Live-cell imaging of the cytoskeleton in elongating cotton fibres

Cotton ( Gossypium hirsutum ) fibres consist of single cells that grow in a highly polarized manner, assumed to be controlled by the cytoskeleton 1 – 3 . However, how the cytoskeletal organization and dynamics underpin fibre development remains unexplored. Moreover, it is unclear whether cotton fibres expand via tip growth or diffuse growth 2 – 4 . We generated stable transgenic cotton plants expressing fluorescent markers of the actin and microtubule cytoskeleton. Live-cell imaging revealed that elongating cotton fibres assemble a cortical filamentous actin network that extends along the cell axis to finally form actin strands with closed loops in the tapered fibre tip. Analyses of F-actin network properties indicate that cotton fibres have a unique actin organization that blends features of both diffuse and tip growth modes. Interestingly, typical actin organization and endosomal vesicle aggregation found in tip-growing cell apices were not observed in fibre tips. Instead, endomembrane compartments were evenly distributed along the elongating fibre cells and moved bi-directionally along the fibre shank to the fibre tip. Moreover, plus-end tracked microtubules transversely encircled elongating fibre shanks, reminiscent of diffusely growing cells. Collectively, our findings indicate that cotton fibres elongate via a unique tip-biased diffuse growth mode. Cotton fibre grows in a protective boll that becomes a barrier for researchers when tracking the growth dynamics. Now, the cytoskeleton in elongating cotton fibres has been visualized by live-cell imaging. The cytoskeleton organization and dynamics shed light on the elongation mode of cotton fibre.