FIGURE 3.
RNA-templated DNA Repair produced Herbicide-resistant Plants. (A) In OsALS Tryptophan (TGG) to leucine (TTG) at amino acid 548 (W548 L) showed high bispyribac sodium (BS) herbicide resistance. The transformed calli were regenerated with BS. The seedlings transformed with cis_anti_humscaff repair template have stronger resistance and are well-established. (B) DNA was extracted from young seedlings and mutations were detected by MfeI digestion. The plants with desired mutations showed digested bands and the control showed an MfeI-resistant undigested band. (C) These amplicons were cloned and Sanger sequenced. The seedlings with the cis_anti_humscaff repair template showed repair (green nucleotides). Red nucleotides show other Indels. (D) Seeds from cis_antis_humanscaff repair template BS-resistant plants were germinated on 1/2MS+0.75 μM BS and showed clear BS resistance. These resistance plants were Sanger sequenced and showed homozygous repair (Supplementary Figure 3).