Anthocyanins: Biosynthesis, Functions, and Applications

344 Pages·2009·7.119 MB·English

by  Jean-Hugues B. Hatier, Kevin S. Gould (auth.), Chris Winefield, Kevin Davies, Kevin Gould (eds.)

Anthocyanins
· ·
Kevin Gould Kevin Davies Chris Winefield
Editors
Anthocyanins
Biosynthesis, Functions, and Applications
123
Editors
KevinGould KevinDavies
School of Biological Sciences CropandFoodResearch
Victoria University of Wellington PalmerstonNorth
Wellington NewZealand
NewZealand [email protected]
[email protected]
ChrisWinefield
LincolnUniversity
Canterbury
NewZealand
winefi[email protected]
ISBN:978-0-387-77334-6 e-ISBN:978-0-387-77335-3
DOI:10.1007/978-0-387-77335-3
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P reface

Amongst the range of naturally occurring pigments, anthocyanins are arguably the
best understood and most studied group. Research into their occurrence, inheritance
and industrial use encompasses hundreds of years of human history, and many
volumes are dedicated to describing the prevalence, type and biosynthesis of
anthocyanins. Only recently have studies begun to explain the reasons for the
accumulation of these red pigments in various tissues of plants. Indeed it has only
been within the last 20 years, coinciding with the ability to genetically manipulate
plants, that we have begun to tease out the multitude of roles that these compounds
play within plants. Alongside these fundamental advances in understanding the
functional attributes of anthocyanins in-planta we are now beginning to realise the
potential of anthocyanins as compounds of industrial importance, both as pigments
in their own right and also as pharmaceuticals.
With this backdrop, the 4th International Workshop on Anthocyanins was
convened in Rotorua, New Zealand in January 2006. The programme was designed
to bring together a wide range of researchers across an array of disciplines to
highlight the increasing importance of these pigments as a research area but also as
chemicals of wider importance to human activity. The chapters in this book represent
a collection of recent and highly relevant reviews prepared by participants in this
symposium who are internationally recognised experts in their respective fields.
The book is divided into 10 chapters that address a wide range of topics including
the proposed roles of anthocyanins in vivo, methods for manipulating the
biosynthesis of anthocyanins to both produce new pigments as well as pigments that
exhibit greater stability, use of plant and microbial cell cultures for large scale
production of anthocyanins for industrial uses and the effectiveness of anthocyanins
as pharmaceutical compounds.
Chapter 1 reviews the current thinking about the role of anthocyanins in leaves,
stems roots and other vegetative organs. The authors point out that current roles for
anthocyanins such as participation in photoprotective, UV-B protective, and
protection from oxidative stresses do not adequately explain the range of spatial and
vi Preface

temporal distributions of anthocyanins observed in plants. They hypothesise that
anthocyanins possess a more indirect role in signalling and developmental regulation
in response to oxidative stress.
The discussion in Chapter 1 is expanded on in Chapter 2, whose authors proceed
to examine the potential roles of anthocyanins in plant/animal interactions. In
particular this chapter reviews current thought and recent experimental evidence on
plants’ use of anthocyanins as visual clues that provide information to animals about
palatability of plant structures and the potential role of anthocyanins in camouflage,
undermining insect crypsis and in mimicry of defensive structures.
The molecular basis for spatial and temporal regulation of anthocyanin
biosynthesis forms the focus of Chapter 3. This review concentrates on the recent
advances in our understanding of the biosynthesis and molecular regulation of this
pathway and how this information has been used in conjunction with recombinant
DNA technologies to manipulate anthocyanin production in plants for both scientific
and commercial applications.
Picking up again the theme of in vivo roles for anthocyanins, Chapter 4 reviews
the role of anthocyanin pigmentation in fruits and adaptive advantages accumulation
of these pigments confers to plants. In particular the author concentrates on the
accumulation of anthocyanin pigments in fruits in response to environmental factors,
seed disperser visual systems and fruit quality parameters. Accumulation of pigments
contributing differing hues to fruit is discussed with respect to the interaction with
animal dispersers and as a measure of fruit ripeness and quality.
Chapter 5 provides an in-depth review of the use of plant cell cultures for the
industrial production of anthocyanins for use as high quality food pigments. A wide
range of plant species are reviewed as to their ability to produce cell cultures capable
of production of anthocyanins in cell culture, the types of cultures obtained and the
pros and cons of using these cell types as production systems for anthocyanins.
Methodology to increase the production of anthocyanins from these cultures, and
limitations of these cultures for anthocyanin production are discussed alongside
potential methods for overcoming production barriers that currently prevent large
scale anthocyanin production from plant derived cell cultures.
Anthocyanin stability and the colour imparted to plant tissues by anthocyanin
accumulation are in part due to the extent and nature of secondary modifications to
the anthocyanin aglycone. Glycosylation, acylation and methylation are especially
important in altering the chemical characteristics of anthocyanins both in vivo and
in vitro. In Chapter 6 the authors review the current advances in our understanding of
the biochemical pathways that lead to these chemical modifications and how this
information may be utilized to modify and stabilize anthocyanins both in plants and
for industrial uses such as those described in Chapter 5 and later in Chapter 9.
While production of anthocyanins in plant cell cultures is well documented,
production of these compounds in microbial cell culture systems is a relatively new
concept. With the increasing knowledge of the biosynthesis of anthocyanins in plant
systems it has become feasible to engineer microbial species to contain a functional
anthocyanin pathway. Chapter 7 reviews current advances in this area and is split
into two sections. The first deals with advances in the metabolic engineering of
bacterial and yeast species for anthocyanin production while the second section
Preface vii

reviews the endogenous biotransformations carried out by host species and provides
a excellent counterpoint to natural modification schemes found in plants that is
reviewed in Chapter 6.
Continuing the theme of industrial application of flavonoids and anthocyanins
developed in earlier chapters, Chapter 8 turns our attention to the utility of this
ubiquitous chemical group in agricultural systems. In particular the roles of
anthocyanins, related flavonoids and their derivatives in forage and forage legume
species is discussed in the context of the value of these compounds in these crops
from an animal health and nutrition perspective. Potential methods for manipulating
the levels of important compound classes are reviewed from both a genetic
modification and traditional breeding standpoint.
Returning to anthocyanins as food colorants, Chapter 9 develops themes outlined
in Chapter 5 with a review of how anthocyanins are currently utilized in the food
industry from sources thorough isolation and analytical methodologies employed.
While industrial utilization of anthocyanins is still to be realized on a wide scale, this
review covers the potential and perspectives for anthocyanins and their derivatives in
food products.
Finally Chapter 10 concludes the book with a discussion of anthocyanins and
other flavonoids as phytochemicals that promote human health. This very relevant
topic is reviewed with a particular emphasis on the interaction of these compounds
with other components of diet to protect and enhance human heath. The chapter
describes how plant cell cultures and research models are being used to increase our
understanding of the complex and multi-faceted roles that interacting phytochemicals
play in the human body, specifically in the context of developing novel insights into
the competing mechanisms of action, bioavailability and distribution in situ.
The editors, Associate Professor Kevin Gould, Dr. Kevin Davies and I hope that
this book will both provide a valuable reference resource and provide inspiration for
new researchers in this exciting and rapidly expanding field.

Dr. Chris Winefield
January 2008
Contents

Preface........................................................................................................................v
Contributors..........................................................................................................xvii
1. Anthocyanin Function in Vegetative Organs.....................................................1
Jean-Hugues B. Hatier and Kevin S. Gould
1.1 Introduction...................................................................................................1
1.2 Anthocyanins and Stress Responses..............................................................2
1.3 Photoprotection.............................................................................................3
1.4 Protection Against Ultraviolet Radiation.......................................................6
1.5 Free Radical Scavenging...............................................................................7
1.6 Paradigm Shift.............................................................................................10
1.7 Modulation of Signalling Cascades: A New Hypothesis.............................10
References............................................................................................................12
2. Role of Anthocyanins in Plant Defence............................................................21
Simcha Lev-Yadun and Kevin S. Gould
2.1 Introduction.................................................................................................21
2.2 Hypotheses..................................................................................................23
2.3 Reluctance to Accept Hypotheses on Defensive Colouration.....................23
2.4 Colour Vision in Animals............................................................................24
2.5 Anthocyanins and Other Red Pigments.......................................................25
2.6 Olfactory Signals.........................................................................................25
2.7 Aposematic Colouration..............................................................................26
2.7.1 Poisonous Plants...............................................................................26
2.7.2 Thorny plants....................................................................................26
x Contents

2.8 Defensive Mimicry......................................................................................29
2.8.1 Mimicry of Dead Leaves..................................................................29
2.8.2 Defensive Thorn Automimicry.........................................................30
2.8.3 Defensive Animal Mimicry by Plants...............................................31
2.8.4 Ant Mimicry.....................................................................................31
2.8.5 Aphid Mimicry.................................................................................32
2.8.6 Mimicry of Aposematic Poisonous Caterpillars...............................32
2.9 Camouflage.................................................................................................33
2.9.1 Whole Plants and Seeds....................................................................33
2.9.2 Variegation in Understory Herbs......................................................34
2.10 Undermining Crypsis of Invertebrate Herbivores.......................................34
2.11 Red Young Leaves Divert Herbivores from More Costly Old Ones..........35
2.12 Signalling by Red Autumn Leaves..............................................................36
2.12.1 General..............................................................................................36
2.12.2 Signalling of Defensive Potential.....................................................36
2.12.3 The “Defence Indication Hypothesis”..............................................38
2.12.4 Aposematism of Red Autumn Leaves..............................................38
2.13 Conclusions and Suggestions for Further Research....................................39
2.13.1 Leaf Colouration and the Level of Risk............................................39
2.13.2 No Defence is Perfect.......................................................................40
2.13.3 Exceptions.........................................................................................41
References............................................................................................................41
3. Modifying Anthocyanin Production in Flowers..............................................49
Kevin M. Davies
3.1 Introduction.................................................................................................49
3.2 Anthocyanin Biosynthesis in Flowers.........................................................50
3.3 Anthocyanins as Flower Pigments..............................................................57
3.4 Regulation of Anthocyanin Production in Flowers.....................................58
3.5 Genetic Modification of Anthocyanin Biosynthesis....................................60
3.5.1 Preventing Anthocyanin Production.................................................62
3.5.2 Increasing Anthocyanin Production by Altering
Biosynthetic Enzyme Activity..........................................................64
3.5.3 Anthocyanins with Unusual Patterns of A- or C-Ring
Hydroxylation...................................................................................65
3.5.4 Generating New Flower Colours by Altering Anthocyanin
B-Ring Hydroxylation......................................................................66
3.5.5 Changing Flower Colour by Altering Anthocyanin
Secondary Modifications..................................................................69
3.5.6 Black Flower Colours.......................................................................70
3.5.7 GM Application of Anthocyanin-related Transcription Factors.............71
3.6 Concluding Comments................................................................................73
Acknowledgments................................................................................................73
References............................................................................................................74
Contents xi

4. Prevalence and Functions of Anthocyanins in Fruits.....................................85
W. J. Steyn
4.1 Introduction.................................................................................................85
4.2 Prevalence of Fruit Colours.........................................................................86
4.3 Developmental Patterns...............................................................................88
4.4 Distribution of Anthocyanins in Fruit..........................................................89
4.5 Environmental Regulation of Colour Development....................................89
4.5.1 Light..................................................................................................90
4.5.2 Temperature......................................................................................90
4.5.3 Other Factors.....................................................................................91
4.6 Anthocyanins in Attraction..........................................................................92
4.6.1 Visual Systems..................................................................................92
4.6.2 Red Fruits..........................................................................................93
4.6.3 Blue and Black Fruits........................................................................94
4.7 Fruit Quality and Composition....................................................................95
4.7.1 Health Benefits.................................................................................95
4.7.2 Nutritional Content and Defensive Strength.....................................96
4.7.3 Maturity............................................................................................97
4.8 Anthocyanin and Fruit Size.........................................................................98
4.9 Photoprotection...........................................................................................98
4.10 Perspectives.................................................................................................99
References..........................................................................................................100
5. Anthocyanin Biosynthesis in Plant Cell Cultures:
A Potential Source of Natural Colourants.....................................................107
Simon Deroles
5.1 Introduction...............................................................................................107
5.1.1 The Anthocyanins...........................................................................107
5.1.2 Plant Cell Cultures..........................................................................109
5.2 Daucus carota (Carrot)..............................................................................111
5.2.1 Types of Anthocyanins...................................................................111
5.2.2 Glucosyltransferases from Carrot Cell Cultures.............................111
5.2.3 Phytohormones...............................................................................112
5.2.4 GA .................................................................................................113
3
5.2.5 Nutrients (Carbon, Nitrogen, Phosphate)........................................114
5.2.6 Elicitation........................................................................................115
5.2.7 Light................................................................................................116
5.2.8 Aggregate Size................................................................................117
5.2.9 Future Strategies for Enhanced Production of Anthocyanins.........117
5.3 Vitis vinifera (Grape).................................................................................118
5.3.1 Types of Anthocyanins...................................................................119
5.3.2 Modification of Anthocyanins in Grape Cell Cultures...................119
5.3.3 Phytohormones...............................................................................119
5.3.4 Nutrients (Carbon, Nitrogen, Phosphate)........................................120
5.3.5 pH, Conditioned Media and Feeder Layers....................................122
5.3.6 Elicitation and Light.......................................................................122
xii Contents

5.3.7 Localisation of Anthocyanins in the Plant Cell...............................123
5.3.8 Physical Parameters........................................................................125
5.3.9 Conclusions.....................................................................................126
5.4 Fragaria ananassa (Strawberry)...............................................................126
5.4.1 Types of Anthocyanins...................................................................126
5.4.2 Phytohormones...............................................................................127
5.4.3 Nutrients (Carbon, Nitrogen, Phosphate)........................................127
5.4.4 Conditioned Medium......................................................................128
5.4.5 Elicitation........................................................................................129
5.4.6 Physical parameters........................................................................130
5.4.7 Conclusions.....................................................................................132
5.5 Ajuga Species............................................................................................132
5.5.1 Types of Anthocyanin.....................................................................132
5.5.2 Phytohomones.................................................................................133
5.5.3 Nutrients (Carbon, Nitrogen, Phosphate)........................................133
5.5.4 Physical Parameters........................................................................134
5.5.5 Conclusions.....................................................................................134
5.6 Ipomea batatas (Sweet Potato)..................................................................134
5.6.1 Types of Anthocyanin.....................................................................135
5.6.2 Phytohormones...............................................................................135
5.6.3 Nutrients (Carbon, Nitrogen, Phosphate)........................................135
5.6.4 Elicitation........................................................................................136
5.6.5 Conclusions.....................................................................................137
5.7 Perilla frutescens.......................................................................................137
5.7.1 Carbon Source and Elicitation........................................................137
5.7.2 Physical Parameters........................................................................137
5.7.3 Conclusions.....................................................................................139
5.8 Vaccinium Species.....................................................................................139
5.8.1 Types of Anthocyanins...................................................................139
5.8.2 Phytohormones...............................................................................140
5.8.3 Media Components and Elicitation.................................................140
5.8.4 Physical Parameters........................................................................141
5.8.5 Conclusions.....................................................................................142
5.9 Other plant Cell Lines...............................................................................142
5.9.1 Aralia Cordata................................................................................142
5.9.2 Bupleurum falcatum........................................................................143
5.9.3 Callistephus chinensis (China Aster)..............................................143
5.9.4 Campanula glomerata....................................................................143
5.9.5 Camptotheca acuminata.................................................................143
5.9.6 Catharanthus roseus.......................................................................144
5.9.7 Centaurea cyanus...........................................................................144
5.9.8 Euphorbia millii..............................................................................145
5.9.9 Fagopyrum esculentum (Buckwheat).............................................145
5.9.10 Glehnia littoralis.............................................................................146
5.9.11 Haplopappus gracilis......................................................................147
5.9.12 Hibiscus sabdariffa (Roselle).........................................................147