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Fig. S1AtGLK1 and AtGLK2 redundantly regulate the Hyaloperonospora arabidopsidis (Hpa) Noco2 resistance in Arabidopsis. One-week-old seedlings were inoculated with 1 × 105/mL Hpa Noco2 spores. At 7 days post-infection (dpi), the spores were quantified from three samples of 25–30 seedlings per sample. The data from one representative experiment are shown and the error bars represent the standard error (SE). Similar trends were observed in three independent biological replicates.

Fig. S2 Polymerase chain reaction (PCR) genotyping of the glk1 glk2 NahG plant. (A) The top gel shows the presence of the dSpm insertion in the glk1.2 mutation detected with the 2bgs2 + Spm5 primer pair at 58 °C after 35 cycles of PCR, and the bottom gel shows the presence of the wild-type (WT) allele for AtGLK1 detected with the GLK1-Promoter-F1 + AtGLK1-R primer pair at 58 °C after 35 cycles of PCR to confirm the homozygous status of the glk1.2 mutation. The asterisk (*) represents nonspecific PCR products. (B) The top gel shows the presence of the dSpm insertion in the glk2.1 mutation detected with the Spm1 + AtGLK2-R primer pair at 62 °C + gradient primer annealing after 35 cycles of PCR, and the bottom gel shows the presence of the WT allele for AtGLK2 detected with the AtGLK2-F + AtGLK2-R primer pair at 62 °C + gradient primer annealing with 35 cycles of PCR to confirm the homozygous status of the glk2.1 mutation. PCR genotyping of glk1.2 and glk2.1 mutations was performed according to Fitter et al. (2002). (C) The presence of the NahG transgene was confirmed with the NahG-F + NahG-R primer pair at 58 °C after 35 cycles of PCR generating a 700-bp product. Aliquots of 5 μL of PCR products from each genotype were separated on a 2% (w/v) agarose gel.

Fig. S3 Polymerase chain reaction (PCR) genotyping of the glk1 glk2 npr1-1 triple mutant. (A) The top gel shows the presence of the dSpm insertion in glk1.2 detected with the 2bgs2 + Spm5 primer pair at 58 °C after 35 cycles of PCR, and the bottom gel shows the presence of the wild-type (WT) allele for AtGLK1 detected with the GLK1-Promoter-F1 + AtGLK1-R primer pair at 58 °C after 35 cycles of PCR to confirm the homozygous status of the glk1.2 mutation. The asterisk (*) indicates nonspecific PCR products. (B) The top gel shows the presence of the dSpm insertion in glk2.1 detected with the Spm1 + AtGLK2-R primer pair at 62 °C + gradient primer annealing with 35 cycles of PCR, and the bottom gel shows the presence of the WT allele for AtGLK2 detected with the AtGLK2-F + AtGLK2-R primer pair at 62 °C + gradient primer annealing after 35 cycles of PCR to confirm the homozygous status of the glk2.1 mutation. Aliquots of 5 μL of PCR product from each genotype were separated on a 2% (w/v) agarose gel. PCR genotyping of glk1.2 and glk2.1 mutations was performed according to Fitter et al. (2002). (C) NPR1 was amplified using the npr1-1 F + npr1-1R dCAPs primer pair at 58 °C and generated a 252-bp product, which was digested with NlaIII (NEB) and separated on a 3% agarose gel. The PCR product was digested in the WT of the NPR1 allele, but not in the npr1-1 mutant allele.

Fig. S4 Polymerase chain reaction (PCR) genotyping of the coi1-16 glk1 glk2 triple mutant. (A) The top gel shows the presence of the dSpm insertion in the glk1.2 mutation detected with the 2bgs2 + Spm5 primer pair at 58 °C primer annealing with 35 cycles of PCR, and the bottom gel represents the presence of the wild-type (WT) allele for GLK1 detected with the GLK1-Promoter-F1 + AtGLK1-R primer pair at 58 °C primer annealing with 35 cycles of PCR to confirm the homozygous status of the glk1.2 mutation. The asterisk (*) represents nonspecific PCR products. (B) The top gel shows the presence of the dSpm insertion in the glk2.1 mutation detected with the Spm1 + AtGLK2-R primer pair at 62 °C + gradient primer annealing with 35 cycles of PCR, and the bottom gel shows the presence of the WT allele for GLK2 detected with the AtGLK2-F + AtGLK2-R primer pair at 62 °C + gradient primer annealing with 35 cycles of PCR to confirm the homozygous status of the glk2.1 mutation. PCR genotyping of the glk1.2 and glk2.1 mutation was performed according to Fitter et al. (2002). (C) COI1 in the WT was detected using the coi1-16 P2 + coi1-16 P4 primer pair at 64 °C primer annealing with 26 cycles of PCR. As coi1-16 is a point mutation, the WT allele for COI1 also generated a PCR product with the coi1-16 P2 + coi1-16 P3 primer pair; however, the coi1-16 P2 + coi1-16 P4 primer pair generated a PCR product only in the WT allele and not in the coi1-16 mutant allele. PCR genotyping of the coi1-16 mutant was performed according to Adams and Turner (2010). Aliquots of 5 μL of PCR products from each genotype were separated on a 2% (w/v) agarose gel.

Fig. S5 Polymerase chain reaction (PCR) genotyping of the coi1-16 35S:AtGLK1 plant. (A) Presence of the genomic GLK1 and cDNA of the inserted AtGLK1 transgene. The GLK1 genomic fragment was detected using the AtGLK1-F + GLK1 genomic-R primer pair at 58 °C after 35 cycles of PCR. (B) Detection of the coi1-16 mutation using the coi1-16 P2 + coi1-16 P3 primer pair at 64 °C after 26 cycles of PCR. Aliquots of 5 mL of PCR products from each genotype were separated on a 2% (w/v) agarose gel. (C) COI1 detected in the wild-type (WT) using the coi1-16 P2 + coi1-16 P4 primer pair at 64 °C after 26 cycles of PCR. Aliquots of 5 mL of PCR products from each genotype were separated on a 2% (w/v) agarose gel. As coi1-16 is a point mutation, the WT allele for COI1 also generated a PCR product with the coi1-16 P2 + coi1-16 P3 primer pair. However, the coi1-16 P2 + coi1-16 P4 primer pair generated PCR products only in the WT, but not in the coi1-16 mutant alleles. PCR genotyping of the coi1-16 mutant was performed according to Adams and Turner (2010).

Table S1 Primer combinations used to genotype mutant and transgenic plants.

Table S2 List of quantitative reverse transcription-polymerase chain reaction (qRT-PCR) primers.

Table S3 Transcript levels of VEGETATIVE STORAGE PROTEIN 2 (VSP2), Thionin 2.1 (Thi2.1), PLANT DEFENSIN 1 and 2 (PDF1 and PDF2) after infection with Botrytis cinerea.

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