1. Volume 26, Issue 14, Pages 1854-1860 (July 2016)
TOR Signaling Promotes Accumulation of BZR1 to Balance Growth with Carbon Availability in Arabidopsis 
Zhenzhen Zhang, Jia-Ying Zhu, Jeehee Roh, Chloé Marchive, Seong-Ki Kim, Christian Meyer, Yu Sun, Wenfei Wang, Zhi-Yong Wang 
Current Biology 
Volume 26, Issue 14, Pages (July 2016)
DOI: /j.cub
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2. Current Biology 2016 26, 1854-1860DOI: (10.1016/j.cub.2016.05.005)
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3. Figure 1
TOR Is Required for Sugar-Promoted Hypocotyl Elongation in the Dark
(A and B) Seedlings and hypocotyl measurement of wild-type (Ler) and gin2-1 seedlings grown on media containing 90 mM mannitol (M), glucose (G), or sucrose (S) for 4 days under light and then 3 days in darkness.
(C and D) Seedlings and hypocotyl measurement of the wild-type (Col-0), tor-es1, and tor-es2 grown in 90 mM mannitol (M) or sucrose (S) with 0 μM estradiol (−ES) or 10 μM estradiol (+ES).
Scale bars represent 1 cm. ∗∗p < 0.01 (Student’s t test). Error bars indicate the SEM (n ≥ 20). See also Figure S1.
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4. Figure 2 TOR Regulates Hypocotyl Elongation through the BR Pathway
(A) qPCR analysis of the mRNA levels of TOR and BZR1 target genes in tor-es treated with the indicated concentrations of estradiol. Seedlings were grown under light for 4 days and then in the dark for 3 days. Error bars indicate the SD (two independent experiments).
(B) qRT-PCR analysis of the expression levels of BZR1 target genes in seedlings grown under light on sugar-free medium (−S) or medium containing 90 mM sucrose (+S) and then either maintained under light (0 hr) or put in the dark for 4 or 12 hr.
(C–F) The wild-type and tor-es were grown on 1/5 Murashige and Skoog (MS) medium with or without 1 μM estradiol (ES) and different concentrations of BL (C and D) or indole-3-acetic acid (IAA) (E and F) in the dark for 6 days.
(G–H) Seedlings of indicated genotypes were grown for 4 days under light on medium containing 90 mM sucrose and then transferred to medium with 0 μM estradiol (−ES) or 10 μM estradiol (+ES) and grown in the dark for 3 days.
(D, F, and H) The ratio between hypocotyl lengths of estradiol-treated and mock-treated seedlings shown in (C), (E), and (G), respectively. n ≥ 20.
Scale bars represent 1 cm. ∗p < 0.05 (Student’s t test). Error bars indicate the SEM (three independent experiments), except in (A). See also Figure S2.
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5. Figure 3
Sugar-TOR Signaling Stabilizes BZR1 Protein in Light-Grown Plants Shifted into Prolonged Darkness
(A and B) Immunoblot analysis of BZR1-CFP protein in seedlings that were grown on medium containing 90 mM mannitol (A) or sucrose (B) under constant light (Light) and then shifted into dark (Dark) for 12 or 24 hr or that after 24 hr in the dark were then treated under light for 12 or 24 hr (Re-light). The upper and lower bands indicate phosphorylated and dephosphorylated BZR1, respectively.
(C) Immunoblot analysis of BZR1-CFP protein after seedlings were grown on sugar-free medium for 10 days and then treated in dark or light for 24 hr; the plants in the dark were then treated in liquid medium with or without 90 mM sucrose for the indicated time.
(D) Immunoblot of BZR1-CFP or bzr1-1D-CFP protein in 10-day-old seedlings grown on sugar-free medium after dark treatment for the indicated time.
(E) Immunoblot of BZR1-CFP or bzr1-1D-CFP in light-grown seedlings on medium containing 90 mM sucrose and 0 μM (−) or 1 μM (+) AZD8055 (AZD) and then treated with darkness for the indicated time.
(F) Light-grown BZR1-CFP or BZR1-CFP/tor-es plants were treated with sucrose-containing medium supplemented with 0 μM estradiol (−ES) or 10 μM estradiol (+ES) in darkness for 12 or 24 hr and then analyzed by immunoblot with Anti-GFP antibody. Histone H3 (H3) was probed as a loading control. CFP/H3, the ratio between the CFP (GFP) and histone H3 signals.
(G) Wild-type (WT), oxTOR (7817), or oxTOR (G548) seedlings were grown on medium without sugar under light for 10 days and then in the dark for the indicated time. BZR1 protein was analyzed by immunoblot with anti-BZR1 antibody. ∗NS, nonspecific bands; BZR1/NS, the ratio between BZR1 and nonspecific signals.
See also Figures S3 and S4 and Table S1.
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6. Figure 4
Loss of TOR Induces BZR1 Degradation through an Autophagy-Dependent and Proteasome-Independent Mechanism
(A and B) BZR1-CFP/tor-es plants were grown on medium containing 90 mM sucrose and 0 μM estradiol (−ES) or 10 μM estradiol (+ES) and then treated with 5 mM 3-MA (A) or 30 μM MG132 (B) in the dark for 12 hr. BZR1-CFP was analyzed by immunoblot with anti-GFP antibody. Histone H3 (H3) or Ponceau S staining was used as a loading control.
(C) The wild-type (Col) and cop1 and bzr1-1D mutants were grown on sugar-free medium under light for 10 days and then kept under light (L-12) or treated by darkness (D-12) for 12 hr. The immunoblot was probed by anti-BZR1 antibody. BZR1/Staining, the ratio between the BZR1 signal and staining.
(D) A model of balancing BR-dependent growth with sugar availability. Sugar activation of TOR inhibits autophagy and stabilizes BZR1, whereas BR signaling activates BZR1 by dephosphorylation, leading to cell elongation. Sugar also promotes BR biosynthesis, probably through TOR, in the dark. The black arrows and bar indicate previously reported mechanisms; the blue arrows indicate findings made in this study.
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