Divergence in Glyphosate Susceptibility between Steinchisma laxum Populations Involves a Pro106Ser Mutation.

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.

Convergent Adaptation of Multiple Herbicide Resistance to Auxin Mimics and ALS- and EPSPS-Inhibitors in Brassica rapa from North and South America.

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.

Reduced Glyphosate Movement and Mutation of the EPSPS Gene (Pro106Ser) Endow Resistance in Conyza canadensis Harvested in Mexico.

Glyphosate has been the most widely used herbicide for decades providing a unique tool, alone or in mixtures, to control weeds on citrus in Veracruz. Conyza canadensis has developed glyphosate resistance for the first time in Mexico. The level and mechanisms of resistance of four resistant populations Rs (R1, R2, R3, and R4) were studied and compared with that of a susceptible population (S). Resistance factor levels showed two moderately resistant populations (R2 and R3) and two highly resistant populations (R1 and R4). Glyphosate translocation through leaves to roots was ∼2.8 times higher in the S population than in the four R populations. A mutation (Pro106Ser) in the EPSPS2 gene was identified in the R1 and R4 populations. Mutation in the target site associated with reduced translocation is involved in increased glyphosate resistance in the R1 and R4 populations; whereas for the R2 and R3 populations, it was only mediated by reduced translocation. This is the first study of glyphosate resistance in C. canadensis from Mexico in which the resistance mechanisms involved are described in detail and control alternatives are proposed.

Non-Target-Site Resistance Mechanisms Endow Multiple Herbicide Resistance to Five Mechanisms of Action in Conyza bonariensis.

The repeated use of herbicides can lead to the selection of multiple resistance weeds. Some populations of Conyza bonariensis occurring in olive groves from southern Spain have developed resistance to various herbicides. This study determined the resistance levels to 2,4-D, glyphosate, diflufenican, paraquat, and tribenuron-methyl in a putative resistant (R) C. bonariensis population, and the possible non-target-site resistance (NTSR) mechanisms involved were characterized. Resistance factors varied as follows: glyphosate (8.9), 2,4-D (4.8), diflufenican (5.0), tribenuron-methyl (19.6), and paraquat (85.5). Absorption of 14C-glyphosate was up to 25% higher in the susceptible (S) population compared to the R one, but 14C-paraquat absorption was similar (up to 70%) in both populations. S plants translocated more than 60% of both 14C-glyphosate and 14C-paraquat toward shoots and roots, while R plants translocated less than 10%. The R population was able to metabolize 57% of the 2,4-D into nontoxic metabolites and 68% of the tribenuron-methyl into metsulfuron-methyl (10%), metsulfuron-methyl-hydroxylate (18%), and conjugate-metsulfuron-methyl (40%). Among the NTSR mechanisms investigated, absorption and translocation could be involved in glyphosate resistance, but only translocation for paraquat. Proofs of the presence of enhanced metabolism as a resistance mechanism were found for tribenuron-methyl and 2,4-D, but not for diflufenican. This research informs the first occurrence of multiple resistance to five herbicide classes (acetolactate synthase inhibitors, 5-enolpyruvylshikimate-3-phosphate synthase inhibitors, photosystem I electron diverters, photosystem II inhibitors, and synthetic auxin herbicides) in C. bonariensis.

Non-Target Site Mechanisms Endow Resistance to Glyphosate in Saltmarsh Aster (Aster squamatus).

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.

Glyphosate ban in Mexico: potential impacts on agriculture and weed management.

Since glyphosate was classified as potentially carcinogenic by the International Agency for Research on Cancer, public debate regarding the environmental impact and health risks from its use has intensified. Almost all regulatory agencies throughout the world have concluded that the judicious use of glyphosate does not pose risks to the environment and human health. However, on the last day of 2020 the Mexican government decreed a ban of this herbicide beginning January, 2024. In current Mexican agriculture there are no safer chemical and/or other weed management technologies that allow for the economical substitution of glyphosate for weed control. Many Mexican weed scientists agree that glyphosate use should be reduced, but not banned outright. This decree could have more negative economic and social consequences as well as environmental and human health risks than benefits, which could compromise the country's food and public security. Crop yields are projected by some to decline by up to 40% with this ban, increasing food prices, making food less accessible to low-income consumers. In addition, a black market for the smuggling and illegal sale of glyphosate is possible. The possible environmental, economic and social impacts caused by the glyphosate ban in Mexico are discussed, emphasizing the impact on weed management. © 2021 Society of Chemical Industry.

Management of Glyphosate-Resistant Weeds in Mexican Citrus Groves: Chemical Alternatives and Economic Viability.

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.

Characterization of three glyphosate resistant Parthenium hysterophorus populations collected in citrus groves from Mexico.

Continuous use of glyphosate in citrus groves in the Gulf of Mexico region has selected for resistant Parthenium hysterophorus L. populations. In this study, the target-site and non-target-site resistance mechanisms were characterized in three putative glyphosate-resistant (GR) P. hysterophorus populations, collected in citrus groves from Acateno, Puebla (GR1 and GR2) and Martínez de la Torre, Veracruz (GR3), and compared with a susceptible population (GS). Based on plant mortality, the GR populations were 9.2-17.3 times more resistant to glyphosate than the GS population. The low shikimate accumulation in the GR population confirmed this resistance. Based on plant mortality and shikimate accumulation, the GR3 population showed intermediate resistance to glyphosate. The GR populations absorbed 15-28% less 14C-glyphosate than the GS population (78.7% absorbed from the applied) and retained 48.7-70.7% of 14C-glyphosate in the treated leaf, while the GS population translocated ~68% of absorbed herbicide to shoots and roots. The GR3 population showed the lowest translocation and absorption rates, but was found to be susceptible at the target site level. The 5-enolpyruvylshikimate-3-phosphate synthase (EPSPS) gene sequence of the GR1 and GR2 populations showed the Pro106-Ser mutation, conferring 19- and 25-times more resistance in comparison to the GS population, respectively. Reduced absorption and impaired translocation conferred glyphosate resistance on the GR3 population, and contributed partially to the resistance of the GR1 and GR2 populations. Additionally, the Pro-106-Ser mutation increased the glyphosate resistance of the last two P. hysterophorus populations.

From tolerance to resistance: mechanisms governing the differential response to glyphosate in Chloris barbata.

BACKGROUND: Susceptibility and the mechanism (s) governing tolerance/resistance to glyphosate were characterized in two putative-glyphosate-resistant Chloris barbata populations (R1 and R2), collected in Persian lime orchards from Colima State, Mexico, comparing them with one non-treated population (referred to as S). RESULTS: Glyphosate doses required to reduce fresh weight or cause mortality by 50% were 4.2-6.4 times higher in resistant populations than in the S population. The S population accumulated 4.3 and 5.2 times more shikimate than the R2 and R1 populations, respectively. There were no differences in 14 C-glyphosate uptake between R and S populations, but the R plants translocated at least 12% less herbicide to the rest of plant and roots 96 h after treatment. Insignificant amounts of glyphosate were metabolized to aminomethyl phosphonate and glyoxylate in both R and S plants. The 5-enolpyruvylshikimate-3-phosphate synthase gene of the R populations contained the Pro106-Ser mutation, giving them a resistance 12 (R2) and 14.7 (R1) times greater at target-site level compared with the S population. CONCLUSION: The Pro106-Ser mutation governs the resistance to glyphosate of the R1 and R2 C barbata populations, but the impaired translocation could contribute to the resistance. These results confirm the first case of glyphosate resistance evolved in this species. © 2018 Society of Chemical Industry.

First confirmation and characterization of target and non-target site resistance to glyphosate in Palmer amaranth (Amaranthus palmeri) from Mexico.

Following the introduction of glyphosate-resistant (GR)-cotton crops in Mexico, farmers have relied upon glyphosate as being the only herbicide for in-season weed control. Continuous use of glyphosate within the same year and over multiple successive years has resulted in the selection of glyphosate resistance in Palmer amaranth (Amarantus palmeri). Dose-response assays confirmed resistance in seven different accessions. The resistance ratio based on GR50 values (50% growth reduction) varied between 12 and 83. At 1000 µM glyphosate, shikimic acid accumulation in the S-accession was 30- to 2-fold higher at compared to R-accessions. At 96 h after treatment, 35-44% and 61% of applied 14C-glyphosate was taken up by leaves of plants from R- and S-accessions, respectively. At this time, a significantly higher proportion of the glyphosate absorbed remained in the treated leaf of R-plants (55-69%) compared to S-plants (36%). Glyphosate metabolism was low and did not differ between resistant and susceptible plants. Glyphosate was differentially metabolized to AMPA and glyoxylate in plants of R- and S-accessions, although it was low in both accessions (<10%). There were differences in 5-enolpyruvylshikimate-3-phosphate synthase (EPSPS) enzyme activity by 50% (I50) between R- and S-accessions. However, no significant differences were found in the basal EPSPS activity (µmol inorganic phosphate µg-1 total soluble protein min-1) between R- and S-accessions. A point mutation Pro-106-Ser was evidenced in three accessions. The results confirmed the resistance of Palmer amaranth accessions to glyphosate collected from GR-cotton crops from Mexico. This is the first study demonstrating glyphosate-resistance in Palmer amaranth from Mexico.

Physiological, morphological and biochemical studies of glyphosate tolerance in Mexican Cologania (Cologania broussonetii (Balb.) DC.).

In recent years, glyphosate-tolerant legumes have been used as cover crops for weed management in tropical areas of Mexico. Mexican cologania (Cologania broussonetii (Balb.) DC.) is an innate glyphosate-tolerant legume with a potential as a cover crop in temperate areas of the country. In this work, glyphosate tolerance was characterized in two Mexican cologania (a treated (T) and an untreated (UT)) populations as being representatives of the species, compared in turn to a glyphosate-susceptible hairy fleabane (S) (Conyza bonariensis (L.) Cronq.) population. Experiments revealed that T and UT Mexican cologania populations had a higher tolerance index (TI), and a lower shikimic acid accumulation and foliar retention than the hairy fleabane S population. Absorption and translocation, leaf morphology and metabolism studies were only carried out in the Mexican cologania T population and the hairy fleabane S population. The latter absorbed 37% more (14)C-glyphosate compared to the Mexican cologania T at 96 h after treatment (HAT). Mexican cologania T translocated less herbicide from the treated leaf to the remainder of the plant than hairy fleabane S. The Mexican cologania T presented a greater epicuticular wax coverage percentage than the hairy fleabane S. This morphological characteristic contributed to the low glyphosate absorption observed in the Mexican cologania. In addition, the Mexican cologania T metabolized glyphosate mainly into AMPA, formaldehyde and sarcosine. These results indicate that the high glyphosate tolerance observed in Mexican cologania is mainly due to the poor penetration and translocation of glyphosate into the active site, and the high glyphosate degradation into non-toxic substances.