Abstract
BackgroundCharacterizing the behaviors of dynamic systems requires capturing them with high temporal and spatial resolution. Owing to its transparency and genetic tractability, the Arabidopsis thaliana root lends itself well to live imaging when combined with cell and tissue-specific fluorescent reporters. We developed a novel 4D imaging method that utilizes simple confocal microscopy and readily available components to track cell divisions in the root stem cell niche and surrounding region for up to 1 week.ResultsUsing this method, we performed a direct measurement of cell division intervals within and around the root stem cell niche. The results reveal a short, steep gradient of cell division rates in proximal stem cells, with progressively more rapid cell division rates from quiescent center (QC), to cells in direct contact with the QC (initials), to their immediate daughters, after which division rates appear to become more homogeneous.ConclusionsThese results provide a baseline to study how perturbations in signaling could affect cell division patterns in the root meristem. This new setup further allows us to finely analyze meristematic cell division rates that lead to patterning.
Highlights
Characterizing the behaviors of dynamic systems requires capturing them with high temporal and spatial resolution
We show that initials in direct contact with the Quiescent Center (QC) do exhibit a distinctly slower rate of division compared to their daughters, which, in turn, showed a slightly lower rate of division than their daughters, forming a short, steep gradient over four cells
We used a well-known commercial platform, but the methods described here should be adaptable to other systems
Summary
Characterizing the behaviors of dynamic systems requires capturing them with high temporal and spatial resolution. Owing to its transparency and genetic tractability, the Arabidopsis thaliana root lends itself well to live imaging when combined with cell and tissue-specific fluorescent reporters. Cells in direct contact with the QC, termed initials, behave like stem cells. Initials divide asymmetrically such that one daughter remains in place next to the QC while the other undergoes multiple transit amplifying divisions [2]. While the optical transparency of the root lends itself well to imaging, the requirements for tracking divisions over a long period are challenging. The indeterminate growth of the root means that its position changes dramatically over the time periods needed to study division patterns
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