defended by Anna CIANCOLINI

250 Pages·2012·8.24 MB·English

PhD THESIS
under Italy-France joint supervision

for the

DOTTORATO DI RICERCA IN ORTOFLOROFRUTTICOLTURA –
XXIV CICLO

awarded by the Università degli Studi della Tuscia, Italy

and the

DOCTORAT DE L’UNIVERSITÉ DE TOULOUSE

awarded by the Institut National Polytechnique de Toulouse, France

Discipline: Sciences des Agroressources

defended by Anna CIANCOLINI
July, 6th 2012

Title: CHARACTERIZATION AND SELECTION OF GLOBE ARTICHOKE AND
CARDOON GERMPLASM FOR BIOMASS, FOOD AND BIOCOMPOUND
PRODUCTION

Research units: Unità Tecnica Sviluppo Sostenibile ed Innovazione del Sistema Agro-Industriale (ENEA),
Dipartimento di Scienze e Tecnologie per l'Agricoltura, le Foreste, la Natura e l'Energia (UNITUS)
Laboratoire de Chimie Agro-industrielle (UMR 1010 INRA/INP-ENSIACET)

Directors of the PhD thesis: Prof. Mario Augusto PAGNOTTA and Prof. Gérard VILAREM

Tutors of the PhD thesis: Dr. Paola CRINO’ and Dr. Marion ALIGNAN

PhD Coordinators: Prof. Alberto GRAIFENBERG and Prof. Michel CAFFAREL

A mio marito, gioia infinita dei miei giorni.
Ai miei genitori, punto fermo del mio essere.
Alla vita che porto dentro con tutto l’amore che posso.

TABLE OF CONTENTS

INTRODUCTION 2

1.1 CYNARA SPP. ORIGIN AND DOMESTICATION 4
1.2 TAXONOMY AND NOMENCLATURE 5
1.3 BOTANICAL DESCRIPTION 6
1.4 BIOLOGY AND ECOLOGY 9
1.5 GEOGRAPHIC DISTRIBUTION AND PRODUCTION 10
1.6 GENETIC RESOURCES 13
1.7 PROPAGATION 15
1.7.1 AGAMIC 15
1.7.2 GAMIC 17
1.8 AGRONOMIC ASPECTS 18
1.8.1 PLANTING 18
1.8.2 FERTILIZATION 18
1.8.3 IRRIGATION 19
1.8.4 WEED CONTROL 19
1.8.5 PARASITES AND PATHOGENS 20
1.8.6 AGRICULTURAL MANAGEMENT OF THE PLANTS 21
1.8.6.1 STALK REMOVAL 21
1.8.6.2 BUD CUTTINGS 21
1.8.6.3 BLEACHING 22
1.9 EDIBLE PRODUCT HARVESTING AND COMMERCIALIZATION 22
1.10 USES 23
1.10.1 TRADITIONAL USES 23
1.10.2 INDUSTRIAL USES 24
1.10.3 PHARMACEUTICAL USES AND MAJOR PHENOLIC COMPOUNDS 27
1.11 FOCUS ON PHENOLIC EXTRACTION TECHNIQUES 34

EXPERIMENTAL PART 38

CHAPTER 1. MORPHOLOGICAL ANALYSIS AND GERMPLASM SELECTION
FOR EDIBLE USE

GLOBE ARTICHOKE 42
1.1 INTRODUCTION 42
1.2 ITALIAN SPRING LANDRACES 44
1.2.1 MATERIALS AND METHODS 44
1.2.1.1 GENETIC RESOURCES AND OPEN FIELD EXPERIMENTS 44
1.2.1.2 MORPHOLOGICAL CHARACTERIZATION USING UPOV DESCRIPTORS 47
1.2.1.3 STATISTICAL ANALYSES 48

1.2.2 RESULTS 50
1.2.3 DISCUSSION 57
1.3 ITALIAN ‘ROMANESCO’ CLONES 58
1.3.1 MATERIALS AND METHODS 58
1.3.1.1 GENETIC RESOURCES AND OPEN FIELD EXPERIMENTS 58
1.3.1.2 MORPHOLOGICAL CHARACTERIZATION USING UPOV DESCRIPTORS 59
1.3.1.3 STATISTYCAL ANALYSES 59
1.3.2 RESULTS 59
1.3.3 DISCUSSION 68

CARDOON 69
1.4 INTRODUCTION 69
1.5 ITALIAN ACCESSIONS 70
1.5.1 MATERIALS AND METHODS 70
1.5.1.1 GENETIC RESOURCES AND OPEN FIELD EXPERIMENTS 70
1.5.1.2 MORPHOLOGICAL CHARACTERIZATION 72
1.5.1.3 STATISTICAL ANALYSES 72
1.5.2 RESULTS 72
1.5.3 DISCUSSION 78

CHAPTER 2. DEVELOPMENT OF POLYPHENOL EXTRACTION TECHNIQUE
FROM CYNARA SPP. BIOMASS

2.1 INTRODUCTION 84
2.2 MATERIALS AND METHODS 85
2.2.1 PLANT MATERIAL, SOLVENTS AND REAGENTS 85
2.2.2 EXTRACTION TECHNIQUES 85
2.2.2.1 SOXHLET EXTRACTION 85
2.2.2.2 MACERATION 86
2.2.2.3 MICROWAVE ASSISTED EXTRACTION 86
2.2.2.4 ACCELERATED SOLVENT EXTRACTION 86
2.2.3 DETERMINATION OF MOISTURE CONTENT 87
2.2.4 COLORIMETRIC ANALYSIS OF TOTAL PHENOLICS 87
2.2.5 HPLC ANALYSIS 87
2.2.6 STATISTICAL ANALYSES 88
2.2.7 EXPERIMENTAL DESIGN FOR MICROWAVE ASSISTED EXTRACTION AND
ACCELERATED SOLVENT EXTRACTION 88
2.3 RESULTS 89
2.3.1 OPTIMIZATION OF MICROWAVE ASSISTED EXTRACTION (MAE) AND
ACCELERATED SOLVENT EXTRACTION (ASE) FOR PHENOLIC COMPOUNDS 89

2.3.2 COMPARISON BETWEEN NOVEL AND TRADITIONAL EXTRACTION METHODS 98
2.3.2.1 COLORIMETRIC ANALYSIS OF TOTAL PHENOLICS 98
2.3.2.2 HPLC ANALYSIS 100
2.4 DISCUSSION 103

CHAPTER 3. BIOMASS AND NUTRACEUTICAL KINETICS EVALUATION IN
GLOBE ARTICHOKE AND CARDOON PLANTS GROWN IN OPEN FIELD

3.1 INTRODUCTION 108
3.2 MATERIALS AND METHODS 109
3.2.1 FIELD EXPERIMENTS 109
3.2.2 AGRO-MORPHOLOGICAL CHARACTERIZATION 109
3.2.3 BIOCHEMICAL CHARACTERIZATION 110
3.2.4 DETERMINATION OF MOISTURE CONTENT 110
3.2.5 SOLVENTS AND REAGENTS 110
3.2.6 EXTRACTION PROCEDURE 111
3.2.7 HPLC ANALYSIS 111
3.2.8 COLORIMETRIC ANALYSIS OF TOTAL PHENOLICS 112
3.2.9 STATISTICAL ANALYSES 113
3.3 RESULTS 113
3.3.1 AGRO-MORPHOLOGICAL CHARACTERIZATION AND PLANT GROWTH
EVALUATION 113
3.3.2 BIOCHEMICAL CHARACTERIZATION AND PHENOLIC COMPOUND KINETICS
EVALUATION 121
3.4 DISCUSSION 130

CHAPTER 4. MORPHOLOGICAL ANALYSIS AND GERMPLASM SELECTION
FOR BIOMASS AND BIOCOMPOUND PRODUCTION

4.1 INTRODUCTION 136
4.2 GLOBE ARTICHOKE: EVALUATION OF BIOMASS AND
BIOCOMPOUND PRODUCTION 137
4.2.1 MATERIALS AND METHODS 137
4.2.1.1 EXPERIMENTAL FIELD AND PLANT MATERIAL 137
4.2.1.2 MORPHOLOGICAL CHARACTERIZATION 138
4.2.1.3 PHENOLIC COMPOUND EXTRACTION AND ANALYSIS 139
4.2.1.3.1 SAMPLE PREPARATION 139
4.2.1.3.2 EXTRACTION PROCEDURE AND ANALYSIS 140
4.2.1.4 STATISTICAL ANALYSES 140
4.2.2 RESULTS 140
4.2.2.1 BIOMETRIC CHARACTERISTICS 140
4.2.2.1.1 BIOMASS PRODUCTION 140