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Published February 22, 2005 | Supplemental Material
Journal Article Open

The early extra petals1 Mutant Uncovers a Role for MicroRNA miR164c in Regulating Petal Number in Arabidopsis

Abstract

Background: MicroRNAs (miRNAs) are small 20–25 nucleotide non-protein-coding RNAs that negatively regulate expression of genes in many organisms, ranging from plants to humans. The MIR164 family of miRNAs in Arabidopsis consists of three members that share sequence complementarity to transcripts of NAC family transcription factors, including CUP-SHAPED COTYLEDON1 (CUC1) and CUC2. CUC1 and CUC2 are redundantly required for the formation of boundaries between organ primordia. The analysis of transgenic plants that either overexpress miR164a or miR164b or express a miRNA-resistant version of CUC1 or CUC2 has shown that miRNA regulation of CUC1 and CUC2 is necessary for normal flower development. A loss-of-function allele of MIR164b did not result in a mutant phenotype, possibly because of functional redundancy among the three members of the MIR164 family. Results: In this study, we describe the characterization of the early extra petals1 (eep1) Arabidopsis mutant, whose predominant phenotype is the formation of extra petals in early-arising flowers. We demonstrate that eep1 is a loss-of-function allele of MIR164c, one of three known members of the MIR164 family. Our analyses of miR164c function and eep1 mir164b double mutants reveal that miR164c controls petal number in a nonredundant manner by regulating the transcript accumulation of the transcription factors CUC1 and CUC2. Conclusions: The data presented in this study indicate that closely related miRNA family members that are predicted to target the same set of genes can have different functions during development, possibly because of nonoverlapping expression patterns.

Additional Information

© 2005 Elsevier Ltd. Received: November 18, 2004. Revised: December 16, 2004. Accepted: December 17, 2004. Published: February 22, 2005. The authors would like to thank Dr. Bobby Williams for identifying the eep1 mutant, as well as Drs. Jeff Long (Salk Institute), John Bowman (University of California, Davis), and Masao Tasaka (Nara Institute) for sharing plasmid vectors and/or seeds. We are also grateful to Drs. Jeff Long and Joseph Ecker (Salk Institute), as well as to members of the Meyerowitz laboratory, for helpful discussions. We thank Dr. Brian Williams for providing tungsten dissection blades, as well as Arnavaz Garda and Alexia Sieber for invaluable technical assistance. The T-DNA insertion line for mir164b-1 was provided by the Salk Institute Genomic Analysis Library. Funding for the SIGnAL indexed insertion mutant collection was provided by the National Science Foundation. This work was supported by grant NSF IBN-0211670 to E.M.M. and by a European Molecular Biology Organisation (EMBO) long-term fellowship (ALTF 460-2003) to P.S.

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