Volume 4, Issue 4, Pages 688-696 (July 2011) Phytochromes Regulate SA and JA Signaling Pathways in Rice and Are Required for Developmentally Controlled Resistance to Magnaporthe grisea  Xie Xian-Zhi , Xue Yan-Jiu , Zhou Jin-Jun , Zhang Bin , Chang Hong , Takano Makoto   Molecular Plant  Volume 4, Issue 4, Pages 688-696 (July 2011) DOI: 10.1093/mp/ssr005 Copyright © 2011 The Authors. All rights reserved. Terms and Conditions

Figure 1 The Number and Distribution of Lesions on Leaves of Rice Plants Inoculated with M. grisea. The Y-axis represents the number of susceptible-type lesions per square centimeter on leaves of the phyAphyBphyC mutant (filled bars) and wild-type (open bars) plants. Averages of at least 10 individual plants with standard errors are shown. The phyAphyBphyC mutant and wild-type rice plants were grown in a greenhouse (28°C day/23°C night) with natural light during August in Tsukuba, Ibaraki, Japan. Similar results were obtained when the plants were grown in December and October (data not shown). Molecular Plant 2011 4, 688-696DOI: (10.1093/mp/ssr005) Copyright © 2011 The Authors. All rights reserved. Terms and Conditions

Figure 2 Accumulation of PR1 Proteins in M. grisea-Infected Leaves. (A) The diagram of time points for spraying blast fungus and harvesting leaves. The wild-type and the phyAphyBphyC plants at the five-leaf stage were sprayed with M. grisea (0 h). The old leaves (the second and third leaves) and new leaves (the fifth leaves) were separately harvested at 20, 24, 48, and 72 h after spraying. (B) Induction of PR1 proteins by M. grisea in the old leaves (OL) or new leaves (NL) of wild-type rice and the phyAphyBphyC mutant. Each lane was loaded with 10 μg protein, and PR1 proteins (PR1) were detected with an anti-tobacco acidic PR1 antibody. Non-specific bands (NS) are shown as loading control. M, Bio-Rad Precision Plus Protein Standards. Molecular Plant 2011 4, 688-696DOI: (10.1093/mp/ssr005) Copyright © 2011 The Authors. All rights reserved. Terms and Conditions

Figure 3 Accumulation of PR1 Proteins in Leaves Treated with JA. (A) The diagram of time points for JA treatment and leaf-harvesting. Wild-type and phyAphyBphyC plants were sprayed at the five-leaf stage with 100 μM cis-, trans-JA (0). Old leaves (the second and third leaves) and new leaves (the fifth leaves) were separately harvested at 24, 48, and 72 h after JA treatment. (B) Induction of PR1 proteins by exogenous JA in the old leaves (OL) or the new leaves (NL) of wild-type and phyAphyBphyC plants. Each lane was loaded with 10 μg protein and PR1 proteins (PR1) were detected with an anti-tobacco acidic PR1 antibody. Non-specific bands (NS) are shown as loading control. M, Bio-Rad Precision Plus Protein Standards. (C) Effect of light on the accumulation of PR1 proteins in the old leaves of wild-type plants treated with JA. The upper panel depicts the dark/light regime for JA treatment and leaf-harvesting. The wild-type plants at the five-leaf stage were sprayed with 100 μM cis-, trans-JA (0). Sprayed plants then were transferred to continuous darkness (dark) for 24 h or grown in light (light) for another 8 h until entering the dark period in the growth chamber. Old leaves were harvested at 8, 24, 48, and 72 h after JA treatment. The lower panel depicts the induction of PR1 proteins by exogenous JA in the old leaves of wild-type plants. Each lane was loaded with 10 μg protein and PR1 proteins (PR1) were detected with an anti-tobacco acidic PR1 antibody. Non-specific bands (NS) are shown as loading control. M, Bio-Rad Precision Plus Protein Standards. Molecular Plant 2011 4, 688-696DOI: (10.1093/mp/ssr005) Copyright © 2011 The Authors. All rights reserved. Terms and Conditions

Figure 4 Accumulation of PR1 Proteins in Leaves Treated with SA. (A) The diagram of time points for SA treatment and leaf-harvesting. Wild-type plant and the phyAphyBphyC mutants were sprayed with 400 mM SA (0). Old and new leaves were separately harvested at 24, 48, and 72 h after spraying with SA. (B) Induction of PR1 proteins by exogenous SA in old leaves (OL) or new leaves (NL) of wild-type plant and the phyAphyBphyC mutant. Each lane was loaded with 10 μg protein and PR1 proteins (PR1) were detected with an anti-tobacco acidic PR1 antibody. Non-specific bands (NS) are shown as loading control. M, Bio-Rad Precision Plus Protein Standards. Molecular Plant 2011 4, 688-696DOI: (10.1093/mp/ssr005) Copyright © 2011 The Authors. All rights reserved. Terms and Conditions

Figure 5 The Basal Transcript Levels of PR1a and PR1b Genes in the Wild-Type and the phyAphyBphyC Mutant. (A) Transcript levels of PR1a and PR1b genes analyzed by RT–PCR. A representative result is shown. (B) Signal intensities of PR1a or PR1b genes relative to ACTIN. The means ± SE obtained from three individual experiments are shown. Old leaves (OL) and new leaves (NL) were separately harvested from the wild-type and the phyAphyBphyC mutant at the five-leaf stage. Molecular Plant 2011 4, 688-696DOI: (10.1093/mp/ssr005) Copyright © 2011 The Authors. All rights reserved. Terms and Conditions

Figure 6 The Basal Transcript Levels of Genes Related to SA-or JA-Dependent Defense Pathways in the Wild-Type and the phyAphyBphyC Mutant. (A) Transcript levels of genes related to SA-or JA-dependent defense pathways as analyzed by RT–PCR. One representative result is shown. (B) Signal intensities of genes relative to ACTIN. The means ± SE obtained from three individual experiments are shown. Old leaves (OL) and new leaves (NL) were separately harvested from wild-type and the phyAphyBphyC mutant at the five-leaf stage. Molecular Plant 2011 4, 688-696DOI: (10.1093/mp/ssr005) Copyright © 2011 The Authors. All rights reserved. Terms and Conditions