Plant tissue culture: techniques and experiments/
Plant tissue culture: techniques and experiments/
Smith, Roberta H.
- 3rd ed.
- Amsterdam: Elsevier, 2013.
- xvi,188p.
Scheduling and Interrelationships of Exercises
History of Plant Cell Culture
Trevor A. Thorpe
Introduction
The Early Years
The Era of Techniques Development
The Recent Past
Cell Behavior
Plant Modification and Improvement
Pathogen-Free Plants and Germplasm Storage
Clonal Propagation
Product Formation
The Present Era
Setup of a Tissue Culture Laboratory
Work Areas
Media Preparation/Culture Evaluation/Record-Keeping Area
Aseptic Transfer Area
Environmentally Controlled Culture Area
Media Components and Preparation
Inorganic Salts
Plant Growth Regulators
Vitamins
Carbohydrates
Hexitols
Gelling Agent
Amino Acids
Antibiotics
Natural Complexes
Media pH
Medium Preparation
VI Contents
4. Explant Preparation 45
Explant Age 46
Season 46
Explant Size 46
Plant Quality 46
Goal 46
Genotype 48
Aseptic Technique 48
Aseptic Germination of Seeds 49
Carrot, Sunflower, Broccoli, Cotton, or Any Seed of Choice 49
Douglas Fir 50
5. Contamination 53
Explant 53
Disinfecting Agents 54
Explant Source 56
Internal Microbial Contamination 56
Explant Pretreatment to Control Microbial Contaminants 57
Personnel 57
Laminar Air Flow Hood 58
Insects 58
Control of Insect Contamination 58
Media 59
Instruments 60
Room Air Handling System 61
6. Callus Induction 63
Callus Initiation 63
Explant Preparation 65
Culture 66
Observations 66
Data and Questions 66
Explant Orientation 67
Explant 67
Observations 68
Establishment of Competent Cereal Cell Cultures 69
Explants 70
Data and Questions 70
Salt Selection In Vitro 70
Explants 71
Data and Questions 72
Growth Curves 72
Cotton 73
Tobacco 74
Cellular Variation from Callus Cultures 76
Contents vil
8.
9.
Explant
76
Data and Questions
76
Regeneration and Morphogenesis
81
Controlled Morphogenesis: Potato Tuberization 82
Explant 83
Data and Questions 84
Somatic Embryogenesis
84
Explant Preparation
86
Procedure 86
Data and'Questions
86
Regeneration of Rice
87
Regeneration
87
Data and Questions
87
Dormancy Requirements of Explants
87
Explant Preparation
88
Questions
89
Primary Morphogenesis: Douglas Fir
89
Explant Preparation
89
Data and Questions
90
Woody Shrubs and Trees 93
Brent H. McCown
Rhododendrons and Azaleas
^5
Explants
97
Advanced Studies
97
Roses
97
Explant
98
Advanced Studies
98
Birch Trees
98
Explant
98
Advanced Studies
99
White Cedar (Arborvitae)
99
Explant
99
Haploid Plants from Anther Culture
103
Datura Anther Culture
105
Explant Preparation
105
Questions
106
African Violet
106
Explant Preparation
107
Observations
107
Root Tip Chromosome Squash Technique
107
Questions
108
Tobacco
108
VIII
10.
Explant Preparation
108
Observations
109
Questions
109
Anther Squash Technique
109
Embryo Rescue
113
Sweet Corn Embryo Culture
114
Preparation and Culture of Corn Embryos
115
Questions
Crabapple and Pear
116
Explant Preparation
Observations
Questions
Meristem Culture for Virus-Free Plants
119
Isolation of the Shoot Apical Meristem
Explant Preparation
Questions
Establishing Virus- and Bacteria-Free Plants
122
Explant Preparation
122
Garlic Propagation from Bulb Scales
123
Explant Preparation
123
Data
123
In Vitro Propagation for Commercial Production of
Ornamentals
127
Boston Fern
Stage 1
Stage 1!
Stage 111
Staghorn Fern
133
Stage 1
Stage II
134
Stage HI
135
Ficus
136
Stage !
136
Stage II
Stage III
137
Kalanchoe
Explant Preparation
138
Data and Questions
138
African Violet
139
Explant Preparation
139
Data and Questions
140
Endangered Species
Pitcher Plants 141
Cactus -|4-|
Explant Preparation 142
13. Protoplast Isolation and Fusion 147
Jungeun Kim Park, Sunghun Park and James E. Craven
Plant Protoplast Isolation and Fusion 148
Supplies 148
Protoplast Liberation 149
Protoplast Purification 150
Vital Staining 151
Protoplast Fusion 151
Yield Determination 15I
Developing a Protoplast Isolation Protocol 152
14. /\grobacfer/t/m-Mediated Transformation of Plants 155
Jungeun Kim Park, Sunghun Park, Qingyu Wu and Stuart Sprague
Petunia or Tobacco Leaf Disk ^ I57
Sequence I57
Preculture 157
Explant Preparation I57
Cocultivation with Agrobacterium 158
Shoot Transformation and Elimination Agrobacterium 158
Rooting and Pretesting For Transformed Shoots 159
Petunia Shoot Apex 160
Sequence 160
Seed Germination 160
Shoot Induction 161
Shoot Apex Transformation 161
Rooting 161
Tobacco Leaf Infiltration 162
Sequence * 162
Crow Agrobacterium 162
Culture Agrobacterium 163
Harvest Agrobacterium 163
Infiltration Tobacco Leaves 163
9789382291763
630 / SMI/P
Scheduling and Interrelationships of Exercises
History of Plant Cell Culture
Trevor A. Thorpe
Introduction
The Early Years
The Era of Techniques Development
The Recent Past
Cell Behavior
Plant Modification and Improvement
Pathogen-Free Plants and Germplasm Storage
Clonal Propagation
Product Formation
The Present Era
Setup of a Tissue Culture Laboratory
Work Areas
Media Preparation/Culture Evaluation/Record-Keeping Area
Aseptic Transfer Area
Environmentally Controlled Culture Area
Media Components and Preparation
Inorganic Salts
Plant Growth Regulators
Vitamins
Carbohydrates
Hexitols
Gelling Agent
Amino Acids
Antibiotics
Natural Complexes
Media pH
Medium Preparation
VI Contents
4. Explant Preparation 45
Explant Age 46
Season 46
Explant Size 46
Plant Quality 46
Goal 46
Genotype 48
Aseptic Technique 48
Aseptic Germination of Seeds 49
Carrot, Sunflower, Broccoli, Cotton, or Any Seed of Choice 49
Douglas Fir 50
5. Contamination 53
Explant 53
Disinfecting Agents 54
Explant Source 56
Internal Microbial Contamination 56
Explant Pretreatment to Control Microbial Contaminants 57
Personnel 57
Laminar Air Flow Hood 58
Insects 58
Control of Insect Contamination 58
Media 59
Instruments 60
Room Air Handling System 61
6. Callus Induction 63
Callus Initiation 63
Explant Preparation 65
Culture 66
Observations 66
Data and Questions 66
Explant Orientation 67
Explant 67
Observations 68
Establishment of Competent Cereal Cell Cultures 69
Explants 70
Data and Questions 70
Salt Selection In Vitro 70
Explants 71
Data and Questions 72
Growth Curves 72
Cotton 73
Tobacco 74
Cellular Variation from Callus Cultures 76
Contents vil
8.
9.
Explant
76
Data and Questions
76
Regeneration and Morphogenesis
81
Controlled Morphogenesis: Potato Tuberization 82
Explant 83
Data and Questions 84
Somatic Embryogenesis
84
Explant Preparation
86
Procedure 86
Data and'Questions
86
Regeneration of Rice
87
Regeneration
87
Data and Questions
87
Dormancy Requirements of Explants
87
Explant Preparation
88
Questions
89
Primary Morphogenesis: Douglas Fir
89
Explant Preparation
89
Data and Questions
90
Woody Shrubs and Trees 93
Brent H. McCown
Rhododendrons and Azaleas
^5
Explants
97
Advanced Studies
97
Roses
97
Explant
98
Advanced Studies
98
Birch Trees
98
Explant
98
Advanced Studies
99
White Cedar (Arborvitae)
99
Explant
99
Haploid Plants from Anther Culture
103
Datura Anther Culture
105
Explant Preparation
105
Questions
106
African Violet
106
Explant Preparation
107
Observations
107
Root Tip Chromosome Squash Technique
107
Questions
108
Tobacco
108
VIII
10.
Explant Preparation
108
Observations
109
Questions
109
Anther Squash Technique
109
Embryo Rescue
113
Sweet Corn Embryo Culture
114
Preparation and Culture of Corn Embryos
115
Questions
Crabapple and Pear
116
Explant Preparation
Observations
Questions
Meristem Culture for Virus-Free Plants
119
Isolation of the Shoot Apical Meristem
Explant Preparation
Questions
Establishing Virus- and Bacteria-Free Plants
122
Explant Preparation
122
Garlic Propagation from Bulb Scales
123
Explant Preparation
123
Data
123
In Vitro Propagation for Commercial Production of
Ornamentals
127
Boston Fern
Stage 1
Stage 1!
Stage 111
Staghorn Fern
133
Stage 1
Stage II
134
Stage HI
135
Ficus
136
Stage !
136
Stage II
Stage III
137
Kalanchoe
Explant Preparation
138
Data and Questions
138
African Violet
139
Explant Preparation
139
Data and Questions
140
Endangered Species
Pitcher Plants 141
Cactus -|4-|
Explant Preparation 142
13. Protoplast Isolation and Fusion 147
Jungeun Kim Park, Sunghun Park and James E. Craven
Plant Protoplast Isolation and Fusion 148
Supplies 148
Protoplast Liberation 149
Protoplast Purification 150
Vital Staining 151
Protoplast Fusion 151
Yield Determination 15I
Developing a Protoplast Isolation Protocol 152
14. /\grobacfer/t/m-Mediated Transformation of Plants 155
Jungeun Kim Park, Sunghun Park, Qingyu Wu and Stuart Sprague
Petunia or Tobacco Leaf Disk ^ I57
Sequence I57
Preculture 157
Explant Preparation I57
Cocultivation with Agrobacterium 158
Shoot Transformation and Elimination Agrobacterium 158
Rooting and Pretesting For Transformed Shoots 159
Petunia Shoot Apex 160
Sequence 160
Seed Germination 160
Shoot Induction 161
Shoot Apex Transformation 161
Rooting 161
Tobacco Leaf Infiltration 162
Sequence * 162
Crow Agrobacterium 162
Culture Agrobacterium 163
Harvest Agrobacterium 163
Infiltration Tobacco Leaves 163
9789382291763
630 / SMI/P