The role of cell division in leaf morphogenesis
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1 Research Collection Doctoral Thesis The role of cell division in leaf morphogenesis Author(s): Wyrzykowska, Joanna Publication Date: 2001 Permanent Link: Rights / License: In Copyright - Non-Commercial Use Permitted This page was generated automatically upon download from the ETH Zurich Research Collection. For more information please consult the Terms of use. ETH Library
2 Diss. ETH No The role of cell division in leaf morphogenesis A dissertadon submitted to the SWISS FEDERAL INSTITUTE OF TECHNOLOGY For the degree of DOCTORof NATURALSCIENCES Presented by Joanna Wyrzykowska MSc of Biology University of Gdahsk, Poland Department of Biology, Geographyand Ocean Studies Born January 1967 in Ciechanow, Poland m äs* m 'tyimßi Accepted on the recommendationof Prof. Dr. N. Amrhein,referent Prof. Dr. U. Grossnikiaus, co-referent Dr. A. Fleming, co-referent (Supervisor) Zürich 2001
3 rv SUMMARY The aim of this study was to better understand the role of cell division in leaf initiation and morphogenesis. To do this, experiments were designed to modulate cell division and observe the influence on leaf development. Two aspects of cell division were the targets of this manipulation: the frequency and the plane of cell division. The strategy taken was to produce transgenic plants containing either components of the cell cycle machinery (for the manipulation cell division frequency) or elements of the cytokinetic machinery (for the manipulation of the plane of cell division) under transcriptional control of a tetracycline inducible promoter. To locally microinduce gene expression in meristem or young leaf primordia a new technique was developed. The genes selected for this approach were Nicta;cycA3;2 (encoding a plant cyclina), Sp;cdc25 (encoding a yeast cdc25 Phosphatase) and Glyma\phragmoplastin (encoding a plant dynamin-like protein). By local application of tetracycline onto meristems or young leaf primordia we aimed to precisely control the local overexpressionof these gene products and, thus, to alter either rate or the orientation of cell division. the proliferation In the meristem, both increased cell proliferation and disruption of the pattern of cell division (indicated both by histological analysis and marker gene expression patterns) did not lead to altered meristem fünction or leaf initiation. However, altered cell proliferation rate on the flank of young leaf primordia dramatically affected final leaf shape, with a transient increase in cell division leading to a final decrease in lamina expansion.conversely, transient Inhibition of cell division in this tissue (via local application of a cyclin dependent kinase inhibitor) led to an increase in lamina expansion. These data show that the role of cell division in plant morphogenesis is context dependent. The meristemdisplays adaptability to altered cell division frequency and orientation whereas there is a certain stage of leaf development when local cell division rate can have a major impact on morphogenesis. Taken in conjunction with other data, the results reported here identify the local cell proliferation rate as potentially a key step in the control of leaf shape. The mechanism by which this mightoccur is discussed.
4 SUMMARY Part of this work has been accepted for publication: Stephane Pien, Joanna Wyrzykowska, Simon McQueen-Mason, Cheryl Smart, and Andrew Fleming. Local expression of expansininduces the entire process of leaf development and modifies leaf shape. PNAS September25, 2001 vol.98 no.20 pages Joanna Wyrzykowska, Stephane Pien, Wen Hui Shen, and Andrew Fleming Manipulation of leaf shape by modulation of cell division Development (in press)
5 VI ZUSAMMENFASSUNG Ziel dieser Arbeit ist es, die Rolle der Zellteilung während der Blattinitiierung und -morphogenese besser zu verstehen. Deshalb wurden Experimenteentworfen, um die Zellteilung zu manipulieren und deren Einfluss auf die Blattentwicklung zu beobachten. Insbesondere zwei Aspekte der Zellteilung waren Ziel dieser Manipulationen: die Zellteilungsrate einerseits und die Zellteilungsebene andererseits. Die eingeschlagene Strategie war es, transgene Pflanzen herzustellen, die entweder Komponentender Zellzyklusmaschinerie(um die Teilungshäufigkeit zu manipulieren) oder der Zytokinesemaschinerie (um die Teilungsebene zu manipulieren) unter der Kontrolle des induzierbaren Tetrazyklinpromotersexprimieren. Zur lokalen Mikroinduktion der Genexpression an Meristemenundjungen Blattprimordienwurde zudem eine neue Technikentwickelt. Die für die beschriebene Strategie ausgewählten Gene waren Nicta;cycA3;2 (welches ein pflanzliches Zyklin kodiert), Sp;cdc25 (welches Hefe cdc25 Phosphatase kodiert) and Glyma;phragmoplastin(welches ein pflanzliches Dynamin-ähnliches Protein kodiert). Durch lokale Applikation von Tetrazyklin auf Meristeme oder junge Blattprimordien erreichten wir eine präzise, lokale Ueberexpression dieser Genprodukte, und somit entweder eine veränderte Zellteilungsrate beziehungsweise Zellteilungsebene. Im Meristem führten weder eine erhöhte Teilungsfrequenz noch die Störung der Zellteilungsmuster (durch histologische und Markergenanalyse nachgewiesen) zu einer veränderten Meristemfunktion oder Blattinitiierung. Hingegen hatte eine verändertezellteilungsrate an der Seite junger Blattprimordieneinen dramatischen Effekt auf die Blattform, wobei eine vorübergehenderhöhte Zellteilungsrate zu einer Abnahme der Laminaexpansion führte. Diese Daten zeigen, dass die Rolle der Zellteilung in der Blattmorphogenese Kontext abhängig ist. Das Meristem zeigt gegenüber geänderter Zellteilungsfrequenzund -ebene hohe Toleranz, während in einer gewissen Stufe der Blattentwicklung die lokale Zellteilungsrate einen ausgeprägten Effekt auf die Blattmorphogenese haben kann. Zusammen mit anderen
6 ZUSAMMENFASSUNG VII Daten, zeigen die hier präsentierten Resultate, dass die Kontrolle der lokalen Zellteilungsrate ein Schlüsselmechanismus in der Ausbildung der Blattform darstellenkönnte. Ein Teil dieser Arbeit ist veröffentlichtworden oder befindet sich zur Zeit im Druck: Stephane Pien, Joanna Wyrzykowska, Simon McQueen-Mason,Cheryl Smart, and Andrew Fleming. Local expression of expansininduces the entire process of leaf development and modifies leaf shape. PNAS September25, 2001 vol.98 no.20 pages Joanna Wyrzykowska, Stephane Pien, Wen Hui Shen, and Andrew Fleming Manipulationof leaf shape by modulationof cell division Development (in press)
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