Cuticular wax biosynthesis of Lycopersicon esculentum and its impact on transpiration barrier properties during fruit development [Elektronische Ressource] / vorgelegt von Jana Leide

Bot2149

Sujets

• Biologie

Informations

• Publié par: julius-maximilians-universitat_wurzburg
• Publié le: 01 janvier 2009
• Nombre de lectures: 26
• Langue: English
• Poids de l'ouvrage: 4 Mo

Extrait

Cuticular Wax Biosynthesis of
Lycopersicon esculentum and Its Impact on
Transpiration Barrier Properties during
Fruit Development



Dissertation zur Erlangung des
naturwissenschaftlichen Doktorgrades
der Bayerischen Julius-Maximilians-Universität Würzburg



vorgelegt von
Jana Leide
aus Grevesmühlen



Würzburg 2008





















Eingereicht am: ………………...……………………………….………….02.12.2008


Mitglieder der Promotionskommission:

Vorsitzender: Prof. Dr. Martin Müller

Gutachter: Prof. Dr. Markus Riederer

Gutachter: Prof. Dr. Wolfram Hartung


Tag des Promotionskolloquiums: ……...…………………….……………….26.02.2009

Doktorurkunde ausgehändigt am: …………………………...........................……………

























Meinen Eltern



















































CONTENTS

INTRODUCTION....…………….……………………………………..………………..………. 1

CHAPTER I..……...………..………………………………………………………….……... 11
Developmental Pattern of Tomato Fruit Wax Accumulation and Its Impact
on Cuticular Transpiration Barrier Properties: Effects of a Deficiency in the
β-Ketoacyl-CoA Synthase LeCER6
RESULTS………..………………………………………………………..……………...….… 11
Cuticular Water Permeability during Development of Tomato Wild Type and LeCER6-
Deficient Fruits..…………………………………..……………………………………………..... 11
Effects of Wax Extraction and Different Removal Techniques…………………………...…….... 13
Cuticular Wax Coverage and Compound Classes…………………………………………...….… 16
Compositional Changes of Specific Wax Constituents during Fruit Development……….…...…. 18
Chemical Composition of the Cutin Polymer Matrix during Fruit Development...…………......... 20
DISCUSSION…………………………………………………………………………….….… 21
Cuticular Water Permeability and Wax Component Accumulation during Tomato
Fruit Development……………………………………………………………………………….... 22
Impact of LeCER6 Deficiency on Cuticular Wax Component Accumulation…...……………….. 24
Contribution of Different Cuticular Layers and Wax Constituents to the Transpiration Barrier..... 25
Impact of the Cutin Matrix……….…………………………………………………………...…... 26
Contribution of Flavonoids…………………………………………………………………..….… 27
Wax Composition and Cuticular Barrier Properties……….………………………………….…... 28

CHAPTER II………………………………………...…..……………………………….…… 30
Transcriptional Changes Related to the Wax Biosynthesis of Tomato during
Fruit Development: Effects of a Deficiency in the β-Ketoacyl-CoA Synthase
LeCER6
RESULTS…………………………………………………………………………………....… 30
Basic Requirements of a Microarray Analysis on Tomato Fruits………..…………………......…. 30
Design of Oligonucleotide Microarray Experiments Related to Cuticular Wax Biosynthesis.…… 33
Examinations of the Transcriptional Activity of Tomato Fruits………………………………..…. 35
Analysis of Transcriptional Changes Due to LeCER6 Deficiency of Tomato Fruits………….….. 38
Temporal Gene Expression Patterns of Tomato Wild Type and LeCER6-Deficient Fruits.…….... 40
Spatial Gene Expression Patterns of Tomato Wild Type and LeCER6-Deficient Fruits…...…….. 44
Validation of Microarray Experiments by Semi-Quantitative RT-PCR Analysis………...…...….. 45
DISCUSSION……………………………………………………………………….…….…… 47
Gene Expression Pattern of β-Ketoacyl-CoA Synthase LeCER6 Related to Cuticular
Wax Biosynthesis……………………….....................................................................................…. 49
Impact of Deficiency in β-Ketoacyl-CoA Synthase LeCER6: Transcriptional Changes
Associated with Cuticular Wax Coverage……………...………………..……………………..…. 51
Transcriptional Activity with Respect to Cuticular Wax Accumulation: Aspects of Wax
Component Biosynthesis and Wax-Related Transport Processes……………………………....…. 52 with Respect to Stress Adaptation: Aspects of Stress Avoidance
and Tolerance…………………….…………………………………………………………….….. 57 with Respect to Fruit Developmental Processes: Aspects of
Maturation, Expansion, Ripening and Senescence…………………………………………......…. 66
Impact of LeCER6: Gene Expression Involved in Cuticular Wax Accumulation.…...…….…….. 73


CONTENTS
CHAPTER III………………………….…………….……………………..……………..… 75
Analysis of Various Parameters of the Cuticular Wax Biosynthesis in Tomato
RESULTS…………………………………………………………………………………….. 75
Impact of LeCER6 Deficiency on Wax Component Accumulation of Tomato Leaves…..…...… 75
Effects of Atmospheric Humidity and Soil Moisture on Fruit Wax Accumulation…………...… 79
Regeneration of Epicuticular Wax Components on Tomato Fruits after Mechanical Removal… 85
DISCUSSION……………………………………………………………………………….... 90
Organ-Specific Character of LeCER6 Deficiency on Cuticular Wax Composition of Tomato…. 90
Dehydration Stress Related Modifications in Wax Component Accumulation of Tomato Fruits. 95
Induction of Wax Component Regeneration by Mechanical Stress of Tomato Fruits................... 99
Aspects of Cuticular Wax Accumulation as Transpirational Water Loss Barrier of Tomato….... 102

CHAPTER IV..………………………………...…................................................................. 104
LeCer6 Gene: Characterization of a Tomato β-Ketoacyl-CoA Synthase
Belonging to the Very-Long-Chain Condensing Enzymes from the Molecular
Level to the Phenotype
RESULTS………………………………………………………………………..………..….. 104
Isolation and Characterization of the LeCer6 Nucleotide Sequence..…........................…. ……... 104
Nucleotide Sequence Alignment of Different Tomato β-Ketoacyl-CoA Synthases..................…. 108
Amino Acid Sequence Alignment of β-Ketoacyl-CoA Synthases from Different Plant
Species…………………………………………………………………………….……….….…. 109
Visual Differentiation of Tomato Wild Type and LeCER6-Deficient Plants….......................….. 111
Development of Tomato Wild Type and LeCER6-Deficient Fruits…......................................…. 112
Relationship of Different Fruit Parameters of the Tomato Model System …............................…. 120
DISCUSSION……………………………………………………………………….…….….. 121
Detailed Genetic Make-up of β-Ketoacyl-CoA Synthase Gene LeCer6 and Homologies
of Its Gene Product a Very-Long-Chain Condensing Enzyme…...……..………………..…….... 122
Changes in Cell-Cell Communication: Pleiotropic Effects of LeCER6 Deficiency on the
Epidermal Barrier Properties of Tomato Plants………....………….……………………........…. 125
Variations between Tomato Wild Type and lecer6 Mutant: Secondary Effects of
LeCER6 Deficiency on Tomato Fruit Development.................................................................…. 130
Comprehensive Character of LeCER6 Deficiency in Tomato Plants…………….……..........….. 140

CHAPTER V…........................................................................................................…….…... 141
Post-Harvest Formation of the Suberized Stem Scar Tissue during Storage of
Tomato Fruits
RESULTS……………………………………………………………………..…………....… 141
Functional Analysis of the Stem Scar Tissue during Storage of Tomato Fruits…........……..…... 141
Chemical Analysis of the Suberized Stem Scar Membrane of Tomato Fruits…........................... 146
Transcriptional Changes at the Stem Scar Tissue during Storage of Tomato Fruits……….……. 147
Content of Endogenous Abscisic Acid in the Stem Scar Tissue during Storage of
Tomato Fruits……………………………………………………………………….……………. 152
Effects of Exogenous ABA Applications at the Stem Scar Tissue during Storage of ………………………………………………………….. 156
DISCUSSION…………………………………………………………………...……………. 157
Wounding of the Tomato Fruit Due to Harvesting Process…..................................................….. 158
Fruit Stem Scar as a Special Tissue of Tomato Fruits…..........................................................….. 162
Impact of the Suberized Stem Scar Tissue on Tomato Fruits……………………………………. 165
Abscisic Acid as a Regulatory Molecule in Formation of the Stem Scar Tissue…..…….. ….….. 168
Contribution of Suberization to the Barrier Properties of Tomato Fruits………….…………….. 171

CONCLUDING REMARKS AND FUTURE PROSPECTS…..........................................….. 173
SUMMARY……...........................................................................................................…. .…….. 175
ZUSAMMENFASSUNG..……………………………………………………...…...............….. 178

CONTENTS
MATERIALS AND METHODS…………………………………………….…..…………..… 181
Plant Material……………………………………………………………………..……...….....… 181
Developmental Categorization of Tomato Fruits ……………………………...........………...… 181
Determination of Fruit and Leaf Surface Area…...……………………………...…..………..…. 182 Fresh Weight, Dry Weight and Water Content…..................................… 182 <