I= A STUDY OF THE CHEMICAL MANIPULATIONS INVOLVED IN THE CLONAL PROPAGATION OF PISTACIA VERA
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Title of Thesis
A STUDY OF THE CHEMICAL MANIPULATIONS INVOLVED IN THE CLONAL PROPAGATION OF PISTACIA VERA

Author(s)
Zaheer Ahmad
Institute/University/Department Details
Institute of Chemistry University of the Punjab, Lahore
Session
1993
Subject
Chemistry
Number of Pages
235
Keywords (Extracted from title, table of contents and abstract of thesis)
PISTACIA VERA, cotyledon, mesocarp, Embryo cultures, Placental Tissues, callus lines

Abstract
Chemical conditions were optimized for Pistacia vera callus growth from cotyledon (mature and immature), leaf (from in vitro grown seedling and 1-year-old plants), root (from in vitro grown seedlings and regenerated from callus), mesocarp, pre-grown embryos, node and internodes. Culture establishment was not achieved with placenta, embryo and peels of cotyledons. Optimum relative growth response of callus was observed in full strength MS medium. 3% sucrose concentration was good for growth of callus and suspension cultures and 2% for embryo growth. Coconut water enhanced callus growth from cotyledon and pre-grown embryos, whereas malt extract and GA3 inhibited the same. Auxin + cytokinin habituation was achieved in a callus line.

Suspension cultures were established directly from leaf, cotyledon and pre-grown embryos and calli grown from cotyledon and embryos. Globular, heart and torpedo shaped embryoids were formed in the suspension cultures. Calli grown with various concentration and combinations of plant growth regulators were resistant to browning for various time periods. Browning in mesocarp cultures was controlled by serial subculturing. Antioxidants, adsorbents, explant presoaking and maize or malt extract did not prevent or delay browning. Same was reduced with the addition of coconut water, decrease in the concentration of MS salt, or incubation at lower temperature. Browning in calli induced and grown from Pistacia vera cotyledons was controlled by using 5 mg/L NAA, 2.5 mg/L Kin and 150 ml/L CW as growth promoters and incubating the cultures at 20 ±1°C.

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33768.39 KB
S. No. Chapter Title of the Chapters Page Size (KB)
1 0 Contents
2688.42 KB
2 1 Introduction 1
1332.65 KB
3 2 Literature review 9
1703.1 KB
  2.1 Pistacia vera -General 9
  2.2 Pistacia vera -In Vitro Culture 10
  2.3 Cotyledon culture 14
  2.4 Callusing from seedling shoot explants 15
  2.5 Embryo cultures 15
  2.6 Callusing from roots regenerated from callus 16
  2.7 Callusing from mesocarp Tissues 16
  2.8 Suspension cultures 16
  2.9 Browning 17
4 3 Materials and methods 18
2030.72 KB
  3.1 Plant Material 18
  3.2 Sterilization procedures 20
  3.3 Growth Regulator stock solution preparations 20
  3.4 Preparation of stock solutions of other additives 21
  3.5 Media used 22
  3.6 Preparation of MS stock solutions 24
  3.7 Preparation of B5, SH and WP stock solutions 25
  3.8 Cell counting 26
  3.9 Cell fresh weight 26
  3.10 Cell dry weight 27
  3.11 Callus index 27
  3.12 Preparation of suspension cultures directly from explants 27
  3.13 Control of browning in callus cultures 28
  3.14 Physical conditions 30
5 4 Results 31
17808.64 KB
  4.1 Surface sterilization of Pistacia vera Seeds 31
  4.2 Callusing from Pistacia vera Cotyledonary explants 33
  4.3 Relative growth response on different media formulations 49
  4.4 Effect of Natural product Extracts on callusing 55
  4.5 Effect of GA 3 on callusing 55
  4.6 Callusing from Pistacia vera embryos 60
  4.7 Callusing from in vitro obtained stem explants 68
  4.8 Callusing from leaf explants 81
  4.9 Callusing from root explants 88
  4.10 Callusing from €œImmature€ cotyledons 93
  4.11 Callusing from Cotyledon peels 93
  4.12 Callusing from measocarp Tissues 98
  4.13 Callusing from Placental Tissues 98
  4.14 Callusing from roots regenerated from callus 102
  4.15 Habituation in callus lines 107
  4.16 Suspension cultures lines 107
  4.17 Suspension cultures from calli induced on explants 112
  4.18 Appropriate carbohydrate source for Pistacia vera suspensions 112
  4.19 Morphogenesis in suspension cultures 114
  4.20 Effect of growth regulators on browning 114
  4.21 Effect of Citric and Ascorbic acids on browning 123
  4.22 Effect of serial sub culturing 127
  4.23 Effect of Light Intensity on Browning 127
  4.24 Effect of sodium thiosulphate on browning 130
  4.25 Effect of sodium ascorbate on browning 130
  4.26 Effect of Charcoals on browning 130
  4.27 Effect of Natural product Extracts on browning 136
  4.28 Effect of Natural product Extracts on browning 136
  4.29 Effect of Lowering in MS Salt concentrations on Browning 139
  4.30 Effect of Na-diethyldithiocarbamate on Browning 139
  4.31 Effect of Polyvinylpolyrrolidone on Browning 146
  4.32 Effect of Explants Presoaking on Browning 146
  4.33 Effect of Additional ZnSO 4 . 7H 2 O on browning 146
  4.34 Effect of Low temperatures on Browning 151
  4.35 Combined Effect of Coconut Water, Selected growth regulators and low Temperature on browning 151
6 5 Discussion 156
4407.39 KB
7 6 Conclusions 177
3766.32 KB
  6.1 Summary 179