Proteome and Metabolome Alterations in Radish (Raphanus sativus L.) Seedlings Induced by Inoculation with Agrobacterium tumefaciens.

Abstract

Infection of higher plants with agrobacteria (Agrobacterium tumefaciens) represents one of the most comprehensively characterized examples of plant-microbial interactions. Incorporation of the bacterial transfer DNA (T-DNA) in the plant genome results in highly efficient expression of the bacterial auxin, cytokinin and opine biosynthesis genes, as well as the host genes of hormone-mediated signaling. These transcriptional events trigger enhanced proliferation of plant cells and formation of crown gall tumors. Because of this, infection of plant tissues with A. tumefaciens provides a convenient model to address the dynamics of cell metabolism accompanying plant development. To date, both early and late plant responses to agrobacterial infection are well-characterized at the level of the transcriptome, whereas only little information on the accompanying changes in plant metabolism is available. Therefore, here we employ an integrated proteomics and metabolomics approach to address the metabolic shifts and molecular events accompanying plant responses to inoculation with the A. tumefaciens culture. Based on the acquired proteomics dataset complemented with the results of the metabolite profiling experiment, we succeeded in characterizing the metabolic shifts associated with agrobacterial infection. The observed dynamics of the seedling proteome and metabolome clearly indicated rearrangement of the energy metabolism on the 10th day after inoculation (d.a.i.). Specifically, redirection of the energy metabolism from the oxidative to the anaerobic pathway was observed. This might be a part of the plant's adaptation response to tumor-induced hypoxic stress, which most likely involved activation of sugar signaling.