An Affordable, Scalable, and Labor-less Experimental Setup for the Vacuum-assisted, Agrobacterium-mediated Transient Expression in Tomato (Solanum lycopersicum)

Several methods are available for use, to deliver the gene of interest into plants. Among these, Agrobacterium-mediated transformation utilizes binary vector systems achieve stable Alas, this process is labor-intensive and time-consuming as several months needed obtain a true transgenic plant. Transient expression (e.g., vacuum-infiltration) were offered an alternative over transformation, specifically overcome time-related drawbacks. However, method requires expensive equipment such vacuum chambers. In study, we report vacuum-infiltration protocol transient reporter gene, modified green fluorescent protein (mGFP), in tomato seedling. With basic experimental setup (including custom-built growth chamber) with affordable equipment, showed that entire leaf can be infiltrated by applying mere 200 millibar 6 minutes, thus achieved plant, evidenced PCR-based detection T-DNA, mGFP both analytically SDS-page analysis, visually images acquired fluorescence microscopy. Furthermore, different Agrobacterium tumefaciens strains tested their efficiency, LBA4404 was most effective strain use vacuum-assisted expression.