Constitutive overexpression of EPSPS by gene duplication is involved in glyphosate resistance in Salsola tragus.

BACKGROUND: Glyphosate-resistant Salsola tragus accessions have been identified in the USA and Argentina; however, the mechanisms of glyphosate resistance have not been elucidated. The goal of this study was to determine the mechanism/s of glyphosate resistance involved in two S. tragus populations (R1 and R2) from Argentina. RESULTS: Both glyphosate-resistant populations had a six-fold lower sensitivity to glyphosate than the S population (i.e. resistance index). No evidence of differential absorption, translocation or metabolism of glyphosate was found in the R1 and R2 populations compared to a susceptible population (S). No 5-enolpyruvylshikimate-3-phosphate synthase (EPSPS) mutations were detected, but S. tragus R1 and R2 plants had ≈14-fold higher EPSPS gene relative copy number compared to the S counterpart. In R1 and R2, EPSPS duplication entailed a greater constitutive EPSPS transcript abundance by approximately seven-fold and a basal EPSPS activity approximately three-fold higher than the S population. CONCLUSION: The current study reports EPSPS gene duplication for the first time as a mechanism of glyphosate resistance in S. tragus populations. The increase of glyphosate dose needed to kill R1 and R2 plants was linked to the EPSPS transcript abundance and level of EPSPS activity. This evidence supports the convergent evolution of the overexpression of the EPSPS gene in several Chenopodiaceae/Amaranthaceae species adapted to drought environments and the role of gene duplication as an adaptive advantage for plants to withstand stress. © 2022 Society of Chemical Industry.

A novel EPSPS Pro-106-His mutation confers the first case of glyphosate resistance in Digitaria sanguinalis.

BACKGROUND: Digitaria sanguinalis has been identified as a species at high risk of evolving herbicide resistance, but thus far, there are no records of resistance to glyphosate. This weed is one of the most common weeds of summer crops in extensive cropping areas in Argentina. It shows an extended period of seedling emergence with several overlapping cohorts during spring and summer, and is commonly controlled with glyphosate. However, a D. sanguinalis population was implicated as a putative glyphosate-resistant biotype based on poor control at recommended glyphosate doses. RESULTS: The field-collected D. sanguinalis population (Dgs R) from the Rolling Pampas has evolved glyphosate resistance. Differences in plant survival and shikimate levels after field-recommended and higher glyphosate doses were evident between Dgs R and the known susceptible (Dgs S) population; the resistance index was 5.1. No evidence of differential glyphosate absorption, translocation, metabolism or basal EPSPS activity was found between Dgs S and Dgs R populations; however, a novel EPSPS Pro-106-His point substitution is probably the primary glyphosate resistance-endowing mechanism. EPSPS in vitro enzymatic activity demonstrated that an 80-fold higher concentration of glyphosate is required in Dgs R to achieve similar EPSPS activity inhibition to that in the Dgs S population. CONCLUSION: This study reports the first global case of glyphosate resistance in D. sanguinalis. This unlikely yet novel transversion at the second position of the EPSPS 106 codon demonstrates the intensity of glyphosate pressure in selecting unexpected glyphosate resistance alleles if they retain EPSPS functionality. © 2022 Society of Chemical Industry.

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.

Genetic Background and Inbreeding Depression in Romosinuano Cattle Breed in Mexico.

The ultimate goal of genetic selection is to improve genetic progress by increasing favorable alleles in the population. However, with selection, homozygosity, and potentially harmful recessive alleles can accumulate, deteriorating genetic variability and hampering continued genetic progress. Such potential adverse side effects of selection are of particular interest in populations with a small effective population size like the Romosinuano beef cattle in Mexico. The objective of this study was to evaluate the genetic background and inbreeding depression in Mexican Romosinuano cattle using pedigree and genomic information. Inbreeding was estimated using pedigree (FPED) and genomic information based on the genomic relationship matrix (FGRM) and runs of homozygosity (FROH) of different length classes. Linkage disequilibrium (LD) was evaluated using the correlation between pairs of loci, and the effective population size (Ne) was calculated based on LD and pedigree information. The pedigree file consisted of 4875 animals born between 1950 and 2019, of which 71 had genotypes. LD decreased with the increase in distance between markers, and Ne estimated using genomic information decreased from 610 to 72 animals (from 109 to 1 generation ago), the Ne estimated using pedigree information was 86.44. The reduction in effective population size implies the existence of genetic bottlenecks and the decline of genetic diversity due to the intensive use of few individuals as parents of the next generations. The number of runs of homozygosity per animal ranged between 18 and 102 segments with an average of 55. The shortest and longest segments were 1.0 and 36.0 Mb long, respectively, reflecting ancient and recent inbreeding. The average inbreeding was 2.98 ± 2.81, 2.98 ± 4.01, and 7.28 ± 3.68% for FPED, FGRM, and FROH, respectively. The correlation between FPED and FGRM was -0.25, and the correlations among FPED and FROH of different length classes were low (from 0.16 to 0.31). The correlations between FGRM and FROH of different length classes were moderate (from 0.44 to 0.58), indicating better agreement. A 1% increase in population inbreeding decreased birth weight by 0.103 kg and weaning weight by 0.685 kg. A strategy such as optimum genetic contributions to maximize selection response and manage the long-term genetic variability and inbreeding could lead to more sustainable breeding programs for the Mexican Romosinuano beef cattle breed.

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.