Extended Data Fig. 8: Stigmatic NO is required for ROS reduction in A. thaliana stigmas during SC and UC.
From: Stigma receptors control intraspecies and interspecies barriers in Brassicaceae
a, b, Confocal and wide-field imaging showing the increase of stigmatic NO at 5 MAP with SC pollen (a) and quantified data of SC-induced NO and pollen hydration in A. thaliana stigmas, both showed NO peaking at ~5 MAP (b). See Fig. 4a. c, d, FER-dependent elevation of NO in A. thaliana stigmas induced by pollen and At-PCP-Bγ .See Fig. 4d, e. e, AtPCP-Bγ was much faster than GST-BrPCP-B3 in increasing NO of A. thaliana roots. f, GST-BrPCP-B3 was more effective in increasing NO of B. rapa roots than that of A. thaliana roots. g, Roots of fer-4 was not responsive to AtPCP-Bγ in inducing NO. h, i, (Left to right) cPTIO scavenged SC-induced NO, suppressed SC-induced ROS reduction, and inhibited SC pollen hydration and tube growth in A. thaliana stigmas. j–n, Relative to WT, noa1 stigmas showed lower NO and higher ROS levels, and slower hydration of SC (WT A. thaliana) pollen and pollen tube growth (j, k, l). Relative to WT, hot5-4 stigmas showed faster NO and lower ROS levels, and faster hydration of SC (WT A. thaliana) pollen and pollen tube growth (j, m, n). o, p, Mutations in NOA1, noa1 (o) and GSNOR, hot5-4 (p), enhanced and reduced, respectively, the reproductive barrier to B. rapa or B. oleracea pollen in A. thaliana stigmas at 1.5 HAP. Scale bars, 200 μm (a, h, j, o, p) and roots (e–g); 50 μm for pollen (h, j). Box plots (a–p): centre line, median; box limits, lower and upper quartiles; dots, individual data points; whiskers, highest and lowest data points. n (in blue), number of stigmas, pollen grains or roots. P values, two-tailed t-tests. Each experiment was repeated at least thrice with consistent results.
