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Plant Cell Advance Online Publication
Published on April 7, 2006; 10.1105/tpc.105.040725

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Received December 25, 2005
Returned for revision March 10, 2006
Accepted March 20, 2006

Endogenous and Synthetic MicroRNAs Stimulate Simultaneous, Efficient, and Localized Regulation of Multiple Targets in Diverse Species

John Paul Alvarez 1, Irena Pekker 2, Alexander Goldshmidt 2, Eyal Blum 2, Ziva Amsellem 2, and Yuval Eshed 2*

1 Department of Plant Sciences, Weizmann Institute of Science, Rehovot, 76100, Israel; School of Biological Sciences, Monash University, Victoria 3800, Australia
2 Department of Plant Sciences, Weizmann Institute of Science, Rehovot, 76100, Israel

* To whom correspondence should be addressed. E-mail: yuval.eshed{at}weizmann.ac.il.

Recent studies demonstrated that pattern formation in plants involves regulation of transcription factor families by microRNAs (miRNAs). To explore the potency, autonomy, target range, and functional conservation of miRNA genes, a systematic comparison between plants ectopically expressing pre-miRNAs and plants with corresponding multiple mutant combinations of target genes was performed. We show that regulated expression of several Arabidopsis thaliana pre-miRNA genes induced a range of phenotypic alterations, the most extreme ones being a phenocopy of combined loss of their predicted target genes. This result indicates quantitative regulation by miRNA as a potential source for diversity in developmental outcomes. Remarkably, custom-made, synthetic miRNAs vectored by endogenous pre-miRNA backbones also produced phenocopies of multiple mutant combinations of genes that are not naturally regulated by miRNA. Arabidopsis-based endogenous and synthetic pre-miRNAs were also processed effectively in tomato (Solanum lycopersicum) and tobacco (Nicotiana tabacum). Synthetic miR-ARF targeting Auxin Response Factors 2, 3, and 4 induced dramatic transformations of abaxial tissues into adaxial ones in all three species, which could not cross graft joints. Likewise, organ-specific expression of miR165b that coregulates the PHABULOSA-like adaxial identity genes induced localized abaxial transformations. Thus, miRNAs provide a flexible, quantitative, and autonomous platform that can be employed for regulated expression of multiple related genes in diverse species.




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