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Title page for etd-0822108-151902


URN etd-0822108-151902 Statistics This thesis had been viewed 2140 times. Download 6 times.
Author Su-Kuei Ching
Author's Email Address No Public.
Department Bioengineering
Year 2007 Semester 2
Degree Master Type of Document Master's Thesis
Language zh-TW.Big5 Chinese Page Count 69
Title Establishment of Agrobacterium-mediated transformation system by multiple shoot clumps of Musa. acuminata cv. ‘Pei Chiao’
Keyword
  • Musa. acuminata cv.
  • Agrobacterium-mediated transformation
  • multiple shoot clumps
  • multiple shoot clumps
  • Agrobacterium-mediated transformation
  • Musa. acuminata cv.
  • Abstract An efficient protocol for the Agrobacterium tumefaciens-mediated transformation of banana (Musa acuminata. cv. ‘Pei Chiao’) is described. Cauliflower-like bud clumps (multiple bud clumps) were co-cultivated with A. tumefaciens EHA105 carrying a plasmid containing β-glucuronidase (GUS) reporter gene. In the absence of kanamycin, the bud clumps discs grew normally and produced shoots. In the presence of 30 mg/L kanamycin, the regeneration of the non-transform disc was inhibited. The growth and regeneration on both transformed and non-transformed discs were completely inhibited, and bleaching was observed under the concentration of 40 mg/L in the medium. In order to increase the regeneration of transformed plant, explants were cultured on growth medium without kanamycin after co-cultivation and putative plants were directly identified by Polymerase Chain Reaction (PCR) assay. Forty-eight out of 138 discs were found to have survived and produced shoots. Twenty-three plants of these were confirmed to contain the GUS transgene by PCR. The GUS expression in transgenic plants was only found in roots but not in leaves. Only four transgenic plants (PCR positive) found GUS expression at different RNA levels by RT-PCR (Reverse Transcriptase PCR) assay. The protocol is useful for the breeding of banana through genetic transformation.
    Advisor Committee
  • Ho-Chin-Wen - advisor
  • Files indicate in-campus access at 5 years and off-campus not accessible
    Date of Defense 2008-07-21 Date of Submission 2008-08-22


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