Abstract
Key message
OsWRKY36 represses plant height and grain size by inhibiting gibberellin signaling.
Abstract
Plant height and grain size are important agronomic traits affecting yield in cereals, including rice. Gibberellins (GAs) are plant hormones that promote plant growth and developmental processions such as stem elongation and grain size. WRKYs are transcription factors that regulate stress tolerance and plant development including height and grain size. However, the relationship between GA signaling and WRKY genes is still poorly understood. Here, we characterized a small grain and semi-dwarf 3 (sgsd3) mutant in rice cv. Hwayoung (WT). A T-DNA insertion in the 5′-UTR of OsWRKY36 induced overexpression of OsWRKY36 in the sgsd3 mutant, likely leading to the mutant phenotype. This was confirmed by the finding that overexpression of OsWRKY36 caused a similar small grain and semi-dwarf phenotype to the sgsd3 mutant whereas knock down and knock out caused larger grain phenotypes. The sgsd3 mutant was also hyposensitive to GA and accumulated higher mRNA and protein levels of SLR1 (a GA signaling DELLA-like inhibitor) compared with the WT. Further assays showed that OsWRKY36 enhanced SLR1 transcription by directly binding to its promoter. In addition, we found that OsWRKY36 can protect SLR1 from GA-mediated degradation. We thus identified a new GA signaling repressor OsWRKY36 that represses GA signaling through stabilizing the expression of SLR1.
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Abbreviations
- TF:
-
Transcription factor
- cv:
-
Cultivar
- OX:
-
Overexpression
- RNAi:
-
RNA interference
- GFP:
-
Green fluorescent protein
- qRT-PCR:
-
Quantitative real-time PCR
- EMSA:
-
Electrophoretic mobility shift assay
References
Ashikari M, Wu J, Yano M, Sasaki T, Yoshimura A (1999) Rice gibberellin-insensitive dwarf mutant gene dwarf 1 encodes the alpha-subunit of GTP-binding protein. Proc Natl Acad Sci USA 96:10284–10289. https://doi.org/10.1073/pnas.96.18.10284
Cai Y et al (2014) Dlf1, a WRKY transcription factor, is involved in the control of flowering time and plant height in rice. PLoS ONE 9:e102529. https://doi.org/10.1371/journal.pone.0102529
Chen LG, Song Y, Li SJ, Zhang LP, Zou CS, Yu DQ (2012a) The role of WRKY transcription factors in plant abiotic stresses. Bba-Gene Regul Mech 1819:120–128. https://doi.org/10.1016/j.bbagrm.2011.09.002
Chen ML et al (2012b) Highly sensitive and quantitative profiling of acidic phytohormones using derivatization approach coupled with nano-LC-ESI-Q-TOF-MS analysis. J Chromatogr B Analyt Technol Biomed Life Sci 905:67–74. https://doi.org/10.1016/j.jchromb.2012.08.005
Chen WW et al (2019) Small Grain and Dwarf 2, encoding an HD-Zip II family transcription factor, regulates plant development by modulating gibberellin biosynthesis in rice. Plant Sci 288:110208. https://doi.org/10.1016/j.plantsci.2019.110208
Foster CE, Martin TM, Pauly M (2010) Comprehensive compositional analysis of plant cell walls (Lignocellulosic biomass) part I: lignin. J Vis Exp 37:e1745. https://doi.org/10.3791/1745
Garcia D, Fitz Gerald JN, Berger F (2005) Maternal control of integument cell elongation and zygotic control of endosperm growth are coordinated to determine seed size in Arabidopsis. Plant Cell 17:52–60. https://doi.org/10.1105/tpc.104.027136
Han M, Kim CY, Lee J, Lee SK, Jeon JS (2014) OsWRKY42 represses OsMT1d and induces reactive oxygen species and leaf senescence in rice. Mol Cells 37:532–539. https://doi.org/10.14348/molcells.2014.0128
Hedden P, Sponsel V (2015) A century of gibberellin research. J Plant Growth Regul 34:740–760. https://doi.org/10.1007/s00344-015-9546-1
Itoh H, Ueguchi-Tanaka M, Sentoku N, Kitano H, Matsuoka M, Kobayashi M (2001) Cloning and functional analysis of two gibberellin 3 beta-hydroxylase genes that are differently expressed during the growth of rice. Proc Natl Acad Sci USA 98:8909–8914. https://doi.org/10.1073/pnas.141239398
Itoh H, Ueguchi-Tanaka M, Sato Y, Ashikari M, Matsuoka M (2002) The gibberellin signaling pathway is regulated by the appearance and disappearance of SLENDER RICE1 in nuclei. Plant Cell 14:57–70. https://doi.org/10.1105/tpc.010319
Itoh H et al (2004) A rice semi-dwarf gene, Tan-Ginbozu (D35), encodes the gibberellin biosynthesis enzyme, ent-kaurene oxidase. Plant Mol Biol 54:533–547. https://doi.org/10.1023/B:Plan.0000038261.21060.47
Jones HD, Smith SJ, Desikan R, Plakidou-Dymock S, Lovegrove A, Hooley R (1998) Heterotrimeric G proteins are implicated in gibberellin induction of a-amylase gene expression in wild oat aleurone. Plant Cell 10:245–254. https://doi.org/10.1105/tpc.10.2.245
Khush GS (1999) Green revolution: preparing for the 21st century. Genome 42:646–655
Koshiba T et al (2013) Characterization of 5-hydroxyconiferaldehyde O-methyltransferase in Oryza sativa. Plant Biotechnol-Nar 30:157–167. https://doi.org/10.5511/plantbiotechnology.13.0219a
Li JA et al (2011) Mutation of rice BC12/GDD1, which encodes a kinesin-like protein that binds to a GA biosynthesis gene promoter, leads to dwarfism with impaired cell elongation. Plant Cell 23:628–640. https://doi.org/10.1105/tpc.110.081901
Li R et al (2015) Prioritizing plant defence over growth through WRKY regulation facilitates infestation by non-target herbivores. Elife 4:e04805. https://doi.org/10.7554/eLife.04805
Li W, Wang H, Yu D (2016) Arabidopsis WRKY transcription factors WRKY12 and WRKY13 oppositely regulate flowering under short-day conditions. Mol Plant 9:1492–1503. https://doi.org/10.1016/j.molp.2016.08.003
Li N, Xu R, Duan P, Li Y (2018) Control of grain size in rice. Plant Reprod 31:237–251. https://doi.org/10.1007/s00497-018-0333-6
Livak KJ, Schmittgen TD (2001) Analysis of relative gene expression data using real-time quantitative PCR and the 2(T)(-Delta Delta C) method. Methods 25:402–408. https://doi.org/10.1006/meth.2001.1262
Luo M, Dennis ES, Berger F, Peacock WJ, Chaudhury A (2005) MINISEED3 (MINI3), a WRKY family gene, and HAIKU2 (IKU2), a leucine-rich repeat (LRR) KINASE gene, are regulators of seed size in Arabidopsis. Proc Natl Acad Sci USA 102:17531–17536. https://doi.org/10.1073/pnas.0508418102
McGinnis KM, Thomas SG, Soule JD, Strader LC, Zale JM, Sun TP, Steber CM (2003) The Arabidopsis SLEEPY1 gene encodes a putative F-box subunit of an SCF E3 ubiquitin ligase. Plant Cell 15:1120–1130. https://doi.org/10.1105/tpc.010827
Miao J et al (2013) Targeted mutagenesis in rice using CRISPR-Cas system. Cell Res 23:1233–1236. https://doi.org/10.1038/cr.2013.123
Miao J et al (2019) Mutation of RGG2, which encodes a type B heterotrimeric G protein gamma subunit, increases grain size and yield production in rice. Plant Biotechnol J 17:650–664. https://doi.org/10.1111/pbi.13005
Murase K, Hirano Y, Sun TP, Hakoshima T (2008) Gibberellin-induced DELLA recognition by the gibberellin receptor GID1. Nature 456:459–415. https://doi.org/10.1038/nature07519
Okuno A et al (2014) New approach to increasing rice lodging resistance and biomass yield through the use of high gibberellin producing varieties. PLoS ONE 9:e86870. https://doi.org/10.1371/journal.pone.0086870
Rushton PJ, Somssich IE, Ringler P, Shen QXJ (2010) WRKY transcription factors. Trends Plant Sci 15:247–258. https://doi.org/10.1016/j.tplants.2010.02.006
Sakamoto T et al (2004) An overview of gibberellin metabolism enzyme genes and their related mutants in rice. Plant Physiol 134:1642–1653. https://doi.org/10.1104/pp.103.033696
Shen H, Liu C, Zhang Y, Meng X, Zhou X, Chu C, Wang X (2012) OsWRKY30 is activated by MAP kinases to confer drought tolerance in rice. Plant Mol Biol 80:241–253. https://doi.org/10.1007/s11103-012-9941-y
Shimono M, Sugano S, Nakayama A, Jiang CJ, Ono K, Toki S, Takatsuji H (2007) Rice WRKY45 plays a crucial role in benzothiadiazole-inducible blast resistance. Plant Cell 19:2064–2076. https://doi.org/10.1105/tpc.106.046250
Spielmeyer W, Ellis MH, Chandler PM (2002) Semidwarf (sd-1), “green revolution” rice, contains a defective gibberellin 20-oxidase gene. Proc Natl Acad Sci USA 99:9043–9048. https://doi.org/10.1073/pnas.132266399
Sun TP (2010) Gibberellin-GID1-DELLA: a pivotal regulatory module for plant growth and development. Plant Physiol 154:567–570. https://doi.org/10.1104/pp.110.161554
Sun Y, Yu D (2015) Activated expression of AtWRKY53 negatively regulates drought tolerance by mediating stomatal movement. Plant Cell Rep 34:1295–1306. https://doi.org/10.1007/s00299-015-1787-8
Tian XJ et al (2017) Transcription factor OsWRKY53 positively regulates brassinosteroid signaling and plant architecture. Plant Physiol 175:1337–1349. https://doi.org/10.1104/pp.17.00946
Ueguchi-Tanaka M, Fujisawa Y, Kobayashi M, Ashikari M, Iwasaki Y, Kitano H, Matsuoka M (2000) Rice dwarf mutant d1, which is defective in the alpha subunit of the heterotrimeric G protein, affects gibberellin signal transduction. Proc Natl Acad Sci USA 97:11638–11643. https://doi.org/10.1073/pnas.97.21.11638
Ueguchi-Tanaka M et al (2005) Gibberellin insensitive dwarf1 encodes a soluble receptor for gibberellin. Nature 437:693–698. https://doi.org/10.1038/nature04028
Updegraff DM (1969) Semimicro determination of cellulose in biological materials. Anal Biochem 32:420–424. https://doi.org/10.1016/s0003-2697(69)80009-6
Wang H, Avci U, Nakashima J, Hahn MG, Chen F, Dixon RA (2010) Mutation of WRKY transcription factors initiates pith secondary wall formation and increases stem biomass in dicotyledonous plants. Proc Natl Acad Sci USA 107:22338–22343. https://doi.org/10.1073/pnas.1016436107
Xie Z, Zhang ZL, Zou XL, Huang J, Ruas P, Thompson D, Shen QJ (2005) Annotations and functional analyses of the rice WRKY gene superfamily reveal positive and negative regulators of abscisic acid signaling in aleurone cells. Plant Physiol 137:176–189. https://doi.org/10.1104/pp.104.054312
Yang L, Zhao X, Yang F, Fan D, Jiang Y, Luo K (2016) PtrWRKY19, a novel WRKY transcription factor, contributes to the regulation of pith secondary wall formation in Populus trichocarpa. Sci Rep 6:18643. https://doi.org/10.1038/srep18643
Yoshida K et al (2013) Engineering the Oryza sativa cell wall with rice NAC transcription factors regulating secondary wall formation. Front Plant Sci 4:383. https://doi.org/10.3389/fpls.2013.00383
Yu YC, Hu RB, Wang HM, Cao YP, He G, Fu CX, Zhou GK (2013) MlWRKY12, a novel Miscanthus transcription factor, participates in pith secondary cell wall formation and promotes flowering. Plant Sci 212:1–9. https://doi.org/10.1016/j.plantsci.2013.07.010
Zhang ZL, Xie Z, Zou XL, Casaretto J, Ho THD, Shen QXJ (2004) A rice WRKY gene encodes a transcriptional repressor of the gibberellin signaling pathway in aleurone cells. Plant Physiol 134:1500–1513. https://doi.org/10.1104/pp.103.034967
Zhang CQ, Xu Y, Lu Y, Yu HX, Gu MH, Liu QQ (2011a) The WRKY transcription factor OsWRKY78 regulates stem elongation and seed development in rice. Planta 234:541–554. https://doi.org/10.1007/s00425-011-1423-y
Zhang Y et al (2011b) A highly efficient rice green tissue protoplast system for transient gene expression and studying light/chloroplast-related processes. Plant Methods 7:30. https://doi.org/10.1186/1746-4811-7-30
Zhou F et al (2013) D14-SCF(D3)-dependent degradation of D53 regulates strigolactone signalling. Nature 504:406–410. https://doi.org/10.1038/nature12878
Acknowledgements
We thank Prof. Gynheung An (Department of Plant Systems Biotech, Kyung Hee University, Yongin, South Korea) for providing the Oswrky36 mutant seeds. We also acknowledge support from the National Key Research and Development Program of China (2017YFD0100404), National Natural Science Foundation of China (31671769), National Transgenic Science and Technology Program (2016ZX08001004-002), and Jiangsu Science and Technology Development Program (BE2018388). Additional support was provided by the Key Laboratory of Biology, Genetics and Breeding of Japonica Rice in the Mid-lower Yangtze River, and Jiangsu Collaborative Innovation Center for Modern Crop Production.
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JMW supervised this research; JL and CLZ performed the experiments and wrote the manuscript; QBL and LJ revised the paper; YKR, XL, RM, RNJ, CLM participated in the experiments; TNY, XJZ, QW were involved in the data discussions; all authors read and approved the final manuscript.
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Lan, J., Lin, Q., Zhou, C. et al. Small grain and semi-dwarf 3, a WRKY transcription factor, negatively regulates plant height and grain size by stabilizing SLR1 expression in rice. Plant Mol Biol 104, 429–450 (2020). https://doi.org/10.1007/s11103-020-01049-0
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DOI: https://doi.org/10.1007/s11103-020-01049-0