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Brassinazole resistant 1 (BZR1)-dependent brassinosteroid signalling pathway leads to ectopic activation of quiescent cell division and suppresses columella stem cell differentiation.

Lee HS, Kim Y, Pham G, Kim JW, Song JH, Lee Y, Hwang YS, Roux SJ, Kim SH - J. Exp. Bot. (2015)

Bottom Line: BR promotes intense nuclear accumulation of BZR1 in the root tip area, and the binding of BZR1 to the promoters of several root development-regulating genes, modulating their expression in the root stem cell niche area.They could also inhibit auxin-dependent distal stem cell differentiation by antagonizing the auxin/WOX5-dependent pathway.In conclusion, BZR1-/BES1-mediated BR signalling pathways show differential effects on the maintenance of root apical meristem activities: they stimulate ectopic QC division while they show opposite effects on the differentiation of distal columella stem cells in a BR concentration- and BZR1-/BES1-dependent manner.

View Article: PubMed Central - PubMed

Affiliation: Division of Biological Science and Technology, Yonsei University, Wonju, 220-710, Republic of Korea.

No MeSH data available.


Related in: MedlinePlus

BR- and BZR1-mediated signalling pathway modulates transcriptional expression of diverse root-regulating genes. (A–C) Quantitative RT-PCR analysis of root-controlling genes in BL-treated Col-0 and BR-related mutant plants. Plants were grown and expression of root-regulating genes was analysed for short-term BL-treated plants (A), long-term BL-treated plants (B), and BL-related mutants (C) as described in Fig. 1G–I. (D) ChIP-qPCR assay of the BZR1- and BES1- binding cis-motif of root-regulating genes. BZR1- and BES1-bound chromatin DNA fragments were isolated from roots of week-old Col-0, pBZR1::BZR1-YFP, and p35S::BES1-GFP plants using anti-BZR1 antibodies. Next, quantitative PCR analysis was performed with the primer sets listed in Supplementary Table S2 as described in Materials and methods. Each primer set amplifies a region of the promoter simplified in Supplementary Figure S2 (available at JXB online) containing the BZR1-binding BRRE sequence (5′-CGTGT/CG-3′, He et al., 2005) and E-box (5′-CANNTG-3′, Yin et al., 2005). Promoters of UBC30 and PP2A genes were used as negative controls for BZR1- and BES1-binding motif. The promoter of DWF4 was used as a positive control (Sun et al., 2010). All ChIP-qPCR analysis was repeated with a minimum of triple biological replicates and the data were statistically analysed by the Student’s t-test.
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Figure 6: BR- and BZR1-mediated signalling pathway modulates transcriptional expression of diverse root-regulating genes. (A–C) Quantitative RT-PCR analysis of root-controlling genes in BL-treated Col-0 and BR-related mutant plants. Plants were grown and expression of root-regulating genes was analysed for short-term BL-treated plants (A), long-term BL-treated plants (B), and BL-related mutants (C) as described in Fig. 1G–I. (D) ChIP-qPCR assay of the BZR1- and BES1- binding cis-motif of root-regulating genes. BZR1- and BES1-bound chromatin DNA fragments were isolated from roots of week-old Col-0, pBZR1::BZR1-YFP, and p35S::BES1-GFP plants using anti-BZR1 antibodies. Next, quantitative PCR analysis was performed with the primer sets listed in Supplementary Table S2 as described in Materials and methods. Each primer set amplifies a region of the promoter simplified in Supplementary Figure S2 (available at JXB online) containing the BZR1-binding BRRE sequence (5′-CGTGT/CG-3′, He et al., 2005) and E-box (5′-CANNTG-3′, Yin et al., 2005). Promoters of UBC30 and PP2A genes were used as negative controls for BZR1- and BES1-binding motif. The promoter of DWF4 was used as a positive control (Sun et al., 2010). All ChIP-qPCR analysis was repeated with a minimum of triple biological replicates and the data were statistically analysed by the Student’s t-test.

Mentions: Total RNA was isolated from root tissues using the Plant RNA Extraction Kit (Intron Biotechnology, Korea) and treated with RQ1 RNase-Free DNase (Promega, USA) according to the manufacturer’s instructions. Quantitative real-time PCR (qRT-PCR) was performed by the SYBR green method using the Applied Biosystems Step One Plus System (Applied Biosystems, USA). The expression of each transcript was normalized against the amount of UBC1 control in each sample. CPD was used as a control for BR-repressed genes. The results were reported as expression relative to the Col-0 mock control (Figs 1G, 1H, 5A, 5B, 6A, and 6B) or Col-0 or En-2 genetic control (Figs 1I, 5C, and 6C). Three to five biological replicates were included in each experiment, and the data were statistically analysed using the Student’s t-test. Primers used for the qRT-PCRs are summarized in Supplementary Table S1 (available at JXB online).


Brassinazole resistant 1 (BZR1)-dependent brassinosteroid signalling pathway leads to ectopic activation of quiescent cell division and suppresses columella stem cell differentiation.

Lee HS, Kim Y, Pham G, Kim JW, Song JH, Lee Y, Hwang YS, Roux SJ, Kim SH - J. Exp. Bot. (2015)

BR- and BZR1-mediated signalling pathway modulates transcriptional expression of diverse root-regulating genes. (A–C) Quantitative RT-PCR analysis of root-controlling genes in BL-treated Col-0 and BR-related mutant plants. Plants were grown and expression of root-regulating genes was analysed for short-term BL-treated plants (A), long-term BL-treated plants (B), and BL-related mutants (C) as described in Fig. 1G–I. (D) ChIP-qPCR assay of the BZR1- and BES1- binding cis-motif of root-regulating genes. BZR1- and BES1-bound chromatin DNA fragments were isolated from roots of week-old Col-0, pBZR1::BZR1-YFP, and p35S::BES1-GFP plants using anti-BZR1 antibodies. Next, quantitative PCR analysis was performed with the primer sets listed in Supplementary Table S2 as described in Materials and methods. Each primer set amplifies a region of the promoter simplified in Supplementary Figure S2 (available at JXB online) containing the BZR1-binding BRRE sequence (5′-CGTGT/CG-3′, He et al., 2005) and E-box (5′-CANNTG-3′, Yin et al., 2005). Promoters of UBC30 and PP2A genes were used as negative controls for BZR1- and BES1-binding motif. The promoter of DWF4 was used as a positive control (Sun et al., 2010). All ChIP-qPCR analysis was repeated with a minimum of triple biological replicates and the data were statistically analysed by the Student’s t-test.
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Figure 6: BR- and BZR1-mediated signalling pathway modulates transcriptional expression of diverse root-regulating genes. (A–C) Quantitative RT-PCR analysis of root-controlling genes in BL-treated Col-0 and BR-related mutant plants. Plants were grown and expression of root-regulating genes was analysed for short-term BL-treated plants (A), long-term BL-treated plants (B), and BL-related mutants (C) as described in Fig. 1G–I. (D) ChIP-qPCR assay of the BZR1- and BES1- binding cis-motif of root-regulating genes. BZR1- and BES1-bound chromatin DNA fragments were isolated from roots of week-old Col-0, pBZR1::BZR1-YFP, and p35S::BES1-GFP plants using anti-BZR1 antibodies. Next, quantitative PCR analysis was performed with the primer sets listed in Supplementary Table S2 as described in Materials and methods. Each primer set amplifies a region of the promoter simplified in Supplementary Figure S2 (available at JXB online) containing the BZR1-binding BRRE sequence (5′-CGTGT/CG-3′, He et al., 2005) and E-box (5′-CANNTG-3′, Yin et al., 2005). Promoters of UBC30 and PP2A genes were used as negative controls for BZR1- and BES1-binding motif. The promoter of DWF4 was used as a positive control (Sun et al., 2010). All ChIP-qPCR analysis was repeated with a minimum of triple biological replicates and the data were statistically analysed by the Student’s t-test.
Mentions: Total RNA was isolated from root tissues using the Plant RNA Extraction Kit (Intron Biotechnology, Korea) and treated with RQ1 RNase-Free DNase (Promega, USA) according to the manufacturer’s instructions. Quantitative real-time PCR (qRT-PCR) was performed by the SYBR green method using the Applied Biosystems Step One Plus System (Applied Biosystems, USA). The expression of each transcript was normalized against the amount of UBC1 control in each sample. CPD was used as a control for BR-repressed genes. The results were reported as expression relative to the Col-0 mock control (Figs 1G, 1H, 5A, 5B, 6A, and 6B) or Col-0 or En-2 genetic control (Figs 1I, 5C, and 6C). Three to five biological replicates were included in each experiment, and the data were statistically analysed using the Student’s t-test. Primers used for the qRT-PCRs are summarized in Supplementary Table S1 (available at JXB online).

Bottom Line: BR promotes intense nuclear accumulation of BZR1 in the root tip area, and the binding of BZR1 to the promoters of several root development-regulating genes, modulating their expression in the root stem cell niche area.They could also inhibit auxin-dependent distal stem cell differentiation by antagonizing the auxin/WOX5-dependent pathway.In conclusion, BZR1-/BES1-mediated BR signalling pathways show differential effects on the maintenance of root apical meristem activities: they stimulate ectopic QC division while they show opposite effects on the differentiation of distal columella stem cells in a BR concentration- and BZR1-/BES1-dependent manner.

View Article: PubMed Central - PubMed

Affiliation: Division of Biological Science and Technology, Yonsei University, Wonju, 220-710, Republic of Korea.

No MeSH data available.


Related in: MedlinePlus