Can Undergrads Make Transgenic Plants?
Comparison of Two Agrobacterium-mediated Gene Transfer Lab Protocols and their Effectiveness at Teaching Concepts of Bacterial Genetics and Biotechnology Anne Y. Tsang, Sabrina Kramer, and Ann C. Smith. Two protocols for the transformation of plants using Agrobacterium tumefaciens were adapted for use in a General Microbiology lab. In protocol 1, Arabidopsis thaliana seedlings are grown, root tissue harvested, and roots soaked in Agrobacterium cultures to allow transfer of an antibiotic resistance gene. The root tissue are induced to form callous tissue and subsequently transferred to media to induce shoot formation and select for transformants. In protocol 2, juvenile Nicotiana benthamiana leaves are infiltrated with the Agrobacterium culture using a 1ml syringe. Plants are observed 48 hours later for expression of a transferred Green Fluorescent Protein gene. Protocol 1 results in the production of a stable transgenic plant, but the procedure takes weeks and requires exacting techniq
