title: EPSPS variability, gene expression, and enzymatic activity in glyphosate-resistant biotypes of Digitaria insularis.
abstract: Weed resistance to herbicides is a natural phenomenon that exerts selection on individuals in a population. In Brazil, glyphosate resistance was recently detected in Digitaria insularis. The objective of this study was to elucidate mechanisms of weed resistance in this plant, including genetic variability, allelism, amino acid substitutions, gene expression, and enzymatic activity levels. Most of these have not previously been studied in this species. D. insularis DNA sequences were used to analyze genetic variability. cDNA from resistant and susceptible plants was used to identify mutations, alleles, and 5-enolpyruvylshikimate-3-phosphate synthase (EPSPS) expression, using real-time quantitative reverse transcription-polymerase chain reaction. In addition, EPSPS activity was measured. We found a decrease in genetic variability between populations related to glyphosate application. Substitutions from proline to threonine and tyrosine to cysteine led to a decrease in EPSPS affinity for the glyphosate. In addition, the EPSPS enzymatic activity was slightly higher in resistant plants, whereas EPSPS gene expression was almost identical in both biotypes, suggesting feedback regulation at different levels. To conclude, our results suggest new molecular mechanisms used by D. insularis to increase glyphosate resistance.
authors.affiliation: Departamento de Ciências Biológicas, Escola Superior de Agricultura "Luiz de Queiroz", Universidade de São Paulo, Piracicaba, SP, Brasil estebangg18@hotmail.com.
authors.forename: E
authors.initials: E
authors.lastname: Galeano
authors.affiliation: Departamento de Ciências Biológicas, Escola Superior de Agricultura "Luiz de Queiroz", Universidade de São Paulo, Piracicaba, SP, Brasil.
authors.forename: A A M
authors.initials: AA
authors.lastname: Barroso
authors.affiliation: Departamento de Ciências Biológicas, Escola Superior de Agricultura "Luiz de Queiroz", Universidade de São Paulo, Piracicaba, SP, Brasil.
authors.forename: T S
authors.initials: TS
authors.lastname: Vasconcelos
authors.affiliation: Grupo Investigación Ciencias Forenses y Salud, Tecnológico de Antioquia Institución Universitaria, Medellín, Antioquia, Colombia.
authors.forename: A
authors.initials: A
authors.lastname: López-Rubio
authors.affiliation: Departamento de Ciências Biológicas, Escola Superior de Agricultura "Luiz de Queiroz", Universidade de São Paulo, Piracicaba, SP, Brasil.
authors.forename: A J P
authors.initials: AJ
authors.lastname: Albrecht
authors.affiliation: Departamento de Ciências Biológicas, Escola Superior de Agricultura "Luiz de Queiroz", Universidade de São Paulo, Piracicaba, SP, Brasil.
authors.forename: R
authors.initials: R
authors.lastname: Victoria Filho
authors.affiliation: Departamento de Ciências Biológicas, Escola Superior de Agricultura "Luiz de Queiroz", Universidade de São Paulo, Piracicaba, SP, Brasil.
authors.forename: H
authors.initials: H
authors.lastname: Carrer
mesh_terms: D051229:3-Phosphoshikimate 1-Carboxyvinyltransferase / Q000235:genetics* / Q000378:metabolism; D019943:Amino Acid Substitution; D031746:Digitaria / Q000187:drug effects / Q000201:enzymology* / Q000235:genetics; D015870:Gene Expression; D005998:Glycine / Q000031:analogs & derivatives* / Q000494:pharmacology; D053209:Herbicide Resistance; D006540:Herbicides / Q000494:pharmacology*; D010802:Phylogeny; D010940:Plant Proteins / Q000235:genetics* / Q000378:metabolism; D011110:Polymorphism, Genetic; D017422:Sequence Analysis, DNA; D000097797:Glyphosate
chemical_list: D051229:3-Phosphoshikimate 1-Carboxyvinyltransferase; D005998:Glycine; D006540:Herbicides; D010940:Plant Proteins; D000097797:Glyphosate