Extended Data Fig. 8: Generating and characterizing multiple transgenic plant lines for dual circuit plants.
From: An orthogonalized PYR1-based CID module with reprogrammable ligand-binding specificity
(a) Ligand-mediated circuit activation in dual sensor Arabidopsis plants. PYR1DIAZI was incorporated into plants harboring the PYR1*AZIN-GFP inducible circuit in response to azinphos-ethyl by floral transformation, obtaining transgenic lines harboring both circuits (L1 and L2). (left) Five day-old seedlings were treated with either mock, diazinon, azinphos-ethyl, and combined diazinon and azinphos-ethyl, each at (500 nM). Seedlings were imaged 48 hours post-treatment (images all corrected for brightness at 40%). (right) Double senor lines were tested for diazinon-mediated activation of the ABA pathway by germination assays in media containing ligands. Germination was recorded after three days. (b) Ligand-mediated circuit activation in dual sensor Arabidopsis plants. PYR1DIAZI was incorporated into plants harboring the PYR1*AZIN-RUBY inducible by crossing and then identifying the homozygous double circuit line in the F2. (left) Seeds were germinated on ½ MS media containing either mock, diazinon, azinphos-ethyl, and combined diazinon and azinphos-ethyl, each at (500 nM) for five days. (right) Double senor lines were tested for diazinon-mediated activation of the ABA pathway by germination assays in media containing ligands. Germination was recorded after three days. Data points represent the % germination from approximately 100 seeds.
