2.5. DNA extraction and molecular studies
DNA was extracted from fresh leaf tissue (∼200 mg) of threeweekold plants with the Puregene DNA isolation kit (Gentra,Minnesota, USA) according to the manufacturer’s instructions.Known primers were used to sequence the psbA gene . Forthe ACCase gene, primers were designed using known sequenceof L. rigidum (DQ184640.1; Supplementary Table S7). Genes wereamplified and PCR products were sequenced to locate the commonpoint mutations that can endow TS resistance. Sequence analysesand alignment were performed using Bioedit software . Theobtained sequences were compared to the known psbA and ACCasesequences of Arabidopsis thaliana and B. distachyon.