Fig. 4: A GA signaling activity map reveals high levels of GA signaling activity in inter-primordia cells in the SAM. | Nature Communications

Fig. 4: A GA signaling activity map reveals high levels of GA signaling activity in inter-primordia cells in the SAM.

From: A quantitative gibberellin signaling biosensor reveals a role for gibberellins in internode specification at the shoot apical meristem

Fig. 4

a Maximum intensity projection of three overlay channels showing the expression of pRPS5a::qmRGA (yellow for VENUS channel and blue for TagBFP channel) and pCLV3::mCherry-NLS (magenta) in the SAM. CZ, central zone; P, primordium. The subprint of P indicates the order of primordia with P1 being the youngest emerged primordium. b–e Maximum intensity projection of two overlay channels (b) showing the ratio expression of pRPS5a::qmRGA (yellow for VENUS channel and blue for TagBFP channel) and of three individual channels (c–e) showing the expression of mRGA-VENUS (c), TagBFP-NLS (d) and pCLV3::mCherry-NLS (e) in the SAM, separately. The experiment was repeated twice with similar results. f Heatmap representation of L1 GA signaling activity averaged from seven SAMs aligned using the CLV3 domain as a reference (see supplemental methods for more details). g, h Quantification of GA signaling activity in central zone (CZ, n = 699 nuclei examined over seven independent SAM samples), inter-primordia region (IPR, n = 1504 nuclei examined over seven independent SAM samples), and primordia (P, n = 1381 nuclei examined over seven independent SAM samples) indicated by different colors as shown in (g). P-values are from one-way ANOVA. i 3D visualization of nlsGPS1 emission ratio in the SAM. Primordia stages are estimated according to morphology. Arrows highlight a higher nlsGPS emission ratio in the boundaries and IPR. The experiment was repeated twice with similar results. j, k Fluorescence distribution in wild-type (Ler) SAMs treated with fluorescein (Fl, (j)) and GA4-Fl (k). l Comparison of the ratio of average fluorescence intensity in the IPR to that in the non-IPR (excluding primordia) between Fl (n = 4 independent plants) and GA4-Fl (n = 5 independent plants) treatment in the SAM. The P-value is from one-way ANOVA with Turkey’s test for multiple comparisons of means. The experiment was repeated twice with similar results. The center lines of boxes in (h, l) show the medians and box limits indicate the 25th and 75th percentiles. Whiskers indicate minima and maxima as determined using the R software. Scale bars = 20 μm.

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