Combination of Synthetic Chemistry and Live-Cell Imaging Identified a Rapid Cell Division Inhibitor in Tobacco andArabidopsis thaliana

Abstract

Cell proliferation is crucial to the growth of multicellular organisms, and thus the proper control of cell division is important to prevent developmental arrest or overgrowth. Nevertheless, tools for controlling cell proliferation are still poor in plant. To develop novel tools, we focused on a specific compound family, triarylmethanes, whose members show various antiproliferative activities in animals. By combining organic chemistry to create novel and diverse compounds containing the triarylmethyl moiety and biological screens based on live-cell imaging of a fluorescently labeled tobacco Bright Yellow-2 (BY-2) culture cell line (Nicotiana tabacum), we isolated (3-furyl)diphenylmethane as a strong but partially reversible inhibitor of plant cell division. We also found that this agent had efficient antiproliferative activity in developing organs of Arabidopsis thaliana without causing secondary defects in cell morphology, and induced rapid cell division arrest independent of the cell cycle stage. Given that (3-furyl)diphenylmethane did not affect the growth of a human cell line (HeLa) and a budding yeast (Saccharomyces cerevisiae), it should act specifically on plants. Taking our results together, we propose that the combination of desired chemical synthesis and detailed biological analysis is an effective tool to create novel drugs, and that (3-furyl)diphenylmethane is a specific antiproliferative agent for plants.

A Novel Biological Agent Cytosinpeptidemycin Inhibited the Pathogenesis of Tobacco Mosaic Virus by Inducing Host Resistance and Stress Response

“…The BY-2 cells are subcultured in MS medium, which are not affected by external temperature, humidity, and light. Therefore, as a model system, the BY-2 cells were widely applied in the studies of plant pathology, plant physiology, and recombinant protein expression. − Virus infection is a complicated process involving the interaction between viruses and host plants. Understanding host responses to antiviral agents is advantageous in the development of effective strategies for virus control.…”

<em>In Vitro</em> Ovule Cultivation for Live-cell Imaging of Zygote Polarization and Embryo Patterning in <em>Arabidopsis thaliana</em>

“…Because the ovule is positioned at the bottom of the dish, an inverted microscope must be used. A confocal microscope is sufficient to monitor the cell division pattern as shown in Figure 3B and Supplemental Videos 2 and 3 8,11 , and a spinning-disk confocal microscope is important to perform long-term imaging without cell damage. In contrast, a two-photon excitation microscopy, which is suitable for deep imaging of living organisms 12 , is necessary to visualize the intracellular dynamics of the zygote (Figure 3A and Supplemental Video 1)…”

Chemical screen of Arabidopsis zygote and proteomics in tobacco BY-2 cells identify general plant cell division inhibitors

“…These strains were imaged as described previously (14). For live cell imaging of tobacco BY-2 cells, we followed a previously described method (8, 32, 45). The fluorescent markers of P. patens and imaging methods have been described previously (36, 37, 46).…”