Abstract
The plant hormone auxin plays a critical role in regulating plant growth and development. Recent advances have been made in the understanding of auxin response pathways, primarily by the characterization of auxin response mutants in Arabidopsis. In addition, microRNAs (miRNAs) have been shown to be critical regulators of genes important for normal plant development and physiology. However, little is known about possible interactions between miRNAs and hormonal signaling during normal development. Here we show that an Arabidopsis microRNA, miR167, which has a complementary sequence to a portion of the AUXIN RESPONSE FACTOR6 (ARF6) and ARF8 mRNAs, can cause transcript degradation for ARF8, but not for ARF6. We report phenotypic characterizations of 35S::MIR167b transgenic lines, and show that severe 35S::MIR167b transgenic lines had phenotypes similar to those of an arf6 arf8 double mutant. The transgenic phenotypes suggest that miR167 may repress ARF6 at the level of translation. We demonstrate that the transgenic plants are defective in all four whorls of floral organs. In the transgenic flowers, filaments were abnormally short, anthers could not properly release pollen, and pollen grains did not germinate. Our results provide an important link between the miRNA-mediated regulatory pathway of gene expression and the auxin signaling network promoting plant reproductive development.
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References
Eckardt NA . New Insights into Auxin Biosynthesis. Plant Cell 2001; 13:101–111.
Kim J, Harter K, Theologis A . Protein-protein interactions among the Aux/IAA proteins. Proc Natl Acad Sci USA 1997; 94:11786–11791.
Ulmasov T, Murfett J, Hagen G, Guilfoyle TJ . Aux/lAA proteins repress expression of reporter genes containing natural and highly active synthetic auxin response elements. Plant Cell 1997; 9:1963–1971.
Liscum E, Reed JW . Genetics of Aux/IAA and ARF action in plant growth and development. Plant Mol Biol 2002; 49:387–400.
Tiwari SB, Wang XJ, Hagen G, Guilfoyle GT . AUX/IAA proteins are active repressors, and their stability and activity are modulated by auxin. Plant Cell 2001; 13:2809–2822.
Llave C, Kasschau KD, Rector MA, Carrington JC . Endogenous and silencing-associated small RNAs in plants. Plant Cell 2002; 14:1605–1619.
Li H, Xu L, Wang H, et al. The Putative RNA-dependent RNA polymerase RDR6 acts synergistically with ASYMMETRIC LEAVES1 and 2 to repress BREVIPEDICELLUS and MicroRNA165/166 in Arabidopsis leaf development. Plant Cell 2005; 17:2157–2171.
Juarez MT, Kui JS, Thomas J, Heller BA, Timmermans MC . microRNA-mediated repression of rolled leaf1 specifies maize leaf polarity. Nature 2004; 428:84–88.
Carrington JC, Ambros V . Role of microRNAs in plant and animal development. Science 2003; 18:336–338.
Aukerman MJ, Sakai H . Regulation of flowering time and floral organ identity by a MicroRNA and its APETALA2-like target genes. Plant Cell 2003; 15(11):2730–41.
Chen XM . A microRNA as a translational repressor of APETALA2 in Arabidopsis flower development. Science 2004; 303:2022–2025.
Palatnik JF, Allen E, Wu X, et al. Control of leaf morphogenesis by microRNAs. Nature 2003; 425:257–263.
Emery JF, Floyd SK, Alvarez J, et al. Radial patterning of Arabidopsis shoots by class III HD-ZIP and KANADI genes. Curr Biol 2003; 13:1768–1774.
Tang G, Reinhart BJ, Bartel DP, Zamore PD . A biochemical framework for RNA silencing in plants. Genes Dev 2003; 17:49–63.
Timmermans MC, Juarez MT, Phelps-Durr TL . A conserved microRNA signal specifies leaf polarity. Cold Spring Harb Symp Quant Biol 2004; 69:409–417.
Kidner CA, Martienssen RA . Spatially restricted microRNA directs leaf polarity through ARGONAUTE1. Nature 2004; 428:81–84.
Mallory AC, Dugas DV, Bartel DP, Bartel B . MicroRNA regulation of NAC-domain targets is required for proper formation and separation of adjacent embryonic, vegetative, and floral organs. Curr Biol 2004; 14:1035–1046.
Mallory AC, Reinhart BJ, Jones-Rhoades MW, et al. MicroRNA control of PHABULOSA in leaf development: importance of pairing to the microRNA 5' region. EMBO J 2004; 23:3356–3364.
Achard P, Herr A, Baulcombe DC, Harberd NP . Modulation of floral development by a gibberellin-regulated microRNA. Development 2004; 131:3357–3365.
Wang JW, Wang LJ, Mao YB, et al. Control of root cap formation by MicroRNA-targeted auxin response factors in Arabidopsis. Plant Cell 2005; 17:2204–2216.
Tian CE, Muto H, Higuchi K, et al. Disruption and overexpression of auxin response factor 8 gene of Arabidopsis affect hypocotyl elongation and root growth habit, indicating its possible involvement in auxin homeostasis in light condition. Plant J 2004; 40:333–343.
Nagpal P, Ellis CM, Weber H, et al. Auxin response factors ARF6 and ARF8 promote jasmonic acid production and flower maturation. Development 2005; 132:4107–4118.
Rhoades MW, Reinhart BJ, Lim LP, Burge CB, Bartel B, Bartel DP . Prediction of plant microRNA targets. Cell 2002; 110:513–520.
Reinhart BJ, Weinstein EG, Rhoades MW, Bartel B, Bartel DP . MicroRNAs in plants. Genes Dev 2002; 16:1616–1626.
Hardtke CS, Berleth T . The Arabidopsis gene MONOPTEROS encodes a transcription factor mediating embryo axis formation and vascular development. EMBO J 1998; 17:1405–1411.
Harper RM, Stowe-Evans EL, Luesse DR, et al. The NPH4 locus encodes the auxin response factor ARF7, a conditional regulator of differential growth in aerial Arabidopsis tissue. Plant Cell 2000; 12:757–770.
Nemhauser JL, Feldman LJ, Zambryski PC, Auxin and ETTIN in Arabidopsis gynoecium morphogenesis. Development 2000; 127:3877–3888.
Okushima Y, Mitina I, Quach HL, Theologis A . AUXIN RESPONSE FACTOR 2 (ARF2): a pleiotropic developmental regulator. Plant J 2005; 43:29–46.
Okushima Y, Overvoorde PJ, Arima K, et al. Functional genomic analysis of the AUXIN RESPONSE FACTOR gene family members in Arabidopsis thaliana: unique and overlapping functions of ARF7 and ARF19. Plant Cell 2005; 17:444–463.
Peragine A, Yoshikawa M, Wu G, Albrecht HL, Poethig RS . SGS3 and SGS2/SDE1/RDR6 are required for juvenile development and the production of trans-acting siRNAs in Arabidopsis. Genes Dev 2004; 18:2368–2379.
Allen E, Xie Z, Gustafson AM, Carrington JC . microRNA-directed phasing during trans-acting siRNA biogenesis in plants. Cell 2005; 121:207–221.
Yoshikawa M, Peragine A, Park MY, Poethig RS . A pathway for the biogenesis of trans-acting siRNAs in Arabidopsis. Genes Dev 2005; 19:2164–2175.
Williams L, Carles CC, Osmont KS, Fletcher JC . A database analysis method identifies an endogenous trans-acting short-interfering RNA that targets the Arabidopsis ARF2, ARF3, and ARF4 genes. Proc Natl Acad Sci USA 2005; 102:9703–9708.
Dugas DV, Bartel B . MicroRNA regulation of gene expression in plants. Curr Opin Plant Biol 2004; 7:512–520.
Wightman B, Ha I, Ruvkun G . Posttranscriptional regulation of the heterochronic gene lin-14 by lin-4 mediates temporal pattern formation in C. elegans. Cell 1993; 75:855–862.
Couzin J . Breakthrough of the year. Small RNAs make big splash. Science 2002; 298:2296–2297.
Okada K, Ueda J, Komaki MK, Bell CJ, Shimura Y . Requirement of the auxin polar transport system in early stages of Arabidopsis floral bud formation. Plant Cell 1991; 3:677–684.
Reinhardt D, Mandel T, Kuhlemeier C . Auxin regulates the initiation and radial position of plant lateral organs. Plant Cell 2000; 12:507–518.
Chen C, Wang S, Huang H . LEUNIG has multiple functions in gynoecium development in Arabidopsis. Genesis 2000; 26:42–54.
Xu L, Xu Y, Dong A, et al. Novel as1 and as2 defects in leaf adaxial-abaxial polarity reveal the requirement for ASYMMETRIC LEAVES1 and 2 and ERECTA functions in specifying leaf adaxial identity. Development 2003; 130:4097–4107.
Huang H, Tudor M, Weiss CA, Hu Y, Ma H . The Arabidopsis MADS-box gene AGL3 is widely expressed and encodes a sequence-specific DNA-binding protein. Plant Mol Biol 1995; 28:549–567.
van den Berg C, Willemsen V, Hendriks G, Weisbeek P, Scheres B . Short-range control of cell differentiation in the Arabidopsis root meristem. Nature 1997; 390:287–289.
Fan LM, Wang YF, Wang H, Wu WH . In vitro Arabidopsis pollen germination and characterization of the inward potassium currents in Arabidopsis pollen grain protoplasts. J Exp BotJ 2001; 52:1603–1614.
Acknowledgements
The authors thank X Gao for assistance with the SEM analysis, L Pi for some experimental protocols, and L Yang for discussion of this work. This research was supported by grants from Shanghai Institutes for Biological Sciences for the Plant Reproductive Development to H Ma and from the Shanghai Scientific Committee to H Huang.
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Ru, P., Xu, L., Ma, H. et al. Plant fertility defects induced by the enhanced expression of microRNA167. Cell Res 16, 457–465 (2006). https://doi.org/10.1038/sj.cr.7310057
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DOI: https://doi.org/10.1038/sj.cr.7310057
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