Fig. 2: Identification, characterization and functional validation of Pm13.

a Schematic representation of transcriptome sequencing analysis of EMS-induced loss-of-function mutants. The de novo assembly of the WT transcriptome was used as a reference. Powdery mildew inoculation of mutants was performed at the M₂ and M₃ generations (IT4). b The structure of the candidate gene of Pm13. Exons are represented as rectangular bars, and introns are shown as black lines between exons. Predicted protein domains are indicated by different colors: red for N-terminal domain of mixed lineage kinase domain-like protein (MLKL_NTD), blue for brace region, orange for serine/threonine kinase domain, gray for unidentified protein region, and black for 5’ UTR and 3’ UTR. Positions of EMS-type point mutations identified are indicated by short bars. c Schematic diagram of two constructs with the CDS of the Pm13 candidate gene MLKL-K driven by the maize Ubi promoter and its native promoter. These constructs were used to transiently transform epidermal cells of susceptible wheat cv. XM44 and stable transform susceptible wheat cv. Fielder, respectively. d The haustorium index of MLKL-K, Pm21, and the empty vector (EV) in transiently transformed cells challenged by isolate E09. Each red dot represents one bombarded leaf (MLKL-K: n = 17, Pm21: n = 17, and EV: n = 18). Haustorium index values are means ± SD from 17 to 18 biological replicates. Error bars represent standard deviations based on 17 to 18 biological replicates. P values were calculated with a two-tailed Student’s t-test. e Responses of seven independent T₀ transgenic lines expressing Pm13 to isolate BgtYZ01 at the adult-plant stage. Line No. 3778 and the recipient Fielder were used as the resistant and susceptible controls. f Responses of the seven corresponding independent T₁ transgenic families to Bgt isolate BgtYZ01 at the seedling stage. Symbols ‘+’ and ‘−’ represent siblings from each family with and without transformed MLKL-K, respectively. The experiment was performed with four plants (three transgene-positive and one negative) selected from ten T₁ seedlings of each family. Source data are provided as a Source Data file.