Stafstrom, Joel P.
M.S. (Master of Science)
Department of Biological Sciences
Agrobacterium mediated transformation can potentially lead to transfer of any fragment of DNA into a plant cell. Pea transformation can be achieved through in vitro techniques. There are certain limitations in using these techniques. For example, tissue culture is quite tedious and time consuming and somaclonal variation frequently occurs during tissue culture. The plants obtained through these methods might be chimeric, sterile or both. In planta transformation methods obviate the need for tissue culture for obtaining transformed plants. There are fewer disadvantages observed in this method when compared to the disadvantages observed while using in vitro techniques. This method is less time consuming and less expensive. We have made an attempt to develop an in planta transformation and regeneration system that would permit the introduction of GUS and BAR genes into Pisum sativum cv. Alaska. GUS activity was observed in some of the transformed shoots that developed from axillary buds (T0 shoots). Injury in the form of removal of a bud along with pricking at second node of pea seedling gave better results in terms of GUS expression when compared to removal of bud alone. Some of the Tx plants showed BASTA resistance and GUS activity. These plants gave rise to T2 seeds. Some T2 seedlings were resistant to BASTA but did not show GUS activity. The vacuum infiltration technique also showed some promising results. In this case, injury was not necessary for plant infection by Agrobacterium. This method is much faster to test the functionality of a construct. This method is suitable for species recalcitrant to tissue culture and/or regeneration because transformed plants can be obtained without going through the step of tissue culture.
Sarup, Vinita B., "Approaches toward in planta transformation of Pisum sativum" (1996). Graduate Research Theses & Dissertations. 1508.
iv, 73 pages
Northern Illinois University
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