Abstract

The Arabidopsis stoma is a specialized epidermal valve made up of a pair of guard cells around a pore whose aperture controls gas exchange between the shoot and atmosphere. Guard cells (GCs) are produced by a symmetric division of guard mother cells (GMCs). The R2R3-MYB transcription factor FOUR LIPS (FLP) and its paralogue MYB88 restrict the division of a GMC to one. Previously, the upstream regions of several core cell cycle genes were identified as the direct targets of FLP/MYB88, including the B-type cyclin-dependent kinase CDKB1;1 and A2-type cyclin CYCA2;3. Here we show that CDKA;1 is also an immediate direct target of FLP/MYB88 through the binding to cis-regulatory elements in the CDKA;1 promoter region. CDKA;1 activity is required not only for normal GMC divisions but also for the excessive cell overproliferation in flp myb88 mutant GMCs. The impaired defects of GMC division in cdkb1;1 1;2 mutants could be partially rescued by a stage-specific expression of CDKA;1. Although targeted overexpression of CDKA;1 does not affect stomatal development, ectopic expression of the D3-type cyclin CYCD3;2 induces GC subdivision, resulting in a stoma with 3-4 GCs instead of the normal two. Co-overexpression of CDKA;1 with CYCD3;2, but not with CYCA2;3, confers a synergistic effect with respect to GC subdivision. Thus, in addition to a role in stomatal formative asymmetric divisions at early developmental stages, CDKA;1 is needed in triggering GMC symmetric divisions at the late stage of stomatal development. However, timely down-regulation of CDKA;1-CYCD3 activity is required for restriction of GC proliferation.

Highlights

  • Guard cell vitality is checked by neutral red staining

  • Neutral red dyes are taken by living cells and stored in vacuoles

  • All GCs are vital in stomata from FAMA:amiRNA-RBR1 (D) and FAMA:CYCD3;2 (E)

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Summary

Introduction

Lane1, probes for binding Element A with His-MYB88 proteins; Lane 2, probes for Element B with His-MYB88 proteins; Lane 3, probes for Element C and D Guard cell vitality is checked by neutral red staining.

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Conclusion
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