Fig. 1: Identification At3g49150/AFF1 for ARF19 hypercondensation.
From: Regulation of AUXIN RESPONSE FACTOR condensation and nucleo-cytoplasmic partitioning
a EMS-mutagenized M2 seeds of arf19-1 35 S:YFP-ARF19 were screened for individuals with increased numbers of YFP-ARF19 condensates using a fluorescence dissecting microscope. Isolate DH8 was backcrossed to the parental line (arf19-1 35 S:YFP-ARF19) and the resultant F2 individuals displaying YFP-ARF19 hypercondensation identified and used for whole-genome sequencing of bulk segregants. b Confocal images of 3d-old Wt (Col-0) and DH8 (aff1-1) seedlings carrying 35 S:YFP-ARF19 (false-colored yellow) with cell walls counterstained with propidium iodide (false-colored magenta). Scale bar = 25 µm. c Confocal images of 3d-old wild type (Wt; Col-0) and aff1-1 seedlings carrying pARF19:ARF19-mVenus, or pARF7:ARF7-YFP (false-colored yellow) with cell walls counterstained with propidium iodide (false-colored magenta). Scale bar = 25 µm. d Time course confocal images showing fusion of condensates in root transition zone cells of 3d-old Wt (Col-0) and aff1-1 seedlings carrying UBQ10:YFP-ARF19. Scale bar = 5 µm. e Condensate circularity measurements after condensate fusion events in 3d-old Wt (Col-0) and aff1-1 seedlings carrying UBQ10:YFP-ARF19 (mean ± SD; n = 20). f Half-condensate FRAP recovery profiles after photobleaching condensates from 3d-old Wt (Col-0) and aff1-1 seedlings in root tip or upper root region carrying UBQ10:YFP-ARF19 (mean ± SD; n = 20). g Map positions of homozygous EMS-caused mutations identified in DH8 bulk segregants. h At3g49150/AFF1 schematic depicting the exons (blue), UTRs (gray), and introns (black). Locations of the aff1-1 point mutation and aff1–2 (Salk_053818), aff1–3 (Salk_083453), and aff1–4 (Sail_427_G06) insertion sites are indicated. AFF1 encodes a putative F-box protein with an N-terminal F-box domain, leucine rich repeat (LRR) region, and C-terminal F-box domain (FBD) motif. Three independent experiments were performed for (b), (c) and (d) with similar results. The source data for (e) and (f) are provided as a Source Data files.
