Fig. 3

Ectopic expression of OsPUB41 induces callose deposition, leads to enhanced expression of JA biosynthesis and response genes, but does not affect expression of SA biosynthesis and response genes in transgenic Arabidopsis lines. Leaves of thirty-days-old OsPUB41 transgenic Arabidopsis plants were infiltrated either with estradiol (inducer) or with DMSO (uninduced). After 12 h, leaves were stained with aniline blue and observed under an epifluorescence microscope. Bright spots in the images represent callose deposits (a). Scale bar represents 50 μm. The graph represents average number of callose deposits per field of view (0.075 mm²) from five-six leaves with ten different fields viewed per leaf in each experiment (b). Error bars represent standard error. Student’s two-tailed t-test for independent means was performed to test for significance (p < 0.05, represented by*). Similar results were obtained in three independent experiments (per transgenic line) and in three independent transgenic lines. Transcript levels of JA (c) and SA (d), biosynthetic and response genes, were measured upon ectopic expression of OsPUB41 in Arabidopsis. AOS: Allene Oxide Synthase (JA biosynthetic gene), PDF1.2a: Plant Defensin, VSP: Vegetative Storage Protein and JAZ: Jasmonate ZIM-Domain (JA response genes). SID2: SA Induction-Deficient 2 (SA biosynthetic gene), PAL2: Phenylalanine Ammonia-Lyase 2 (SA biosynthetic gene), NPR1: Nonexpresser of PR1 (SA response gene), PR1: Pathogenesis Related gene 1 (SA response gene) and PR5: Pathogenesis Related gene 5 (SA response gene). AtUbq5 was used as an internal control in qPCR. Three biological repeats were performed for each independent transgenic line. Similar results were obtained in three independent transgenic lines. Student’s two-tailed t-test for independent means was performed on delta C_t values to test for significance (p < 0.05)