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  • TITLE
  • DECLARATION
  • CERTIFICATE
  • DEDICATION
  • ACKNOWLEDGEMENT
  • CONTENTS
  • ABBREVIATIONS
  • 1. Introduction and Review of Literature
  • INTRODUCTION
  • REVIEW OF LITERATURE
  • 1. Importance of Natural products in anticancer therapy.
  • 1.1 Anti-tumour compounds from plants.
  • PLATE-Mechanism of DNA strand breakage and religation by Topoisomerase
  • 1.2 Plant Biotechnology for the production of alkaloids
  • 1.3. Strategies to improve product yield from plant tissue culture
  • 1.3.1. Screening and Selection
  • 1.3.2. Influence of growth phases
  • 1.3.3. Differentiation and culture type.
  • 1.3.4 Organogenesis
  • 1.3.5 Hairy root culture
  • 1.3.6. Culture conditions
  • 1.3.7 Elicitation
  • 1.3.8. Precursor feeding
  • 1.3.9 Biotransformation
  • 1.3.10 Inducers.
  • 1.3.11 Immobilization
  • 1.3.1 2 Permeabilization
  • 1.3.13 Biosynthesis, enzymology and regulation
  • 1.3.14 Alkaloid storage compartment
  • 1.3.15 Genetic modification
  • 1.3.16 Combination of treatments
  • 1.3.1 7 Large-scale production
  • 2. Materials And Methods
  • 2.1 Plant Material
  • 2.2 Chemicals
  • 2.3 Instruments
  • 2.4 Explants and Surface sterilization.
  • 2.5 Preparation of nutrient medium
  • 2.6 Culture conditions
  • 2.7 Establishment of suspension culture
  • 2.8 Determination of fresh weight
  • 2.9 Determination of dry weight.
  • 2.10 Determination of packed cell volume
  • 2.11 Estimation of growth rates of culture.
  • 2.12 Effect of growth hormones on callus induction and organogenesis
  • 2.13 Hairy root cultures.
  • 2.14 Elicitation
  • 2.15 Two phase culture
  • 2.16 Permeabilization
  • 2.17 Preparation of plant extract.
  • 2.18.Isolation and crystallization of Camptothecin (CPT)
  • 2.19 TLC analysis.
  • 2.20 Melting point
  • 2.21 UV - Visible absorption spectra.
  • 2.22 High performance liquid chromatography (HPLC)
  • 2.23 Electron spray mass spectrometry -
  • 2.24 Topoisomerase I and I1 inhibition assay
  • 2.25 Statistical analysis
  • 3. Comparison of camptothecin production from intact plants and callus culture of Ophiorrhiza mungos
  • 3.1 Materials and Methods
  • 3.2 Results
  • Plate 2- Figure: 1 Ophiorrhiza mungos mother plant
  • Plate 2- Figure: 2 Ophiorrhiza mungos compact callus
  • Plate 2- Figure: 3 Ophiorrhiza mungos friable callus
  • Plate 3- Figure: 1 Ophiorrhiza mungos rhizogenesis
  • Plate 3- Figure: 2 Ophiorrhiza mungos rooted callus
  • Mass Spectra of Standard Camptothecin
  • Mass spectra of isolated camptothecin
  • HPLC Profiles of Selected Samples
  • DISCUSSION
  • 4. Multiple shoot cultures - a viable alternative for camptothecin production
  • 4.1. Materials and Methods.
  • 4.2 Results
  • Plate 4- Figure: 1 Ophiorrhiza mungos shoot regeneration
  • Plate 4- Figure: 2 Ophiorrhiza mungos multiple shoots
  • Plate 4- Figure: 3 Ophiorrhiza mungos multiple shoots
  • Plate 4- Figure: 4 Ophiorrhiza mungos multiple shootsuspension culture
  • HPLC Profiles of Selected Samples
  • Amala Cancer Research Centre-Kerala
  • 4.3 Discussion
  • 5. An attempt to enhance camptothecin production by precursor feeding, elicitation Permeabilization and two- phase culture.
  • 5.1 Materials and Methods
  • 5.2 Results
  • HPLC Profiles of Selected Samples
  • DISCUSSION
  • 6. Comparative evaluation of camptothecin production in callus, cell suspension culture, root culture and somatic embryos of Nothapodytes foetida
  • 6.1 Materials and Methods.
  • 6.2 Result
  • Plate 5- Figure: 1 Nothapodytes foetida mother plant
  • Plate 5- Figure: 2 Nothapodytes foetida- callus
  • Plate 5- Figure: 3 Nothapodytes foetida- cell suspension culture
  • Plate 6- Figure: 1 Nothapodytes foetida- root regeneration
  • Plate 6- Figure: 2 Nothapodytes foetida- somatic embryogenesis
  • Plate 6- Figure: 3 Nothapodytes foetida- mature somatic embryo
  • Plate 6- Figure: 4 Nothapodytes foetida- regenerated roots
  • Plate 6- Figure: 5 Nothapodytes foetida- seedling developed from somatic embryo
  • HPLC Profiles of Selected Samples
  • 6.3 Discussion
  • 7. Summary and Conclusion
  • Future prospectus
  • BIBILIOGRAPHY