Characterisation of selected Arabidopsis aldehyde dehydrogenase genes: role in plant stress physiology and regulation of gene expression [Elektronische Ressource] / vorgelegt von Tagnon Degbedji MIssihoun

Tagnon Dègbédji Missihoun

Sujets

• Biologie

Informations

• Publié par: rheinische_friedrich-wilhelms-universitat_bonn
• Publié le: 01 janvier 2011
• Nombre de lectures: 51
• Langue: Deutsch
• Poids de l'ouvrage: 6 Mo

Extrait

Characterisation of selected Arabidopsis
aldehyde dehydrogenase genes: role in plant
stress physiology and regulation of gene
expression

Dissertation

Zur
Erlangung des Doktorgrades (Dr. rer. nat.)
der
Mathematisch-Naturwissenschaftlichen Fakultät
der
Rheinischen Friedrich-Wilhelms-Universität Bonn

vorgelegt von

Tagnon Dègbédji MISSIHOUN

aus
Cotonou, Benin

Bonn, November 2010


Angefertigt mit Genehmigung der Mathematisch-
Naturwissenschaftlichen Fakultät
der Rheinischen Friedrich-Wilhelms-Universität Bonn





Gedruckt mit Unterstützung des Deutschen Akademischen
Austauschdienstes








1. Referentin: Prof. Dr. Dorothea Bartels

2. Koreferent: Priv. Doz. Dr. Hans-Hubert Kirch


Tag der Promotion: 22. Februar 2011

Erscheinungsjahr: 2011

II
DECLARATION

I hereby declare that the whole PhD thesis is my own work, except where explicitly
stated otherwise in the text or in the bibliography.






Bonn, November 2010 ------------------------------------
Tagno D.MISHOUN














III
DEDICATION

To

My wife: Fabienne TOSSOU-MISSIHOUN and our
kids Floriane S. Jennifer and Sègnon Anges-
Anis


My parents: Lucrèce KOTOMALE and Dadjo
MISSIHOUN


My sister and brothers: Mariette, Marius, Ricardo, Renaud, Ulrich


And my dearest aunts and uncles: Hoho, Rebecca, Cyriaque, Dominique,
Alphonsine







IV
CONTENTS

ABBREVIATIONS ...............................................................................................................................................X
FIGURES AND TABLESXIII
SUMMARY............................ 1
1. INTRODUCTION........ 3
1.1 Climate changes and environmental stress...................................................................................... 3
1.2 Plant stress and mechanisms of tolerance........................................................................................ 3
1.3 Gene products related to abiotic stress ............................................................................................ 4
1.3.1 Regulatory pathways of stress-related gene expression in plants............................................... 5
1.3.1.1 Osmotic/oxidative stress signalling ....................................................................................... 5
2+1.3.1.2 Ca -dependent signalling...................................................................................................... 6
1.3.2 ABA signalling........................................................................................................................... 7
1.3.2.1 ABA metabolism ................................................................................................................... 7
1.3.2.2 ABA perception..................................................................................................................... 7
1.3.2.3 ABA signal transduction........................................................................................................ 8
1.3.3 Stress inducible proteins and other compounds........................................................................ 10
1.3.3.1 LEA proteins....................................................................................................................... 10
1.3.3.2 Compatible solutes............................................................................................................... 11
1.3.3.2.1 Mannitol, D-ononitol and sorbitol ................................................................................ 11
1.3.3.2.2 Trehalose ...................................................................................................................... 12
1.3.3.2.3 Sucrose ......................................................................................................................... 12
1.3.3.2.4 Fructans ........................................................................................................................ 13
1.3.3.2.5 Proline .......................................................................................................................... 13
1.3.3.2.6 Glycine betaine............................................................................................................. 14
1.3.3.3 Small RNAs 15
1.3.3.4 Reactive Oxygen Species (ROS) ......................................................................................... 15
1.3.3.5 Aldehydes and the peroxidation of membrane lipids........................................................... 16
1.3.3.6 Aldehyde dehydrogenases (ALDHs) as ROS-detoxifying enzymes.................................... 17
1.3.3.7 Aldehyde dehydrogenase genes........................................................................................... 18
1.3.3.8 Betaine aldehyde dehydrogenases ....................................................................................... 19
1.3.3.9 Aminoaldehyde dehydrogenases and the polyamine metabolism........................................ 20
1.4 Objectives of the study..................................................................................................................... 20
2. MATERIALS AND METHODS.............................................................................................................. 22
2.1 Materials........................................................................................................................................... 22
2.1.1 Plant materials .......................................................................................................................... 22
2.1.2 Chemicals.. 22
2.1.3 DNAs, vectors and bacteria...................................................................................................... 22
2.1.3.1 cDNAs.. 22
2.1.3.2 Vectors................................................................................................................................. 23
V
2.1.3.2.1 pJET1.2......................................................................................................................... 23
2.1.3.2.2 pBT10-GUS.................................................................................................................. 23
2.1.3.2.3 pRTL2-GUS vector ...................................................................................................... 23
2.1.3.2.4 pGJ280.......................................................................................................................... 23
2.1.3.2.5 pET28a 23
2.1.3.2.6 pBIN19 and pROK2 ..................................................................................................... 24
2.1.3.2.7 pPG-Tkan ..................................................................................................................... 24
2.1.3.3 Bacteria................................................................................................................................ 24
2.1.4 Enzymes and DNA-marker 24
2.1.5 Software, programs and online tools ........................................................................................ 25
2.1.6 Machines and other devices...................................................................................................... 25
2.1.7 Membranes 26
2.1.8 Kits............ 26
2.1.9 Media, buffers and solutions .................................................................................................... 26
2.1.9.1 Media.... 26
2.1.9.2 Buffers and solutions ........................................................................................................... 27
2.2 Methods............................................................................................................................................. 28
2.2.1 Growth conditions .................................................................................................................... 28
2.2.1.1 Seed culture and plant growth ............................................................................................. 28
2.2.1.2 Growth of microorganisms .................................................................................................. 29
2.2.2 Primers...... 29
2.2.3 Extraction of nucleic acids ....................................................................................................... 31
2.2.3.1 Extraction of genomic DNA from A. thaliana..................................................................... 31
2.2.3.2 Plasmid DNA mini-prep (Birnboim and Doly 1979; Sambrook et al. 1989)....................... 31
2.2.3.3 Purification and precipitation of DNA................................................................................. 32
2.2.3.4 Extraction of DNA fragments from agarose gels................................................................. 32
2.2.3.5 Extraction of total RNAs from A. thaliana.......................................................................... 32
2.2.4 Qualitative and quantita