The transcriptome, extracellular proteome and active secretome of agroinfiltrated N. benthamiana uncover an expanded protease repertoire

ABSTRACT: Background: The infiltration of disarmed Agrobacterium tumefaciens into leaves of Nicotiana benthamiana (agroinfiltration) is a widely-adopted procedure to produce and study proteins in plant science, for instance to determine subcellular localization and protein-protein interactions. Agroinfiltration also facilitates quick and safe production of biopharmaceuticals. However, the agroinfiltration expression platform is hampered by unintended proteolysis, some of which might be associated with an elevated immune response. Results: Here, we studied the transcriptome, extracellular proteome and active secretome of agroinfiltrated leaves over a time course, with and without the p19 silencing inhibitor. Remarkably, p19 expression had little effect on the leaf transcriptome and no effect on the extracellular proteome. 25% of the detected transcripts changed in abundance upon agroinfiltration, associated with a gradual upregulation of immunity and a downregulation of photosynthesis. By contrast, 70% of the extracellular proteins increase in abundance and for some proteins this coincides with increased efficiency of extracellular delivery. The protease repertoire of N. benthamiana is expanded compared to other model plants and consists of 1245 proteases and protease homologs. We detect transcripts encoding for 975 proteases, identified extracellular peptides from 196 proteases and monitored 17 active extracellular Ser and Cys proteases in agroinfiltrated leaves. Conclusions: Analysis of agroinfiltrated leaves at transcript, protein and activity levels uncover transcriptional and post-transcriptional/translational reprogramming upon agroinfiltration, associated with substantial increase of the extracellular proteome, and a complex, expanded protease repertoire. This dataset increases our understanding of the plant response to agroinfiltration and indicates ways to improve a key expression platform for both plant science and molecular farming.

OTHER RELATED OMICS DATASETS IN: PRJNA390722

INSTRUMENT(S): LTQ Orbitrap Elite

ORGANISM(S): Agrobacterium Fabrum Str. C58 Nicotiana Benthamiana

SUBMITTER: Farnusch Kaschani  

LAB HEAD: Farnusch Kaschani

PROVIDER: PXD006708 | Pride | 2017-10-24

REPOSITORIES: pride