In plant cytokinesis, de novo formation of a cell plate evolving into the new cell wall partitions the cytoplasm of the dividing cell. In our earlier chemical genomics studies, we identified and characterized the small molecule endosidin-7, that specifically inhibits callose deposition at the cell plate, arresting late-stage cytokinesis in arabidopsis. Endosidin-7 has emerged as a very valuable tool for dissecting this essential plant process. To gain insights regarding its mode of action and the effects of cytokinesis inhibition on the overall plant response, we investigated the effect of endosidin-7 through a nuclear magnetic resonance spectroscopy (NMR) metabolomics approach. In this case study, metabolomics profiles of arabidopsis leaf and root tissues were analyzed at different growth stages and endosidin-7 exposure levels. The results show leaf and root-specific metabolic profile changes and the effects of endosidin-7 treatment on these metabolomes. Statistical analyses indicated that the effect of endosidin-7 treatment was more significant than the developmental impact. The endosidin-7 induced metabolic profiles suggest compensations for cytokinesis inhibition in central metabolism pathways. This study further shows that long-term treatment of endosidin-7 profoundly changes, likely via alteration of hormonal regulation, the primary metabolism of arabidopsis seedlings. Hormonal pathway-changes are likely reflecting the plant’s responses, compensating for the arrested cell division, which in turn are leading to global metabolite modulation. The presented NMR spectral data are made available through the Metabolomics Workbench, providing a reference resource for the scientific community.
Digital Object Identifier (DOI)
The work was supported by the U.S. National Science Foundation MCB-1818219 award to G.D., and U.S. Department of Agriculture award CA-D-PLS-2132-H to G.D and in part by the U.S. National Institutes of Health grant SC3-GM125546 to V.V.K.
All the NMR spectral data were deposited in Metabolomics Workbench (accession ST001478) under project DOI: 10.21228/M85T2G.
Wilkop, Thomas E.; Wang, Minmin; Heringer, Angelo; Singh, Jaideep; Zakharov, Florence; Krishnan, Viswanathan V.; and Drakakaki, Georgia, "NMR Spectroscopy Analysis Reveals Differential Metabolic Responses in Arabidopsis Roots and Leaves Treated with a Cytokinesis Inhibitor" (2020). Physiology Faculty Publications. 160.
S1 Fig. Representative NMR spectra of roots and leaves (10-day old plants), and simulated spectra based on the metabolites that are significantly alerted due to endosidin-7 treatment. https://doi.org/10.1371/journal.pone.0241627.s001
pone.0241627.s002.pdf (686 kB)
S2 Fig. Statistical analyses of the arabidopsis metabolome without endosidin-7 treatment. https://doi.org/10.1371/journal.pone.0241627.s002
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S3 Fig. Evaluation of the PLS-DA model. https://doi.org/10.1371/journal.pone.0241627.s003
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S4 Fig. PCA analysis of the data corresponding to PLS-DA analysis shown in Figs 2A and 2B and S2C. https://doi.org/10.1371/journal.pone.0241627.s004
pone.0241627.s005.pdf (293 kB)
S1 Table. Significantly changed leaves and root metabolites upon endosidin-7 treatment. https://doi.org/10.1371/journal.pone.0241627.s005
pone.0241627.s006.pdf (320 kB)
S2 Table. Quantification of metabolite level changes upon endosidin-7 treatment in leaves and roots. https://doi.org/10.1371/journal.pone.0241627.s006
pone.0241627.s007.pdf (702 kB)
S3 Table. Quantification of metabolite level modulations in leaves and roots during seedling development. https://doi.org/10.1371/journal.pone.0241627.s007