Table Of ContentMOLECULAR BIOLOGY & BIOTECHNOLOGY OF THE GRAPEVINE
MOLECULAR BIOLOGY
& BIOTECHNOLOGY
OF THE GRAPEVINE
edited by
KALLIOPI A. ROUBELAKIS-ANGELAKIS
Professor of Plant Physiology and Biotechnology,
Department of Biology, University of Crete,
Heraklion, Greece
and
President of the Federation of European Societies of Plant Physiology
SPRINGER-SCIENCE+BUSINESS MEDIA, B.V.
A C.I.P. Catalogue record for this book is available from the Library of Congress.
ISBN 978-94-017-2310-7 ISBN 978-94-017-2308-4 (eBook)
DOI 10.1007/978-94-017-2308-4
Printed an acid-free paper
An Rights Reserved
© 2001 Springer SciencetBusiness Media Dordrecht
Originally published by Kluwer Academic Publishers in 200 1
Softcover reprint of the hardcover 1s t edition 200 1
No part of the material protected by this copyright notice may be reproduced or
utilized in any form or by any means, electronic or mechanical,
including photocopying, recording or by any information storage and
retrieval system, without written permis sion from the copyright owner.
To the Memory of my Parents
Apostolos and Maria Roubelakis
PROLEGOMENA
Research in Plant Biology, in the pre-molecular era, dealt mostly with work at the
organismallevel. The molecular era has opened new avenues in our understanding of
the secrets of life. Molecular Biology and Biotechnology have emerged as the
crossing-point of basic biological sciences, such as Biochemistry, Cellular Biology,
Genetics, Microbiology, and Physiology. The use of molecular techniques and other
analytical instrumentation has increasingly contributed to further understanding' how,
when and where' physiological phenomena occur in organisms. Non-molecular plant
biotechnological methods, such as the plant tissue culture techniques, have been
developed during the past decades whereas the advances in Plant Molecular Biology
have been used for the development of molecular biotechnological application; they
have been based upon the non-molecular developments ..
Grapevine is one of the most widely cultivated plant woody species. As with most
wooc(y plant species, and also some cereals and legumes, Molecular Biology and
Biotechnology have had progressed at a slower pace, due to several obstacles, which
have had to be overcome. In any case, it is now that substantial progress has been
made and useful information has been accumulated in the literature.
During the last decade, more than 100 genes have been characterized from grape
vine and several genomic and chloroplastic microsatellite sequences have been
deposited in the Genbanks. These genes encode for enzymes mediating synthesis and
transport of sugars, polyp henols and pigments, organic acids, amino acids and
polyamines, as well as for proteins related to biotic and abiotic stresses and to cell wall
structure. Furthermore, protocols for non-molecular and molecular biotechnol-ogical
applications for grapevine have been published.
In an effort to collect and present the available information on Grapevine Molecular
Biology and Biotechnology, 51 scientists from 10 countries jointly worked for the
preparation of this Book. It is intended to be used as a reference-book by researchers,
graduate and undergraduate students, viticulturists, biotechnological companies and
any scientist, who is interested in the Molecular Biology and Biotechnology of
Grapevine.
Sincere thanks are due to all worldWide-leading scientists in their field, who have
contributed and especially for their impeccable collaboration during the preparation
Vlll
of this Volume; to Mrs Mary Papadakis-Savvopoulos for editorial assistance; to Miss
Maria Mandelenakis for secretarial assistance; to Mr Nikolaos Papadoyannakis for
his endless and devoted work during the preparation oft he ready-to-camera material;
to Kluwer Academic Publishers for the publication of the Book. Last but lot least to
my husband, Andreas Angelakis, for his continuous encouragement and patience.
Herak/ion, Crete, Greece
January 2001
Kalliopi A. Roubelakis-Angelakis
University ofC rete, Greece
CONTENTS
Contributing Authors xxi
Chapter 1
MOLECULAR BIOLOGY OF SUGAR AND ANTHOCYANIN
ACCUMULA TION IN GRAPE BERRIES 1
P.K. Boss and C. Davies
1. Introduction 1
2. The Molecular biology of sugar transport and accumulation in grape 2
2.1. Grape sucrose transporters 5
2.2. Grape monosaccharide transporters 6
2.3. Grape invertases 9
2.4. Future directions 12
3. Anthocyanins 13
3.1. Grape anthocyanins 13
3.2. The anthocyanin biosynthesis pathway 14
3.2.1. Introduction 14
3.2.2. The structural genes 14
3.2.3. Genes involved in pathway regulation 17
3.3. Grape anthocyanin gene expression 18
3.3.1. Anthocyanin gene expression in grapevine seedlings 18
3.3.2. Anthocyanin gene expression in berry skins during development 18
3.3.3. Anthocyanin gene expression in red and white grapes 21
3.5. ManipUlating grapevine anthocyanins 24
3.5.1. Total anthocyanins 24
3.5.2. Specific anthocyanins 25
4. Summary 27
Acknowledgments 28
References 28
Chapter 2
GRAPE BERRY ACIDITY 35
N. Terrier and C. Romieu
1. Introduction 35
2. Changes in acidity during berry development 36
2.1. Evolution pattern of berry composition 36
2.2. Organic acid metabolic pathways in grape berries 38
x
2.2.1. Organic acid synthesis 38
2.2.2. The induction of malate respiration during ripening 38
2.2.3. Aerobic fermentation and malate breakdown 40
3. Compartmentation of organic acids in grape berries 41
3.1. Vacuolar proton pumps 41
3.1.1. Molecular structure 41
3.1.2. Thermodynamic properties 42
3.1.3. Enzymic properties 43
3.1.4. Two pumps on the same membrane 44
3.2. Organic acid accumulation 46
3.3. Vacuolar transport and pH variation 49
3.3.1. Proton pumps 49
3.3.2. Secondary transport 51
3.3.3. Vacuolar content efflux 51
References 52
Chapter 3
NITROGEN ASSIMILATION IN GRAPEVINE 59
K.A Loulakakis and K.A Roubelakis-Angelakis
1. Introduction 59
2. Nitrogen assimilation 60
2.1. Reduction of nitrate 60
2.2. Ammonium assimilation 63
2.2.1. Glutamine synthetase 65
2.2.2. Glutamate synthase 68
2.2.3.Glutamate dehydrogenase 71
3. Regulation of ammonia assimilating enzymes in grapevine by nitrogen
source 77
4. Future perspectives 80
References 80
Chapter 4
MOLECULAR BIOLOGY AND BIOCHEMISTRY OF PROLINE
ACCUMULATION IN DEVELOPING GRAPE BERRIES 87
R. van Heeswijck, AP. Stines, J. Grubb, I. Skrumsager Mpller and P.B. Hpj
1. Introduction 87
2. Amino acid composition of grape berries 87
3. The influence of grape berry proline on fermentation 91
xi
4. Proline accumulation in plants 91
5. Pathways of proline biosynthesis 92
5.1. The glutamate pathway of proline biosynthesis 92
5.2. The Ornithine pathway of proline biosynthesis 94
5.3. Genes encoding P5CS and OAT are expressed in grape berry tissue 94
6. Vvp5cs gene expression during grape berry development 97
7. Other factors which could affect proline accumulation in grape berries 99
7.1. Ammonium and glutamine metabolism 99
7.2. Arginine metabolism and regulation of OAT 100
7.3. Proline degradation 101
7.4. Protein accumulation 102
8. Conclusions 103
Acknowledgments 104
References 104
ChapterS
POLYAMINES IN GRAPEVINE 109
K.A. Paschalidis, A. Aziz, L. Geny, N.!. Primikirios and
K.A. Roubelakis-Angelakis
1. Introduction 109
2. Biosynthesis of polyamines 110
3. Endogenous polyamines in grapevine organs 112
3.1. Polyamines in various grapevine organs ll2
3.2. Spatial and temporal free and conjugated polyamine distribution in
grapevine leaves ll2
3.3. Polyamines and berry development 116
3.3.1. Polyamine oxidase activities and diaminopropane contents during
floral development in grapevine 116
3.3.2. Hydroxycinnamic acid amines in flowers and berries of grapevine 118
4. ADC enzyme activity and transcript levels in developing grapevine organs 119
5. Polyamines and disorders of grape berry development 121
5.1. Polyamines and fruit set 121
5.2. Polyamines and abnormal development of berry (shot grape berries) 122
5.3. Polyamine metabolism in relation to flower and fruitlet abscission 122
5.3.1. Polyamines and abscission potential 124
5.3.2. Polyamines counteract abscission 125
5.3.3. Polyamine biosynthesis and abscission 128
5.3.4. Polyamine catabolism and abscission 129
xii
5.3.5. Photodependance of polyamine levels and abscission 130
5.3.6. Modulation of carbohydrate and amino acid levels by polyamines 130
6. Polyamines and stress 133
6.1. Free polyamines, ADC enzyme activity and transcript levels in
grapevine cell suspension cultures under different treatments 133
6.2. Free polyamine titers and stress adaptation 136
6.3. Polyamines and potassium nutrition 140
6.3. Polyamines and biotic stress (Botrytis cinerea) 142
References 144
Chapter 6
PHYSIOLOGICAL ROLE AND MOLECULAR ASPECTS OF GRAPEVINE
STILBENIC COMPOUNDS 153
L. Bavaresco and C. Fregoni
1. Introduction 153
2. Plant disease resistance mechanisms 153
3. Phytoalexins and biotic/abiotic elicitors 154
4. Grapevine induced stilbenes 155
4.1. First evidence of stilbenes in grapevine 155
4.2. Biotic elicitors 157
4.2.1. Botrytis cinerea 158
4.2.2. Plasmopara viticola 161
4.2.3. Phomopsis viticola 162
4.2.4. Rhizopus stolonifer 162
4.2.5. Bacteria 163
4.3. Abiotic elicitors 163
4.3.1. UV irradiation 163
4.3.2. Aluminum chloride 165
4.3.3. Ozone 165
4.3.4. Wounding 165
4.3.5. Fosetyl-Al 166
4.3.6. Other chemicals 166
4.4. Stilbene glycosides in Vitis 166
4.5. Cultural factors affecting induced stilbene synthesis 167
4.5.1. Fertilizer supply 167
4.5.2. Rootstock 168
5. Stilbenes in soft tissues of field grow grapevines 168
6. Grapevine constitutive stilbenes 169
7. Stilbenes in the wine 170
8. Molecular and biotechnological aspects of stilbene synthesis in grapevine 171
