Journal: The Plant Journal
Article Title: Redox buffering and H 2 O 2 orchestrate the vegetative development of Marchantia polymorpha
doi: 10.1111/tpj.70317
Figure Lengend Snippet: Functional characterization of HyPer7 as H 2 O 2 sensor in 3 DAG gemmae. (A) Plate reader measurements of HyPer7 fluorescence intensity in 3 DAG gemmae. After 10 min, gemmae were submersed in medium with different H 2 O 2 concentrations (0.25–10 mM) for 10 min, followed by recovery measurements for 100 min. Error bars represent standard deviation, calculated from n ≥ 6. Values were normalized to the 488/405 ratio of the first time point. (B) Microscopic images of HyPer7 fluorescence in 3 DAG transgenic gemmae exposed to the H 2 O 2 series. The rainbow color scheme depicts the detected HyPer7 ratio 488/405 range from low (blue) to high (red) H 2 O 2 concentrations. Ratiometric images visualize spectral changes in response to increasing H 2 O 2 concentrations ( n > 7). Scale bars, overview: 100 μm, insets: 50 μm. Images are maximum intensity Z ‐stack projections processed using RRA software. Insets represent single slice images of corresponding overview Z ‐stack projections. Arrows indicate higher oxidized meristematic zones, disappearing after 2 mM treatments. (C) HyPer7 ratios from the meristematic region (mr) and surrounding differentiated tissue (dt) were determined using representative images ( n = 7) shown in (B). (D) Phenotypic analysis of WT plants after cultivation for 3 days, followed by 10 min H₂O₂ treatment and recovery for 4 days. Plants treated with 10 mM H₂O₂ were non‐viable and excluded from further analysis. Scale bar: 500 μm. (E) Quantitative assessment of the optical surface area of 7‐day‐old WT plants after H₂O₂ treatments ( n > 35). Data are represented as mean ± standard deviation. Statistical significance was determined using the single ANOVA test followed by Tukey ( P < 0.05).
Article Snippet: A ratiometric analysis was performed by calculating the 405/488 nm ratio for each pixel using the Matlab‐based program Redox Ratio Analysis (RRA), resulting in a false color ratiometric image according to Fricker ( ).
Techniques: Functional Assay, Fluorescence, Standard Deviation, Transgenic Assay, Software
MathWorks Inc is a verified supplier 
