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Most plant biotechnology laboratories utilize Agrobacterium for production of transgenic plants, and most of the transgenic plants that are in farmer's fields today were generated using Agrobacterium-mediated transformation. Although production of transgenic crop plants is now routine and is often automated, improvements in the efficiency of transgenic plant production are still needed for basic research in crop improvement and in the production of commercial transgenic plants. The standard protocol for production of transgenic plants involves the inoculation of specific target plant tissue with Agrobacterium, co-culture of the bacterium with the plant tissue for 2-3 days, washing of the plant tissue to remove the bacterium, and finally culture of the plant tissue on a medium containing antibiotics to remove or kill the residual Agrobacterium. This standard protocol has been in place since the advent of plant transformation and no one has evaluated our newly discovered and protected technology involving use of a low inoculum of Agrobacterium with long co-culture periods.
This protocol would be useful to plant biotechnology companies involved in producing transgenic plants or biosimilars. The market for biosimilars is large and growing tremendously.
Researchers at The Ohio State University, led by Dr. Finer, have established a protocol using low inoculation levels of Agrobacterium with a long co-culture period. This new approach, which we have termed LILC for Low Inoculum/Long Co-culture, has applications to the transformation targets that are currently being used, and also could allow the use of alternate transformation targets that were not previously considered. Since the low inoculum approach allows a long term co-culture period, targets that would either be sensitive to or overtaken by this bacterium could now be used. Some example of alternate targets include seeds, seedlings, shoot meristems, floral meristems, whole plants, pollen, flowers, and other tissues, which were previously not considered or resulted in very low transformation rates.