Transient expression efficiency of a novel Agrobacterium strain in soybean leaf tissue

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Agroinfiltration is an efficient tool used to rapidly assess plant gene function by transiently expressing DNA of interest. In agroinfiltration, we take advantage of a complex and naturally-occurring DNA transformation mechanism belonging to a bacterium from the genus, Agrobacterium. Agrobacterium cells are forced into plant tissue in order to introduce and express desired genes in the plant. Agroinfiltration is commonly utilized for gene function studies such as gene silencing or overexpression. Despite wide use of agroinfiltration in a number of plant species, the technique only recently became available for soybean (Glycine max), and efficiency improvements are needed. In agroinfiltration, variation in transient expression has been observed among Agrobacterium strains; therefore, a novel strain may provide improvements. As compared to commonly used laboratory strains of Agrobacterium, a newly acquired strain, JTND, has been characterized as possessing higher transient expression rates in soybean explants and embryogenic tissue cultures following infiltration. Thus, this strain may also exhibit a higher rate of transient expression in agroinfiltrated soybean seedlings. The JTND strain was disarmed to remove plant tumor inducing genes, and the disarmed strain was named SBHT. In this work, we aim to assess the transient expression rate of SBHT in agroinfiltrated leaf tissue from soybean seedlings. Transient expression rates from agroinfiltrated soybean primary leaf tissue will be compared by image analysis of the expression of a reporter gene, β-glucuronidase (GUS), for SBHT and two check strains which exhibit the current highest measured transient expression levels in soybean. Three experimental replicates will be completed and evaluated. Evaluation of one replicate will be presented at the Denman Forum. If shown to give higher transient expression rates than previously evaluated Agrobacterium strains, SBHT will provide researchers with an improved Agrobacterium strain for use in soybean gene function studies.


Agriculture/Environmental Science: 3rd Place (The Ohio State University Denman Undergraduate Research Forum)


transient expression, Agrobacterium, agroinfiltration, reporter gene, GUS staining