Endoplasmic Microtubules Connect the Advancing Nucleus to the Tip of Legume Root Hairs, but F-Actin is Involved in Basipetal Migration (original) (raw)
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Microtubules regulate tip growth and orientation in root hairs ofArabidopsis thaliana
Plant Journal, 1999
The polarized growth of cells as diverse as fungal hyphae, pollen tubes, algal rhizoids and root hairs is characterized by a highly localized regulation of cell expansion confined to the growing tip. In apically growing plant cells, a tipfocused [Ca 2⍣ ] c gradient and the cytoskeleton have been associated with growth. Although actin has been established to be essential for the maintenance of elongation, the role of microtubules remains unclear. To address whether the microtubule cytoskeleton is involved in root hair growth and orientation, we applied microtubule antagonists to root hairs of Arabidopsis. In this report, we show that depolymerizing or stabilizing the microtubule cytoskeleton of these apically growing root hairs led to a loss of directionality of growth and the formation of multiple, independent growth points in a single root hair.
Positioning of Nuclei in Arabidopsis Root Hairs: An Actin-Regulated Process of Tip Growth
THE PLANT CELL ONLINE, 2002
In growing Arabidopsis root hairs, the nucleus locates at a fixed distance from the apex, migrates to a random position during growth arrest, and moves from branch to branch in a mutant with branched hairs. Consistently, an artificial increase of the distance between the nucleus and the apex, achieved by entrapment of the nucleus in a laser beam, stops cell growth. Drug studies show that microtubules are not involved in the positioning of the nucleus but that subapical fine F-actin between the nucleus and the hair apex is required to maintain the nuclear position with respect to the growing apex. Injection of an antibody against plant villin, an actin filament-bundling protein, leads to actin filament unbundling and movement of the nucleus closer to the apex. Thus, the bundled actin at the tip side of the nucleus prevents the nucleus from approaching the apex. In addition, we show that the basipetal movement of the nucleus at root hair growth arrest requires protein synthesis and a functional actin cytoskeleton in the root hair tube.
Journal of Plant Physiology, 1996
Using indirect immunofluorescence, the system of radiating endoplasmic microtubules (REMTs) within intact cells of meristematic root tissues of maize has been examined throughout the cell cycle, paying special attention to its relationship with the pre-and post-mitotic nuclei with which it is associated. At early interphase, REMTs are not uniformly disposed around the nucleus but grow out from faint, though easily recognizable, perinuclear foci. During S and G2 phases, REMTs increase in number and have a close association with the assembly of the preprophase band (PPB) MT array. Later, when the cortical part of the PPB disintegrates, the REMTs align along the nuclear surface, predicting the long axis of the future mitotic spindle. In contrast to naturally wall-less cells, or to cells with perturbed cell walls, these pre-mitotic, as well as the subsequently formed post-mitotic cells display symmetrical rearrangements of their REMTs around the nuclear surface. Mitotic cells sectioned in the median plane show a symmetrical quadripolar MT organization which is obvious at all stages of mitosis. The symmetrical redistributions of the REMTs which occur during the cell cycle are perturbed, or even prevented, by treatments with chemical or with physical anti-MT agents. Nuclei of cells so treated accumulate REMTs, but fail to redistribute them symmetrically. As a result, the pre-and post-mitotic nuclei of root cells treated with anti-MT agents resemble, with respect to their REMTs, the corresponding nuclei of wall-less plant cells, or of cells which have perturbed cell walls. The dynamic REMTs which connect the pre-and post-mitotic nuclei with the cell periphery are suggested as being involved in sensing the position of dividing cells within the intact plant organ. This property of REMTs enables them to spatially control the sequential alignment of cell division planes of immobile walled plant cells which underlies the morphogenesis of higher plant organs.
The Journal of cell …, 1990
To investigate the spatial relationship between the nucleus and the cortical division site, epidermal cells were selected in which the separation between these two areas is large. Avoiding enzyme treatment and air drying, Datura stramonium cells were labeled with antitubulin antibodies and the threedimensional aspect of the cytoskeletons was reconstructed using computer-aided optical sectioning. In vacuolated cells preparing for division, the nucleus migrates into the center of the cell, suspended by transvacuolar strands. These strands are now shown to contain continuous bundles of microtubules which bridge the nucleus to the cortex. These nucleus-radiating microtubules adopt different configurations in cells of different shape. In elongated cells with more or less parallel side walls, oblique strands radiating from the nucleus to the long side walls are presumably unstable, for they are progressively realigned into a transverse disc (the phragmosome) as broad, cortical, preprophase bands (PPBs) become tighter. The phragmosome and the PPB are both known predictors of the division plane and our observations indicate that they align simultaneously in elongated epidermal cells. These observations suggest another hypothesis: that the PPB may contain microtubules polymerized from the nuclear surface. In elongated cells, the majority of the radiating microtubules, therefore, come to anchor the nucleus in the transverse plane, consistent
In this study we compare the contributions of Factin and microtubules to tip growth in filamentous cells of the moss Physcomitrella patens. In tip growth, expansion seems to be restricted to the hemispherical apical dome. Cytoskeletal elements have been suspected, from drug studies, to be involved in this but electron microscopy has generally not confirmed the presence of an apical cytoskeleton. However, in a previous immunofluorescence study we reported that microtubules could be seen to focus upon the apical dome in tip cells of the moss P. patens. In the present investigation F-actin has also been detected at the apices of these cells. Anti-cytoskeletal drugs were therefore used to differentiate between the roles of actin filaments and microtubules in tip growth. At high concentrations (30//M), the herbicide cremart de-polymerized microtubules and caused tip swelling. F-actin was still present under such conditions but its fragmentation by cytochalasin D suppressed this herbicide-induced swelling. On its own, cytochalasin D arrested tip growth without causing swollen tips. At lower concentrations, cremart disorganized microtubules rather than causing their complete depolymerization. Under these conditions, new but swollen growing points were initiated along the filament. The addition of taxol to cremarttreated filaments tended to reduce swelling and to re-polarize outgrowth. With particular combinations of these drugs, multiple lateral outgrowths were initiated in the vicinity of the nucleus. It is concluded: (1) that F-actin is present at the tips of Physcomitrella caulonemal apical cells; (2) that unfragmented F-actin is necessary for outgrowth; (3) that even disorganized microtubules permit some degree of outgrowth but that an unperturbed distribution of axial microtubules, focussing upon an apex, is essential in order to impose tubular shape and directionality upon expansion.
THE PLANT CELL ONLINE, 2003
Plant cells expand by exocytosis of wall material contained in Golgi-derived vesicles. We examined the role of local instability of the actin cytoskeleton in specifying the exocytosis site in Arabidopsis root hairs. During root hair growth, a specific actin cytoskeleton configuration is present in the cell's subapex, which consists of fine bundles of actin filaments that become more and more fine toward the apex, where they may be absent. Pulse application of low concentrations of the actin-depolymerizing drugs cytochalasin D and latrunculin A broadened growing root hair tips (i.e., they increased the area of cell expansion). Interestingly, recovery from cytochalasin D led to new growth in the original growth direction, whereas in the presence of oryzalin, a microtubule-depolymerizing drug, this direction was altered. Oryzalin alone, at the same concentration, had no influence on root hair elongation. These results represent an important step toward understanding the spatial and directional regulation of root hair growth.
Microtubules and Microfilaments in Cell Morphogenesis in Higher Plants
Current Biology, 2002
Microtubules and microfilaments play important roles in cell morphogenesis. The picture emerging from drug studies and molecular-genetic analyses of mutant higher plants defective in cell morphogenesis shows that the roles played by them remain the same in both tip-growing and diffuse-growing cells. Microtubules are important for establishing and maintaining growth polarity whereas actin microfilaments deliver the materials required for growth to specified sites. The recent cloning of several cell morphogenesis genes has revealed that conserved mechanisms as well as novel signal transduction pathways spatially organize the plant cytoskeleton.
In the presence of cytokinin, undetermined side branch initials of the moss, Physcomitrella patens, are induced to form buds and then leafy shoots rather than to develop as tip-growing filaments. This represents a transition between the two modes of plant cell expansion-tip growth and uniform intercalary growth. The organization of microtubules in filaments is different from that in leafy shoots and can be traced back to the influence of phytohormones on side branch initials. Microtubules either focus at a particular region (as in tip-growing cells) or in the presence of high levels of cytokinin form swollen bud initials in which microtubules are more diffusely organized. Higher levels of cytokinin are capable of destabilizing tip microtubules in caulonemal filaments. Although caulonemata are not normally target cells, this implies that cytokinin may exert its morphogenetic effects by altering microtubule organization. IR tip-growing filaments, interphase microtubules trace a meandering course through the cytoplasm towards the tip and are not for the main part associated with the plasma membrane as are cortical arrays. There is no pre-prophase band of microtubules to indicate the future division plane, even though the oblique division plane is known to be precisely controlled relative to environmental factors. This microtubule cycle contrasts with cells of the leafy shoots that develop from buds: in these, the interphase array is cortical, consisting of flat-pitched microtubular helices that do not focus upon a growing tip. It is now shown that pre-prophase bands occur at this stage. The absence of bands does not readily correlate with imprecise control of the division plane. Instead, it is proposed that the ability to form pre-prophase bands depends upon the arrangement of microtubules in the preceding interphase array. Ways in which bands might be formed are discussed and the generality of these ideas is tested by observations on higher plant cells.
PLANT PHYSIOLOGY, 2002
To investigate the configuration and function of microtubules (MTs) in tip-growing Medicago truncatula root hairs, we used immunocytochemistry or in vivo decoration by a GFP linked to a MT-binding domain. The two approaches gave similar results and allowed the study of MTs during hair development. Cortical MTs (CMTs) are present in all developmental stages. During the transition from bulge to a tip-growing root hair, endoplasmic MTs (EMTs) appear at the tip of the young hair and remain there until growth arrest. EMTs are a specific feature of tip-growing hairs, forming a three-dimensional array throughout the subapical cytoplasmic dense region. During growth arrest, EMTs, together with the subapical cytoplasmic dense region, progressively disappear, whereas CMTs extend further toward the tip. In full-grown root hairs, CMTs, the only remaining population of MTs, converge at the tip and their density decreases over time. Upon treatment of growing hairs with 1 m oryzalin, EMTs disappear, but CMTs remain present. The subapical cytoplasmic dense region becomes very short, the distance nucleus tip increases, growth slows down, and the nucleus still follows the advancing tip, though at a much larger distance. Taxol has no effect on the cytoarchitecture of growing hairs; the subapical cytoplasmic dense region remains intact, the nucleus keeps its distance from the tip, but growth rate drops to the same extent as in hairs treated with 1 m oryzalin. The role of EMTs in growing root hairs is discussed. ; fax 31-317-485005.