herbicide groups in three different species of the genus Lolium in South America represents a significant management challenge.

The excessive use of glyphosate by Brazilian citrus growers leads to a vicious and unsustainable circle: Increasing the glyphosate use and the selection pressure of resistant/tolerant weeds, as well as the phytointoxication of the crop. In addition, there is speculation on the consequences of using glyphosate and the studies are not conclusive. Therefore, this study aimed to evaluate the glyphosate management in citrus orchards by assessing its effects in a 5-year field experiment using different doses and application frequencies. Here, we determine the weed control levels, the orchard growth and fruit yield, as well as the economic viability of the treatments. Higher weed control was observed more often with the increasing frequency of glyphosate application, and occasionally with increasing doses. However, some species predominated even at high glyphosate usage, such as BIDPI (Bidens pilosa (L.)), RAPRA (Raphanus raphanistrum (L.)), and ERICA (Conyza canadensis (L.) Cronquist). Phytotoxicity symptoms were demonstrated up to the fourth year of the orchard, and onwards the plants no longer expressed them. This was a highlight, since there was a decrease over time in growth (up to 5.3 m3) and fruit yield (up to 36.3 t ha−1), with losses that reached −56% of the total income. This is the first report to demonstrate that the increase in glyphosate usage can occasionally increase weed control, but it can also decrease orchard development and its financial viability.

Of the six-glyphosate resistant weed species reported in Mexico, five were found in citrus groves. Here, the glyphosate susceptibility level and resistance mechanisms were evaluated in saltmarsh aster (Aster squamatus), a weed that also occurs in Mexican citrus groves. The R population accumulated 4.5-fold less shikimic acid than S population. S plants hardly survived at 125 g ae ha−1 while most of the R plants that were treated with 1000 g ae ha−1, which suffered a strong growth arrest, showed a vigorous regrowth from the third week after treatment. Further, 5-enolpyruvylshikimate-3-phosphate basal and enzymatic activities did not diverge between populations, suggesting the absence of target-site resistance mechanisms. At 96 h after treatment, R plants absorbed ~18% less glyphosate and maintained 63% of the 14C-glyphsoate absorbed in the treated leaf in comparison to S plants. R plants metabolized twice as much (72%) glyphosate to amino methyl phosphonic acid and glyoxylate as the S plants. Three non-target mechanisms, reduced absorption and translocation and increased metabolism, confer glyphosate resistance saltmarsh aster. This is the first case of glyphosate resistance recorded for A. squamatus in the world.

Species of Phalaris have historically been controlled by acetyl-coenzyme A carboxylase (ACCase)-inhibiting herbicides; however, overreliance on herbicides with this mechanism of action has resulted in the selection of resistant biotypes. The resistance to ACCase-inhibiting herbicides was characterized in Phalaris brachystachys, Phalaris minor, and Phalaris paradoxa samples collected from winter wheat fields in northern Iran. Three resistant (R) biotypes, one of each Phalaris species, presented high cross-resistance levels to diclofop-methyl, cycloxydim, and pinoxaden, which belong to the chemical families of aryloxyphenoxypropionates (FOPs), cyclohexanediones (DIMs), and phenylpyrazolines (DENs), respectively. The metabolism of 14C-diclofop-methyl contributed to the resistance of the P. brachystachys R biotype, while no evidence of herbicide metabolism was found in P. minor or P. paradoxa. ACCase in vitro assays showed that the target sites were very sensitive to FOP, DIM, and DEN herbicides in the S biotypes of the three species, while the R Phalaris spp. biotypes presented different levels of resistance to these herbicides. ACCase gene sequencing confirmed that cross-resistance in Phalaris species was conferred by specific point mutations. Resistance in the P. brachystachys R biotype was due to target site and non-target-site resistance mechanisms, while in P. minor and P. paradoxa, only an altered target site was found.

Abstract Glyphosate has been used for more than 15 years for weed management in citrus groves in the Gulf of Mexico, at up to 3–4 applications per year. Goosegrass (Eleusine indica (L.) Gaertn.) control has sometimes failed. In this research, the mechanisms governing three goosegrass biotypes (Ein-Or from an orange grove, and Ein-Pl1 and Ein-Pl2 from Persian lime groves) with suspected resistance to glyphosate were characterized and compared to a susceptible biotype (Ein-S). Dose-response and shikimate accumulation assays confirmed resistance of the resistant (R) biotypes. There were no differences in glyphosate absorption, but the R biotypes retained up to 62–78% of the herbicide in the treated leaf at 96 h after treatment (HAT), in comparison to the Ein-S biotype (36%). The 5-enolpyruvylshikimate-3-phosphate synthase (EPSPS) activity in the Ein-Or and Ein-S biotypes was over 100-fold lower than the Ein-Pl1 and Ein-Pl2 ones. The latter showed a high EPSPS-basal activity, a mutation at Pro-106-Ser position in the EPSPS gene, and EPSPS overexpression. The EPSPS basal and EPSPS overexpression were positively correlated. The R goosegrass biotypes displayed poor glyphosate translocation. Furthermore, this grassweed showed, for the first time, two mechanisms at the target-site level (Pro-106-Ser mutation + EPSPS overexpression) acting together simultaneously against glyphosate.

Eleusine indica is a highly competitive and difficult-to-control plant in annual and perennial crops. In Colombia, broad-spectrum herbicides, such as paraquat and glyphosate, have begun to present poor levels of control for this weed. The multiple resistance to glyphosate and paraquat, the increase in herbicide performance with adjuvants (Retenol® and Trend® 90), and alternative herbicides were evaluated in a resistant (R) population of E. indica collected in rice fields, which is rotated with herbicide-resistant (HR) crops. Based on plant mortality, the R population was 9.8 and 7.2 times more resistant than susceptible (S) plants to glyphosate and paraquat, respectively. R plants accumulated 4.2 less shikimic acid and had at least 70% less electrolyte leakage than S plants when they were exposed to glyphosate and paraquat, respectively. Both adjuvants increased the foliar retention of herbicides. In addition, adjuvants also increased the performance of glyphosate effectiveness between 22% and 58% and that of paraquat from 61% to 100%. Alternative herbicides (atrazine, clethodim, imazamox, diuron, flazasulfuron, glufosinate, oxyfluorfen, quizalofop, and tembotrione) provided high levels of control in both populations of E. indica. This is the first case of multiple resistant E. indica confirmed in Colombia. Adjuvants improved the leaf retention and efficacy of glyphosate and paraquat. In summary, the alternative herbicides evaluated in this study should be adopted by Colombian farmers and provide additional herbicide modes-of-action to combat future resistance.

Conyza bonariensis (L.) Cronquist is one of the main glyphosate-resistant weeds in no-till fields in Brazil. Here, the seed quality of glyphosate-resistant (R) and -susceptible (S) C. bonariensis biotypes, collected from different sites, was evaluated under stressful conditions by different seed tests. Glyphosate resistance was confirmed by dose-response and shikimate accumulation assays. The resistance factors were 6.9 (R1/S1), 4.5 (R2/S2), and 5.8 (R3/S3). Biotypes S1, S2, and S3 accumulated 2.7, 2.4, and 2.8 times more shikimic acid than biotypes R1, R2, and R3, respectively. Stress-free seed viability and germination potential ranged from 39% to 57% and from 37% to 57%, respectively, with no difference between R and S biotypes within each collection site. Seed incubation at 8 °C over 7 days (cold test) promoted greater germination in S biotypes (54% to 79%) compared to R ones (28% to 39%). In the accelerated aging tests (incubation at 42 °C over 48 hours), the germination decreased in both S (11% to 27%) and R (6% to 16%) biotypes. In the high-temperature stress tests, there were no differences in germination within biotypes at 35 and 45 °C; however, at 60 °C, the germination of the S1, R1, S2, R2, S3, and R3 biotypes was reduced by approximately 51%, 54%, 63%, 59%, 40%, and 30%, respectively. Under non-stressful conditions, germination potential and seed viability were similar in R and S biotypes; however, under cold or heat stress conditions, R biotypes reduced their germination rates, revealing that glyphosate resistance causes a fitness penalty in C. bonariensis at the seed level. However, because seed viability was not determined after experiments, it cannot be stated that such reduction in germination was due to the death or only a dormant phase of the seeds.

Wild poinsettia (Euphorbia heterophylla L.) is a difficult-to-control weed in soybean production in Brazil that has developed resistance to herbicides, including acetolactate synthase inhibitors. We investigated the potential fitness cost associated to the Ser-653-Asn mutation that confers imazamox resistance in this weed. Plant height, leaf and stem dry weight, leaf area and seed production per plant as well as the growth indices of specific leaf area, leaf area ratio, relative growth rate and net assimilation in F2 homozygous resistant (R) and susceptible (S) wild poinsettia progenies were pairwise compared. S plants were superior in most of the traits studied. Plant heights for S and R biotypes, recorded at 95 days after planting (DAP), were 137 and 120 cm, respectively. Leaf areas were 742 and 1048 cm2 in the R and S biotypes, respectively. The dry weights of leaves and stems in the S plants were 30 and 35%, respectively, higher than in the R plants. In both biotypes, the leaves had a greater share in dry weight at early development stages, but from 50 DAP, the stem became the main contributor to the dry weight of the shoots. The R biotype produced 110 ± 4 seed plant−1, i.e., 12 ± 3% less seeds per plant than that of the S one (125 ± 7 seed plant−1). The growth indices leaf area ratio and specific leaf area were generally higher in the S biotype or similar between both biotypes; while the relative growth rate and net assimilation rate were punctually superior in the R biotype. These results demonstrate that the Ser-653-Asn mutation imposed a fitness cost in imazamox R wild poinsettia.

Forest restoration in Brazil has gained relevance in the country’s environmental agenda, due to the need for forest recovery of large liabilities of existing forests and participation in several international vegetation restoration agreements. However, forest restoration management faces challenges, it being necessary to create a database of species-level performances to increase the success of these projects. The objective was to evaluate the survival and growth of five Atlantic Forest native species (Anadenanthera macrocarpa; Ceiba speciosa; Cytharexyllum myrianthum; Hymenaea courbaril; and Peltophorum dubium) in plastic bags (1177 cm3) and tubes (180 cm3). Ninety seedlings (18 of each species) were planted per container. Plant performance in the field consisted of evaluating the increase in the diameter and height of seedlings of the native forest species. Diameter at soil level (DSL) and plant height (H) were measured at 42 months after transplanting, and the monthly periodic increments (MPI) of the DSL and H were calculated. Plant survival (SV) of seedlings was affected by the type of container, registering the highest SV rates in those planted in plastic bags. Cytharexyllum myrianthum and H. courbaril presented high SV rates in tubes. The growth rate of the species at 42 months differed according to the containers tested. Cytharexyllum myrianthum presented the lowest SV rates (16.7–27.8%), regardless of the container used in this experiment. Ceiba speciosa was sensitive to the reduction in size of the container, showing low SV in tubes (27%) compared with plastic bags (61%); i.e., this species did not tolerate conditions with root growth restriction. Anadenathera macrocarpa and H. courbaril showed no differences in SV, regardless of the container used. The results assist the production of native species of the Atlantic Forest, reinforcing the need to understand performances in the field at the species level.

Glyphosate is the main tool for weed management in Brazilian citrus orchards, where weeds, such as Conyza bonariensis and Digitaria insularis, have been found with resistance to this herbicide. Field prospections have allowed the identification of a possible new case of glyphosate resistance. In this work, the susceptibility levels to glyphosate on three Amaranthus viridis L. populations, with suspected resistance (R1, R2, and R-IAC), collected in citrus orchards from the São Paulo State, Brazil, as well as their accumulation rates of shikimic acid, were determined. The fresh weight of the susceptible population (S) was reduced by 50% (GR50) with ~30 g ea ha−1 glyphosate, while the GR50 values of the R populations were between 5.4 and 11.3 times higher than that for S population. The LD50 (herbicide dose to kill 50% of individuals of a weed population) values of the S population were ≤150 g ea ha−1 glyphosate, while the LD50 of the R populations ranged from 600 to 920 g ea ha−1. Based on the reduction of fresh weight and the survival rate, the R1 population showed the highest level of glyphosate resistance, which had GR50 and LD50 values of 248 and 918 g ea ha−1 glyphosate, respectively. The S population accumulated 240 µg shikimic acid at 1000 µM glyphosate, while the R1, R2, and R-IAC populations accumulated only 16, 43, and 33 µg shikimic acid, respectively (between 5.6 to 15 times less than the S population). Enzyme activity assays suggested that at least one target site-type mechanism was involved in resistance. This result revealed the first report of glyphosate resistance in A. viridis reported in the world.

A failure of the EPSPS-inhibiting herbicide glyphosate to control several populations of Hordeum murinum subsp. leporinum (or H. murinum) occurred in southern Spain after more than fifteen applications in both crop (olive, orchards, and citrus) and non-crop (dry areas, roadsides and ditches) areas. Eight out of 18 populations studied were resistant (R) to glyphosate with R factors higher than four based on GR50. These populations also had the highest values of LD50 and the lowest levels of shikimic acid accumulation. Two adjuvants tested increased glyphosate efficacy in both susceptible (S) and R populations thanks to better spray foliar retention. Moreover, PS I-, PS II-, and ACCase-inhibiting herbicides, in pre- or post-emergence, proved to be the best chemical alternatives with different sites of action (SoA) to control both S and glyphosate-R populations. This study represents the first report worldwide of glyphosate resistance in H. murinum found in very different crop and non-crop areas from southern Spain. To design chemical strategies to implement integrated weed management programs for glyphosate-R H. murinum, both adjuvants and herbicides with alternative SoA as well as application timings should be considered.

The use of herbicides with different modes of action is the primary strategy used to control weeds possessing resistance to a single mechanism of action (MOA). However, this practice can lead to selection for generalist resistance mechanisms and may cause resistance to all MOAs. In this research, we characterized the resistance to diquat/paraquat (bipyridiliums) in an Epilobium ciliatum biotype (R1) collected in an olive orchard from Chile, where alternatives herbicides (2,4-D, glyphosate, glufosinate, flazasulfuron and pyraflufen-ethyl) with different MOAs were used, but they have also showed failure in controlling this species. Because the resistance/susceptibility patterns of the R1 biotype to glufosinate, 2,4-D and pyraflufen-ethyl were not clear, a recurrent resistance selection was carried out in field and greenhouse using these herbicides on R1 plants for three generations (R2 biotype). One biotype that was never treated with herbicides (S) was included as control. Results indicated that the S biotype was controlled at the field dose of all herbicides tested. The biotype R1 exhibited resistance to diquat, paraquat and flazasulfuron and natural tolerance to glyphosate. The R2 biotype displayed resistance to glufosinate, 2,4-D and pyraflufen-ethyl with LD50 (herbicide dose to kill 50% of plants) values higher than field doses in all assays. Physiological and biochemical studies determined the resistance to diquat of the R1 biotype, which was due to impaired translocation. The resistance to flazasulfuron in the R1 and R2 biotypes was confirmed by the low sensitivity of the acetolactate synthase (ALS) activity compared to the S biotype. The similar accumulation of shikimate in treated S, R1, and R2 plants with glyphosate supported the existence of innate tolerance to this herbicide in E. ciliatum. Resistance to glufosinate, 2,4-D and pyraflufen-ethyl in the R2 biotype, acquired after recurrent selection, was determined by low sensitivity of the glutamine synthetase, low accumulation of ethylene and protoporphyrinogen IX oxidase, respectively, in comparison to the S biotype. Epilobium ciliatum from Chilean olive orchards had resistance to only two MAOs (photosystem I and ALS inhibitors), but resistance to five MOAs could occur in the next cropping seasons, if alternatives to weed management, other than herbicides, are not included.

In Brazil, infestations of crop areas with glyphosate-resistant (GR) sourgrass (Digitaria insularis (L.) Fedde) biotypes has risen significantly, increasing crop production costs. Glyphosate efficacy on three biotypes (GO, BA and MT) of sourgrass with suspected resistance was evaluated. A susceptible biotype (MG) was used as the control. The results confirmed that the MG and GO biotypes were susceptible to glyphosate (control > 90%). The MG biotype exhibited growth reduction and mortality by 50% (GR50 and LD50, respectively) with mean glyphosate doses of 243.7 and 431.6 g ae ha-1. The resistance index of the biotypes with suspected resistance ranged from 2.8 to 6.1 in relation to GR50 and between 1.4 to 26.7 in relation to LD50. The glyphosate susceptibility ranking of the sourgrass biotypes was MG < GO < MT < BA. The MT and BA biotypes demonstrated high glyphosate resistance levels, and the GO biotype had a high potential to develop resistance. Farmers should avoid the application of glyphosate overdoses to minimize the selection pressure on weeds.

The Chloris genus is a C4 photosynthetic species mainly distributed in tropical and subtropical regions. Populations of three Chloris species occurring in citrus orchards from central Cuba, under long history glyphosate-based weed management, were studied for glyphosate-resistant status by characterizing their herbicide resistance/tolerance mechanisms. Morphological and molecular analyses allowed these species to be identified as C. ciliata Sw., Chloris elata Desv., and Chloris barbata Sw. Based on the glyphosate rate that causes 50% mortality of the treated plants, glyphosate resistance (R) was confirmed only in C. elata, The R population was 6.1-fold more resistant compared to the susceptible (S) population. In addition, R plants of C. elata accumulated 4.6-fold less shikimate after glyphosate application than S plants. Meanwhile, populations of C. barbata and C. ciliata with or without glyphosate application histories showed similar LD50 values and shikimic acid accumulation rates, demonstrating that resistance to glyphosate have not evolved in these species. Plants of R and S populations of C. elata differed in 14C-glyphosate absorption and translocation. The R population exhibited 27.3-fold greater 5-enolpyruvyl shikimate-3-phosphate synthase (EPSPS) activity than the S population due to a target site mutation corresponding to a Pro-106-Ser substitution found in the EPSPS gene. These reports show the innate tolerance to glyphosate of C. barbata and C. ciliata, and confirm the resistance of C. elata to this herbicide, showing that both non-target site and target-site mechanisms are involved in its resistance to glyphosate. This is the first case of herbicide resistance in Cuba.

Multiple mechanisms of resistance to glyphosate are exhibited by populations of Lolium spp. worldwide. Association of resistance with growth and reproductive fitness is an important predictor for long-term success of glyphosate-resistant (R) versus glyphosate-susceptible (S) biotypes. Numerous studies were conducted on R- and S-biotypes of Italian ryegrass (Lolium multiflorum) and perennial ryegrass (L. perenne) to characterize the underlying mechanism(s) of glyphosate resistance and associate this with growth and reproductive fitness. L. perenne expressed both altered uptake and translocation as well as a genetic change at 106-Pro to –Ser, This pattern for two resistance mechanisms is unique. L. multiflorum also exhibited altered uptake and translocation as well as duplication of EPSPS gene copies. Reduced plant biomass and height for R-versus S-biotypes of both species was evident over two growing seasons. This resulted in S- versus R- L. multiflorum producing up to 47 and 38% more seeds in 2014 and 2015, respectively. S- L. perenne produced up to 20 and 30% more seeds in 2014 and 2015, respectively. Both non-target site and target-site mechanisms of glyphosate resistance can render Lolium spp. at a competitive disadvantage. This has long-term implications for the success of glyphosate-resistant plants in the absence of selection pressure.

Leptochloa virgata (L.) P. Beauv. is an annual weed common in citrus groves in the states of Puebla and Veracruz, Mexico limiting their production. Since 2010, several L. virgata populations were identified as being resistant to glyphosate, but studies of their resistance mechanisms developed by this species have been conducted. In this work, three glyphosate-resistant populations (R8, R14 and R15) collected in citrus orchards from Mexico, were used to study their resistance mechanisms comparing them to one susceptible population (S). Dose-response and shikimic acid accumulation assays confirmed the glyphosate resistance of the three resistant populations. Higher doses of up to 720 g ae ha-1 (field dose) were needed to control by 50% plants of resistant populations. The S population absorbed between 7 and 13% more 14C-glyphosate than resistant ones, and translocated up to 32.2% of 14C-glyphosate to the roots at 96 h after treatment (HAT). The R8, R14 and R15 populations translocated only 24.5, 26.5 and 21.9%, respectively. The enzyme activity of 5-enolpyruvyl shikimate-3-phosphate synthase (EPSPS) was not different in the S, R8 and R14 populations. The R15 Population exhibited 165.9 times greater EPSPS activity. Additionally, this population showed a higher EPSPS basal activity and a substitution in the codon 106 from Proline to Serine in the EPSPS protein sequence. EPSPS gene expression in the R15 population was similar to that of S population. In conclusion, the three resistant L. virgata populations show reduced absorption and translocation of 14C-glyphosate. Moreover, a mutation and an enhanced EPSPS basal activity at target-site level confers higher resistance to glyphosate. These results describe for the first time the glyphosate resistance mechanisms developed by resistant L. virgata populations of citrus orchards from Mexico.

In 2014 hairy beggarticks (Bidens pilosa L.) has been identified as being glyphosate-resistant in citrus orchards from Mexico. The target and non-target site mechanisms involved in the response to glyphosate of two resistant populations (R1 and R2) and one susceptible (S) were studied. Experiments of dose-response, shikimic acid accumulation, uptake-translocation, enzyme activity and EPSPS gene sequencing were carried out in each population. The R1 and R2 populations were 20.4 and 2.8-fold less glyphosate sensitive, respectively, than the S population. The resistant populations showed a lesser shikimic acid accumulation than the S population. In the latter one, 24.9% of 14C-glyphosate was translocated to the roots at 96 h after treatment; in the R1 and R2 populations only 12.9 and 15.5%, respectively, was translocated. Qualitative results confirmed the reduced 14C-glyphosate translocation in the resistant populations. The EPSPS enzyme activity of the S population was 128.4 and 8.5-fold higher than the R1 and R2 populations of glyphosate-treated plants, respectively. A single (Pro-106-Ser), and a double (Thr-102-Ile followed by Pro-106-Ser) mutations were identified in the EPSPS2 gene conferred high resistance in R1 population. Target-site mutations associated with a reduced translocation were responsible for the higher glyphosate resistance in the R1 population. The low-intermediate resistance of the R2 population was mediated by reduced translocation. This is the first glyphosate resistance case confirmed in hairy beggarticks in the world.

Glyphosate is a cheap herbicide that has been used to control a wide range of weeds (4−6 times/year) in citrus groves of the Gulf of Mexico; however, its excessive use has selected for glyphosate-resistant weeds. We evaluated the efficacy and economic viability of 13 herbicide treatments (glyphosate combined with PRE- and/or POST-emergence herbicides and other alternative treatments), applied in tank-mixture or sequence, to control glyphosate-resistant weeds in two Persian lime groves (referred to as SM-I and SM-II) of the municipality of Acateno, Puebla, during two years (2014 and 2015). The SM-I and SM-II fields had 243 and 346 weeds/m2, respectively, composed mainly of Bidens pilosa and Leptochloa virgata. Echinochloa colona was also frequent in SM-II. The glyphosate alone treatments (1080, 1440, or 1800 g ae ha−1) presented control levels of the total weed population ranging from 64% to 85% at 15, 30, and 45 d after treatment (DAT) in both fields. Mixtures of glyphosate with grass herbicides such as fluazifop-p-butyl, sethoxydim, and clethodim efficiently controlled E. colona and L. virgata, but favored the regrowth of B. pilosa. The sequential applications of glyphosate + (bromacil + diuron) and glufosinate + oxyfluorfen controlled more than 85% the total weed community for more than 75 days. However, these treatments were between 360% and 390% more expensive (1.79 and 1.89 $/day ha−1 of satisfactory weed control, respectively), compared to the representative treatment (glyphosate 1080 g ae ha−1 = USD $29.0 ha−1). In practical and economic terms, glufosinate alone was the best treatment controlling glyphosate resistant weeds maintaining control levels >80% for at least 60 DAT ($1.35/day ha−1). The rest of the treatments, applied in tank-mix or in sequence with glyphosate, had similar or lower control levels (~70%) than glyphosate at 1080 g ae ha−1. The adoption of glufosiante alone, glufosinate + oxyfluorfen or glyphosate + (bromacil + diuron) must consider the cost of satisfactory weed control per day, the period of weed control, as well as other factors associated with production costs to obtain an integrated weed management in the short and long term.

The introduction of glyphosate-resistant (GR) crops revolutionized weed management; however, the improper use of this technology has selected for a wide range of weeds resistant to glyphosate, referred to as superweeds. We characterized the high glyphosate resistance level of an Amaranthus hybridus population (GRH)—a superweed collected in a GR-soybean field from Cordoba, Argentina—as well as the resistance mechanisms that govern it in comparison to a susceptible population (GSH). The GRH population was 100.6 times more resistant than the GSH population. Reduced absorption and metabolism of glyphosate, as well as gene duplication of 5-enolpyruvylshikimate-3-phosphate synthase (EPSPS) or its overexpression did not contribute to this resistance. However, GSH plants translocated at least 10% more 14C-glyphosate to the rest of the plant and roots than GRH plants at 9 h after treatment. In addition, a novel triple amino acid substitution from TAP (wild type, GSH) to IVS (triple mutant, GRH) was identified in the EPSPS gene of the GRH. The nucleotide substitutions consisted of ATA102, GTC103 and TCA106 instead of ACA102, GCG103, and CCA106, respectively. The hydrogen bond distances between Gly-101 and Arg-105 positions increased from 2.89 Å (wild type) to 2.93 Å (triple-mutant) according to the EPSPS structural modeling. These results support that the high level of glyphosate resistance of the GRH A. hybridus population was mainly governed by the triple mutation TAP-IVS found of the EPSPS target site, but the impaired translocation of herbicide also contributed in this resistance.

Amino acid substitutions that confer herbicide-resistance may cause fitness costs in mutant plants at unfavorable levels in contrast to wild-species. The fitness costs in three Lolium rigidum populations (AH3 (Ile-2041-Asn) and BO2 (Ile-1781-Leu) as resistant (R) to clodinafop-propargyl, an ACCase (acetyl-CoAcarboxylase) inhibitor, carrying the mutations 1781 and 2041, respectively, and HF as susceptible (S)) were studied during 2014 and 2016. The germination rates and percentages of the three L. rigidum populations, and competition between them and Triticum aestivum using substitution series experiments were assessed. The BO2 and AH3 populations showed resistance to clodinafop-propargyl due to mutations in their ACCase genes. The germination rate for L. rigidum decreased as the sowing depth increased, with the lowest germination rate being found at 8 cm. AH3 and HF populations presented higher seed germination under water and NaCl salinity stress, but no fitness cost variations were observed among these R populations under optimal growth conditions. Diverse germination responses to light conditions were observed between the S and R L. rigidum populations. The highest germination percentage was observed in the HF population at the two-week lighting + two-week darkness regime. The comparison of relative yield total and relative crowding coefficient showed that T. aestivum was more competitive than L. rigidum. However, among ACCase-resistant L. rigidum populations, AH3 population was the most competitive presenting no fitness costs. This R population was more competitive than the S (HF) one under competitive conditions. These results show that fitness costs in the R L. rigidum populations vary according to the specific mutation at the ACCase gene that confers resistance to clodinafop-propargyl. In conclusion, mutations occurring at the 2041 position in the ACCase gene caused fitness costs, but those occurring at the 1781 position did not generate fitness costs for L. rigidum. Therefore, non-chemical methods should be considered unfavorable for resistant populations of this species.