I= IN VITRO REGENERATION & PLASTID TRANSFORMATION OF HIGH VALUE CROP PLANTS
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Title of Thesis
IN VITRO REGENERATION & PLASTID TRANSFORMATION OF HIGH VALUE CROP PLANTS

Author(s)
Muhammad Usman
Institute/University/Department Details
Institute of Horticultural Sciences/ University of Agriculture, Faisalabad
Session
2007
Subject
Horticultural Sciences
Number of Pages
191
Keywords (Extracted from title, table of contents and abstract of thesis)
in vitro regeneration, plastid transformation, crop plants, somatic embryogenesis, plant regeneration, tomato, direct shoot regeneration, carrot, edible plants

Abstract
Different cultivars of edible crops like tomato and carrot were explored for development of efficient plant regeneration system. Efficient direct shoot regeneration system was developed from leaf disc in tomato cv. Moneymaker on medium containing MS salts, ImgL-1 each of zeatin and lAA along with B5 vitamins comparable to efficient regeneration system available in tobacco. Occurrence of hyperhydricity ruled out the use of phytagel in the plant growth and development medium in carrot. Use of B5 vitamins in combination with 20gL-1 of sucrose and MS salts (MSIII medium) gave better plant growth with more number of highly regenerable stem explants (3-5) per plant. Provision of stress treatment to the calli induced under light conditions by reducing MS salts and sucrose in the medium (HT2) markedly enhanced (10 fold) its regeneration potential and yielded 20-22 embryos compared to 2-3 embryos per calli segment through conventional means of somatic embryogenesis. Among carrot cultivars, cv. T-29 was screened out for its better regeneration potential and adoptability to the indigenous environment. No regeneration could have been obtained from resistant plant material on the selective medium despite using direct regeneration system confirming the hypothesis of low regeneration potential in tomato on selective medium. However, excellent embryogenesis and plant regeneration was obtained in the bombarded calli on selective medium (MSIII) after stress providing pretreatment on HT4 medium. Use of reporter gene encoding fluorescent protein facilitated facile and early detection of transgenic sectors of gfp in somatic embryos and leaves of regenerated transgenic plants on medium containing spectinomycin (200-400mgL-1). Complex PRC analysis confirmed the transgene integration in to the plastome. In, future, the established plastid transformation system in edible crop will facilitate the cost effective production of edible vaccines by reducing the purification cost required when using non-edible crops like tobacco.

Download Full Thesis
5009.59 KB
S. No. Chapter Title of the Chapters Page Size (KB)
1 0 Contents
327.51 KB
2 1 Introduction 1
176.79 KB
3 2 Review Of Literature 8
1309.67 KB
  2.1 Somatic Embryogenesis And Plant Regeneration Studies 88
  2.2 Plastid Transformation In Higher Plants 33
4 3 Materials Methods 58
419.35 KB
  3.1 In Vitro Regeneration Studies 58
  3.2 Development Of Plastid Transformation Vectors 65
5 4 Results 77
1262.14 KB
  4.1 In Vitro Regeneration Studies 77
  4.2 Development Of Direct Shoot Regeneration From Leaf Disc In Tomato Cultivars 77
  4.3 Development Of Somatic Embryogenesis And Plant Regeneration In Carrot 78
6 5 Discussion 123
514.96 KB
  5.1 Tomato
  5.2 Carrot 15
7 6 Results 134
85.86 KB
  6.1 Plastid Transformation Of Edible Plants 134
  6.2 Plastid Transformation Of Tomato 134
  6.3 Plastid Transformation Of Carrot 135
8 7 Discussion 153
1161.18 KB
  7.1 Summary 158
  7.2 Literature Cited 162
  7.3 Appendices 182