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Plant and Cell Physiology Advance Access originally published online on January 19, 2007
Plant and Cell Physiology 2007 48(3):391-404; doi:10.1093/pcp/pcm008
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© The Author 2007. Published by Oxford University Press on behalf of Japanese Society of Plant Physiologists. All rights reserved. For permissions, please email: journals.permissions@oxfordjournals.org

Overexpression of miR165 Affects Apical Meristem Formation, Organ Polarity Establishment and Vascular Development in Arabidopsis

Gong-Ke Zhou1, Minoru Kubo2, Ruiqin Zhong1, Taku Demura2 and Zheng-Hua Ye1,*

1Department of Plant Biology, University of Georgia, Athens, GA 30602, USA
2RIKEN Plant Science Center, Yokohama, Kanagawa, 230-0045 Japan

*Corresponding author: E-mail, zhye{at}plantbio.uga.edu; Fax, +1-706-542-1805.


  Abstract

The class III homeodomain leucine-zipper (HD-ZIP III) genes are thought to be targets of microRNAs (miRNAs) 165 and 166, but it is not known whether all the developmental processes affected by mutations of the HD-ZIP III genes could be recapitulated by an alteration in the expression of miR165 and miR166. Previous work showed that overexpression of miR166 by activation tagging results in down-regulation of the ATHB-9/PHV, ATHB-14/PHB and ATHB-15 genes, and concomitantly causes an enlargement of shoot apical meristems (SAMs) and an enhancement in vascular development. Here we demonstrated that overexpression of miR165 causes a drastic reduction in the transcript levels of all five HD-ZIP III genes in Arabidopsis. The miR165 overexpressors display prominent phenotypes reminiscent of loss-of-function mutants of rev phb phv and rev/ifl1, including loss of SAM, alteration of organ polarity, abnormal formation of carpels, inhibition of vascular development and aberrant differentiation of interfascicular fibers. Global gene expression analysis revealed a link between miR165 overexpression and altered expression of genes involved in auxin signaling and vascular development. Our results demonstrate that overexpression of miR165 recapitulates the phenotypes caused by loss-of-function mutations of HD-ZIP III genes, such as loss of SAM, altered organ polarity and defects in development of vascular tissues and interfascicular fibers.

Keywords: Arabidopsis thaliana - Homeodomain proteins - Meristem - MicroRNA - Organ polarity - Vascular development

Abbreviations: CaMV, cauliflower mosaic virus; DIG, digoxigenin; HD-ZIP, homeodomain leucine-zipper protein; RT–PCR, reverse transcription–PCR; SAM, shoot apical meristem

(Received December 20, 2006; Accepted January 11, 2007)
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