Susceptible Biotypes Research Articles

EPSPS amplification in glyphosate‐resistant spiny amaranth (Amaranthus spinosus): a case of gene transfer via interspecific hybridization from glyphosate‐resistant Palmer amaranth (Amaranthus palmeri)

Amaranthus spinosus, a common weed of pastures, is a close relative of Amaranthus palmeri, a problematic agricultural weed with widespread glyphosate resistance. These two species have been known to hybridize, allowing for transfer of glyphosate resistance. Glyphosate-resistant A. spinosus was recently suspected in a cotton field in Mississippi. Glyphosate-resistant A. spinosus biotypes exhibited a fivefold increase in resistance compared with a glyphosate-susceptible biotype. EPSPS was amplified 33-37 times and expressed 37 times more in glyphosate-resistant A. spinosus biotypes than in a susceptible biotype. The EPSPS sequence in resistant A. spinosus plants was identical to the EPSPS in glyphosate-resistant A. palmeri, but differed at 29 nucleotides from the EPSPS in susceptible A. spinosus plants. PCR analysis revealed similarities between the glyphosate-resistant A. palmeri amplicon and glyphosate-resistant A. spinosus. Glyphosate resistance in A. spinosus is caused by amplification of the EPSPS gene. Evidence suggests that part of the EPSPS amplicon from resistant A. palmeri is present in glyphosate-resistant A. spinosus. This is likely due to a hybridization event between A. spinosus and glyphosate-resistant A. palmeri somewhere in the lineage of the glyphosate-resistant A. spinosus plants. Published 2014. This article is a U.S. Government work and is in the public domain in the USA.

Confirmation and Control of Triazine and 4-Hydroxyphenylpyruvate Dioxygenase-Inhibiting Herbicide-Resistant Palmer Amaranth (Amaranthus palmeri) in Nebraska

Palmer amaranth is a difficult-to-control broadleaf weed that infests corn and soybean fields in south-central and southwestern Nebraska and several other states in the United States. The objectives of this research were to confirm triazine and 4-hydroxyphenylpyruvate dioxygenase (HPPD)-inhibiting herbicide-resistant Palmer amaranth in Nebraska and to determine sensitivity and efficacy of POST-applied corn herbicides for control of resistant and susceptible Palmer amaranth biotypes. Seeds from a putative HPPD-resistant Palmer amaranth biotype from Fillmore County, NE were collected from a seed corn production field in fall 2010. The response of Palmer amaranth biotypes to 12 rates (0 to 12×) of mesotrione, tembotrione, topramezone, and atrazine was evaluated in a dose–response bioassay in a greenhouse. On the basis of the values at the 90% effective dose (ED90) level, the analysis showed a 4- to 23-fold resistance depending upon the type of HPPD-inhibiting herbicide being investigated and susceptible biotype used for comparison. This biotype also had a 9- to 14-fold level of resistance to atrazine applied POST. Results of a POST-applied herbicide efficacy study suggested a synergistic interaction between atrazine and HPPD-inhibiting herbicides that resulted in > 90% control of all Palmer amaranth biotypes. The resistant biotype had a reduced sensitivity to acetolactate synthase inhibiting herbicides (halosulfuron and primisulfuron), a photosystem-II inhibitor (bromoxynil), and a protoporphyrinogen oxidase inhibitor (fluthiacet-methyl). Palmer amaranth biotypes were effectively controlled (≥ 90%) with glyphosate, glufosinate, and dicamba, whereas 2,4-D ester provided 81 to 83% control of the resistant biotype and > 90% control of both susceptible biotypes.

Glyphosate influence on the physiological parameters of Conyza bonariensis biotypes

The objective of this study was to determine changes in gas exchange and inhibition of EPSPs, based on the accumulation of shikimic acid in horseweed biotypes resistant and sensitive to glyphosate. Two experiments were conducted in a factorial model. The first one evaluated horseweed biotypes (one resistant and one susceptible to glyphosate), and herbicide rates (0 and 1,080 g a.e. ha ¹) applied on the weed. In the second experiment, the horseweed biotypes (susceptible and resistant to glyphosate) were evaluated in five periods as following: 0, 3, 7, 10, and 14 days after herbicide application (DAH). The photosynthetic rate, transpiration, carboxylation efficiency, and water efficiency were determined using an infrared gas analyzer (IRGA), and shikimic acid concentration by HPLC. The application of glyphosate damaged the photosynthetic parameters of the susceptible biotype, causing complete inhibition of the photosynthetic rate, transpiration rate, carboxylation efficiency and water use efficiency, starting from the 7 DAH. On the other hand, total inhibition of the photosynthetic parameters was not observed for the resistant biotype. Shikimic acid accumulation occurred in both biotypes after glyphosate application but the susceptible biotype had the highest concentrations, indicating greater sensitivity of the enzyme EPSPs. The accumulation of shikimic acid in the resistant biotype indicates that the mechanism of resistance is not related to the total insensitivity of the enzyme EPSPs to glyphosate and/or that other resistance mechanisms may be involved.

Herbicidal activity on acetolactate synthase‐resistant barnyardgrass (Echinochloa crus‐galli) in Arkansas, USA

The intensive use of the acetolactate synthase (ALS)‐inhibiting herbicides, imazethapyr, penoxsulam and bispyribac‐sodium, in imidazolinone‐resistant (Clearfield) rice increases the risk of the evolution of ALS‐resistant barnyardgrass. In 2009, imazethapyr failed to control barnyardgrass that was collected from a field in Arkansas, USA, following the failure of the herbicide in 2008. A greenhouse experiment was conducted to confirm and document the level of resistance of the biotype against three ALS‐inhibiting herbicides that currently are labeled in rice. The level of control of the resistant biotype at the labeled rate of bispyribac‐sodium of 35 g ai ha−1 was 10%, penoxsulam at 22 g ai ha−1 was 0% and imazethapyr at 70 g ai ha−1 was 25%. The level of mortality of the susceptible biotype was 100% with all the herbicides at the labeled rate. The dose needed to kill 50% of the resistant plants was 49 g ha−1 of bispyribac‐sodium, 254 g ha−1 of penoxsulam and 170 g ha−1 of imazethapyr. For the susceptible biotype, bispyribac‐sodium at 6 g ha−1, penoxsulam at 10 g ha−1 and imazethapyr at 12 g ha−1 killed 50% of the treated plants. Based on these findings, it was confirmed that a barnyardgrass population has evolved cross‐resistance to three ALS‐inhibiting herbicides in rice culture in Arkansas. Furthermore, an experiment was conducted to determine if the ALS‐resistant biotype could be controlled using other mechanisms of action. The results indicated that propanil, a photosystem II inhibitor, and quinclorac, a synthetic auxin, failed to control the resistant biotype at the labeled rates, whereas all the other evaluated herbicides provided effective control of both biotypes.

Distribution of Herbicide-Resistant Johnsongrass (Sorghum halepense) in Arkansas

In 2008, a population of johnsongrass collected from a soybean field near West Memphis, AR, in Crittenden County was confirmed resistant to glyphosate. This was the first documented case of glyphosate-resistant johnsongrass in Arkansas. The purpose of this study was to determine the geographical distribution of glyphosate-resistant johnsongrass in Arkansas crops and screen for resistance to additional herbicides. A total of 141 johnsongrass accessions were collected from 14 counties in Arkansas in the fall of 2008, 2009, and 2010 and screened for resistance to four of the most commonly used POST herbicides for johnsongrass control—imazethapyr, glyphosate, clethodim, and fluazifop. One accession potentially resistant to glyphosate (J12) and another with apparent resistance to imazethapyr (J14) were further evaluated in a dose–response experiment. The lethal dose required to kill 50% of the plants from the putative glyphosate-resistant and imazethapyr-resistant accessions was higher than that of a biotype known to be susceptible to these herbicides. The J12 accession had an LD50of 1,741 g ae ha−1glyphosate, which was 8.5-fold greater than the susceptible biotype. The J14 accession had an LD50of 73 g ai ha−1imazethapyr, which was 3.7-fold greater than the LD50of the susceptible biotype. All other accessions were effectively controlled by the four evaluated herbicides. Widespread herbicide-resistant johnsongrass was not found in Arkansas, although accession J12 was resistant to glyphosate and J14 resistant to imazethapyr.

Does the resistance to glyphosate herbicide affect the competitive ability of ryegrass with soybean?

The objective this work was to investigate the competitive ability between resistant and susceptible ryegrass biotypes and of these with soybean crop. Four experiments were carried under greenhouse, in a completely randomized design with four replications, in 2011 and 2012. Treatments were arranged in additive series and replacement series assay. In each series, the proportions among ryegrass resistant and susceptible plants related to soybean were: 100:0, 75:25, 50:50, 25:75 e 0:100. Leaf area and shoot dry mass were evaluated. Competitiveness statistical analysis consisted in applying diagrams to the replacement series and alternative interpretations of the competitiveness indexes. The soybean crop had equivalent competitiveness to the susceptible ryegrass biotype and inferior to the resistant biotype, while the biotypes, both susceptible and resistant to glyphosate, present equivalent competitive ability. In general, the intraspecific competition is more harmful to ryegrass when in competition with soybean, while interspecific competition is predominant for culture.

Impact of non-target-site-resistance on herbicidal activity of imazamox on black¬grass ( Alopecurus myosuroides Huds.) in comparison to other ALS-graminicides

A black-grass (Alopecurus myosuroides Huds.) resistance-monitoring conducted by BASF in 2010 - 2012 revealed a high number of accessions with resistance against imazamox. However, application of imazamox- based products in a winter crop was limited to winter beans in France and United Kingdom only until the introduction of the Clearfield ® -production system in autumn 2012 in winter oilseed rape. It is therefore assumed that the resistance mechanisms were probably selected by the frequent use of ACCase- and ALS- inhibitors in winter crop rotations during the last 2 decades. Resistance level for each product-biotype combination was calculated according the R-classification system (S, R?, RR, RRR) by directly comparing the product performance on a biotype versus untreated control. Majority of resistant biotypes did not show a target-site mutation at the known codon Pro197 or Trp574. In order to better evaluate the impact of Non- Target-Site-Resistance (NTSR) on the activity of BEYOND (imazamox), ATLANTIS WG (mesosulfuron+iodosulfuron) and ABAK (pyroxsulam), biotypes who have shown an ALS-target-site mutation were removed from further analysis. At the dose rate of 35 g ai/ha BEYOND provided good activity on susceptible biotypes of black-grass almost matching up with ATLANTIS WG and ABAK. However, activity of BEYOND declined stronger on biotypes classified as R? or RR for that product, while ATLANTIS WG and ABAK hardly showed any decline in control on this group of biotypes when applied at the recommended dose rate. It is assumed that the underlying NTSR- mechanism is not effective enough yet to confer resistance to ATLANTIS WG and ABAK, but on BEYOND. In contrast, biotypes classified as R? for ATLANTIS WG did show a stronger impact on the activity of BEYOND and ABAK then of ATLANTIS WG. These differences in control level probably do translate into differences in selection pressure as well.

Evaluation of 2,4-D Amine, Glyphosate, 2,4-D Amine plus Glyphosate DMA and 2,4-D Choline/Glyphosate DMA for Their Efficacy on Glyphosate Susceptible and Resistant Canada Fleabane Populations

The 2,4-D choline/glyphosate DMA formulation has reduced drift and volatility compared to the amine or ester formulation of 2,4-D and therefore is advantageous compared to a tank mix of 2,4-D amine or ester with glyphosate. The objective of this research was to compare the control of glyphosate susceptible and glyphosate resistant Canada fleabane with 2,4-D choline/glyphosate DMA with 2,4-D amine, glyphosate, and a tank mix of 2,4-D amine and glyphosate. Ten rates of 2,4-D amine (0 - 6708 g∙ae∙ha⁻¹), glyphosate (0 - 7052 g∙ae∙ha⁻¹), a tank mix of glyphosate plus 2,4- D amine (0 - 7052 g∙ae∙ha⁻¹ + 0 - 6708), and 2,4-D choline/glyphosate DMA (0 - 13760 g∙ae∙ha⁻¹) were examined in the greenhouse for the control of two susceptible (GS) and two resistant to gly- phosate (GR) Canada fleabane biotypes. The tank mix of 2,4-D amine plus glyphosate and 2,4-D choline/glyphosate DMA provided equivalent control of the GR Canada fleabane biotypes at 35 days after the application (DAA). The 2,4-D choline/glyphosate DMA treatment was more effica- cious than the tank mix on the GS biotypes. Glyphosate (880 g∙ae∙ha⁻¹) provided 50% and 100% control of the resistant and susceptible biotypes, respectively. The 2,4-D choline/glyphosate DMA formulation and the tankmix of 2,4-D amine and glyphosate provided similar control of GR Canada fleabane. * Corresponding author.

Distribution of Glyphosate- and Thifensulfuron-Resistant Palmer Amaranth (Amaranthus palmeri) in North Carolina

Glyphosate resistance in Palmer amaranth was first confirmed in North Carolina in 2005. A survey that year indicated 17 and 18% of 290 populations sampled were resistant to glyphosate and thifensulfuron, respectively. During the fall of 2010, 274 predetermined sites in North Carolina were surveyed to determine distribution of Palmer amaranth and to determine if and where resistance to fomesafen, glufosinate, glyphosate, and thifensulfuron occurred. Palmer amaranth was present at 134 sites. When mortality for each biotype was compared to a known susceptible biotype for each herbicide within a rate, 93 and 36% of biotypes were controlled less by glyphosate (840 g ae ha−1) and thifensulfuron (70 g ai ha−1), respectively. This approach may have underestimated resistance for segregating populations due to lack of homogeneity of the herbicide resistance trait and its contribution to error variance. When mortality and visible control were combined, 98% and 97% of the populations were resistant to glyphosate and the ALS inhibitor thifensulfuron, respectively, and 95% of the populations expressed multiple resistance to both herbicides. This study confirms that Palmer amaranth is commonly found across the major row crop production regions of North Carolina and that resistance to glyphosate and ALS-inhibiting herbicides is nearly universal. No resistance to fomesafen or glufosinate was observed.

Herbicidas alternativos para o controle de Raphanus raphanistrum L. resistente ao herbicida metsulfurom metílico

O controle de plantas daninhas dicotiledôneas, em culturas de inverno, é realizado basicamente com herbicidas inibidores da enzima ALS. O uso intensivo desses herbicidas em áreas cultivadas com trigo tem proporcionado pressão de seleção nas populações de Raphanus raphanistrum resultando em biótipos resistentes a esses herbicidas. Objetivou-se com esse trabalho determinar a existência de resistência cruzada e múltipla aos herbicidas inibidores de ALS e alternativas de controle químico dessa planta daninha resistente ao metsulfurom metílico. Os tratamentos resultaram da combinação de dois biótipos de R. raphanistrum resistente (biótipo R) e suscetível (biótipo S), com 12 herbicidas: glifosato, atrazina, bentazona, 2,4-D, sulfometurom metílico + clorimurom etílico, clorimurom etílico, cloransulam metílico, iodosulfurom, nicossulfurom, imazetapir, metsulfurom metílico, imazapic + imazapir e mais um tratamento controle sem aplicação. Aplicou-se a dose recomendada de cada herbicida, para o controle da espécie, no estádio de desenvolvimento de 3 a 4 folhas verdadeiras. Foram analisados a eficiência de controle e redução de massa seca da parte aérea. O biótipo S foi suscetível aos herbicidas inibidores da ALS e aos demais mecanismos de ação testados. O biótipo R apresentou resistência cruzada aos grupos químicos imidazolinonas, sulfoniluréias e triazolpirimidinas variando o grau de sensibilidade entre esses grupos. O biótipo R não foi diagnosticado com resistência múltipla, e os herbicidas glifosato, bentazona, atrazina e 2,4-D demonstraram serem eficazes no seu controle, podendo utilizá-los como alternativas para o manejo químico dessa planta daninha.

Dose-response curves of Lolium multiflorum biotypes resistant and susceptible to clethodim

Chemical control with herbicides, especially glyphosate, is the main method used to control ryegrass. However, the repeated use of glyphosate has selected resistant ryegrass biotypes. Thus, the ACCase inhibitor herbicides have become the main alternative to control glyphosate-resistant biotypes, being widely used by farmers in Rio Grande do Sul. Repeated use of ACCase inhibitors, in turn, have selected ryegrass biotypes resistant to this herbicide mechanism. Thus, the objective of this study was to evaluate the response of ryegrass biotypes to different clethodim rates by dose-response curves. Increasing doses (0, 12, 24, 48, 72, 96, 144 and 192 g a.i. ha-1) of the herbicide clethodim were applied at the 3-4 ryegrass leaf stage. The variables control at 14 and 28 days after treatment (DAT) and shoot dry weight were evaluated. The data were fitted by nonlinear regression log-logistic and C50 and GR50 were calculated based on the equation. The resistance factor was obtained by the ratio of C50 or GR50 of the resistant biotype by matching the susceptible biotype. Based on the equation parameters, the doses of GR50 64.7 and 234.5 g a.i. ha-1 clethodim and C50 11.2 and 172.1 g a.i. ha-1 clethodim were obtained, at 28 DAT for the susceptible and resistant biotypes, respectively. The ryegrass biotype denominated Cotril is resistant to clethodim, being controlled with a dose 15.3 times greater than that of the susceptible biotype, and a 50% reduction of this biotype occurs with a dose 3.62 times higher than that of the susceptible one.

Adaptive value of ryegrass biotypes with low-level resistance and susceptible to the herbicide fluazifop and competitive ability with the wheat culture

Ryegrass is the main weed in wheat crop, causing yield loss due to competition by environmental resources. The objectives of this study were to estimate the fitness cost of ryegrass biotypes with low-level resistance and susceptible to fluazifop and to investigate the relative competitive ability of these biotypes between themselves and against the crop. Thus, fitness cost and competitive ability experiments were conducted under greenhouse conditions. For the fitness cost experiments, the low-level resistant ryegrass biotypes and those susceptible to fluazifop were used. For competitive ability, the treatments were arranged in a replacement series, with five proportions of the wheat cultivar FUNDACEP Horizonte and the low-level resistant and susceptible ryegrass biotypes 100:0, 75:25, 50:50, 25:75 and 0:100. Competitive analysis was carried out through diagrams applied to the replacement experiments and use of relative competitiveness indices. Variables evaluated were plant height, in the fitness cost experiment, and leaf area and shoot dry biomass in both experiments. The ryegrass biotypes show overall similar fitness cost and competitive ability. The wheat cultivar FUNDACEP Horizonte is superior in competitive ability to the ryegrass biotype with low-level resistance and equivalent to the susceptible biotype.

Qualitative/quantitative strategy for the determination of glufosinate and metabolites in plants

A simple method for the simultaneous determination of glufosinate and its metabolites in plants based on liquid chromatography-ultraviolet (LC-UV) absorption detection after derivatization with fluorenylmethoxycarbonyl chloride (FMOC-Cl) of some analytes to facilitate separation is reported here. Nonavailable standard metabolites were identified by LC-TOF/mass spectrometry (MS), which also confirmed all target analytes. Ultrasound-assisted extraction was used for sample preparation (power of 70 W and duty cycle of 0.7 s/s for 10 min) with subsequent evaporation of the extractant, reconstitution and filtration as the cleanup/concentration step prior to derivatization, and chromatographic separation and detection at 270 nm for underivatized analytes and 340 nm for those that were derivatized. The chromatographic analysis was completed in 40 min using a Luna® column (C18 phase). The analytical characteristics of the method were linear dynamic range of the calibration curves within 0.047-700 μg/mL with a regression coefficient (rc) of 0.999 for glufosinate, 0.077-700 μg/mL with a rc of 0.998 for N-acetyl-glufosinate, and 0.116-600 μg/mL with a rc of 0.998 for 3-(methylphosphinico)propanoic acid. The precision for the determination of glufosinate (studied at two levels, 0.1 and 5 μg/mL) was 2.7 and 6.0 % for repeatability and 4.7 and 7.2 % for within-laboratory reproducibility, respectively. Identification and confirmatory analysis of the presence of glufosinate and metabolites in the extracts from treated plants was carried out by LC-TOF/MS in high-resolution mode for the precursor ion. The method was validated by analyzing wheat (Triticum aestivum) samples (resistant and susceptible biotypes) treated with 300 g of glufosinate/ha following conventional agronomical practices.

Physiological tests for early detection of rigid ryegrass (Lolium rigidum Goud.) resistance to fenoxaprop-P

The study aimed to determine the usefulness of isothermal calorimetry and FT-Raman spectroscopy for the early evaluation of rigid ryegrass resistance to fenoxaprop-P ethyl (active ingredient one of aryloxyphenoxypropionate herbicides). The calorimetric measurements were done on the 4-day-old seedlings of susceptible and resistant biotypes of rigid ryegrass (Lolium rigidum Goud.) for 72 h, at 20 °C. It was observed that the specific thermal power–timecurves of the susceptible and resistant biotypes growing on water (control) were qualitatively similar. Herbicides changed the shape of the specific thermal power–time curves of both biotypes. Furthermore, the total specific thermal energy was significantly higher for the seedlings of resistant biotype, growing both on water or herbicide, as compared to the susceptible ones. The analysis of the seedlings’ endosperm, conducted using FT-Raman spectroscopy, showed a weaker intensity of the bands in the spectra derived from the resistant biotype. Differences in the specific thermal power–time curves and FT-Raman spectra between susceptible and resistant biotypes growing on water indicate that the sensitive and resistant biotypes are metabolically and chemically different already in the early stages of the seedling growth. We conclude that isothermal calorimetry and FT-Raman spectroscopy are efficient tools for the early detection of rigid ryegrass resistance to fenoxaprop-P ethyl.

Identification and mRNA expression profile of glutamate receptor-like gene in quinclorac-resistant and susceptible Echinochloa crus-galli

Animal ionotropic glutamate receptors (iGluRs) function as Ca2+ ion channels during excitatory neurotransmission in nerve cells. Here, a glutamate receptor-like gene (GLR) was identified and characterized from a plant — Echinochloa crus-galli. The GLR gene was designated EcGLR1 with GenBank no: JX518597. It has a 2793bp open reading frame predicted to encode a 101.7kDa protein. Sequence alignment showed that EcGLR1 is a GLR homologue. Its expression in response to quinclorac treatment was assessed by real-time PCR in near-isogenic lines of quinclorac-resistant (R) and susceptible (S) biotypes of E. crus-galli. The expression of EcGLR1 in the seedling leaf and root at least increased 5 times in the S plants and 22 times in the R plants after exposure to quinclorac. In the adult plant leaves, roots and stems, its expression increased 11–14 times in the S plants and 23–25 times in the R plants after quinclorac stimulation. In the seed, its expression was 4 times less in the S plants than that in the R plants, but after treatment, the levels all increased by about 24 times in the two biotypes. EcGLR1 expression was 1–4 times greater in the R plants than in that in the S plants, and after treatment by quinclorac, the difference increased to a ratio of 4 to 9. Its expression was higher in all tissues tested of R biotypes than in that of S plants before or after quinclorac treatment. The results of this study provide basic information for the further research of function of the EcGLR1 in resistance to quinclorac in E. crus-galli.

Germination and growth of Fimbristylis miliacea biotypes resistant and susceptible to acetolactate synthase-inhibiting herbicides

Weed biotypes resistant and susceptible to herbicides may have differences in their adaptive values. The aims of this study were to compare, under controlled and non-competitive condition, the growth analysis, germination features and seed weight of Fimbristylis miliacea (FIMMI) biotypes resistant and susceptible to acetolactate synthase (ALS) inhibiting herbicides. Experiments were conducted in a greenhouse and in a laboratory from October 2008 to February 2010. Two resistant biotypes (FIMMI 10 and FIMMI 12) and one susceptible biotype (FIMMI 13) were used for the studies. For the study on growth analysis, the treatments were arranged in a completely randomized experimental design with four replications and sampled at 21, 28, 35, 42, 49, 56, 69 days after emergence (DAE) and at flowering stage. For the studies on germination speed, germination and seed weight, the indexes for germination speed, percentage of germination at different temperatures and seed weight of the biotypes were determined. The results showed that the resistant biotype FIMMI 12 shows differences in all variables compared to the resistant biotype FIMMI 10 and compared to the susceptible biotype FIMMI 13, only for the evaluation at flowering. The susceptible biotype FIMMI 13 showed a higher germination speed index and higher germination rate when compared with the resistant biotypes. On the other hand, the resistant biotypes FIMMI 10 and FIMMI 12 showed higher seed weight.

Low level resistance of goosegrass (Eleusine indica) to glyphosate in Rio Grande do Sul-Brazil

In the last growing seasons, goosegrass (Eleusine spp.) control failures have been observed following application of glyphosate on Roundup Ready® soybean in Rio Grande do Sul (RS) - Brazil, suggesting this species' resistance to the herbicide. Thus, the objectives of this study were to identify the occurrence of goosegrass resistance to the herbicide glyphosate in RS; and to determine the predominant species of the genus Eleusine, as well as the LD50 and GR50 of the suspected resistant biotypes. Two experiments were conducted under greenhouse conditions: one to identify the biotypes resistant to glyphosate, and the other, a dose-response curve experiment, as well as a study of the botanical characteristics of the species. In the first experiment, 39 biotypes were tested, mainly Eleusine indica, collected with suspected resistance to glyphosate. The glyphosate dose was 2,160 g e.a. ha-1, and the control was evaluated at 28 days after treatment. All biotypes were effectively controlled,with the biotypes from the municipality of Boa Vista do Incra showing greater tolerance. Two biotypes suspected of resistance (12.1 and 12.3) and a susceptible biotype in a dose-response experiment were tested at the following doses: 0, 135, 270, 540, 1,080, 1,620, and 2,160 g e.a. ha-1. The results of this experiment showed that biotype 12.1 does not present resistance to glyphosate and biotype 12.3 has a low level resistance since it is effectively controlled by the herbicide at the maximum dose.

Rapid diagnosis of resistance to imidazolinone herbicides in barnyardgrass (Echinochloa crus-galli) and control of resistant biotypes with alternative herbicides

The resistance of barnyardgrass (Echinochloa crus-galli) to imidazolinone herbicides is a worldwide problem in paddy fields. A rapid diagnosis is required for the selection of adequate prevention and control practices. The objectives of this study were to develop expedite bioassays to identify the resistance to imidazolinone herbicides in barnyardgrass and to evaluate the efficacy of alternative herbicides for the post-emergence control of resistant biotypes. Three experiments were conducted to develop methods for diagnosis of resistance to imazethapyr and imazapyr + imazapic in barnyardgrass at the seed, seedling and tiller stages, and to carry out a pot experiment to determine the efficacy of six herbicides applied at post-emergence in 13 biotypes of barnyardgrass resistant to imidazolinones. The seed soaking bioassay was not able to differentiate the resistant and susceptible biotypes. The resistance of barnyardgrass to imidazolinones was effectively discriminated in the seedlings and tiller bioassays seven days after incubation at the concentrations of 0.001 and 0.0001 mM, respectively, for both imazethapyr and imazapyr + imazapic. The biotypes identified as resistant to imidazolinones showed different patterns of susceptibility to penoxsulam, bispyribac-sodium and pyrazosulfuron-ethyl, and were all controlled with profoxydim and cyhalofop-butyl. The seedling and tiller bioassays are effective in the diagnosis of barnyardgrass resistance to imidazolinone herbicides, providing an on-season opportunity to identify the need to use alternative herbicides to be applied at post-emergence for the control of the resistant biotypes.

Resistance Mechanism to Tribenuron-Methyl in White Mustard (Sinapis alba) from Southern Spain

Tribenuron-methyl has been used widely for the last 15 yr to control white mustard in cereal crops from southern Spain. Since 2007, several cases of tribenuron-methyl resistance have been reported in wheat fields. Greenhouse and laboratory studies were conducted to characterize the mechanism of suspected tribenuron-methyl resistance in a white mustard biotype (hereafter AR16) from Malaga (southern Spain). Under greenhouse conditions, the dose (g ai ha−1) inhibiting fresh weight by 50% (ED50) was 5.20 and 0.57 for the AR16and AR3(known susceptible) biotypes, respectively. With the use of14C-tribenuron-methyl, absorption and translocation from treated leaves were similar between biotypes. Thin-layer chromatography indicated foliar metabolism of tribenuron-methyl was low in both R and S biotypes. Assays on the binding affinity of tribenuron-methyl on acetolactate synthase (ALS) revealed enzyme activity was reduced by 50% (I50value) at 638.7 and 0.23 nM for the AR16and AR3biotypes, respectively. This resulted in 2,777-fold greater resistance to tribenuron-methyl for the AR16compared to AR3biotype. Sequencing the gene encoding ALS, a proline/serine amino-acid substitution, was detected in position 197 of the A domain. Based on these results, it is concluded that tribenuron-methyl resistance in the AR16biotype is due to a target-site mutation in the ALS enzyme, resulting in a lack of affinity to tribenuron-methyl.

Caracterización de la resistencia de Amaranthus quitensis a tres familias de herbicidas

Amaranthus quitensis is an annual species from South America that reproduces by seed. In Argentina its distribution covers central and northern country and is a major weed of summer crops (soybean, corn and sunflower) Pampa area. In the central region (Province of Cordoba and Santa Fe) flaws in its control were detected, when using different herbicide families (imidazolinone, sulfonylurea and triazolopyrimidines). Therefore, the aim of this study was to characterize the resistance at the biochemical and molecular level of biotypes of A. quitensis from four localities along this region. Specifically we used different herbicides inhibitors Aceto-Lactate Synthase (ALS), an enzyme involved in the synthesis of a group of aminoacids, and compared them with a susceptible biotype. Specific objectives were to determine the geographical origin of the resistance to each of these three herbicides and to assess the effect of different doses on ALS activity in vitro. Results, based on the observed genetic variability and on the sequences of the als genes, indicated that the analyzed populations were originated independently and not from a common mutation event that later dispersed to the different sites. These results allowed for a deeper knowledge at the biochemical and molecular levels of the resistance mechanisms of five ecotypes of A. quitensis to different herbicides that have the ALS enzyme as a target.