Proper plant development in a closed ecosystem under weightlessness will be crucial for the success of future space missions. To supplement spaceflight experiments, such conditions of microgravity are modelled on Earth using a two-axial (2A) clinorotation, and in several fundamental studies resulted in the data on proteome and metabolome adjustments, embryo development, cell cycle regulation, etc. Nevertheless, our understanding of the cytoskeleton responses to the microgravity is still limited. In the present work, we study the adjustment of actin microfilaments (MFs) and microtubules (MTs) in Arabidopsis thaliana (L.) Heynh. seedlings under 2A clinorotation. Modelled microgravity resulted in not only the alteration of seedlings phenotype, but also a transient increase of the hydrogen peroxide level and in the cytoskeleton adjustment. Using GFP-fABD2 and Lifeact-Venus transgenic lines, we demonstrate that MFs became 'scattered' in elongating root and hypocotyl cells under 2A clinorotation. In addition, in GFP-MAP4 and GFP-TUA6 lines the tubulin cytoskeleton had higher fractions of transverse MTs under 2A clinorotation. Remarkably, the first static gravistimulation of continuously clinorotated seedlings reverted MF organisation to a longitudinal one in roots within 30 min. Our data suggest that the 'scattered' organisation of MFs in microgravity can serve as a good basis for the rapid cytoskeleton conversion to a 'longitudinal' structure under the gravity force.