Motility is essential for apicomplexan parasites to infect their hosts. In a three-dimensional (3D) environment, the apicomplexan parasite Toxoplasma gondii moves along a helical path. The cortical microtubules, which are ultra-stable and spirally arranged, have been considered to be a structure that guides the long-distance movement of the parasite. Here, we address the role of the cortical microtubules in parasite motility, invasion and egress by utilizing a previously generated mutant (dubbed \'TKO\') in which these microtubules are destabilized in mature parasites. We found that the cortical microtubules in ∼80% of the non-dividing (i.e. daughter-free) TKO parasites are much shorter than normal. The extent of depolymerization was further exacerbated upon commencement of daughter formation or cold treatment, but parasite replication was not affected. In a 3D Matrigel matrix, the TKO mutant moved directionally over long distances, but along trajectories that were significantly more linear (i.e. less helical) than those of wild-type parasites. Interestingly, this change in trajectory did not impact either movement speed in the matrix or the speed and behavior of the parasite during entry into and egress from the host cell.

Ultraviolet-B (UV-B) radiation is one of the important environmental factors limiting plant growth. Both abscisic acid (ABA) and microtubules have been previously reported to be involved in plant response to UV-B. However, whether there is a potential link between ABA and microtubules and the consequent signal transduction mechanism underlying plant response to UV-B radiation remains largely unclear. Here, by using sad2-2 mutant plants (sensitive to ABA and drought) and exogenous application of ABA, we saw that ABA strengthens the adaptive response to UV-B stress in Arabidopsis thaliana (A. thaliana). The abnormal swelling root tips of ABA-deficient aba3 mutants demonstrated that ABA deficiency aggravated the growth retardation imposed by UV-B radiation. In addition, the cortical microtubule arrays of the transition zones of the roots were examined in the aba3 and sad2-2 mutants with or without UV-B radiation. The observation revealed that UV-B remodels cortical microtubules, and high endogenous ABA can stabilize the microtubules and reduce their UV-B-induced reorganization. To further confirm the role of ABA on microtubule arrays, root growth and cortical microtubules were evaluated after exogenous ABA, taxol, and oryzalin feeding. The results suggested that ABA can promote root elongation by stabilizing the transverse cortical microtubules under UV-B stress conditions. We thus uncovered an important role of ABA, which bridges UV-B and plants\' adaptive response by remodeling the rearrangement of the cortical microtubules.

Terrestrial plants respond to and resist gravitational force. The response is termed \"gravity resistance\", and centrifugal hypergravity conditions are efficient for investigating its nature and mechanism. A functional screening of Arabidopsis T-DNA insertion lines for the suppression rate of elongation growth of hypocotyls under hypergravity conditions was performed in this study to identify the genes required for gravity resistance. As a result, we identified PEPTIDYL-tRNA HYDROLASE II (PTH2). In the wild type, elongation growth was suppressed by hypergravity, but this did not happen in the pth2 mutant. Lateral growth, dynamics of cortical microtubules, mechanical properties of cell walls, or cell wall thickness were also not affected by hypergravity in the pth2 mutant. In other words, the pth2 mutant did not show any significant hypergravity responses. However, the gravitropic curvature of hypocotyls of the pth2 mutant was almost equal to that of the wild type, indicating that the PTH2 gene is not required for gravitropism. It is suggested by these results that PTH2 is responsible for the critical processes of gravity resistance in Arabidopsis hypocotyls.

Coumarin is a phytotoxic natural compound able to affect plant growth and development. Previous studies have demonstrated that this molecule at low concentrations (100 µM) can reduce primary root growth and stimulate lateral root formation, suggesting an auxin-like activity. In the present study, we evaluated coumarin\'s effects (used at lateral root-stimulating concentrations) on the root apical meristem and polar auxin transport to identify its potential mode of action through a confocal microscopy approach. To achieve this goal, we used several Arabidopsis thaliana GFP transgenic lines (for polar auxin transport evaluation), immunolabeling techniques (for imaging cortical microtubules), and GC-MS analysis (for auxin quantification). The results highlighted that coumarin induced cyclin B accumulation, which altered the microtubule cortical array organization and, consequently, the root apical meristem architecture. Such alterations reduced the basipetal transport of auxin to the apical root apical meristem, inducing its accumulation in the maturation zone and stimulating lateral root formation.

BACKGROUND: Cortical microtubules regulate cell expansion by determining cellulose microfibril orientation in the root apex of Arabidopsis thaliana. While the regulation of cell wall properties by cortical microtubules is well studied, the data on the influence of cell wall to cortical microtubule organization and stability remain scarce. Studies on cellulose biosynthesis mutants revealed that cortical microtubules depend on Cellulose Synthase A (CESA) function and/or cell expansion. Furthermore, it has been reported that cortical microtubules in cellulose-deficient mutants are hypersensitive to oryzalin. In this work, the persistence of cortical microtubules against anti-microtubule treatment was thoroughly studied in the roots of several cesa mutants, namely thanatos, mre1, any1, prc1-1 and rsw1, and the Cellulose Synthase Interacting 1 protein (csi1) mutant pom2-4. In addition, various treatments with drugs affecting cell expansion were performed on wild-type roots. Whole mount tubulin immunolabeling was applied in the above roots and observations were performed by confocal microscopy.
RESULTS: Cortical microtubules in all mutants showed statistically significant increased persistence against anti-microtubule drugs, compared to those of the wild-type. Furthermore, to examine if the enhanced stability of cortical microtubules was due to reduced cellulose biosynthesis or to suppression of cell expansion, treatments of wild-type roots with 2,6-dichlorobenzonitrile (DCB) and Congo red were performed. After these treatments, cortical microtubules appeared more resistant to oryzalin, than in the control.
CONCLUSIONS: According to these findings, it may be concluded that inhibition of cell expansion, irrespective of the cause, results in increased microtubule stability in A. thaliana root. In addition, cell expansion does not only rely on cortical microtubule orientation but also plays a regulatory role in microtubule dynamics, as well. Various hypotheses may explain the increased cortical microtubule stability under decreased cell expansion such as the role of cell wall sensors and the presence of less dynamic cortical microtubules.

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The typical scale cells (TSCs) of Marchantia paleacea Bert, contain a well-developed cortical microtubule (Mt) cytoskeleton, particularly below the anticlinal walls and also display complete but broad preprophase-prophase Mt bands (PMBs). In contrast, the cortical cytoskeleton of the inner thallus cells (ITCs) is less developed than that of TSCs and the PMBs are incomplete. The latter consist of one to four separate Mt bundles which lie on the cytokinetic plane, but do not form a complete Mt ring. In both cell types PMB formation precedes or keeps pace with the activation of the polar Mt-organizing centres (MTOCs) and nuclear shaping. The Mts in the PMBs are more numerous and densely packed at the cell edges than on the cell face. The polar MTOCs persist up to late prophase-prometaphase. Afterwards, the spindle Mts are focused on several minipoles, where endoplasmic reticulum is localized. In postcytokinetic cells the cortical Mts first reappear on the daughter wall surface. Our findings suggest that: (a) The formation of complete or incomplete PMBs in TSCs and ITCs of M. paleacea is related to differences in the development of the interphase cortical Mt arrays, (b) The cell edges are able to form or at least arrange the Mts of the PMB. (c) Tight mature PMBs like those found in flowering plant cells are not formed in the cells examined in the present study. (d) The final orientation of the cell plate is controlled by the PMB cortical zone. (e) The cytoplasm abutting on the postcytokinetic daughter wall has the ability to assemble cortical Mts.

This light- and electron-microscope study of four species of Sphagnum reveals that stem elongation involves meristematic activities unique to the group and hitherto unrecognized. The internal tissue of the mature stem arises by the concerted activity of an apical (primary) and a subapical (secondary) meristem. The primary meristem comprises the immediate derivatives of the single apical cell. Following a small number of divisions, the primary derivatives differentiate into highly vacuolate parenchymatous cells with a storied arrangement. Subsequently, the large vacuoles are replaced by numerous small vacuoles and the cells then divide repeatedly, by transverse septa, producing files of about nine short cells. Finally, ninefold elongation of these secondary cells is responsible for extension growth of the main stem below the mature capitulum. An early step in primary differentiation is the confinement of pre-existing plasmodesmata to distinct pitted areas. Further enlargement of the cells during primary and secondary differentiation involves the thickening of non-pitted wall areas, followed by expansion and thinning out, while the pitted areas remain virtually unchanged. A cortical array of microtubules is regularly found in association with non-pitted wall areas, while the unexpanded pitted areas are associated with smooth endoplasmic reticulum showing continuity with desmotubules. Though sharing much the same cytology as the conducting cells in bryoid mosses, in terms of their development the central stem cells in Sphagnum are not homologous with those of other mosses. The unique mode of stem development may be an important factor in the ecological success of Sphagnum.

IQ67-domain (IQD) proteins, first identified in Arabidopsis and rice, are plant-specific calmodulin-binding proteins containing highly conserved motifs. They play a critical role in plant defenses, organ development and shape, and drought tolerance. Driven by comprehensive genome identification and analysis efforts, IQDs have now been characterized in several species and have been shown to act as microtubule-associated proteins, participating in microtubule-related signaling pathways. However, the precise molecular mechanisms underpinning their biological functions remain incompletely understood. Here we review current knowledge on how IQD family members are thought to regulate plant growth and development by affecting microtubule dynamics or participating in microtubule-related signaling pathways in different plant species and propose some new insights.

Upon water uptake and release of seed dormancy, embryonic plant cells expand, while being mechanically constrained by the seed coat. Cortical microtubules (CMTs) are key players of cell elongation in plants: their anisotropic orientation channels the axis of cell elongation through the guidance of oriented deposition of load-bearing cellulose microfibrils in the cell wall. Interestingly, CMTs align with tensile stress, and consistently, they reorient upon compressive stress in growing hypocotyls. How CMTs first organise in germinating embryos is unknown, and their relation with mechanical stress has not been investigated at such an early developing stage.
Here, we analysed CMT dynamics in dormant and non-dormant Arabidopsis seeds by microscopy of fluorescently tagged microtubule markers at different developmental time points and in response to abscisic acid and gibberellins. We found that CMTs first appear as very few thick bundles in dormant seeds. Consistently, analysis of available transcriptome and translatome datasets show that limiting amounts of tubulin and microtubule regulators initially hinder microtubule self-organisation. Seeds imbibed in the presence of gibberellic acid or abscisic acid displayed altered microtubule organisation and transcriptional regulation. Upon the release of dormancy, CMTs then self-organise into multiple parallel transverse arrays. Such behaviour matches the tensile stress patterns in such mechanically constrained embryos. This suggests that, as CMTs first self-organise, they also align with shape-derived tensile stress patterns.
Our results provide a scenario in which dormancy release in the embryo triggers microtubule self-organisation and alignment with tensile stress prior to germination and anisotropic growth.