I= DISEASE ERADICATION THROUGH TISSUE CULTURE AND GENETIC TRANSFORMATION STUDIES IN BANANA (MUSA)
Pakistan Research Repository Home
 

Title of Thesis
DISEASE ERADICATION THROUGH TISSUE CULTURE AND GENETIC TRANSFORMATION STUDIES IN BANANA (MUSA)

Author(s)
AISH MUHAMMAD
Institute/University/Department Details
Department of Biochemistry/ Arid Agriculture University. Rawalpindi
Session
2008
Subject
Biochemistry
Number of Pages
136
Keywords (Extracted from title, table of contents and abstract of thesis)
disease eradication, tissue culture, genetic transformation, banana, in vitro multiplication

Abstract
Tissue culture and genetic transformation in banana (Musa) cv. Basrai was studied. In vitro cultures were established from field grown suckers. Shoot tips were sterilized with sodium hypochlorite (NaGCI) and at higher concentrations of sodium hypochlorite browning of tissues increased. After culture initiation BBTV was indexed by double antibody sandwich (DAS) enzyme linked immuno sorbant assay (ELISA) and polymerase chain reaction (PCR) and 97.4 percent cultures were found BBTV free. During in vitro multiplication, BAP affected the shoot multiplication significantly and maximum shoot number (5.44/explant) was achieved on liquid medium having 4.0 mg/l BAP 1.0 mg/l IAA. When kinetin was used instead of BAP keeping all other parameters same, the maximum shoot number was 4.33/explant. Above optimal concentrations of cytokinins lowered the multiplication rate and callus like swelling was observed at the base of shoot tips. It was also observed from the results that auxins (IAA) did not affect shoot multiplication but increased shoot length up to 5.5 cm against 1.4 cm in control. During in vitro multiplication it was observed that multiplication behavior of each shoot tip was different on the same medium and under similar culture conditions. Multiplication rate declined after 3rd sub-culturing however shoot tips which initially had higher rate of multiplication continued this behavior in succeeding sub-culturings. During in vitro rooting of banana shoots, use of IAA and IBA (1 mg/l) resulted in an increase in the root number and length to 8.6 and 6.8 cm respectively. During hardening of in vitro raised plants, maximum plants survived(88.7 percent) in green house when covered with polythene. When tissue culture raised plants were compared with conventionally propagated plants in the field, it was observed that tissue culture plants were taller than suckers (8.1 percent), produced more suckers (31 percent), time to flowering was reduced (2.8 percent) and produced more fingers/bunch (9 percent). For callus induction and regeneration different explant sources were used but callus and regeneration was observed from male flowers. During plant transformation studies, shoot tips were injured either by biolistic gene gun or bisected longitudinally into two halves. Transgenic banana plants having marker gene were achieved when bisected shoot tips were co-cultivated with Agrobacterium tumefaciens EHA 105 containing pCAMBIA 1301. Transgenic banana plants were selected on MS. Medium containing 50 mg/l hygromycin. A part of hygromycin gene was amplified by PCR and three out of five surviving plants were found positive for hygromycin gene.

Download Full Thesis
22701.98 KB
S. No. Chapter Title of the Chapters Page Size (KB)
1 0 Contents
1864.68 KB
2 1 Introduction 1
684.96 KB
3 2 Review Of Literature 5
3918.85 KB
  2.1 Tissue Culture 5
4 3 Materials And Methods 28
3226.21 KB
  3.1 Tissue Culture 28
  3.2 Genetic Engineering 44
5 4 Results And Discussion 51
9462.74 KB
  4.1 Sterilization Explants 51
  4.2 Effect Of Media On Culture Initiation 56
  4.3 In Vitro Multiplication 60
  4.4 Effect Of Explants On In Vitro Multiplication Of Banana 73
  4.5 In Vitro Rooting Of Banana Shoots 77
  4.6 Hardening Of In Vitro Raised Banana Plants 81
  4.7 Comparison Of In Vitro Raised And Conventionally Propagated Banana Plants In The Field 86
  4.8 Callus Induction And Regeneration 92
  4.9 Genetic Transformation 98
6 5 Summary
3735.12 KB
  5.1 Listerature Cited 115
  5.2 Appendices 134