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Journal: Proceedings of the National Academy of Sciences of the United States of America
Article Title: Gibberellin-deactivating GA2OX enzymes act as a hub for auxin–gibberellin cross talk in Arabidopsis thaliana root growth regulation
doi: 10.1073/pnas.2425574122
Figure Lengend Snippet: Auxin signaling directly regulates GA2OX genes. ( A ) Root tips of nlsGPS2 and ga2ox heptuple nlsGPS2 treated for 5 h with 50 nM IAA or mock. ( B ) Quantification of the nlsGPS2 emission ratio corresponding to GA levels along the longitudinal axis of nlsGPS2 _line1 and ga2ox heptuple nlsGPS2 _line1 treated for 5 h with 50 nM IAA or mock. n ≥ 10. ( C ) Quantification of nlsGPS2 emission ratio in the meristematic (MZ) or elongation zone (EZ) of nlsGPS2 _line1 and ga2ox heptuple nlsGPS2 _line1 treated for 5 h with 50 nM IAA or mock n ≥ 10. ( D ) Growth rate of ga2ox heptuple and Col-0 treated with 5 nM IAA for 12 h. Normalized to the mock-treated growth rate. n ≥ 12. ( E ) Growth rate of pG1090::XVE ≫ mCherry-GA2OX8 and Col-0 treated with 5 nM IAA for 12 h. Normalized to the mock-treated growth rate. All seedlings grown on estradiol. n ≥ 9. ( F ) Growth rate of ga2ox heptuple and Col-0 treated with 1.5 μM L-Kyn and 10 μM PEO-IAA for 4 h. n ≥ 14. ( G ) Quantification of YFP-NLS along the longitudinal axis of pGA2OX8::YFP-NLS roots in plants treated for 5 h with 100 μM IAA/mock on the shoot. n ≥ 5. ( H ) Quantification of YFP-NLS along the longitudinal axis of pGA2OX8::YFP-NLS roots 16h after decapitation. n ≥ 6. ( I ) Root tips of pGA2OX8::YFP-NLS pretreated for 24 h with 1.5 μM L-kyn or mock and subsequently treated for 5 h with 100 μM IAA/mock on the shoot. The EZ is indicated by a white line. Quantification of YFP-NLS signal intensity in the EZ. n ≥ 6. ( J ) Root tips of pGA2OX8::mCherry-GA2OX8 pretreated for 24 h with 1.5 μM L-kyn or mock and subsequently treated for 5 h with 100 μM IAA/mock on the shoot. Quantification of mCherry signal intensity in the EZ. The outlines of the root are highlighted. n ≥ 15. ( K ) Root tips of pGA2OX6::mVenus-GA2OX6 pretreated for 24 h with 1.5 μM L-kyn or mock and subsequently treated for 5 h with 100 μM IAA/mock on the shoot. Quantification of mVenus signal intensity in the EZ. The outlines of the root are highlighted. n ≥ 5. ( L ) Quantification of nlsGPS2 emission ratio of nuclei in the MZ or EZ of nlsGPS2 line treated for 24 h with 1.5 μM L-kyn and subsequently treated for 5 h with 100 μM IAA/mock on the shoot. ( M ) A simplified scheme summarizing auxin-modulated regulation of GA2OX genes in the root EZ. The asterisks indicate statistically significant differences based on one-way ANOVA followed by Tukey HSD or Kruskal–Wallis test followed by post hoc Dunn’s test ( C ) (* P < 0.05, ** P < 0.01, and *** P < 0.001). Error bars in boxplots are CI and in line graphs are SD. The EZ in A , I , J , and K is indicated by a white line. (Scale bar, 50 μm.)
Article Snippet: Estradiol (Sigma, 20 mM stock in DMSO, 2,5 μM working concentration), Indole Acetic Acid (IAA, Sigma, 10 mM stock in 96% ethanol),
Techniques: