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Title of Thesis

Shagufta Naz
Institute/University/Department Details
Department of Botany/ University of the Punjab
Number of Pages
Keywords (Extracted from title, table of contents and abstract of thesis)
chickpea, callogenesis, organogenesis, legumes, callus formation, multiple shoot formation, somatic embryogenesis, bap

The problem of regeneration through callus culture in chickpea has been a serious challenge to the workers in this field and a little progress toward the development of in vitro regeneration system has been made so far. In present study attempts were made develop protocols for efficient regeneration system through organogenesis, somatic embryogenesis and multiple shoot formation. A biochemical correlation of phenolics vis-à-vis growth inhibition and inefficient differentiation response was also investigated. Present study highlighted the morphogenetic response of calluses derived from different explants i.e., cotyledon, node , leaf, shoot and root apices on selected organogenic media. Morphogenetic response was successfully induced through organogenesis, embryogenesis and multiple shoot formation. Efficient embryogenesis response was induced both direct and indirect pathways from various explants but further growth was remained the major obstacle in obtaining the whole plants. An efficiently and highly reproducible regeneration system was developed through multiple shoot formation from apical shoot and nodal segments. Although decline efficient organ differentiation was triggered through callus tissues but a gradual decline in morphogenic activity and sustainability of differentiated organs was observed in successive subcultures. The important detrimental factor indemnified for poor survival response was the initiation of browning and necrosis of differentiated organs. From main to 3rd subculture the survival percentage of differentiated tissues was 100 which declined to zero at 5th subculture. This browning and decrease in growth of callus tissue has been associated with the production of phenolic compound. A Gradual increase in total phenolics was noticed in all successive subcultures. A detailed account of quantitative and qualitative analysis of total phenols and phenol related enzymes clearly indicated their potential role in growth inhibition and browning.

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3078.51 KB
S. No. Chapter Title of the Chapters Page Size (KB)
1 0 Contents
144.5 KB
2 1 Introduction 1
50.36 KB
3 2 Literatures Review 5
495.37 KB
  2.1 Tissue Culture Studies In Legumes 5
  2.2 The Chickpea Plant 18
  2.3 Tissue Cultural Studies In Chickpea 26
  2.4 Biochemical Investigation Of Growth Inhibition 33
4 3 Materials And Methods 49
219.95 KB
  3.1 Callus Formation Organogenesis, Multiple Shoot Formation And Somatic Embryogenesis 49
  3.2 Biochemical Of Total Phenolics From Main To Successive Subcultures In Calli Derived From Different Explants 56
5 4 Callus Formation And Organogenesis 68
549.45 KB
  4.1 Callus Formation 76
  4.2 Effect Of Different Media Sub-Culturing On Callus Growth 84
  4.3 Organogenesis 93
  4.4 Effect Of Remedial Measures On Elimination Of Browning 103
  4.5 Discussion 106
6 5 Multiple Shoot Formation Results 114
327.69 KB
  5.1 Effect Of Bap On Multiple Shoot Formation From Shoot Apices 118
  5.2 Effect Of Bap On Multiple Shoot Formation From Shoot Apices 118
  5.3 Multiplication Of Regenerated Shoots 121
  5.4 Rooting Of Regenerated Shoots 123
  5.5 Discussion 127
7 6 Somatic Embryogenesis Results 132
245.62 KB
  6.1 Direct Somatic Embryogenesis 134
  6.2 Indirect Somatic Embryogenesis 134
  6.3 Effect Of Auxin And Auxin-Cytokinin Interaction On Somatic Embryogenesis 134
  6.4 Percentage Of Directed And Indirect Embryogenesis On Selected Media 141
  6.5 Differentiation Of Somatic Embryoids 14
  6.6 Discussion 148
8 7 Biochemical Investigation Of Growth Inhibition In Non Morphogenic And Morphogenic Calluses 152
1003.51 KB
  7.1 Total Phenolic Content( Mg/G Callus) In Non- Morphogenic And Morphogenic Calluses 152
  7.2 Specific Activity Of Phenylalanie Ammonia- Lyase (Pal) (N-Moles Of Trans- Cinnamic Acid/ œg Protein) 155
  7.3 Specific Activity Of Polyphenol Oxidase ( Units/ œg Protein) 159
  7.4 Specific Activity Of Peroxidases (Units/ œg Protein) In Non- Morphogenic And Morphogenic Calluses 183
  7.5 Qualitative Study Of Total Phenolics In Nonmorphogenic Calluses 166
  7.6 Electrophoretic Study Of Perosidases In Non Morphogenic Calluses 183
  7.7 Discussion 195
  7.8 Summary 203
  7.9 References 210