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Article

First Report of Resistance to Glyphosate in Several Species of the Genus Echinochloa in Argentina

by
Eduardo Cortés
1,
Ana Schneider
2,
Elisa Panigo
2,
Mariel Perreta
2,
Rafael De Prado
3 and
Ignacio Dellaferrera
2,*
1
Facultad de Ciencias Agrarias, Universidad Nacional del Litoral, Esperanza 3080, Argentina
2
ICiAgro Litoral (UNL-CONICET)—MEP (Laboratorio de Análisis y Modelización de la Estructura de Plantas Cultivadas y Malezas), R.P. Kreder 2805, Esperanza 3080, Argentina
3
Departamento de Química Agrícola y Edafología, Universidad de Córdoba, 14014 Córdoba, Spain
*
Author to whom correspondence should be addressed.
Agronomy 2023, 13(5), 1219; https://doi.org/10.3390/agronomy13051219
Submission received: 14 December 2022 / Revised: 21 December 2022 / Accepted: 22 December 2022 / Published: 26 April 2023
(This article belongs to the Special Issue Recent Insights in Herbicide Biochemistry, Herbicide Resistance and Weed Management)
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Abstract

:
The genus Echinochloa consists of about 33 species worldwide; some of these are weeds that are very difficult to control in Argentina, and only E. colona was reported as resistant to glyphosate. The objective of this work is to determine if one or more populations of E. colona, E. crus-galli, E. oryzoides, and E. chacoensis are resistant to or less susceptible to glyphosate. Between 2015 and 2017, seeds of different Echinochloa populations were collected from the provinces of Córdoba, Santa Fe, Buenos Aires, and Entre Ríos, all from fields with a history of at least 10 consecutive years of glyphosate application and complaints from farmers due to failures in control. With these populations, survival, dose–response, and shikimic acid quantification tests were carried out to determine their level of susceptibility to glyphosate. The results obtained allow us to report the first worldwide case of resistance to glyphosate in populations of E. crus-galli, E. oryzoides, and E. chacoensis and expand the information on E. colona.

1. Introduction

The genus Echinochloa consists of about 33 species worldwide, including subspecies and varieties [1], while in Argentina, seven species are present [2], but few of these are reported as weeds. Instead, and also as weeds, the two most widespread species that are found both globally and in Argentina are Echinochloa colona (L.) Link and Echinochloa crus-galli (L.) P. Beauv. Additionally, Echinochloa chacoensis Michael ex Renvoize and Echinochloa oryzoides (Ard.) Fritsch are present in Argentina as weeds in various crops [3], particularly in soybean and corn crops [4].
Worldwide, they are found in crops such as rice, sorghum, sugarcane, cotton, peanuts, etc. [5]. There four Echinochloa species behave similar to weeds and compete strongly with many crops. E. colona reduces soybean yield by 12 [kg/ha] to 27 [kg/ha] per percent of cover by this weed [4,6] and E. crus-galli can reduce rice yield by about 25% in heavy infestations [7].
In Argentina, from 2010 to the present, the genus Echinochloa recorded a sustained development in the occupied area, reaching 35% of the agricultural area in 2021 and positioning itself as the fourth most important weed [8]. In particular, E. colona has increased its presence and abundance as a result of the continuous use of glyphosate [9].
All species of the genus Echinochloa are C4 [10] annuals, with emergence in spring and summer. They reproduce by seed and coexist in agricultural areas. The plants in this genus are very diverse, and their taxonomy is confusing [11,12]. Only E. chacoensis has ligules; the other three species do not have ligules or auricles. The inflorescence of E. crus-galli is an erect pyramidal panicle consisting of 15 to 30 pseudo-spikes. The panicle of E. colona is erect and has a greater number of reproductive organs arranged on lateral branches, between 5 and 15 of them. E. oryzoides has contracted pointed inflorescences with 6–15 short first-order branches, and E. chacoensis has a contracted panicle with densely packed spikelets on the branches [13,14,15]. Various ecotypes, high seed production, short seed dormancy, rapid growth, competitive potential, allelopathic interaction, and resistance to various herbicides make these weeds a more adaptable and persistent challenge in various agroecosystems [5,6].
Regarding their chemical control, worldwide failures are registered in different populations of the Echinochloa genus. The first resistant population of E. colona was confirmed in 2007 in Australia [7]. The Weed Science Society of America declares cases of glyphosate resistance in the population of Echinochloa sp. for the United States as of 2008 and in Argentina as of 2009 [7,8].
Herbicide resistance is an evolutionary and ecological process. The dynamics of this process are influenced by (1) weed characteristics, which further include the morphological and genetic factors associated with the weed; (2) herbicide characteristics, which further include the mode or site of action of herbicide; and (3) finally the operational factors, i.e., the prevalent farm practices [16].
A total of 83 cases of eight species and cultivars of the genus Echinochloa have been reported worldwide with resistance to various groups of herbicides, including E. colona; Echinochloa crus-galli (L.) P. Beauv. var. crus-galli; Echinochloa crus-galli var. formosensis Ohwi; Echinochloa crus-galli var. zelayensis (Humb., Bonpl. & Kunth) Hitchc.; Echinochloa crus-pavonis (Kunth) Schult; Echinochloa erecta (Pollacci) Pignatti; E. oryzoides and Echinochloa phyllopogon (Stapf) Stapf ex Kossenko (=E. oryzicola) [17]. E. crus-galli var. crus-galli presents the highest number of cases of resistance and together with E. colona, they are the two most widespread species in extensive crops. In all species, resistance occurs mainly in rice crops, but there are also some cases in maize, soybeans, sorghum, cotton, grapes, orchards, watermelons, fallow land, fences, and roadsides [18].
For these various species, there are no previous publications that confirm glyphosate herbicide resistance worldwide [18]. Additionally, there has not been a comparative study of the susceptibility of the different species of Echinochloa in Argentina [19].
For this reason, this study aims to determine whether populations of E. colona, E. crus-galli, E. oryzoides, and E. chacoensis that currently infest different annual crops in Argentina are resistant or less susceptible to glyphosate.

2. Materials and Methods

Plant Material:
To determine the susceptibility to glyphosate, an initial of nine populations of E. colona, seven populations of E. crus-galli, six populations of E. oryzoides, and three populations of E. chacoensis were examined for possible resistance.
Between 2015 and 2017, seeds of the genus Echinochloa were collected from the provinces of Córdoba, Santa Fe, Buenos Aires, and Entre Ríos; all populations proceed from cultivated fields with a history of at least 10 consecutive years of glyphosate application and complaints from farmers of control failures.
Seeds were collected from 25 fields. In each place, only one species is observed dominating the community of gramineous weeds. One hundred whole plants were taken and bulked into a single sample from each field, for the determination of glyphosate resistance. In all these fields, it was known that Echinochloa sp. plants had survived multiple glyphosate applications with reduced phytotoxic effects. Samples were stored at room temperature (~l8 °C) in the laboratory until sowing. The determination of the species was made using the keys of Flora Argentina [11]. Table 1 details the origin of each population along with the abbreviated name of each one.
Screening (survival):
A hundred seeds of each specie and population were cleaned and then sown in 500 [cm3] pots, which contained approximately 720 [g] of typical Esperanza Series Argiudol soil (loam-clay-silty texture; 66.7-28.7-4.7). Then they were placed in a growth room at a temperature of 28/18 [°C] (day and night), with a photoperiod of 16 h, a light intensity of 850 [μmol m−2 s−1], and 60–80% relative humidity (as suggested for summer spring cycle species [20]).
When the seedlings had four leaves a discriminant dose of 600 [g a.i./ha] of glyphosate (36 [% p/v a.i.]) were applied. All applications were carried out with a laboratory spray chamber equipped with a Magnojet 8001 flat fan nozzles, calibrated to spray 175 [L/ha] at a pressure of 275 [kPa].
Before application (at plant emergence) and 21 days after application of glyphosate (plant survived), the number of plants present per pot was determined. A plant was considered alive if a meristem was green [21]. The % survival is determined as (plant survival)/(plants emergence) × 100.
The populations were classified into three groups according to their survival rate: (i) resistant, if more than 20% of the plants survived the herbicide treatment, (ii) in development of resistance if 6–19% of the plants survived, and (iii) susceptible if less than 5% survived to herbicide treatment [22].
Dose Response (vegetal material):
Populations of all Echinochloa species (Table 1) with suspected low sensitivity to glyphosate were cultivated; in addition, populations of known high sensitivity were also cultivated for comparison.
They were germinated on a moistened filter paper in Petri dishes, and then transplanted to one-liter pots and placed in a growth room under the same conditions mentioned in the previous trial.
For each combination of dose and population, 10 replicates were used, and the test was performed in a completely randomized design. To compare the susceptibilities of the different populations, three dose–response trials were conducted, for which increasing doses of glyphosate 0 X, ¼ X, ½ X, X, 2 X, 4 X, and 8 X will be applied, with X (1080 g a.i./ha). All applications will be carried out with the laboratory spray chamber previously described.
The fresh weight was determined per pot 21 days after application. These weights were expressed as a percentage of the untreated control, and statistical analysis was performed using the drc package [23] in R. The data by species and population are adjusted to a logistic model represented by the function of 4 parameters (1), where GR50 is the dose that produces the average response between the upper limit d and the lower limit c. Parameter b marks the inflection around GR50 [24,25].
F r e s h   w e i g h t = c + d − c 1 + e x p b l o g h e r b i c i d e   d o s e − l o g G R 50
The dose of herbicide that inhibits growth by 50 and 80% concerning the untreated control (GR50 and GR80) was also determined for each population. The susceptibility differences were also evaluated using the resistance factor (RF) to be determined as GR50 (resistant)/GR50 (susceptible).
Shikimic Acid Accumulation:
Individuals from all populations of the before-mentioned species, under the same growth conditions described above, were sprayed with a glyphosate solution containing 600 [g a.i./ha] under the same specific conditions as the previous test. The last expanded leaves of plants of all species were harvested for shikimic acid extraction 24, 48, 72, and 96 h after application. The foliar tissues (50 [mg] fresh weight) were homogenized, and the samples were frozen at −40 [°C].
Subsequently, the protocol described by Singh and Shaner [26] was followed. The absorption of shikimic acid was optically measured at 382 [nm] using a Biotraza 722 spectrophotometer (Biotraza, China). Standard curves are determined using untreated plants and known shikimic acid concentration solutions. Both experiments were repeated 3 times with 10 replicates per treatment.

3. Results

3.1. Survival

All the plants used in the survival test showed different symptoms, but all produced shoots and viable seeds.
In E. colona, of the total populations, five of them (Ec02, Ec03, Ec06, Ec12, and Ec16) did not show resistance; only one presented resistance in evolution (Ec05), and the rest of the populations (Ec07, Ec14, and Ec04) show survival between 25 and 94% of individuals at the use dose. In E. crus-galli, only Ecg13 did not show resistance (susceptible), Ecg12 presented resistance in evolution, and the remaining populations (Ecg06, Ecg07, Ecg14, Ecg15, and Ecg16) exceeded 20% survival, with values ranging between 25 and 96%.
For E. oryzoides, four of the six populations (Eo02, Eo03, Eo07, and Eo08) had very high survival values between 75 and 87%, qualifying them as resistant. On the other hand, two populations (Eo04 and Eo06) did not survive the application of glyphosate. In addition, finally, the survival test for E. chacoensis shows that only one population (Ech02) has a survival rate higher than 20%, the other two were populations proved to be susceptible.

3.2. Dose Response Trials

In the four species, it was observed that the biomass of the aerial part of the populations decreases as the herbicide dose increases, but the GR50 differs significantly between the different populations of each species.

3.2.1. Echinochloa colona

As can be seen in Table 2 and Figure 1, the highest GR50 is in the Ec04 and Ec14 populations. The dose–response experiments confirmed that the Ec12 and Ec16 populations were not significantly different from the Ec03 population, while the rest of the populations were different from the Ec03 population.
The regression parameters also indicate that the Ec04 and Ec07 populations have most expressed this susceptibility variation, while the rest are less susceptible to dose variation; this is quantified by the inflection point around e (regression parameter b) (Table 2).
The Ec04 and Ec14 populations presented a RF of 3.66 to 3.68 compared to the Ec03, although their GR80 was between 30 and 35% lower than the dose taken as field use (1080 [g a.i./ha]). In addition, populations Ec07, Ec06, Ec05, and Ec02 presented an RF of 1.35 to 2.32 with respect to Ec03, and their GR80 was between 2 and 3 times below the use dose previously considered.

3.2.2. Echinochloa crus-galli

The populations Ecg06, Ecg07, Ecg14, and Ecg15 have GR50 values higher than the rest (Table 3 and Figure 2), and it was determined that these populations were significantly different from the Ecg12 population (considered susceptible). On the other hand, the Ecg13 and Ecg16 populations were not significantly different from the Ecg12.
In addition, the regression parameters indicate that the Ecg07, Ecg15, and Ecg14 populations are the ones that have expressed this susceptibility variation the most, while the Ecg13 population was the least susceptible to the dose variation; this is quantified by the inflection point around GR50 (parameter b of the regression) (Table 3).
The Ecg06, Ecg07, Ecg14, and Ecg15 populations presented an RF of 6.91 to 13.08 compared to Ecg12, and their GR80 was above the dose of use (1080 [g a.i./ha]) between 1.5 and 2.5 times. Ecg13 and Ecg16 populations presented an RF of 1.25 to 1.45 with respect to Ecg12, in agreement with their GR80, which was well below the previously considered use dose.

3.2.3. Echinochloa oryzoides

GR50 was the highest in the populations Eo02, Eo03, Eo07, and Eo08 (Table 4 and Figure 3), while it was intermediate in the Eo04 population. Evaluating the susceptibility to the herbicide glyphosate in the dose–response trials in the different populations, it was determined that Eo02, Eo03, Eo07, and Eo08 were significantly different from population Eo06 (considered susceptible) (Table 4).
The regression parameters also indicate that the populations Eo02, Eo03, Eo07, and Eo08 are the populations that have most expressed this variation in susceptibility, while Eo06 was less susceptible to the variation in dose, as quantified by the point of inflection around e (parameter b of the regression) (Table 4).
Unlike the other species evaluated, in E. oryzoides, all populations presented an RF from 2.28 to 14.49 with respect to Eo06, and their GR80 was above the dose of use (1080 [g a.i./ha]) between 1.5 and 12 times. The RF values in this species are much higher compared to all other species.

3.2.4. Echinochloa chacoensis

The GR50 differs significantly between the populations, being the highest in the population Ech02 (Table 5 and Figure 4), while it was intermediate in the Ech03 population.
Evaluating the susceptibility to glyphosate in the dose–response trials in the different populations, it was determined that Ech02 and Ech03 were significantly different from the population Ech01 (considered susceptible). This specie presented an RF of 1.53 to 2.70 compared to Ech01, but its GR80 was well below the dose of use (1080 [g a.i./ha]), between three and four times the same.

3.3. Shikimic Acid

The accumulation of shikimic acid in the nine populations of E. colona in leaves after the application of 600 [g a.i./ha] of glyphosate was quantified in [mg] of shikimic acid per gram of leaf. Additionally, 33% of them increased the level of shikimic acid as time went by, as can be seen in Figure 5. Populations Ec03, Ec12, and Ec16 showed an increase at 48 h post application, respectively, reaching maximum values of 2.50 and 2.60 [mg] shikimic acid/[g] leaf at 96 h post application.
Carrying out particular contrasts, it was found that the populations Ec03, Ec12, and Ec16 accumulate values significantly different from the rest of the times after 48 h (p = 0.0003 for 48 h and p > 0.0001 for 72 and 96 h).
In populations Ec02, Ec04, Ec05, Ec06, and Ec07, these increases were not significant and did not exceed 0.50 [mg] of shikimic acid per gram of leaf.
In the populations of E. crus-galli, 33% of them (Ecg12, Ecg13, and Ecg16) increased the levels of shikimic acid after 48 h, as can be seen in Figure 6. In the populations Ecg06, Ecg07, Ecg14, and Ecg15, these increases were not significant and did not exceed 0.50 [mg] of shikimic acid per gram of leaf. While Ecg12, Ecg13, and Ecg16 inhibited their metabolic pathway and reached maximum values of 1.59, 2.30, and 2.53 [mg] shikimic acid/[g] leaf, respectively, at 96 h post application.
Carrying out particular contrasts, it was found that the Ecg13 and Ecg16 populations accumulate shikimic acid after 48 h, presenting values significantly different from the rest of the time (p = 0.0037; p = 0.0009; and p > 0.0001 for 48, 72, and 96 h, respectively).
E. oryzoides presents only one population (Eo06) that increases shikimic acid levels above 0.50 [mg/g] of leaf after 96 h (Figure 7), while populations Eo02, Eo03, Eo04, Eo07, and Eo08; these increases were not significant. The metabolic pathway of the Eo06 population reaches its maximum value of 2.12 [mg] shikimic acid/[g] leaf at 96 h post application.
The contrasts confirm that the Eo06 population accumulates shikimic acid only after 96 h, presenting values significantly different from the rest of the times (p > 0.0001).
In E. chacoensis, the 3 evaluated populations increase the levels of shikimic acid above 0.50 [mg/g] of leaf after 72 h (Figure 8), but the Ech02 population decreases at 96 h below the value considered as a limit. The Ech01 and Ech03 populations and their metabolic pathways reach their maximum value of [mg] shikimic acid/[g] leaf at 96 h post application.
The contrasts prove that the populations Ech01 and Ech03 accumulate shikimic acid only after 96 h, presenting values that are significantly different from the rest of the time (p > 0.0001).

4. Discussion

The survival tests carried out with the different populations of Echinochloa analyzed show differences between them. The analyzed samples were divided into three categories according to the percentage of survival observed [22] with the discriminant dose of 600 [g a.i./ha].
In the evaluated populations of the genus Echinochloa, thirteen of them presented high survival and can be considered resistant; another group made up of two populations presented with developing resistance; and the remaining ten populations presented survival values of less than 5%, which makes them susceptible.
The survival values found in this work for E. colona are similar to those of Alarcón-Reverte et al. [28] or Alarcón-Reverte et al. [29], with values ranging from 4 to 80% in populations of E. colona from California, although in this experiment the discriminating dose was 620 [g a.i./ha]. In populations from southern and southwestern Australia, Asaduzzaman et al. [30] found that in eighteen populations of E. colona exposed to a glyphosate dose of 540 [g a.i./ha], two of them survived at greater than 80% and 87%, respectively.
Regarding the dose–response experiments, differences in susceptibility to the herbicide glyphosate can be seen in all populations of the four species studied.
Based on the estimated resistance factor (RF), which is taken as a reference value to discriminate between resistant and susceptible populations [31], in the populations of E. colona Ec04, Ec07, and Ec14, they presented an RF greater than 2; therefore, they are considered populations resistant to glyphosate and the rest of the populations (Ec02, Ec03, Ec05, Ec12, and Ec16) as susceptible.
Studies carried out on E. colona by various authors in other countries [28,32,33] confirm that the population used has differences in susceptibility to this herbicide; however, the values of RF and GR50 calculated vary between the different authors.
Gaines et al. [32] in Australia, reported an RF 5.6 times higher than the susceptible population. Nguyen et al. [34], in another region of Australia, determined an RF of 2 to 12 times higher compared to the susceptible biotype. In Greece, Travlos et al. [35] obtained populations with an RF that was between 3.6 and 12.4. Closer to the values found in this work in E. colona are those found in the populations of California (USA), analyzed by Morran et al. [33], who found RF between 2.1 and 6.3. In other populations of southwestern and southern Australia, Asaduzzaman et al. [30] found populations that were 2.5 to 5 times more resistant than their corresponding susceptible phenotypes. In other populations of northern Australia, Mahajan et al. [36] evaluated populations that showed RF values between 6.7 and 15.1 in relation to susceptible populations.
The E. colona population with the highest RF in this study required 610 [g a.i./ha], similar to California populations that required 736 [g a.i./ha], but far from the 1116, 1289, 1187, and 3064 [g a.i./ha] required by populations in southern and southeastern Australia to reduce GR50. They were also very different from populations from northern Australia, with 1086 to 2339 [g a.i./ha] or 1220 [g a.i./ha] in the case of E. colona populations from Greece.
In E. crus-galli, the resistant populations (whose RF is greater than 2) are Ecg06, Ecg07, Ecg14, and Ecg15, whereas the susceptible populations are Ecg12, Ecg13, and Ecg16. In the populations declared resistant, the GR50 values of E. crus-galli are about three times those of E. colona, so it is important to recognize the species and act accordingly.
Experiences carried out in other countries show considerable evidence that these two species have the propensity to develop resistance to most groups of herbicides. Therefore, efforts to manage and control herbicide resistance in this species must be diligently continued [37].
In E. oryzoides, Eo06 is a population susceptible to glyphosate, and the rest (Eo02, Eo03, Eo04, Eo07, and Eo08) are resistant since their RFs are in a range of 2.28 to 14.49. In E. chacoensis, only the Ech02 population is resistant, while the others (Ech01 and Ech03) are susceptible.
In the work of Vázquez-García et al. [38], they classified susceptibility to glyphosate based on the RF value obtained, where populations with an RF value of less than 4 are susceptible, between 4 and 10 are resistant, and above 11 are very resistant. Using this classification, it is only of interest to determine the populations that meet the "very resistant" criterion, namely the Ecg15 population (RF of 13.089) in E. crus-galli and the Eo08 population (RF of 14.49) in E. oryzoides.
Shikimic acid accumulated after exposure to glyphosate (at a dose of 600 [gr a.i./ha]) in all susceptible populations of the different Echinochloa species. This accumulation suggests that glyphosate is successfully reaching the chloroplast and that at least part of the EPSPS enzyme present is sensitive to the herbicide. Here, differences were also found between the 4 species analyzed regarding the accumulation of shikimic acid over the time evaluated. In addition, it allows discrimination between the different levels of susceptibility to glyphosate in different populations.
In E. colona and E. chacoensis, 66% of the populations had their metabolic pathways inhibited (Ec2, Ec3, Ec5, Ec6, Ec12, Ec16, Ech01, and Ech03, respectively). In E. crus-galli, this percentage was 40% (Ecg12, Ecg13, and Ecg16), and in E. oryzoides, it was only 17% (Eo06). While in the rest of the populations, there was no inhibition of the metabolic pathway, which is consistent with what was explained by Singh and Shaner [26], who explained that the lack of accumulation of shikimate in plants treated with glyphosate can be taken as an indication that the plant is resistant to this herbicide.
The results obtained agree with what was expressed in the work of Alarcon-Reverte et al. [28] and Morran et al. [33], which also determined in E. colona that the accumulation of shikimic acid is greater in susceptible populations than in resistant ones. Such accumulation over time (24 h after application) in susceptible populations can lead to more than twice the concentration of shikimic acid than in resistant populations. At 96 h, these shikimic acid values can be up to fivefold.
Finally, if we take into account the classification between resistant and susceptible weeds that we described in the different experiments, we consider that a population is a biotype resistant to a herbicide when it has more than 20% survival, a resistance factor (RF) greater than 2, and does not have an accumulation of shikimic acid greater than 0.5 [mg/g] of a leaf.
The proportion of populations resistant to glyphosate within the genus Echinochloa clarifies the situation initially raised in the first reports of resistance [9]. Both the resistant populations and the less susceptible ones explain the current position as the fourth most important weed species in Argentina. A change in the strategy for the control of the species of the genus will be necessary for good agronomic management.

5. Conclusions

Summarizing the three tests carried out in this work, the following populations are considered resistant to glyphosate: Ec04, Ec07, and Ec14 of E. colona, Ecg06, Ecg07, Ecg14, and Ecg15 of E. crus-galli, Eo02, Eo03, Eo04, Eo07, and Eo08 of E. oryzoides, and Ech02 of E. chacoensis.
The populations susceptible to glyphosate are Ec02, Ec03, Ec05, Ec06, Ec12, and Ec16 for E. colona, Ecg12, Ecg13, and Ecg16 for E. crus-galli, only Eo06 for E. oryzoides, and Ech01, and Ech03 for E. chacoensis.

Author Contributions

Conceptualization: E.C. and I.D.; formal analysis: E.C., R.D.P. and I.D.; funding acquisition and project administration: I.D.; investigation: E.C., A.S., E.P. and M.P.; methodology: E.P. and I.D.; writing—original draft, E.C., R.D.P. and I.D.; writing—review and editing, E.C., A.S., E.P., R.D.P., M.P. and ID. All authors have read and agreed to the published version of the manuscript.

Funding

This research was funded by project PIBAA 2022-2023.

Conflicts of Interest

The authors declare no conflict of interest.

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Figure 1. Dose response curves corresponding to the different populations of Echinochloa colona. Each point is the average of three experiments. These curves were calculated using the model proposed by Streibig and Kudsk [27].
Figure 1. Dose response curves corresponding to the different populations of Echinochloa colona. Each point is the average of three experiments. These curves were calculated using the model proposed by Streibig and Kudsk [27].
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Figure 2. Dose–response curves corresponding to the different populations of Echinochloa crus-galli. Each point is the average of three experiments. These curves were calculated using the model proposed by Streibig and Kudsk [27].
Figure 2. Dose–response curves corresponding to the different populations of Echinochloa crus-galli. Each point is the average of three experiments. These curves were calculated using the model proposed by Streibig and Kudsk [27].
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Figure 3. Dose–response curves corresponding to the different populations of Echinochloa oryzoides. Each point is the average of three experiments. These curves were calculated using the model proposed by Streibig and Kudsk [27].
Figure 3. Dose–response curves corresponding to the different populations of Echinochloa oryzoides. Each point is the average of three experiments. These curves were calculated using the model proposed by Streibig and Kudsk [27].
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Agronomy 13 01219 g004 550
Figure 4. Dose response curves corresponding to the different populations of Echinochloa chacoensis. Each point is the average of three experiments. These curves were calculated using the model proposed by Streibig and Kudsk [27].
Figure 4. Dose response curves corresponding to the different populations of Echinochloa chacoensis. Each point is the average of three experiments. These curves were calculated using the model proposed by Streibig and Kudsk [27].
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Figure 5. Shikimic acid accumulation from 24 to 96 h after glyphosate application on E. colona plants.
Figure 5. Shikimic acid accumulation from 24 to 96 h after glyphosate application on E. colona plants.
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Figure 6. Shikimic acid accumulation from 24 to 96 h after glyphosate application on E. crus-galli plants.
Figure 6. Shikimic acid accumulation from 24 to 96 h after glyphosate application on E. crus-galli plants.
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Figure 7. Shikimic acid accumulation from 24 to 96 h after glyphosate application on E. oryzoides plants.
Figure 7. Shikimic acid accumulation from 24 to 96 h after glyphosate application on E. oryzoides plants.
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Figure 8. Shikimic acid accumulation from 24 to 96 h after glyphosate application on E. chacoensis plants.
Figure 8. Shikimic acid accumulation from 24 to 96 h after glyphosate application on E. chacoensis plants.
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Table
Table 1. Populations of the genus Echinochloa used.
Table 1. Populations of the genus Echinochloa used.
PopulationSpeciesCrop in Which it Was FoundLocation (Town)ProvinceGPS LocationYears of
Agriculture		Date of Harvest
Ec02E. colonaSoybean-CornManfrediCórdoba−63.76988
−31.84076		202015
Ec03E. colonaSoybean-CornColonia MarinaCórdoba−62.41800
−31.21785		202017
Ec04E. colonaSoybean-CornFreyreCórdoba−62.0710
−31.17283		202015
Ec05E. colonaSoybean-CornLuxardoCórdoba−62.25967
−31.27033		102015
Ec06E. colonaSoybean-CornEstación LuxardoCórdoba62.09588
−31.31240		202017
Ec07E. colonaSoybean-CornColonia CastelarSanta Fe62.13270
−31.65483		202015
Ec12E. colonaSoybean-CornSan FranciscoCórdoba−62.18506
−31.39578		202016
Ec14E. colonaSoybean-CornAriasCórdoba−62.45239
−33.62968		152017
Ec16E. colonaSoybean-CornEl ParaísoBuenos Aires−59.96448
−33.58911		152017
Ecg06E. crus-galliSoybean-CornColonia MasciasSanta Fe−60.00472
−30.78352		102016
Ecg07E. crus-galliSoybean-CornSaladero CabalSanta Fe−60.05598
−30.88797		102016
Ecg12E. crus-galliSoybeanSanta AnitaEntre Ríos−58.86091
−32.15184		102016
Ecg13E. crus-galliRiceVilla ElisaEntre Ríos−58.38793
−32.17088		152016
Ecg14E. crus-galliCornChajaríEntre Ríos−57.95368
−30.76185		152016
Ecg15E. crus-galliSoybeanSan SalvadorEntre Ríos−58.54736
−31.64929		152016
Ecg16E. crus-galliNot agriculturalC. del UruguayEntre Ríos−58.24531
−32.42552		----2016
Eo02E. oryzoidesSoybean-CornColonia MascíasSanta Fe−60.02201
−30.83483		152015
Eo03E. oryzoidesNot agriculturalColonia MascíasSanta Fe−59.99370
−30.83492		-----2015
Eo04E. oryzoidesSoybean-CornColonia MascíasSanta Fe−60.10391
−30.75559		152016
Eo06E. oryzoidesRiceColonia MascíasSanta Fe−60.1063
−30.79600		152016
Eo07E. oryzoidesSoybean-CornColonia MascíasSanta Fe−60.1063
−30.67708		152016
Eo08E. oryzoidesSoybean-CornColonia MascíasSanta Fe−60.20002
−30.81921		152016
Ech01E. chacoensisSoybean-CornColonia MascíasSanta Fe−59.99849
−30.83808		102016
Ech02E. chacoensisSoybean-CornColonia MascíasSanta Fe−60.12071
−30.8313		102016
Ech03E. chacoensisNot agriculturalColonia MascíasSanta Fe−60.08638
−30.7233		102016
Table
Table 2. Logistic adjusted model parameters (3 parameters, GR50 as parameter and GR80 estimated); the doses required a reduction of 50% and 80% of the fresh weight, respectively, parameter e corresponds to the GR50 parameters c, d, and b that were defined in formula (1); fixed lower limit at 0.
Table 2. Logistic adjusted model parameters (3 parameters, GR50 as parameter and GR80 estimated); the doses required a reduction of 50% and 80% of the fresh weight, respectively, parameter e corresponds to the GR50 parameters c, d, and b that were defined in formula (1); fixed lower limit at 0.
PopulationbdGR50seRF 1p-Value *GR80se
Ec022.5855851.065524224.27623.8251.350.0218383.3961.27
Ec033.7263381.008724165.99318.1601.00-240.8026.87
Ec0410.744250.746329610.12330.6273.68<0.0001694.1578.72
Ec052.5697050.957845292.05340.4731.76<0.0001500.9076.89
Ec065.1473920.933955300.98319.1481.81<0.0001394.0138.05
Ec0724.221640.871925385.12239.3612.32<0.0001407.8122.27
Ec123.2940160.984736175.34923.1881.060.7428267.1029.37
Ec145.7083620.926718608.01724.1873.66<0.0001775.1547.18
Ec161.6353260.985857187.77737.3131.130.5632438.3384.83
1 RF: resistance factor. * The value of p is the level of probability of significance for each factor for the model. The Lack of Fit test indicates that the regression accurately describes the data in the table (p = 0.0177).
Table
Table 3. Parameters of the logistic adjusted model (3 parameters, GR50 as parameter and GR80 estimated); the doses required for a reduction of 50% and 80% of the fresh weight, respectively, parameter e corresponds to the GR50 parameters c, d, and b that were defined in formula (1); fixed lower limit at 0.
Table 3. Parameters of the logistic adjusted model (3 parameters, GR50 as parameter and GR80 estimated); the doses required for a reduction of 50% and 80% of the fresh weight, respectively, parameter e corresponds to the GR50 parameters c, d, and b that were defined in formula (1); fixed lower limit at 0.
PopulationbdGR50seRF 1p-Value *GR80se
Ecg062.65791.137884.881138.4866.91<0.00011491.045359.492
Ecg071.59593.014984.309241.8057.72<0.00012347.048600.342
Ecg122.40597.351127.40820.4511.00-226.7744.102
Ecg1317.32987.430185.437127.8371.450.62128200.88111.082
Ecg141.77698.2691039.178172.7558.15<0.00012267.936524.427
Ecg154.07097.4051667.596190.31313.09<0.00012344.365615.555
Ecg164.145102.580160.41017.1141.260.18881224.11929.17
1 RF: resistance factor. * The value of p is the level of probability of significance for each factor for the model. The Lack of Fit test indicates that the regression accurately describes the data in the table (p = 0.586).
Table
Table 4. Parameters of the logistic adjusted model for Echinochloa oryzoides (3 parameters, GR50 as parameter and GR80 estimated); the doses required for a reduction of 50% and 80% of the fresh weight, respectively, parameter e corresponds to the GR50 parameters c, d, and b that were defined in formula (1); fixed lower limit at 0.
Table 4. Parameters of the logistic adjusted model for Echinochloa oryzoides (3 parameters, GR50 as parameter and GR80 estimated); the doses required for a reduction of 50% and 80% of the fresh weight, respectively, parameter e corresponds to the GR50 parameters c, d, and b that were defined in formula (1); fixed lower limit at 0.
PopulationbdGR50seRF 1p-Value *GR80se
Eo021.40095.4493.006.63615.2448.530.00038090.581.798.65
Eo030.98898.0073.312.02856.8059.390.002313463.873.820.10
Eo041.984112.915804.93117.4472.280.00451619.15450.23
Eo064.295104.756352.5445.9431-486.8383.65
Eo071.28296.9553.070.23981.3198.710.000019055.233.455.12
Eo081.48896.6585.107.501.050.11514.490.0000112969.333.078.01
1 RF: resistance factor. * The value of p is the level of probability of significance for each factor for the model. The Lack of Fit test indicates that the regression accurately describes the data in the table (p = 0.586).
Table
Table 5. Parameters of the logistic adjusted model for Echinochloa chacoensis (3 parameters, GR50 as parameter and GR80 estimated); the doses required for a reduction of 50% and 80% of the fresh weight, respectively, parameter e corresponds to the GR50 parameters c, d, and b that were defined in formula (1); fixed lower limit at 0.
Table 5. Parameters of the logistic adjusted model for Echinochloa chacoensis (3 parameters, GR50 as parameter and GR80 estimated); the doses required for a reduction of 50% and 80% of the fresh weight, respectively, parameter e corresponds to the GR50 parameters c, d, and b that were defined in formula (1); fixed lower limit at 0.
PopulationbdGR50seRF 1p-Value *GR80se
Ech011.70899.4599.6713.131.00-224.42031.045
Ech024.00486.91269.3621.082.70<0.0001380.78539.791
Ech032.62299.93152.4412.631.530.00101258.62327.930
1 RF: resistance factor. * The value of p is the level of probability of significance for each factor for the model. The Lack of Fit test indicates that the regression accurately describes the data in the table.
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Cortés, E.; Schneider, A.; Panigo, E.; Perreta, M.; De Prado, R.; Dellaferrera, I. First Report of Resistance to Glyphosate in Several Species of the Genus Echinochloa in Argentina. Agronomy 2023, 13, 1219. https://doi.org/10.3390/agronomy13051219

AMA Style

Cortés E, Schneider A, Panigo E, Perreta M, De Prado R, Dellaferrera I. First Report of Resistance to Glyphosate in Several Species of the Genus Echinochloa in Argentina. Agronomy. 2023; 13(5):1219. https://doi.org/10.3390/agronomy13051219

Chicago/Turabian Style

Cortés, Eduardo, Ana Schneider, Elisa Panigo, Mariel Perreta, Rafael De Prado, and Ignacio Dellaferrera. 2023. "First Report of Resistance to Glyphosate in Several Species of the Genus Echinochloa in Argentina" Agronomy 13, no. 5: 1219. https://doi.org/10.3390/agronomy13051219

APA Style

Cortés, E., Schneider, A., Panigo, E., Perreta, M., De Prado, R., & Dellaferrera, I. (2023). First Report of Resistance to Glyphosate in Several Species of the Genus Echinochloa in Argentina. Agronomy, 13(5), 1219. https://doi.org/10.3390/agronomy13051219

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