Occurrence and expression of ellarinacin-like BGCs in Brachypodium and wheat ancestral species. (A) Wheat ellarinacin BGC is conserved in wheat wild ancestors A. tauschii and wild emmer wheat (T. turgidum subsp. dicoccoides), and homologous to a BGC identified on chromosome 3 of B. distachyon. Gray lines link between wheat and B. distachyon BlastP reciprocal best hits. (B) Semiquantitative RT-PCR of B. distachyon Chr.3 clustered genes. ntc, no template control; 2s, seedling shoot (2 d old); 2r, seedling root; 14st, young plant stem base (14 d old); 14l, young plant leaf; 60sp, mature plant spike (60 d old); 60r, mature plant root; 60l, mature plant leaf; 60st, mature plant stem base. (C) Quantitative real-time PCR of brachynacin BGC genes in B. distachyon plants infected with FHB or FRR. Con, control (noninfected); dpi, days post infection. Relative quantification values (in fold change) indicate means of three biological replicates ± SEM. Asterisks denote t test statistical significance of differential expression. *P < 0.05, **P < 0.01, ***P < 0.001.