Date of Award
2013
Degree Type
Thesis
Degree Name
Master of Science (MS)
Department
Forestry and Natural Resources
First Advisor
Paula Pijut
Committee Chair
Paula Pijut
Committee Member 1
Charles Michler
Committee Member 2
Peter Goldsbrough
Abstract
Hypocotyls isolated from mature embryos germinated on Murashige and Skoog (MS) medium supplemented with 22.2 µM 6-benzyladenine (BA) and 0.5 µM thidiazuron (TDZ) were transformed using Agrobacterium tumefaciens strain EHA105 harboring the pBI121-8D2 vector containing the neomycin phosphotransferase (nptII) gene and a full length, codon-optimized Bacillus thuringiensis Cry8Da protoxin gene (cry8D2). Explants were transformed in a bacterial suspension with 100 µM acetosyringone using 90 s sonication and 10 min vacuum-infiltration. Four independent lines of transformed shoots were selectively regenerated on MS medium with 22.2 µM BA, 0.5 µM TDZ, 50 mg L-1 adenine sulfate, 10% coconut water, 30 mg L-1 kanamycin, and 500 mg L-1 timentin. The presence of cry8D2 and nptII were confirmed by polymerase chain reaction for all four transgenic lines. Transgenic white ash microshoots are being multiplied for use in future feeding trials to determine potential efficacy of the cry8D2 gene against emerald ash borer attack.
Recommended Citation
Palla, Kaitlin Joy, "Developing White Ash with Resistance to the Emerald Ash Borer" (2013). Open Access Theses. 82.
https://docs.lib.purdue.edu/open_access_theses/82
Included in
Forest Sciences Commons, Natural Resources Management and Policy Commons, Plant Biology Commons