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TITLE
DECLARATION
CERTIFICATE
ACKNOWLEDGEMENT
ABBREVIATIONS
CONTENTS
LIST OF TABLES
LIST OF FIGURES
1. INTRODUCTION
1.1 The Richness of Indian Biodiversity
1.1.1 The South Indian Perspective
1. 2 Role of Biotechnology in Conservation
1. 3 Plants Proposed in the Present Study
l. 3. 1 Adenia hondala (Gaertn.) de Willde
1. 3. 2 Baliospermum montanum Muell-Arg.
1. 4 Objectives of the Present Study
2. REVIEW OF LITERATURE
2. 1 Conservation attempts on Adenia hondala and Baliospermum montanum.
2. 2 Physiochemical studies
2. 3 Conservation of Rare and Endangered Plants using in vitro methods.
2. 4 Plant Tissue Culture
2. 4. 1 Culture medium
2. 4. 2 Role of Plant Growth Regulators
2. 4. 3 Additives
2. 4.4 Explant
2. 4. 5 Axillary Bud Multiplication
2. 5 Callus Culture and Plant Regeneration.
2. 6 In vitro Root Induction and Development
2. 7 Somatic Embryogenesis
2. 8 Somatic Embryogenesis - Germination
2. 9 Acclimatization and Field Establishment
2. 10 Histological Studies
2. 11 Histochemical and Histoenzymological Studies
2. 12 Biochemical Studies
2.13 Physiochemical Analysis
2. 14 Isozymes Studies
3. MATERIALS AND METHODS
3. 1 Source and Choice of Plant materials
3. 1. 1 Adenia hondala (Gaertn.) de Wilde
3. 1. 2 Baliospermum montanum Muell. Arg.
3. 2 Establishment of Cultures
3. 2. 1 Glassware and instruments
3. 2. 2 Basal Media
3. 2. 3 Preparations of Culture Media
3. 2. 4 Sterilization
3. 2. 5 Explant Preparations and Surface Sterilization
3. 2. 6 Inoculation and Incubation
3. 2. 7 Subculture
3. 2. 8 Experiments & Observations
3. 3 Direct Regeneration
3. 3. 1 Explants
3. 3. 2 Culture medium
3. 3. 3 Callus Proliferation Scoring
3. 3. 4 Data Presentation
3. 4 Callogenesis
3. 4. 1 Explants
3. 4. 2 Culture medium
3. 4. 3 Subculture
3. 4. 4 Data collection and Presentation
3. 4. 4. 1 Callusing Frequency
3. 4. 4. 2 Growth measurements
3. 4. 4. 3 Callus lindex.
3. 4. 4. 4 Nature of Callus
3. 5 Indirect Regeneration
3. 5. 1. Explants
3. 5. 2 Culture medium
3. 5. 3 Data presentation
3. 6 Root Initiation
3. 7 Somatic embryo Induction
3.7. 1 Explants
3. 7. 2 Culture medium
3. 7. 3 Culture Conditions
3. 8 Somatic embryo Maturation
3. 8. 1 Carbon source
3. 8. 2 Data collection and Presentation
3. 8. 3 Growth measurements
3. 9 Hardening and Potting.
3. 10 Statistical Analysis
3. 11 Histology, Histochemistry and Histoenzymology
3.11.1 Collection and Fixation of Samples
3. 11. 2 Sectioning
3. 11. 3 Staining
3. 11. 4 Photomicrography
3. 11. 5 Presentation of Results
3. 12 Biochemical Quantification
3. 12. 1 Sample Collection
3. 12. 2 Estimation of Proteins
3. 12. 3 Estimation of Soluble Sugars
3. 12. 4 Estimation of Starch
3. 12. 5 Presentation of Results
3. 13 Quality analysis
3. 13. 1 Chromatographic Separation of Constituents
3. 13. 1. 1 Sample Collection
3. 13. 1. 2 Thin Layer Chromatography
3.13.1.3 Result Analysis
3. 13. 2 Phytochemical Tests.
3. 13. 2. 1 Sample preparation
3. 13. 2. 2 Extraction and Isolation of Saponins
3. 13. 2. 3 Data presentation
3. 14 Physiochemical Analysis
3. 14. 1 Sample Collection
3. 14. 2 Preparation of Sample
3. 14. 3 Solvent Extractions
3. 14. 4 Determination of Alcohol Soluble Extractive
3. 14. 5 Determination of Water Soluble Extractive
3. 14. 6 Determination of Etl er Soluble Extractive
3. 14. 7 Determination of chloroform Soluble Extractive
3.15 Ash Tests
3. 15. 1 Determination of Total Ash
3. 15. 2 Determination of Acid Insoluble Ash.
3. 15. 3 Determination of Water Soluble Ash.
3. 15. 4 Presentation of Results
3. 16 Isozyme Studies
3. 16. 1 Sample Preparation
3. 16. 2 Staining
3. 16. 3 Presentation of Results
4. RESULTS
4.1 ADENIA HONDALA (GARETN.) DE WILLD
4.1.1 Selection of basal media
4.1.2 Caulogenesis
4.1.2.1 Effect of the position of nodal segments on direct shooting
4.1.2.2 General Response of explants
4.1.2.3 Effect of plant growth regulators
4.1.2.4 Enhancement of shoot induction by additive supplementation
4.1.2.5 Comparative effect of Additives
4.1.2.6 Effect of different strength of MS medium
4.1.3 Callogenesis
4.1.3.1 Effect of Plant Growth Regulators (PGR)
4.1.3.2 Comparative analysis of optimum treatments
4.1.3.3 Morphology of calli
4.1.3.4 Development of callus
4.1.4 Indirect Organogenesis
4.1.4.1 General response of calli
4.1.4.2 Effect of BAP on shoot induction
4.1.4.3 Effect of Kn on shoot induction
4.1.4.4 Effect of 2iP on shoot induction
4.1.4.5 Optimization of BAP and Kn
4.1.4.6 Optimization of BAP and 2iP
4.1.4.7 Enhancement of shoot induction by additive supplementation
4.1.4.8 Effect of different strength of MS medium
4.1.4.9 Development of Shoots by Indirect Organogenesis
4. 1.5 Rhizogenesis
4.1.5.1 Effect of Plant growth regulators
4.1.5.2 Effect of different strength of MS medium
4.1.5.3 Nature of Roots.
4.1.6 Somatic embryogensis
4.1.6.1 Direct Somatic Embryogenesis
4.1.6.2 Indirect Somatic Embryogenesis
4.1.7 Acclimatization and Planting out
4.1.8 Influence of explanting period
4.1.9 Histology
4.1.9.1 Histology of direct shoot induction
4.1.9.2 Ontogeny of callus
4.1.9.3 Histology of Non organogenic callus
4.1.9.4 Histology of Organogenic callus
4.1 9.5 Histology of root inducing calli
4.1.9.6 Ontogeny of direct somatic embryos
4.1.9.7 Ontogeny of Indirect somatic embryos
4.1.10 Histochemistry
4.1.10.1 Histochemical Localization in stem and root of Adenia hondala
4.1.10.2 Histochemical localization in the non-organogenic calli
4.1.10.3 Histochemical localization in organogenic callus
4.1.11 Biochemical Estimations
4.1.11.1 Metabolite mobilization in various cultures of Adenia hondala
4.1.12 Quality Analysis
4.1.12.1 Chromatographic separation of constituents
4.1.12.2 Phytochemical studies
4.1.12.3. Thin Layer Chromatography
4.1.12.4 Physiochemical standards analysis
4.1.13 Isozyme Studies
4.1.13.1 Peroxidase
4.1.13.2 Esterase
TABLES 1-111
Table - 1 Chemical composition of various nutrient media used
Table - 2 Media Strength, Plant Growth Regulators and other supplements tried in cultures of Adenia hondala
Table - 3 Media Strength, Plant Growth Regulators and other Supplements tried in cultures of Baliospermum montanum
Table - 4 Staining methods followed for general Histology, Histochemistry and Histoenzymology
Table - 5 Frequency of establishment and general response of various explants on different basal media
Table - 6 Effect of different nodal segments on direct shooting in Adenia hondala
Table - 7 Effect of different PGR on leaf, node and inter node for direct shoot induction in Adenia hondala
Table - 8 Effect of BAP on shoot induction from nodal segments in Adenia hondala on MS medium
Table - 9 Effect of Kn on shoot induction from nodal segments in Adenia hondala on MS medium
Table - 10 Effect of 2iP on shoot induction from nodal segments in Adenia hondala on MS medium
Table - 11 Effect of BAP + IAA on shoot induction from nodal segments in Adenia hondala on MS medium
Table - 12 Effect of BAP + NAA on shoot induction from nodal segments in Adenia hondala on MS medium
Table - 13 Effect of BAP + 2, 4-D on shoot induction from nodal segments in Adenia hondala on MS medium
Table - 14 Effect of Kn + IAA on shoot induction from nodal segments in Adenia hondala on MS medium
Table - 15 Effect of Kn + NAA on shoot induction from nodal segments in Adenia hondala on MS medium
Table - 16 Effect of Kn + 2, 4-D on shoot induction from nodal segments in Adenia hondala on MS medium
Table - 17 Effect of 2iP + IAA on shoos induction from nodal segments in Adenia hondala on MS medium
Table - 18 Effect of 2iP + NAA on shoat induction from nodal segments in Adenia hondala on MS medium
Table - 19 Effect of 2iP + 2, 4-D on• shoat induction from nodal segments in Adenia hondala on MS medium
Table - 20 Effect of BAP, Kn and 2iPo? 1 shoot induction in Adenia hondala on MS medium
Table - 21 Effect of Coconut water or, shoot induction from nodal segments of Adenia hondala in the optimized. medium
Table - 22 Effect of CAP on shoot induction from nodal segments of Adenia hondala •in the optimized medium
Table - 23 Effect of Biotin on shoot induction from nodal segments of Adenia hondala in the optimized medium
Table- 24 Effect of GA3 on shoot induction from nodal segments of Adenia hondala in the optimized medium
Table - 25 Effect of ABA on shoot induction from nodal segments in Adenia hondala in the optimized medium
Table - 26 Effect of MS strength on shoot induction from nodal segments of Adenia hondala in the optimized medium
Table - 27 Effect of IAA on callus induction from leaf explants of Adenia hondala on MS medium
Table - 28 Effect of IAA on callus induction from inter nodal segments of Adenia hondala on MS medium
Table - 29 Effect of IAA on callus induction from nodal segments of Adenia hondala on MS medium
Table - 30 Effect of NAA on callus induction from leaf explants of Adenia hondala on MS medium
Table - 31Effect of NAA on callus induction from intemodal segments of Adenia hondala on MS medium
Table - 32 Effect of NAA on callus induction from nodal segments of Adenia hondala on MS medium
Table - 33 Effect of 2, 4-D on callus induction from leaf explants of Adenia hondala on MS medium
Table - 34 Effect of 2, 4-D on callus induction from internodal segments of Adenia hondala on MS medium
Table 35 Effect of 2, 4-D on callus induction from nodal segments of Adenia hondala on MS medium
Table - 36 Effect of IAA + BAP on callus induction from leaf explants of Adenia hondala on MS medium
Table- 37 Effect of IAA + BAP on callus induction from internodal segments of Adenia hondala on MS medium
Table 38 Effect of IAA+ BAP on callus induction from nodal segments of Adenia hondala on MS medium
Table 39 Effect of IAA + Kn on callus induction from leaf explants of Adenia hondala on MS medium
Table - 40 Effect of IAA + Kn on callus induction from internodal segments of Adenia hondala on MS medium
Table - 41 Effect of IAA + Kn on callus induction from nodal segments of Adenia hondala on MS medium
Table - 42 Effect of IAA + 2iP on callus induction from leaf explants of Adenia hondala on MS medium
Table - 43 Effect of IAA + 2iP on callus induction from internodal explants of Adenia hondala on MS medium.
Table - 44 Effect of IAA + 2iP on callus induction from nodal segments of Adenia hondala on MS medium
Table - 45 Effect of NAA + BAP on callus induction from leaf explants of Adenia hondala on MS medium
Table - 46 Effect of NAA + BAP on callus induction from internodal segments of Adenia hondala on MS medium
Table - 47 Effect of NAA + BAP on callus induction from nodal segments of Adenia hondala on MS medium
Table - 48 Effect of NAA + Kn on callus induction from leaf explants of Adenia hondala on MS medium
Table - 49 Effect of NAA + Kn on callus induction from internodal segments of Adenia hondala on MS medium
Table - 50 Effect of NAA + Kn on call _is induction from nodal segments of Adenia hondala on MS medium
Table - 51 Effect of NAA ± 21P on callus induction from leaf explants of Adenia hondala on MS medium
Table - 52 Effect of NAA + 2iP on callus induction from internodal segments of Adenia hondala on MS medium
Table - 53 Effect of NAA + 2iP on callus induction from nodal segments of Adenia hondala on MS medium
Table - 54 Effect of 2, 4- D + BAP on callus induction from leaf explants of Adenia hondala on MS medium
Table - 55 Effect of 2, 4 D + BAP on callus induction from internodal segments of Adenia hondala on MS medium
Table - 56 Effect of 2, 4 D + BAP on callus induction from nodal segments of Adenia hondala on MS medium
Table -57 Effect of 2, 4 D + Kn on callus induction from leaf explants of Adenia hondala on MS medium
Table -58 Effect of 2, 4 D + Kn on callus induction from internodal segments of Adenia hondala on MS medium
Table -59 Effect of 2, 4 D + Kn on callus induction from nodal segments of Adenia hondala on MS medium
Table - 60 Effect of 2, 4 D + 2iP on callus induction from leaf explants of Adenia hondala on MS medium
Table - 61 Effect of 2, 4 D + 2iP on callus induction from internodal segments of Adenia hondala on MS medium
Table - 62 Effect of 2, 4 D + 2iP on callus induction from nodal segments of Adenia hondala on MS medium
Table - 63 Response of various PGR combinations on Indirect organogenesis in Adenia hondala
Table - 64 Effect of BAP on shoot induction in Adenia hondala on MS medium
Table - 65 Effect of Kn on shoot induction in Adenia hondala on MS medium
Table - 66 Effect of 2iP on shoot induction in Adenia hondala on MS medium
Table - 67 Optimization of BAP and Kn combinations for indirect organogenesis in Adenia hondala on MS medium
Table - 68 Optimization of BAP + 2iP combinations for indirect organogenesis in Adenia hondala on MS medium
Table - 69 Effect of Coconut water on indirect organogenesis in Adenia hondala in the optimized medium
Table - 70 Effect of CAP on indirect organogenesis in Adenia hondala in the optimized medium
Table - 71 Effect of TDZ on indirect organogenesis in the optimized medium
Table - 72 Effect of GA3 on indirect organogenesis in Adenia hondala in the optimized medium
Table - 73 Effect of ABA on indirect organogenesis in Adenia hondala in the optimized medium
Table - 74 Effect of IBA on rhizogenesis in Adenia hondala on MS medium
Table - 75 Effect of IBA on rhizogenesis in Adenia hondala on MS medium
Table. - 76 Effect of IAA on rhizogenesis in Adenia hondala on MS medium
Table - 77 Effect of different strength of MS on rhizogenesis in Adenia hondala on MS medium
Table - 78 Somatic embryogenesis in Adenia hondala
Table - 79 Effect of NAA + BAP on direct somatic embryogenesis from leaf explants of Adenia hondala on MS medium
Table - 80 Effect of coconut water on somatic embryo induction from leaf explants of Adenia hondala on MS medium
Table - 81 Effect of GA3 on somatic embryo induction from leaf explants of Adenia hondala on MS medium
Table - 82 Effect of TDZ on embryo induction from leaf explants of Adenia hondala in the optimized medium
Table - 83 Effect of ABA on embryo induction from leaf explants of Adenia hondala in the optimized medium
Table - 84 Effect of CH on embryo induction from leaf explants of Adenia hondala in the optimized medium
Table - 85 Effect of MS strength from leaf explants of Adenia hondala in the optimized medium
Table - 86 Effect of NAA + BAP on Indirect somatic embryogenesis in Adenia hondala on MS medium
Table - 87 Effect of 2, 4-D + Kn on Indirect Somatic Embryogenesis in Adenia hondala on MS medium
Table - 88 Effect of coconut water on embryo induction in Adenia hondala on MS medium
Table - 89 Effect of GA3 on embryo induction in Adenia hondala on MS medium
Table - 90 Effect of TDZ on embryo induction in Adenia hondala on MS medium
Table - 91 Effect of ABA on embryo induction in Adenia hondala on MS medium
Table - 92 Effect of CH on embryo induction in Adenia hondala on MS medium
Table - 93 Effect of MS strength in Adenia hondala on MS medium
Table - 94 Effect of PGR on embryo maturation in Adenia hondala on MS medium
Table - 95 Optimisation of NAA + BAP on embryo maturation in Adenia hondala on MS medium
Table - 96 Effect of CW on embryo maturation in Adenia hondala on MS medium
Table - 97 Effect of ABA on embryo maturation in Adenia hondala on MS medium
Table - 98 Effect of Sucrose on embryo maturation in Adenia hondala on MS medium
Table - 99 Effect of GA3 on embryo maturation in Adenia hondala on MS medium
Table - 100 Effect of MS strength on embryo maturation in Adenia hondala on MS medium
Table - 101 Evaluation of different planting substrates for acclimatisation of in vitro shoots of adenia hondala
Table - 102 Influence of explanting period on culture establishment of adenia hondala on ms supplements with 0.5 mgl-1 bap + 0.5 mgl-1 kn + 1 mgl-l2ip +10% CW
Table.- 103 Intensity of metabolite and enzyme localization in stem and root of Adenia hondala
Table - 104 Intensity of metabolite and enzyme localization in the Non organogenic call] of Adenia hondala
Table - 105 Intensity of metabolite and enzyme localization in the organogenic callus of Adenia hondala
Table - 106: Qualitative chemical examination of various extracts obtained by successive solvent extraction of stem of Adenia hondala
Table - 107: Qualitative chemical examination of: various extracts obtained by successive solvent extraction of leaves of Adenia hondala
Table - 108: Qualitative chemical examination of various extracts obtained by successive solvent extraction of tubers of Adenia hondala
Table - 109: Qualitative chemical examination of various extracts obtained by successive solvent extraction of regenerated shoots of Adenia hondala
Table - 110: Qualitative chemical examination of various extracts obtained by successive solvent extraction of non organogenic calli of Adenia hondala
Table - 111 Analysis of physiochemical standards in in viva plant and regenerated plants of Adenia hondala.
FIGURES 1-26
Fig. 1 Adenia hondala (Gaertn) de Willde.
Fig. 2 Multiple shoot induction directly on nodal segments of Adenia hondala
Fig. 3 Induction of callus in Adenia hondala
Fig. 4 Induction and elongation of shoot buds in Adenia hondala from callus
Fig. 5 In vitro root induction on regenerated plants of Adenia hondala
Fig. 6 Somatic embryogenesis in Adenia hondala
Fig. 7 Hardening of the regenerated plants of Adenia hondala in solicit
Fig. 8 Histology of shoot regeneration in Adenia hondala.
Fig. 9-Histology of call] of Adenia hondala
Fig. 10 Histological studies of different stages of shoot regeneration from organogenic calli of Adenia hondala
Fig. 11 Ontogeny of direct somatic embryogenesis from leaf explants of Adenia hondala
Fig. 12. Ontogeny of indirect somatic embryogenesis from friable calli of Adenia hondala
Fig. 13 Localization of metabolites and enzymes in the root tubers of Adenia hondala
Fig. 14 Localization of metabolites and enzymes in the. non-organogenic calli of Adenia hondala
Fig. 15 Localization of metabolites and enzymes in the organogenic calli of Adenia hondala
Fig. 16 Thin layer chromatography of Adenia hondala
Fig. 17. Froth formation showing indicative test for saponins and Thin layer chromatography showing saponins specific spot
Fig. 18 Electrophoretogram showing isozymes of peroxidase and esterases in explants and in vitro cultures of Adenia hondala
Fig. 19 Zymogram and schematic representation of various developmental stages in cultures of Adenia hondala
Fig. 20 Comparison of Optimum treatments of PGR combination on callogenesis from leaf explants of Adenia hondala.
Fig. 21 Comparison of Optimum treatments of PGR combination on callogenesis from internodal explants of Adenia hondala
Fig. 22 Comparison of Optimum treatments of PGR combination on callogenesis from nodal explants of Adenia hondala
Fig. 23 Metabolite mobilization in direct organogenic cultures of Adenia hondala
Fig. 24 Metabolite mobilization in organogenic callus cultures of Adenia hondala
Fig. 25 Metabolite mobilization in embryogenic cultures of Adenia hondala
Fig. 26 Metabolite mobilization in non-organogenic cultures of Adenia hondala
4.2 BALIOSPERMUM MONTANUM Muell. Arg
4.2.1 Selection of Basal Media
4. 2. 2 Caulogensis
4.2.2.1 Effect of position of rodal segments on direct shooting
4.2.2.2 General response of explants
4.2.2.3 Direct Regeneration from nodal segments.
4.2.2.3.1 Effect of Plant Growth Regulator
4.2.2.3.2. Enhancement of shoot induction by additive supplementation
4.2.2.4 Direct Regeneration from leaf explants.
4.2.2.4.1 Optimization of Kn and IAA
4.2.2.4.2. Enhancement of shoot induction by additive supplementation
4.2.3. Callogenesis
4.2.3.1 Effect of Plant Growth Regulators.
4.2.3.2 Comparative analysis of optimum treatments in callogenesis
4.2.3.3 Morphology of calli
4.2.3.4 Development of callus;
4.2.4 Indirect organogenesis
4.2.4.1 General response of calli
4.2.5 Rhizogenesis
4.2.5.1 Effect of Plant growth regulators
4.2.5.2 Nature of Roots.
4.2.6 Somatic Embryogenesis
4.2.6.1 Optimization of NAA and Kn
4.2.6.2 Optimization of IAA and TDZ
4.2.6.3 Enhancement of embryo induction by additive supplementation
4.2.6.4 Embryo maturation
42.6.5 Optimization of Kn + NAA
4.2.6.6 Optimization of TDZ + NAA
4.2.6.7 Enhancement of embryo maturation by additive supplementation.
4.2.7 Acclimatization and Planting out
4.2.8 Comparative study of efficiency of regeneration pathway
4.2.9 Histology
4.2.9.1 Direct regneration from nodal segments
4.2.9.2 Ontogeny of callus
4.2.9.3 Ontogeny of direct organogenesis from leaf explants
4.2.9.4 Ontogeny of somatic embryogenesis.
4.2.10 Histochemistry
4.2.10.1 Histochemical and Histoenzymological localization in direct organogenic cultures
4.2.10.2 Histochemical and Histoenzymological localization in root forming callus
4.2. 10.3 Histochemical and Histoenzymological localization in non organogenic callus
4.2.10.4 Histochemical and Histoenzymological localization in nodular callus
4.2.11 Metabolite mobilization in various cultures of B. montanum
4.2.12 Quality Analysis
4.1 12.1 Chromatographic separation of constituents
4.1 12. 2 Phytochemical studies
4.2.12.3 Thin Layer Chromatography
4.2.12.4 Physiochemical standards analysis
4.2.13 Isozymes studies
4.2.13.1 Peroxidase
4.2.13.2 Esterase
TABLES 112 - 204
Table - 112 General response of various explants of Baliospermum montanum on different basal media (after 30 days)
Table - 113 Effect of different nodal segment explants on direct shooting in Baliospermum montanum on MS medium
Table - 114 Effect of different PGR on leaf, node and inter node explants for direct shoot induction in Baliospermum montanum on MS medium
Table - 115 Effect of BAP on direct organogenesis from nodal segments in Baliospermum montanum -on MS medium
Table - 116 Effect of Kn on direct organogenesis from nodal segments in Baliospermum montanum on MS medium
Table - 117 Effect of 2iP on direct organogenesis from nodal segments in Baliospermum montanum on MS medium
Table - 118 Effect of BAP + NAA on in vitro shoot induction from nodal segments in Baliospennum montanum on MS medium
Table - 119 Effect of BAP + 2, 4-D on in vitro shoot induction from nodal segments in Baliospermum montanum on ISIS medium
Table - 120 Effect of Kn + NAA on in vitro shoot induction from nodal segments in Baliospermum montanum on MS medium
Table - 121 Effect of Kn + 2, 4-D on in vitro shoot induction from nodal segments in Baliospermum montanum on MS medium
Table - 122 Effect of 2iP +NAA on in vitro shoot induction from nodal segments in Baliospermum montanum on MS medium
Table - 123 Effect of 2iP +2, 4-D on in vitro shoot induction from nodal segments in Baliospermum montanum on MS medium
Table - 124 Effect of BAP, Kn and 2iP on shoot induction from nodal segments in Baliospermum montanum on MS medium
Table - 125 Effect of coconut water or. shoot induction from nodal segments in Baliospermum montanum on MS medium
Table - 126 Effect of CAP on shoot induction from nodal segments in Baliospermum montanum on MS medium
Table -127 Effect of Biotin on shoot induction from nodal segments in Baliospermum montanum on MS medium
Table - 128 Effect of GA3 on shoot induction from nodal segments in Baliospermum montanum on MS medium,
Table - 129 Effect of ABA on shoot induction from nodal segments in Baliospermum montanum on MS.medium
Table - 130 Effect of Kn + IAA on shoot induction on leaf explants of Baliospermum montanum on MS medium
Table - 131 Effect of Coconut water on shoot induction from leaf explants of Baliospermum montanum in optimized medium
Table - 132 Effect of CAP on shoot induction from leaf explants of Baliospermum montanum in optimized medium
Table - 133 Effect of Biotin on shoot induction from leaf explants of Baliospermum montanum in optimized medium
Table - 134 Effect of GA3 on shoot induction from leaf explants of Baliospermum montanum in optimized medium
Table - 135 Effect of ABA on shoot induction from leaf explants of Baliospermum montanum in optimized medium
Table - 136 Effect of IAA on callogenesis from Leaf explants of Baliospermum montanum on MS medium
Table - 137 Effect of IAA on callogenesis from internodal segments of Baliospermum montanum on MS medium
Table - 138 Effect of IAA on callogenesis from nodal segments of Baliospermum montanum on MS medium
Table - 139 Effect of NAA on callogenesis from leaf explants of Baliospermum montanum on MS medium
Table - 140 Effect of NAA on callogenesis from inter nodal segments of Baliospermum montanum on MS medium
Table - 141 Effect of NAA on callogenesis from nodal segments of Baliospermum montanum on MS medium
Table - 142 Effect of 2, 4-D on callogenesis from leaf explants Baliospermum montanum on MS medium
Table - 143 Effect of 2, 4-D on callogenesis from inter nodal segments Baliospermum montanum on MS medium
Table - 144 Effect of 2, 4-D on callogenesis from nodal segments Baliospermum montanum on MS medium
Table - 145 Effect of IAA + BAP on callogenesis from leaf explants of Baliospermum montanum on MS. medium
Table - 146 Effect of IAA + BAP on callogenesis from internodal segments of Baliospermum montanum on MS medium
Table - 147 Effect of IAA + BAP on callogenesis from nodal segments of Baliospermum montanum on MS medium
Table - 148 Effect of IAA + Kn on callogenesis leaf explants of Baliospermum montanum on MS medium
Table - 149 Effect of IAA + Kn on internodal segments of Baliospermum montanum on MS medium
Table - 150 Effect of IAA + Kn on nodal segments of Baliospermum montanum on MS medium
Table - 151 Effect of IAA + 2iP on leaf explants of Baliospermum montanum on MS medium
Table - 152 Effect of IAA + 2iP on internodal segments of Baliospermum montanum on MS medium
Table - 153 Effect of IAA + 2iP on nodal segments of Baliospennum montanum on MS medium
Table - 154 Effect of NAA + BAP on leaf explants of Baliospermum montanum on MS medium
Table - 155 Effect of NAA + BAP on internode segments of Baliospermum montanum on MS medium
Table - 156 Effect of NAA + BAP on nodal segments of Baliospermum montanum on MS medium
Table - 157 Effect of NAA + Kn on leaf explants of Baliospermum montanum on MS medium
Table -158 Effect of NAA + Kn on internodal segments of Baliospermum montanum on MS medium
Table -159 Effect of NAA + Kn on nodal segments -of Baliospermum montanum on MS medium
Table -160 Effect of NAA + 2iP on leaf segments of Baliospermum montanum on MS medium
Table - 161 Effect of NAA + 2iP on internodal segments of Baliospermum montanum on MS medium
Table -162 Effect of NAA + 2iP on nodal segments of Baliospermum montanum on MS medium
Table - 163 Effect of 2, 4 D + BAP on callus induction from leaf explants of Baliospermum montanum on MS medium.
Table - 164 Effect of 2, 4 D + BAP on internodal segments of Baliospermum montanum on MS medium
Table 165 Effect of 2, 4 D + BAP on nodal segments of Baliospermum montanum on MS medium
Table - 166 Effect of 2, 4 D + Kn on leaf explants of Baliospermum montanum on MS medium
Table - 167 Effect of 2, 4 D + Kn on internodal segments of Baliospermum montanum on MS medium
Table - 168 Effect of 2, 4 D + Kn on nodal segments of Baliospermum montanum on MS medium
Table - 169 Effect of 2, 4 D + 2iP on leaf explants of Baliospermum montanum on MS medium
Table - 170 Effect of 2, 4 D +2iP on internodal segments of Baliospermum montanum on MS medium
Table - 171 Effect of 2, 4 D + 2iP on nodal segments of Baliospermum montanum on MS medium
Table - 172 Indirect Organogenesis In Baliospermum montanum
Table - 173 Effect of IBA on root induction in Baliospermum montanum on MS medium
Table - 174 Effect of NAA on root induction in Baliospermum montanum on MS medium
Table - 175 Effect of IAA on root induction in Baliospermum montanum on MS medium
Table - 176 Effect of MS on root induction in Baliospermum montanum
Table - 177 Somatic Embryogenesis In Baliospermum montanum
Table - 178 Effect of NAA + TDZ on somatic embryo induction in Baliospermum montanum on MS medium
Table -179 Effect of NAA + Kn on somatic embryo induction in Baliospermum montanum on MS medium
Table - 180 Effect of CW on embryo induction in Baliospermum montanum on MS medium
Table - 181 Effect of CAP on embryo induction in Baliospermum montanum on MS medium
Table - 182 Effect of Biotin on embryo induction in Baliospermum montanum on MS medium
Table - 183 Effect of GA3 on embryo induction in Baliospermum montanum on MS medium
Table - 184 Effect of MS strength with control media in Baliospermum montanum on MS medium
Table - 185 Effect of PGR on embryo maturation in Baliospermum montanum on MS medium
Table - 186 Effect of Kn + NAA on embryo maturation in Baliospermum montanum
Table - 187 Effect of TDZ + NAA on embryo maturation in Baliospermum montanum
Table - 188 Effect of coconut water an embryo maturation in Baliospermum montanum in optimized media
Table - 189 Effect of CAP on embryo maturation in Baliospermum montanum in optimized media
Table - 190 Effect of GA3 on embryo maturation in Baliospermum montanum in optimized media
Table - 191 Effect of ABA on embryo maturation in Baliospermum montanum in optimized media
Table - 192 Effect of sucrose on embryo maturation in Baliospermum montanum in optimized medium
Table - 193 Effect of MS strength on embryo maturation in Baliospermum montanum in optimized medium
Table - 194 Evaluation of different planting substrates for acclimatization of in vitro shoots of Baliospermum montanum
Table - 195 Effect of subculture on micropropagation of Adenia hondala and Baliospermum montanum
Table - 196 Intensity of metabolite and enzyme localization during direct organogenesis in Baliopsermum montanum
Table - 197 Intensity of metabolite and enzyme localization in root forming callus of Baliopsermum montanum
Table - 198 Intensity of metabolite and enzyme localization in non-organogenic callus of Baliospermum montanum
Table - 199 Intensity of metabolite and enzyme localization in nodular calli of Baliopsermum montanum
Table - 200: Qualitative chemical examination of various extracts obtained by successive solvent extraction of leaves of Baliospermum montanum
Table - 201 Qualitative chemical examination of various extracts obtained by successive solvent extraction of the roots of Baliopsermum montanum Table - 202 Qualitative chemical examination of various extracts obtained by successive solvent extraction of the leaves of regenearated shoots of Baliospermum montanum
Table - 202 Qualitative chemical examination of various extracts obtained by successivesolvent extraction of the leaves of regenerated shoots of Baliospemum montanum
Table - 203 Qualitative chemical examination of various extracts obtained by successive solvent extraction of the leaves of regenearated shoots of Baliospermum montanum
Table - 204 Physiochemical Analysis of different samples of Baliospermum monatnum
FIGURES 27 - 52
Fig. 27 Baliospermum montanum Muell. Arg.
Fig. 28 Direct regeneration from nodal segments in Baliospermum montanum
Fig. 29 Direct regeneration from leaf segments of Baliospermum montanum
Fig. 30 White compact calli derived from abaxial portion of Baliospermum montanum
Fig. 31 Callus induction from leaf explants of Baliospermum montanum
Fig. 32 Rhizogensis in calli derived from various explants of Baliospermum montanum
Fig. 33 Indirect somatic embryogenesis in Baliospermum montanum
Fig. 34 Rooting of the plantlets of Baliospermum montanum obtained through regeneration pathways other than somatic embryogenesis
Fig. 35 Hardening of regenerated shoots of Baliospermum montanum
Fig. 36 Histology of shoot development in Baliospermum montanum
Fig. 37 General histology of the regenerated shoot of Baliospermum montanum. L.S of leaf showing calli.
Fig. 38 Stages in the development of somatic embryogenesis in Baliospermum montanum
Fig. 39 T.S. of root inducing calli of Baliospermum montanum showing localization of metabolites and enzymes.
Fig. 40 T.S. of non-organogenic callus showing metabolites and enzyme deposition in Baliospermum montanum
Fig. 41 T.S. of nodular calli of Baliospermum montanum showing metabolite and enzyme localization.
Fig. 42 Thin layer chromatography of Baliospermum montanum
Fig. 43 Test for saponins in in vivo roots of Baliospermum montanum and thin layer chromatography showing saponin specific spot formation.
Fig. 44 Electrophoretogram showing isozymes of peroxidase and esterases in explants and in vitro cultures of Baliospermum montanum
Fig. 45 Zymogram and schematic representation of various developmental stages in cultures of Baliospermum montanum
Fig. 46 Comparison of Optimum treatments of PGR combination on callogenesis from leaf explants of Baliospermum montanum.
Fig. 48 Comparison of Optimum treatments of PGR combination on callogenesis from nodal explants of Baliospermum montanum
Fig.49 Metabolite mobilization in direct organogenic cultures of Baliospermum montanum
Fig. 50 Metabolite mobilization in root forming callus of Baliospermum montanum
Fig. 51 Metabolite mobilization in non organogenic cultures of Baliospermum montanum
Fig. 52 Metabolite mobilization in nodular callus cultures of Baliospermum montanum
5. DISCUSSION
5. 1 Selection of Basal media
5. 2 Micropropagation through direct regeneration
5.2.1 Explant selection and response
5.2.2 Position of the Node
5.2.3 Influence of Plant growth Regulators
5. 2.3. 1 Nodal cultures
5. 2.3. 2 Basal Callus Formation
5.2.3.3 Role of Subculture in multiplying Shoot number
5.2.3.4 Leaf Cultures
5.2.4. Effect of Additives in Direct Regeneration
5.2.5 Influence of Explanting Period
5.3 Callogenesis
5.4 Indirect Organogenesis
5.5 Rooting
5. 6 Somatic Embryogenesis
5. 7 Acclimatization of Micropropagated Shoots
5.8 Histology
5.9 Histochemistry and Histoenzymology
5.10 Metabolite mobilization
5.11 Phytochemical analysis
5.12 Isozyme studies
6. SUMMARY
REFERENCES