Extended Data Fig. 5: De-esterified pectin induces FER-dependent NO production.
From: FERONIA controls pectin- and nitric oxide-mediated male–female interaction
a, Exudates from pollinated wild-type, but not fer-4, pistils induced NO at the filiform apparatus region (arrows) of wild-type ovules. b, Sonicated PGA (100 μg) electrophoresed in a 26% polyacrylamide gel47. Lanes represent fractions eluted from a diethylaminoethyl column by the indicated salt gradient. The last fraction approximated PGA before sonication. Oligogalacturonic-acid-sized25 fragments would be ahead of the RuR-stained materials on the basis of comparison with a previous publication47. Active species for ovular NO induction remains to be determined. c, Representative quantification of PGA-induced NO response based on filiform apparatus:synergid cell signal ratio, as described in Extended Data Fig. 3a. Data plots summarize the quantification data (left dot plot) and data from direct scoring (right histogram), showing comparable results. Data are average ± s.d., n, number of ovules. d, cPTIO suppressed PGA induced accumulation of NO at the filiform apparatus. cPTIO was added together with the DAF-DA dye. Data are average ± s.d. n, number of pistils. e, PGA treatment of wild-type and fer-4 ovules showed NO in wild-type (arrow), but not fer-4, filiform apparatus. f, PGA treatment of wild-type, fer-4 and FER–GFP-complemented fer-4 seedling roots showed FER dependence for PGA-induced root NO. Under equal image acquisition conditions (but without added DAF-DA), FER–GFP signal was negligible (last data bar). Signals were quantified from equal areas of interest (white box). Data are average ± s.d., n, number of seedlings. P values were obtained by two-tailed t-tests. Scale bars, 50 μm. All observations were representative of three independent experiments. Box plots: centre line, median; box limits, lower and upper quartiles; dots, individual data points; whiskers, highest and lowest data points.
