Abstract
Brassinosteroids (BRs) control virtually every aspect of plant growth and development. BRs act alone or in combination with other signals. To identify the signaling components that interact with BRs, we screened for mutants that suppress the dwarf phenotypes of brassinosteroid insensitive 1-5 (bri1-5) using an overexpression mutagenesis method. We established a mutant population by introducing a cDNA library in which cDNA was overexpressed under a constitutive promoter into Arabidopsis bri1-5 plants, which lacked a functional brassinosteroid (BR) receptor. One of the mutants, dubbed ‘bri1-5 with long petioles’ (blp), was selected based on its suppression phenotype. blp contained a chimeric DNA consisting of the 3’ half of PHYB, a 2-bp insertion, and a part of the chloroplast ribosomal RNA gene. Re-introduction of the chimeric DNA into bri1-5 recapitulated the blp phenotype. Prompted by the phenotypic similarity between blp and phyB, we examined both the transcript and protein levels of PHYB in the mutants. The levels were lower in blp 35Spro:PHYB than in 35Spro:PHYB plants, suggesting that introduction of the chimeric gene interfered with the stability of PHYB transcripts. Genome-wide screening for a specific target phenotype resulted in the finding that overexpression of the 3’ half of PHYB in the sense direction can cause a loss-of-function phenotype, and that PHYB plays an important role in BR signaling. Our results validate overexpression mutagenesis as a method to identify the function of Arabidopsis genes.
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References
Chen M, Schwabb R, Chory J (2003) Characterization of the requirements for localization of phytochrome B to nuclear bodies. Proc Natl Acad Sci USA 100:14493–14498
Choe S, Dilkes BP, Gregory BD, Ross AS, Yuan H, Noguchi T, Fujioka S, Takatsuto S, Tanaka A, Yoshida S, Tax FE, Feldmann KA (1999a) The Arabidopsis dwarf1 mutant is defective in the conversion of 24-methylenecholesterol to campesterol in brassinosteroid biosynthesis. Plant Physiol 119:897–907
Choe S, Fujioka S, Noguchi T, Takatsuto S, Yoshida S, Feldmann KA (2001) Overexpression of DWARF4 in the brassinosteroid biosynthetic pathway results in increased vegetative growth and seed yield in Arabidopsis. Plant J 26:573–582
Choe S, Schmitz RJ, Fujioka S, Takatsuto S, Lee MO, Yoshida S, Feldmann KA, Tax FE (2002) Arabidopsis brassinosteroidinsensitive dwarf12 mutants are semidominant and defective in a glycogen synthase kinase 3 beta-like kinase. Plant Physiol 130:1506–1515
Choe S, Tanaka A, Noguchi T, Fujioka S, Takatsuto S, Ross AS, Tax FE, Yoshida S, Feldman KA (2000) Lesions in the sterol Δ7 reductase gene of Arabidopsis cause dwarfism due to a block in brassinosteroid biosynthesis. Plant J 21:431–443
Choe S, Dilkes BP, Fujioka S, Takatsuto S, Sakurai A, Feldmann KA (1998) The DWF4 gene of Arabidopsis encodes a cytochrome P450 that mediates multiple 22 alpha-hydroxylation steps in brassinosteroid biosynthesis. Plant Cell 10:231–243
Choe S, Noguchi T, Fujioka S, Takatsuto S, Tissier CP, Gregory BD, Ross AS, Tanaka A, Yoshida S, Tax FE, Feldmann KA (1999b) The Arabidopsis dwf7/ste1 mutant is defective in the Δ7 sterol C-5 desaturation step leading to brassinosteroid biosynthesis. Plant Cell 11:207–221
Chung Y, Choe S (2013) The Regulation of Brassinosteroid Biosynthesis in Arabidopsis. Crit Rev Plant Sci 32:396–410
Chung Y, Maharjan PM, Lee O, Fujioka S, Jang S, Kim B, Takatsuto S, Tsujimoto M, Kim H, Cho S, Park T, Cho H, Hwang I, Choe S (2011) Auxin stimulates DWARF4 expression and brassinosteroid biosynthesis in Arabidopsis. Plant J 66:564–578
Clouse SD (2011) Brassinosteroid Signal Transduction: From Receptor Kinase Activation to Transcriptional Networks Regulating Plant Development. Plant Cell 23:1219–1230
Clouse SD, Sasse JM (1998) Brassinosteroids: Essential regulators of plant growth and development. Annu Rev Plant Physiol Plant Mol Biol 49:427–451
Devlin PF, Halliday KJ, Harberd NP, Whitelam GC (1996) The rosette habit of Arabidopsis thaliana is dependent upon phytochrome action: Novel phytochromes control internode elongation and flowering time. Plant J 10:1127–1134
Kikis EA, Oka Y, Hudson ME, Nagatani A, Quail PH (2009) Residues Clustered in the Light-Sensing Knot of Phytochrome B are Necessary for Conformer-Specific Binding to Signaling Partner PIF3. PLoS Genet 5:e1000352
Kim B, Jeong YJ, Corvalan C, Fujioka S, Cho S, Park T, Choe S (2014a) Darkness and gulliver2/phyB mutation decrease the abundance of phosphorylated BZR1 to activate brassinosteroid signaling in Arabidopsis. Plant J 77:737–747
Kim B, Kwon M, Jeon J, Schulz B, Corvalan C, Jeong YJ, Choe S (2014b) The Arabidopsis gulliver2/phyB mutant exhibits reduced sensitivity to brassinazole. J Plant Biol 57:20–27
Kim TW, Wang ZY (2010) Brassinosteroid Signal Transduction from Receptor Kinases to Transcription Factors. Annu Rev Plant Biol 61:681–704
Kircher S, Kozma-Bognar L, Kim L, Adam E, Harter K, Schafer E, Nagy F (1999) Light quality-dependent nuclear import of the plant photoreceptors phytochrome A and B. Plant Cell 11:1445–1456
Li J, Wen JQ, Lease KA, Doke JT, Tax FE, Walker JC (2002) BAK1, an Arabidopsis LRR receptor-like protein kinase, interacts with BRI1 and modulates brassinosteroid signaling. Cell 110:213–222
Li JM, Chory J (1997) A putative leucine-rich repeat receptor kinase involved in brassinosteroid signal transduction. Cell 90:929–938
Li JM, Nagpal P, Vitart V, McMorris TC, Chory J (1996) A role for brassinosteroids in light-dependent development of Arabidopsis. Science 272:398–401
Li JM, Nam KH, Vafeados D, Chory J (2001) BIN2, a new brassinosteroid-insensitive locus in Arabidopsis. Plant Physiol 127:14–22
Nagatani A (2010) Phytochrome: structural basis for its functions. Curr Opin Plant Biol 13:565–570
Nam KH, Li JM (2002) BRI1/BAK1, a receptor kinase pair mediating brassinosteroid signaling. Cell 110:203–212
Noguchi T, Fujioka S, Choe S, Takatsuto S, Yoshida S, Yuan H, Feldmann KA, Tax FE (1999) Brassinosteroid-insensitive dwarf mutants of Arabidopsis accumulate brassinosteroids. Plant Physiol 121:743–752
Oka Y, Matsushita T, Mochizuki N, Quail PH, Nagatani A (2008) Mutant Screen Distinguishes between Residues Necessary for Light-Signal Perception and Signal Transfer by Phytochrome B. PLoS Genet 15:e1000158
Perez-Perez JM, Ponce MR, Micol JL (2002) The UCU1 Arabidopsis gene encodes a SHAGGY/GSK3-like kinase required for cell expansion along the proximodistal axis. Dev Biol 242:161–173
Quail PH (2002) Phytochrome photosensory signalling networks. Nat Rev Mol Cell Biol 3:85–93
Reed JW, Nagpal P, Poole DS, Furuya M, Chory J (1993) Mutations in the Gene for the Red Far-Red Light Receptor Phytochrome-B Alter Cell Elongation and Physiological-Responses Throughout Arabidopsis Development. Plant Cell 5:147–157
Sakamoto K, Nagatani A (1996) Nuclear localization activity of phytochrome B. Plant J 10:859–868
Sasse JM (2003) Physiological actions of brassinosteroids: An update. J Plant Growth Regul 22:276–288
Sun Y, Fan XY, Cao DM, Tang W, He K, Zhu JY, He JX, Bai MY, Zhu S, Oh E, Patil S, Kim TW, Ji H, Wong WH, Rhee SY, Wang ZY (2010) Integration of brassinosteroid signal transduction with the transcription network for plant growth regulation in Arabidopsis. Dev Cell 19:765–777
Szekeres M, Nemeth K, KonczKalman Z, Mathur J, Kauschmann A, Altmann T, Redei GP, Nagy F, Schell J, Koncz C (1996) Brassinosteroids rescue the deficiency of CYP90, a cytochrome P450, controlling cell elongation and de-etiolation in arabidopsis. Cell 85:171–182
Tsukaya H, Kozuka T, Kim GT (2002) Genetic control of petiole length in Arabidopsis thaliana. Plant Cell Physiol 43:1221–1228
Wagner D, Koloszvari M, Quail PH (1996) Two small spatially distinct regions of phytochrome B are required for efficient signaling rates. Plant Cell 8:859–871
Wagner D, Tepperman JM, Quail PH (1991) Overexpression of Phytochrome-B Induces a Short Hypocotyl Phenotype in Transgenic Arabidopsis. Plant Cell 3:1275–1288
Wang ZY, Bai MY, Oh E, Zhu JY (2012) Brassinosteroid Signaling Network and Regulation of Photomorphogenesis. Annu Rev Genet 46:701–724
Wang ZY, Nakano T, Gendron J, He JX, Chen M, Vafeados D, Yang YL, Fujioka S, Yoshida S, Asami T, Chory J (2002) Nuclear-localized BZR1 mediates brassinosteroid-induced growth and feedback suppression of brassinosteroid biosynthesis. Dev Cell 2:505–513
Yeh KC, Lagarias JC (1998) Eukaryotic phytochromes: Lightregulated serine/threonine protein kinases with histidine kinase ancestry. Proc Natl Acad Sci USA 95:13976–13981
Yin YH, Wang ZY, Mora-Garcia S, Li JM, Yoshida S, Asami T, Chory J (2002) BES1 accumulates in the nucleus in response to brassinosteroids to regulate gene expression and promote stem elongation. Cell 109:181–191
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Jeong, Y.J., Park, S., Suh, S.J. et al. Overexpression of the 3’ half of the PHYB partially suppresses dwarfism in the brassinosteroid-insensitive bri1-5 mutant. J. Plant Biol. 59, 83–91 (2016). https://doi.org/10.1007/s12374-016-0513-6
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DOI: https://doi.org/10.1007/s12374-016-0513-6