Smashing Barriers in Biolistic Plant Transformation.
A foundation of modern biotechnology is the ability to stably introduce foreign DNA into an organism. The two most widely used methods, Agrobacterium-mediated transformation and biolistics, are both steeped in a rich history of creative exploration into the molecular unknown. Agrobacterium research accelerated in the early 1970s, particularly with the discovery of the large Ti (tumor-inducing) plasmid of Agrobacterium that contained a region of transfer DNA (T-DNA). Culturing plant calli in autoclaved jelly jars, and long before the advent of PCR, Southern blots were first used to show that T-DNA fragments could stably integrate into the nuclear genome (Chilton et al., 1980; Chilton, 2001). On the other hand, the first manufactured biolistic “gene gun” was an actual gun; it shot a blank .22 caliber cartridge loaded with DNA-coated tungsten shards to integrate foreign DNA into the nuclear genome. While it has long been known that biolistic transformation violently integrates DNA in a largely random, unpredictable and imprecise way, the cellular mechanisms of damage repair and successful integration remain a complicated issue to disentangle.