Figure 3

AVR-Pita₁₇₆ is an elicitor. (A) AVR-Pita polypeptides tested in the transient assay. The white region indicates the putative secretory signal sequence, the gray region indicates the putative pro-protein domain and the hatched region indicates the putative protease motif. The black region indicates the putative mature protein. The number of amino acids missing from the N-terminus is indicated. GUS activity is indicated by '+', whereas decreased GUS activity is indicated by '-'. (B) Representative rice seedlings showing GUS activity. Two-leaf Pi-ta (Yashiro-mochi and YT14) and pi-ta (Nipponbare and YT16) seedlings were co-bombarded with 35S/Adh1-6::AVR-Pita₁₇₆ and 35S::uidA. Leaves were assayed histochemically for GUS activity and cleared in 70% ethanol to visualize GUS staining. (C) RNA gel blot analysis of AVR-Pita expression in the transient assay. YT14 (Pi-ta) and YT16 (pi-ta) were co-bombarded with the 35S/Adh1-6::AVR-Pita₁₇₆ and 35S::uidA plasmids. Leaf tissue was harvested 2 days after bombardment. Poly(A)⁺ mRNA was then extracted, blotted to Hybond-N and hybridized with a radiolabeled AVR-Pita₁₇₆ probe. The AVR-Pita transcript is indicated. Similar loading was verified before blotting by visualizing mRNA in the gel stained with ethidium bromide.