Biotechnology

Preface
List of Contributors
1. Biotechnology for Medicinal Plants: Research Need
J.M. Merillon and K.G. Ramawat
I. Introduction
II. Medicinal Plants Exploration
III. Resources: Wild and Cultivated Plants
IV. Role of Biotechnological Approaches
Development of Seed Material for Domestication
Improvement — Somaclonal Variation and Genetic
Engineering
Understanding Metabolic Pathways
New Compounds/Derivatives Production
Scaling-up Technology Through Bioreactors
Plant Molecular Farming
Metabolomics
V. Conclusions
References
VI. Further Readings
2. Secondary Plant Products in Nature
K.G. Ramawat
I. Introduction
II. Alkaloids
A. Definition
B. Biosynthesis of Alkaloids
C. Occurrence of Alkaloids
D. Distribution in Animal Kingdom and Lower Plants
E. Classifications of Alkaloids
F. Biological Functions of Alkaloids
III. Other Metabolites
A. Plant Amines
B. Non-protein Amino Acids in Plants
C. Quinones
D. Resins
E. Steroids
F. Phenylpropanoids
G. Polyisoprene (Rubber)
H. Phenolics
I. Flavours and Colours
J. Insecticides of Plant Origin
IV. Conclusions
References
V. Further Readings
3. Factors Affecting the Production of
Secondary Metabolites
K.G. Ramawat and Meeta Mathur
I. Introduction
II. Culture Conditions
The Environment
III. Culture Initiation
IV. Time-course Study of Growth and
Secondary Metabolite Production
V. Optimization
A. Physical Factors
B. Effect of Nutrients
Use of Elicitors
VII. Selection of Clones
A. Variability in Field-grown Materials
B. State of Differentiation
C. Producer/Non-producer Cells
D. Plating and Selection
VIII. Clonal Stability
IX. Flow Cytometry
X. Optimization and Selection
XI. Selection Parameters
XII. Assessment of Somaclonal Variation
XIII. Prospects
References
XIV. Further Readings
4. Production of Food Additives
G.A. Ravishankar, N. Bhagyalakshmi and S. Ramachandra Rao
I. Introduction
II. Colours
A. Anthocyanins
B. Betalaines
C. Crocin and Crocetins
D. Capsaicin and Other Capsaicinoids
III. Flavours
A. Vanilla
B. Garlic and Onion
IV. Sweeteners
A. Steviosides
B. Thaumatin
V. Safety Aspects
References
5. Production of Insecticides
Amita Pal
I. Introduction
II. Botanical Insecticides
A. Phytoecdysterones
B. Azadirachtin
C. Rotenoids
D. Pyrethrins
E. Nicotine and Anabasine
F. Quassin
G. Limonene
H. Ryania
I. Sabadilla
X. Optimization and Selection
XI. Selection Parameters
XII. Assessment of Somaclonal Variation
XIII. Prospects
References
XIV. Further Readings
4. Production of Food Additives
G.A. Ravishankar, N. Bhagyalakshmi and S. Ramachandra Rao
I. Introduction
II. Colours
A. Anthocyanins
B. Betalaines
C. Crocin and Crocetins
D. Capsaicin and Other Capsaicinoids
III. Flavours
A. Vanilla
B. Garlic and Onion
IV. Sweeteners
A. Steviosides
B. Thaumatin
V. Safety Aspects
References
5. Production of Insecticides
Amita Pal
I. Introduction
II. Botanical Insecticides
A. Phytoecdysterones
B. Azadirachtin
C. Rotenoids
D. Pyrethrins
E. Nicotine and Anabasine
F. Quassin
G. Limonene
H. Ryania
I. Sabadilla
VI. Accumulation of Alkaloids
A. Uptake and Storage
B. Permealization
C. Site of Alkaloid Accumulation
D. Phenylalkylamines (Ephedra Alkaloids)
E. Pyridine Alkaloids
F. Tropane Alkaloids
G. Quinolizidine and Pyrrolizidine Alkaloids
H. Isoquinoline Alkaloids
I. Quinoline (Cinchona) Alkaloids
J. Monoterpene Indole Alkaloids
K. Purine Alkaloids
L. Ruta Alkaloids
VII. Conclusions
VIII. Prospects
References
8. Production of Steroids and Saponins
Anupama Wagle, G.D. Kelkar and M.R. Heble
I. Introduction
II. Saponins
A. Dioscorea
B. Agave
C. Glycyrrhiza
D. Aesculus
E. Panax
III. Cardiac Glycosides
A. Digitalis
B. Strophanthus
C. Urginea
IV. Other Steroids
A. Withania somnifera
B. Holarrhena
C. Solanum
V. Conclusions
References
9. Understanding the Regulatory Mechanism
of Secondary Metabolite Production
J.M. Merillon and K.G. Ramawat
I. Introduction
II. Plant Growth Regulators, Elicitors, Sugars and Signal
Transduction
III. Protein and RNA
IV. Genetic Manipulations
A. Choice of Gene
B. Manipulation of Biosynthetic Pathways
V. Conclusions
References
10. Production of Secondary Metabolites by Bioconversion
Niesko Pras and Herman J. Woerdenbag
I. Introduction
II. General Principles of Bioconversion
III. Systems Applied for Bioconversion
A. Freely Suspended Plant Cells
B. Immobilized Plant Cells
C. Enzyme Preparations
IV. Kinetics of Immobilized Systems
V. Bioconversion of Water-insoluble Precursors
VI. Bioconversion of Synthetic Precursors
VII. New Developments
VIII. Final Consideration
References
11. Genetic Transformation for Production of
Secondary Metabolites
Sumita Jha
I. Introduction
II. Genetic Manipulation of Natural-Product Pathways
III. Genetic Transformation for Development of
Transformed Organ Cultures using Agrobacterium
A. Relationship between Cell Organization,
Differentiation and Secondary Metabolism
B. Organ Cultures
C. Transformed Organ Cultures
D. Agrobacterium and Transformation Process
IV. Applications of Transformed Root Cultures
V. Experimental Procedure for Production of
Transformed Hairy Roots in Artemisia annua Linn.
VI. Experimental Procedure for Production of Transformed
Shooty Teratomas in Coleus forskohlii Briq.
VII. Prospects
Acknowledgement
References
VIII. Further Reading
12. Large-scale Production in Bioreactors
J.M. Merillon
I. Introduction
II. Plant Cell Physiology
III. Plant Cell Culture Processes
A. Introduction
B. Batch Cultivation
C. Multi-stage Batch Cultivation
D. Feed-Batch Cultivation
E. Continuous Cultivation
F. Semicontinuous Cultivation
G. Immobilized Cell Cultivation
IV. Bioreactor System Design and Operation
A. Sterilization
B. Oxygen Supply
C. Bioreactor Types
V. Applications
A. Production of Secondary Metabolites
B. Organ Culture
C. Hairy Root Culture
D. Commercialization
V. Conclusions
References
13. Production of Ergot Alkaloids from Claviceps
Arun Kumar and R. Raj Bhansali
I. Introduction
II. Historical Background
III. Natural Occurrence
IV. Alkaloids
A. Formation and Production
B. Chemical Properties
C. Physical Properties
D. Pharmacological Properties
E. Chemistry
F. Biosynthesis
V. Artificial Culture
VI. Conclusion
References
14. Lichen Products
Wanda Quilhot, Cecilia Rubio and Ernesto Fernandez
I. Introduction
II. Chemistry of Lichen Substances
III. Role of Lichen Metabolites in the Lichen Thallus
IV. Biological
A. Activity of Lichen Extracts and Lichen Compounds
B. Anticancer-Antiviral Activity
C. Inhibitory Enzyme and Antioxidant Activity
D. Analgesic, Anti-inflammatory,
Antipyretic and Other Medicinal
E. Antiprotozoa Activity
F. Antifeedant Activity
G. Phototoxicidal and Other Activity in Plants
H. Herbicidal Activity
I. Plant Growth Activities
J. Perfumes
V. The Future
References
15. Chinese Herbal Drug Industry: Past,
Present and Future
Lixin Zhang and Wei Jia
I. Introduction
II. The Development of the Chinese Herbal Drug Industry
III. Modernization of TCM
A. The Status of Medicinal Plant Resources in China
B. Status of Chinese Prepared Medicinal Herbs
C. Status of TCM Manufacturing Enterprises
D. Status of Plant Extracts
E. Status of Pre-clinic and Clinical Research
IV. The Internationalization of TCM
A. Open Dialogue and Mutual Understanding
Among All Parties Involved in Health Care System
B. Deciphering the Preventive Nature of TCM
C. Respect and Protect Intellectual Property
D. Safety Issues of TCM Need to be Seriously Evaluated
E. The Small Scale and Limited Capability of
Herbal Drug Enterprises
V. Closing Remarks
Acknowledgments
References
16. Secondary Metabolites in in vitro Cultures of Ruta
graveolens L. and Ruta graveolens ssp. divaricata
(Tenore) Gams
Halina Ekiert and Franz Ch. Czygan
I. Distribution and Chemistry of the Investigated Plants
A. Ruta graveolens L. (garden rue = common rue)
B. Ruta graveolens ssp. divaricata (Tenore) Gams
II. Groups of Secondary Metabolites Investigated in vitro —
Their Chemical Characteristics, Distribution in Plant
Kingdom and Biological Properties
A. Linear Furanocoumarins
B. Phenolic Acids
C. Arbutin
III. Production of the Investigated Metabolites in
Plant in vitro Cultures
A. Linear Furanocoumarins
B. Phenolic Acids
C. Arbutin
IV. Studies from Our Laboratory
A. Linear Furanocoumarins and other Coumarin
Compounds Isolated as Main Fractions from Biomass
Cultured in vitro
B. Accumulation of Linear Furanocoumarins
C. Accumulation of Phenolic Acids
D. Biotransformation of Hydroquinone into Arbutin
V. Conclusions and Prospects
VI. Experimental
A. Origin of Cultures Under Study
B. Initial Stationary Liquid Cultures
C. Experimental Cultures
Acknowledgement
References
17. Antimicrobial and Cytotoxic Activity of Extracts and
Secondary Metabolites Obtained from Plants and
Lichens of Patagonia Austral.
R.D. Enriz, M.L. Freile, E. Correche and M.J. Gomez-Lechon
I. Introduction
A. Natural Products as Sources for Lead Structures
B. Cell Culture Systems in Toxicity Testing
II. Techniques, Material and Methods
A. Plant Material
B. Extraction and Isolation of Test Compounds
C. Microorganisms and Media
D. Antimicrobial Assays
E. Antifungal in vivo Assay
F. Antifungal Activity
G. Application of the Test Compounds
H. Acute Toxicity Test
I. Toxicity of Berberine to Embryo-larval
Stages of Bufo arenarum (H). Test Conditions
III. In vitro/in vivo Antimicrobial Activity of Aqueous
Extracts and of Berberine Isolated from Berberis
heterophilla
IV. Antibacterial, Cytotoxic and Apoptotic Effects on
Hepatocytes of Secondary Metabolites Obtained from
Lichens of Patagonia
A. Antibacterial and Cytotoxic Activity
B. Apoptotic Activity
V. Concluding Remarks
Acknowledgements
References
18. Tools and Techniques for the Study of
Plant Tissue Culture
K.G. Ramawat and Meeta Mathur
I. Microscopy
II. Micrometry
III. Cell Counting by Haemocytometer
IV. Autoradiography
V. Spectrophotometry
A. UV-VIS Spectrophotometry
B. Fluorescence Spectrophotometry
VI. Centrifugation
A. Analytical Model
B. Preparative Model
C. Application
D. Methods of Separation
VII. Gel Filtration
VIII. Chromatography
A. Column Chromatography
B. Paper Chromatography
C. Thin-layer Chromatography
D. Adsorption Chromatography
E. lon-exchange Chromatography
F. Affinity Chromatography
G. Gas Liquid Chromatography (GLC)
H. High Performance Liquid Chromatography (HPLC)
IX. Further Readings
19. Practicals
K.G. Ramawat
I. Callus and Cell Culture
A. Initiation of Callus
B. Initiation of Suspension Culture
C. Fresh Weight and Dry Weight Determinations
II. Cell Viability Determination using Fluorescein Diacetate
III. Diosgenin Estimation using Colorimeter
IV. Solasodine Quantification
V. Indole Alkaloid Production by Catharanthus roseus
VI. TLC of Opium Alkaloids
VII. Other Alkaloids
VIII. Qualitative Tests
A. Demonstration of Presence of Sterols
B. Leuco-anthocyanin Test
C. Saponin Test
D. Phenol Test
E. Flavonoid Test
IX. Further Readings
X. Annexures
Index