Effect of Agrobacterium-derived Cytokinin on Nicotiana benthamiana Cells

Agrobacterium tumefaciens-based transient assays have become a common tool in researches regarding protein localization and gene expression in cells. However, the cellular organization and organelle morphology in Agrobacterium-infiltrated cells has not been examined in detail. The objective of this study was to characterize the effect of Agrobacterium tumefaciens-based transient assay on the cell from short-term bacterial infestation as well as to identify the factor responsible for these changes.

The impact of GV3101 infiltration on chloroplast behaviour and morphology in Nicotiana benthamiana was evaluated. Our experiments confirmed that GV3101 consistently induces stromules or 'stroma-filled-tubules' and alters plastid position relative to the nucleus. These were the result of a strain-dependent secretion of cytokinin and its accumulation in the plant tissue. The production of the hormone by the bacteria was found to be dependent on the presence of a trans-zeatin synthase gene (tzs) located on the Ti plasmid of GV3101. Bacteria-derived cytokinins also affected both soluble sugar level and starch accumulation.

The impact of the bacteria can be minimized by utilizing Agrobacterium strains without the tzs gene, however, the alterations can not be completely avoided using this strategy.

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