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CLASP balances two competing cell division plane cues during leaf development

Nature Plants volume 8, pages 682–693 (2022)Cite this article

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Abstract

Starting as small, densely packed boxes, leaf mesophyll cells expand to form an intricate mesh of interconnected cells and air spaces, the organization of which dictates the internal surface area of the leaf for light capture and gas exchange during photosynthesis. Despite their importance, little is known about the basic patterns of mesophyll cell division, and how they contribute to cell and intercellular space organization. To address this, we tracked divisions within individual cell lineages in three dimensions over time in Arabidopsis spongy mesophyll. We found that early on, successive cell division planes switch their orientation such that each new cell wall intersects the previous at a right angle, creating a new multi-cell junction (the intersection of three or more cells). These junctions then open to create intercellular spaces. During subsequent enlargement of the spaces, the division planes of the surrounding cells show an increasing tendency to tilt in the direction of their adjacent intercellular spaces. This disrupts the alternating pattern, and by extension, halts the initiation of new multi-cell junctions and intercellular spaces, but allows the expansion of existing spaces. Both division patterns are specified before mitosis by the orientation of interphase cortical microtubules, which gradually narrow to form a preprophase band in the same orientation to establish the future plane of cell division. In the absence of the microtubule-associated protein CLASP, the early alternating division plane and microtubule patterns are compromised, whereas space-oriented divisions are exacerbated. This results in large distortions of the topological relations between cells and intercellular spaces, as well as changes in their relative abundance. Our data reveal the existence of two competing cell division mechanisms that are balanced by CLASP to specify the distribution of cells and intercellular spaces in spongy mesophyll tissue.

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Fig. 1: Alternating cell divisions drive intercellular space formation in wild-type spongy mesophyll.
Fig. 2: Intercellular space-oriented cell divisions.
Fig. 3: Organization of interphase MTs predicts the subsequent cell divisions.
Fig. 4: CLASP localizes to cell edges and is required for the establishment of perpendicular PCAs.
Fig. 5: CLASP is required for the alternating division pattern.
Fig. 6: Cell division patterns in wild-type and clasp-1 spongy mesophyll cells.
Fig. 7: Influence of clasp-1 cell division defects on cell and intercellular space topologies in spongy mesophyll.
Fig. 8: Cell division plane defects in clasp-1 lead to a less organized topology of cell–cell contacts.

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Data availability

The plasmids, Arabidopsis lines and confocal stacks used in this study are available from the corresponding author upon request. Source data are provided with this paper.

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Acknowledgements

We would like to thank D. McEvoy (University of Saskatchewan) for a critical reading of the manuscript. This work was supported by the Natural Sciences and Engineering Research Council (NSERC) Discovery Grant 298264-2009, the University of Saskatchewan new faculty start-up funds and the University of Saskatchewan Biology Department graduate student scholarships.

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  1. Department of Biology, The University of Saskatchewan, College of Arts and Science, Saskatoon, Saskatchewan, Canada

    Liyong Zhang & Chris Ambrose

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L.Z. and C.A. conceptualized the project. L.Z. developed the methodologies and performed the experiments. L.Z. and C.A. analysed the data and wrote the manuscript. C.A. acquired the funding.

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Correspondence to Chris Ambrose.

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Zhang, L., Ambrose, C. CLASP balances two competing cell division plane cues during leaf development. Nat. Plants 8, 682–693 (2022). https://doi.org/10.1038/s41477-022-01163-5

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