Extended Data Fig. 1: Generation and analysis of FolSvp2 transformants.
From: Plant PR1 rescues condensation of the plastid iron-sulfur protein by a fungal effector
a, Expression profile of FolSvp2 in Fol with or without two-week-old tomato root incubation. The data are presented as the means ± SEs of three independent replicates. b, Genomic DNA was analyzed via PCR to determine which strains were harboring GFP, FolSvp2-GFP, or a K205 mutant (FolSvp2Q-GFP and FolSvp2R-GFP) under the RP27 constitutive promoter. The Histone H4 (FOXG_09402) was used as a positive PCR control. c, Secretion of FolSvp2 in the presence or absence of tomato roots. Conidia of strains harboring FolSvp2-GFP were inoculated into 5% liquid YEPD in the presence or absence of tomato roots. Forty hours after inoculation, proteins were extracted from mycelia or culture supernatants and probed with α-GFP and α-Actin. Coomassie brilliant blue (CBB) or silver staining was used to determine protein loading in each lane. d, Functional validation of the secreted signal peptide (SP) of FolSvp2. The yeast strains were cultured on CMD-W or YPRAA medium for 3 days. TTC was added to the culture supernatant to measure the enzymatic activity for reducing TTC to the red TPF. e, Secretion of FolSvp2 proteins in the presence of tomato roots. Protein extraction and identification were performed as described in (c). f, Homologous recombination-based deletion of FolSvp2. g, PCR analysis to identify the ∆FolSvp2 (KO1, KO9, KO26 and KO37) knockout and wild-type (WT) strains with the primer pairs FolSvp2-out-F/R, FolSvp2-in-F/R and H4-qRT-F/R. h, PCR analysis to identify the WT, ∆FolSvp2, complementation (∆FolSvp2-C) and K205 mutant (∆FolSvp2-CQ, ∆FolSvp2-CR) strains with the native promoter. The primer pairs FolSvp2-pQB-F/R, FolSvp2-pQB-F/pYF11-cGFP-R, FolSvp2-out-F/R and H4-qRT-F/R were used as described above. i, Mycelial growth of the WT, ∆FolSvp2 (∆FolSvp2-KO1), ∆FolSvp2-C, ∆FolSvp2-CQ and ∆FolSvp2-CR mutant strains on the PDA media, complete media (CM), and minimal media (MM) after 5 days of cultivation. Scale bars = 2 cm. j, Mycelial diameter and conidial production of the indicated strains in the indicated media or at the indicated times. No significant difference (ns) was indicated according to multiple comparison tests (mean ± SEM, n = 3). The experiments were performed three times with similar results obtained.
