Aluminium effects on microtubule organization in dividing root-tip cells of Triticum turgidum. I. Mitotic cells

The effects of aluminium (Al) on dividing root-tip cells of Triticum turgidum were investigated with tubulin immunolabelling and electron microscopy. Aluminium affects the mechanisms controlling the organization of microtubule (MT) cytoskeleton, as well as tubulin polymerization, and induces the following aberrations in mitotic cells. (1) It delays the MT disassembly during mitosis, resulting in the persistence of preprophase MT bands in the late prophase cells, the presence of prophase spindles in prometaphase cells, and a disturbance in the shortening of kinetochore MT bundles in anaphase cells. (2) It interferes with the self-organization process of MTs into bipolar systems, inhibiting the formation of prophase and metaphase spindles. (3) Aluminium induces the formation of atypical MT arrays, which in the immunofluorescent specimens appear as ring-like tubulin aggregations in the cortical cytoplasm of the preprophase/prophase cells and as endoplasmic tubulin bundles in prophase and metaphase/anaphase cells; abnormal preprophase MT bands are assembled, consisting of atypical cortical and endoplasmic MT bundles, the latter clearly lining the nuclear envelope on the preprophase MT band plane. (4) It disorders the chromosome movements carried out by the mitotic spindle. In addition, after prolonged Al treatments chromatin condensation is inhibited. The outcome is greatly disturbed organization and function of the mitotic apparatus, as well as inhibition of cells from entering mitosis. This study shows that the MT cytoskeleton is a target site of Al toxicity in mitotic root-tip cells of T. turgidum . The possible mechanisms by which Al exerts its toxicity on MT organization and function are discussed.     

Studies on the Development of the Air Pores and Air Chambers of Marchantia paleacea: 1. Light Microscopy

The ontogeny of the air pores and air chambers of Marchantiapaleacea begins with the schizogenous development of protodermalintercellular spaces of the initial apertures, and is completedwith the formation of the air pores and giant sub-epidermalair chambers bearing numerous photosynthetic filaments. Intercellularspace formation commences from the thallus surface and proceedsinwards to the first internal layer of cells. The cells amongwhich spaces develop do not originate from one mother cell.Spaces are formed only in the regions of the intersection ofthe anticlinal walls of three, four, or sometimes more successivederivatives of S₁, S₃ and S₄ segments of the apical cell, oneor two of which have been divided periclinally and the restanticlinally. Protodermal intercellular spaces appear in mostor all the corners of these cells, the anticlinal walls of whichexhibit an opposite disposition. The S₁, S₂, S₃ and S₄ segmentsare produced by definite divisions of a five-sided apical celland by a series of divisions give rise to initial cells of theinternal layers of the thallus and initial cells of the protodermaland sub-protodermal layers. The concept of a quiescent apicalcell cannot be accepted, since dividing apical cells have beenobserved, and the pattern of wall disposition of the thallusapex cannot be explained without the active participation ofthe apical cell. The air chambers are apparently of exogenous origin. They resultfrom the broadening of the bottom of the initial apertures bythe coordination of the rate of anticlinal divisions and growthof the protodermal and sub-protodermal cells surrounding theintercellular spaces of the initial apertures. The ontogenyof the pore rings starts at an advanced stage of air chamberformation not from a mother cell but from the cells which surroundthe closed entrance of the air chamber, by a shift of the planeof division from anticlinal to periclinal. Before the periclinaldivisions a new axis of growth perpendicular to the thallussurface is established in the mother cells of the pore. By a polarized growth into the air chamber followed by periclinaldivisions, the cells of the floor form initial cells of thephotosynthetic filaments. The latter divide again to form singleor branched photosynthetic filaments. Marchantia paleacea, air pore, air chamber     

Pre-prophase Microtubule Band and Local Wall Thickening in Guard Cell Mother Cells of Some Leguminosae

An examination of leaf protodermal cells from 20 species belongingto the family Leguminosae (15 Lotoideae, three Mimosoideae andtwo Caesalpinioideae) revealed: (a) the organization of pre-prophasemicrotubule bands (PMBs), and (b) that the regions of the anticlinalwalls of the guard cell mother cells (GMCs) lined by the PMBbecome detectably thickened. These local thickenings are depositedduring the presence of pre-prophase microtubules which persistup to late prophase. During deposition of the thickenings theGMCs exhibit a significant dictyosome and endoplasmic reticulum(ER) activity. Smooth as well as coated dictyosome vesiclesseem to fuse preferentially with the plasmalemma of the PMBcortical zone. The cell plate of the symmetrical division ofthe GMC meets the parental walls at the middle of the thickeningswith surprising accuracy. It intersects the middle of the corticalcytoplasmic region traversed previously by the PMB. The observations favour the view that: (a) the deposition ofthe local wall thickenings in the cortical site of the planeof the future cytokinesis in GMCs must be a general featureof the Leguminosae, (b) in these plants the guard cell walldifferentiation commences in GMCs and (c) the PMB is a cytoplasmicstructure appearing extensively in vegetative cells of higherplants. Pre-prophase microtubule band, wall thickening, guard cell mother cell, Leguminosae     

Phosphorylation of a p38‐like MAPK is involved in sensing cellular redox state and drives atypical tubulin polymer assembly in angiosperms

Reactive oxygen species (ROS) imbalance is a stressful condition for plant cells accompanied by dramatic changes in tubulin cytoskeleton. Here, evidence is provided that alterations in ROS levels directly interfere with the phosphorylation state of a p38‐like MAPK in the angiosperms Triticum turgidum and Arabidopsis thaliana. Both oxidative stress generators and chemicals inducing ROS scavenging or decreasing ROS production resulted in the accumulation of a phospho‐p46 protein similar to p38‐MAPK. Importantly, the rhd2 A. thaliana mutants exhibited a remarkable increase in levels of phospho‐p46. The presence of the p38‐MAPK inhibitor SB203580 attenuated the response to ROS disturbance, prevented microtubule disappearance and resulted in a dramatic decrease in the number of atypical tubulin polymers. Moreover, in roots treated simultaneously with substances inducing ROS overproduction and others resulting in low ROS levels, phospho‐p46 levels and the organization of tubulin cytoskeleton were similar to controls. Collectively, our experimental data suggest, for the first time in plants, that p46 functions as a putative sensor of redox state, the activation of which initiates downstream signalling events leading to microtubule disruption and subsequent assembly of atypical tubulin polymers. Thus, p46 seems to participate in perception of ROS homeostasis disturbance as well as in cellular responses to redox imbalance.