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We thank Ana Rapp for technical assistance, Jose Leon (Instituto de Biologia Molecular y Celular de Plantas, Consejo Superior de Investigaciones Cientificas, Valencia, Spain) and the TRANSPLANTA consortium for contributing to obtain MUTE and FAMA over-expressing lines, Mark Curtis (Institute of Plant Biology and Zurich-Basel Plant Science Centre, University of Zurich, Switzerland) for the vectors used in construction of the dual-inducible system, and the Nottingham Arabidopsis Stock Centre for providing Col-0 seeds. This work was supported by grants from the Spanish Government (BIO2007-60276 and BIO2012-33952 to C.F. and M.M., BFU2009-9783 to C.G., and CSD2007-00057 TRANSPLANTA to C.F.), the Castilla la Mancha Government (PAI07-0036-3278 to M.M. and pre- and postdoctoral grants to M.T. and I.B.), and an institutional grant from the Fundacion Ramon Areces to the Centro de Biologia Molecular Severo Ochoa.

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Timely expression of the Arabidopsis stoma-fate master regulator MUTE is required for specification of other epidermal cell types

Publicat a:Plant Journal. 75 (5): 808-822 - 2013-09-01 75(5), DOI: 10.1111/tpj.12244

Autors: Trivino, Magdalena; Martin-Trillo, Mar; Ballesteros, Isabel; Delgado, Dolores; de Marcos, Alberto; Desvoyes, Benedicte; Gutierrez, Crisanto; Mena, Montana; Fenoll, Carmen;

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Epidermal differentiation in Arabidopsis thaliana aerial organs involves stomatal lineage development. Lineages derive from meristemoids, which arise from asymmetric divisions of protodermal cells. Each meristemoid divides repeatedly in an inward spiral before it transits to a guard mother cell (GMC) that produces the stoma, leaving a trail of surrounding stomatal lineage ground cells (SLGCs) that eventually differentiate into endoreplicated pavement cells. MUTE is a bHLH transcription factor that is expressed in late meristemoids and drives their transition to GMCs. Loss-of-function mute mutants are stomata-less dwarf plants with arrested lineages, in which stunted putative SLGCs surround a halted meristemoid. We analysed MUTE functions using a chemically inducible system for mute-3 complementation based on conditional MUTE expression in its normal domain. Continuous induction from germination produced stomata-bearing, normal-sized plants with viable mute-3 seeds. In 2-week-old mute-3 cotyledons, meristemoids appeared to retain their identity and synchronously formed stomata in response to induced MUTE expression. However, arrested SLGCs were not complemented: many produced stomata, leading to stomatal clusters, and others remained unexpanded and diploid. In contrast, non-lineage pavement cells, which are under-endoreplicated in mute-3, expanded and increased their ploidy level upon induction, showing that the lack of response of SLGCs is specific to this arrested cell type. Leaf phenotypic mosaics include wild-type lineages and adjacent mute-3 lineages, whose meristemoids and putative SLGCs remained arrested, indicating that the role of MUTE in SLGC fate is strictly lineage-autonomous. These results show that timely MUTE expression is essential to prevent stomatal fate in SLGCs and to promote their differentiation as pavement cells.

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