Kinesin-4 optimizes microtubule orientations for responsive tip growth guidance in moss.

Tip-growing cells of, amongst others, plants and fungi secrete wall materials in a highly polarized fashion for fast and efficient colonization of the environment. A polarized microtubule cytoskeleton, in which most microtubule ends are directed toward the growing apex, has been implicated in directing growth. Its organizing principles, in particular regarding maintenance of network unipolarity, have remained elusive. We show that a kinesin-4 protein, hitherto best known for a role in cytokinesis, suppresses encounters between antiparallel microtubules. Without this activity, microtubules hyper-aligned along the growth axis and increasingly grew away from the apex. Cells themselves displayed an overly straight growth path and a delayed gravitropic response. This result revealed conflicting systemic needs for a stable growth direction and an ability to change course in response to extracellular cues. Thus, the use of selective inhibition of microtubule growth at antiparallel overlaps constitutes a new organizing principle within a unipolar microtubule array.

Mechano-chemical regulation of complex cell shape formation: Epidermal pavement cells-A case study.

All plant cells are encased by walls, which provide structural support and control their morphology. How plant cells regulate the deposition of the wall to generate complex shapes is a topic of ongoing research. Scientists have identified several model systems, the epidermal pavement cells of cotyledons and leaves being an ideal platform to study the formation of complex cell shapes. These cells indeed grow alternating protrusions and indentations resulting in jigsaw puzzle cell shapes. How and why these cells adopt such shapes has shown to be a challenging problem to solve, notably because it involves the integration of molecular and mechanical regulation together with cytoskeletal dynamics and cell wall modifications. In this review, we highlight some recent progress focusing on how these processes may be integrated at the cellular level along with recent quantitative morphometric approaches.

Long-term live-cell imaging techniques for visualizing pavement cell morphogenesis.

Recent advancements in microscopy and biological technologies have allowed scientists to study dynamic plant developmental processes with high temporal and spatial resolution. Pavement cells, epidermal cells found on leaf tissue, form complex shapes with alternating regions of indentations and outgrowths that are postulated to be driven by the microtubule cytoskeleton. Given their complex shapes, pavement cells and the microtubule contribution towards morphogenesis have been of great interest in the field of developmental biology. Here, we focus on two live-cell imaging methods that allow for early and long-term imaging of the cotyledon (embryonic leaf-like tissue) and leaf epidermis with minimal invasiveness in order to study microtubules throughout pavement cell morphogenesis. The methods described in this chapter can be applied to studying other developmental processes associated with cotyledon and leaf tissue.