Date of Award


Degree Type


Degree Name

Master of Science (MS)


Forestry and Natural Resources

First Advisor

Paula Pijut

Committee Chair

Paula Pijut

Committee Member 1

Charles Michler

Committee Member 2

Peter Goldsbrough


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.