Cytochemical Localization of Calcium in Cap Cells of Primary Roots of Zea mays L.

The distribution of calcium (Ca) in caps of vertically- andhorizontally-oriented roots of Zea mays was monitored to determineits possible role in root graviresponsiveness. A modificationof the antimonate precipitation procedure was used to localizeCa in situ. In vertically-oriented roots, the presumed graviperceptive(i.e., columella) cells were characterized by minimal and symmetricstaining of the plasmalemma and mitochondria. No precipitatewas present in plasmodesmata or cell walls. Within 5 min afterhorizontal reorientation, staining was associated with the portionof the cell wall adjacent to the distal end of the cell. Thisasymmetric staining persisted throughout the onset of gravicurvature.No staining of lateral cell walls of columella cells was observedat any stage of gravicurvature, suggesting that a lateral flowof Ca through the columella tissue of horizontally-orientedroots does not occur. The outermost peripheral cells of rootsoriented horizontally and vertically secrete Ca through plasmodesmata-likestructures in their cell walls. These results are discussedrelative to proposed roles of root-cap Ca in root gravicurvature. Key words: Antimonate, calcium, columella cell, peripheral cell, root gravitropism, Zea mays L.     

Defective Secretion of Mucilage is the Cellular Basis for Agravitropism in Primary Roots of Zea mays cv. Ageotropic

Root caps of primary, secondary, and seminal roots of Z. mayscv. Kys secrete large amounts of mucilage and are in close contactwith the root all along the root apex. These roots are stronglygraviresponsive. Secondary and seminal roots of Z. mays cv.Ageotropic are also strongly graviresponsive. Similarly, theircaps secrete mucilage and closely appress the root all alongthe root apex. However, primary roots of Z. mays cv. Ageotropicare non-responsive to gravity. Their caps secrete negligibleamounts of mucilage and contact the root only at the extremeapex of the root along the calyptrogen. These roots become graviresponsivewhen their tips are coated with mucilage or mucilage-like materials.Peripheral cells of root caps of roots of Z. mays cv. Kys containmany dictyosomes associated with vesicles that migrate to andfuse with the plasmalemma. Root-cap cells of secondary and seminal(i.e. graviresponsive) roots of Z. mays cv. Ageotropic are similarto those of primary roots of Z. mays cv. Kys. However, root-capcells of primary (i.e. non-graviresponsive) roots of Z. mayscv. Ageotropic have distended dictyosomal cisternae filled withan electron-dense, granular material. Large vesicles full ofthis material populate the cells and apparently do not fusewith the plasmalemma. Taken together, these results suggestthat non-graviresponsiveness of primary roots of Z. mays cv.Ageotropic results from the lack of apoplastic continuity betweenthe root and the periphery of the root cap. This is a resultof negligible secretion of mucilage by cells along the edgeof the root cap which, in turn, appears to be due to the malfunctioningof dictyosomes in these cells. Corn, dictyosomes, mucilage, root gravitropism, Zea mays cv. Ageotropic, Zea mays cv. Kys     

Root Gravitropism in a Cultivar of Zea mays Whose Columella Cells Contain Starch-deficient Amyloplasts

Starch occupies 4.2 per cent of the volume of plastids in calyptrogencells in primary roots of Zea mays L. cv. vp-7 wild type. Plastidsin calyptrogen cells are distributed randomly around large,centrally located nuclei. The differentiation of calyptrogencells into columella cells is characterized by cellular enlargementand the sedimentation of plastids to the bottom of the cells.Although sedimented plastids in columella cells do not containsignificantly more starch than those in calyptrogen cells, primaryroots are graviresponsive. The onset of root gravicurvatureis not associated with a significant change in the distributionof plastids in columella cells. These results indicate thatin this cultivar of Z. mays (1) the sedimentation of plastidsin columella cells is not based upon their increased densityresulting from increased starch content alone, (2) starch-ladenamyloplasts need not be present in columella cells for rootsto be graviresponsive, and (3) the onset of root gravicurvaturedoes not require a major redistribution of plastids in columellacells. Columella cell, gravitropism (root), plastids, root cap, Zea mays     

Cell Displacement Through the Columella of the Root Cap of Zea mays L

Exposing roots of Zea mays to a solution of caffeine for 1 hinduces a small population of binucleate cells in the meristem.The progress of the binucleate cell population was then followed,in time, as it was displaced along the length of the cap columella.Since this method of marking cells seems to have no effect onthe subsequent pattern of cell proliferation in the cap meristem,the movement of the binucleate cells through the cap is inferredto be similar to the movement of cells in an undisturbed cap.The binculeate cells that persist in the cap are believed tobe cells that were engaged in their final mitosis at the timeof the caffeine treatment, so the time that it takes for themto appear at the edge of the cap is a measure of the periodfor which a cell is contained in the non–dividing portionof the tissue before being lost from the cap surface. In rootsof Zea grown at 22 °C cells take about 7 days to reach thetip of the cap columella and about 2 to 3 days to reach theflanks of the cap following their displacement from the capmeristem. Zea mays, root cap, cell displacement, binucleate cells     

A MORPHOMETRIC ANALYSIS OF CELLULAR DIFFERENTIATION IN THE ROOT CAP OF ZEA MAYS

In order to quantify the ultrastructural changes associated with cellular differentiation, we have performed a morphometric analysis of the ultrastructure of the calyptrogen, columella, and peripheral cells of the root cap of Zea mays. The relative volumes of the nucleus, nucleolus, and mitochondria in the protoplasm gradually decrease as a cell moves through the root cap. The relative volume of plastids increases 240% during the differentiation of calyptrogen cells into columella cells. This increase is transient, however, since the relative volume of plastids as well as starch in plastids decreases markedly as columella cells differentiate into peripheral cells. Dictyosomes and spherosomes increase more gradually than plastids, peaking in relative volume in the innermost peripheral cells (PCI). The relative volume of the vacuome decreases as calyptrogen cells differentiate into columella cells, after which it increases during the differentiation of peripheral cells. By the time the outermost peripheral cells (PCIII) are sloughed from the cap, the relative volume of the vacuome has almost tripled. These results indicate that each cell type comprising the root cap of Zea mays is characterized by a distinctive ultrastructure. Furthermore, the ultrastructural changes associated with the differentiation of these cells are organelle specific. The results of this study are discussed relative to the function of the various cell types of the root cap.     

Characterizing Pathways by Which Gravitropic Effectors Could Move from the Root Cap to the Root of Primary Roots of Zea mays

Plasmodesmata linking the root cap and root in primary rootsZea mays are restricted to approx. 400 protodermal cells borderingapprox. 110000 µm² of the calyptrogen of the root cap.This area is less than 10% of the cross-sectional area of theroot-tip at the cap junction. Therefore, gravitropic effectorsmoving from the root cap to the root can move symplasticallyonly through a relatively small area in the centre of the root.Decapped roots are non-responsive to gravity. However, decappedroots whose caps are replaced immediately after decapping arestrongly graviresponsive. Thus, gravicurvature occurs only whenthe root cap contacts the root, and symplastic continuity betweenthe cap and root is not required for gravicurvature. Completelyremoving mucilage from the root tip renders the root non-responsiveto gravity. Taken together, these data suggest that gravitropiceffectors move apoplastically through mucilage from the capto the root. Calyptrogen, open meristem, protoderm, root cap, root gravitropism, Zea mays     

CELLULAR VOLUME AND TISSUE PARTITIONING IN CAPS OF PRIMARY ROOTS OF ZEA MAYS

Cellular and tissue volumes were measured in caps of primary roots of Zea mays. There is an 850% increase in cellular volume as cellular function changes from that of being meristematic (i.e., calyptrogen cells) to graviperception (i.e., columella cells), and a 22% increase in cellular volume during the functional transition from graviperception to the production and secretion of mucilage. Cellular volume does not change significantly after cells cease mucilage production and are sloughed from the cap. Root caps of Z. mays allocate 7.5% of their volume for regeneration, 14.9% for graviperception, 24.3% for the transition of function from graviperception to mucilage production and secretion, and 38.7% for the production and secretion of mucilage. The remaining 14.5% of the cap volume is comprised of cells being sloughed from the cap.     

The Locations and Amounts of Endogenous lons and Elements in the Cap and Elongating Zone of Horizontally Oriented Roots of Zea mays L: An Electron-probe EDS Study

We used quantitative electron-probe energy-dispersive x-raymicroanalysis to localize endogenous Na, Cl, K, P, S, Mg andCa in cryofixed and freeze-dried cryosections of the cap (i.e.the putative site of graviperception) and elongating zone (i.e.site of gravicurvature) of horizontally oriented roots of Zeamays. Ca, Na, Cl, K and Mg accumulate along the lower side ofcaps of horizontally oriented roots. The most dramatic asymmetriesof these ions occur in the apoplast, especially the mucilage.We could not detect any significant differences in the concentrationsof these ions in the central cytoplasm of columella cells alongthe upper and lower sides of caps of horizontally-oriented roots.However, the increased amounts of Na, Cl, K and Mg in the longitudinalwalls of columella cells along the lower side of the cap suggestthat these ions may move down through the columella tissue ofhorizontally-oriented roots. Ca also accumulates (largely inthe mucilage) along the lower side of the elongating zone ofhorizontally-oriented roots, while Na, P, Cl and K tend to accumulatealong the upper side of the elongating zone. Of these ions,only K increases in concentration in the cytoplasm and longitudinalwalls of cortical cells in the upper vs lower sides of the elongatingzone. These results indicate that (1) gravity-induced asymmetriesof ions differ significantly in the cap and elongating zoneof graviresponding roots, (2) Ca accumulates along the lowerside of the cap and elongating zone of graviresponding roots,(3) increased growth of the upper side of the elongating zoneof horizontally-oriented roots correlates positively with increasedamounts of K in the cytoplasm and longitudinal walls of corticalcells, and (4) the apoplast (especially the mucilage) may bean important component of the pathway via which ions move ingraviresponding rots of Zea mays. These results are discussedrelative to mechanisms for graviperception and gravicurvatureof roots. Corn, gravitropism (root), ions, x-ray microanalysis, Zea mays     

Image Analysis of Maize Root Caps--Estimating Cell Numbers from 2-D Longitudinal Sections

The cap of the primary root of maize produces several thousandborder cells that are shed from the outside of the cap eachday. Border cell production is important in the penetrationof soil by roots, and in influencing the activity of both beneficialand pathogenic organisms in the rhizosphere. To improve understandingof the dynamics of border cell production, it is desirable toknow the number of cells in different parts of the root cap.An image analysis procedure was used to quantify cell dimensionsand locations in the median longitudinal section of maize (Zeamays L.) root caps. Calculations based on root symmetry werethen used to estimate the number of cells in 3-dimensions. Ourestimation procedure was tested initially using regular arraysof identical square and hexagonal shapes to represent cells.It was then tested using two different tissues showing analogousarrays: a transverse section through the maize root cap junction,and a transverse section through a barley root. Good linearcorrelations were obtained between the number of cells estimatedand the number of cells actually counted in the microscope.The numbers of cells in the whole maize root cap (8870 ±390) were then estimated from longitudinal sections. These numbersof cap cells agreed with values that had been estimated formaize by other methods. In the first tier of the cap meristem,ten-times more meristematic cells were located in the cap flanks(>500 cells) than were present in the columella portion.Similarly, only 7% of cells in the outermost layer of the rootwere associated with the columella region of the cap, a fractionwhich compared well with previous measurements of sloughed cellsextracted from rhizosphere sand. This present technique canbe applied to estimate the numbers of cells in any cylindricallysymmetrical tissue from two-dimensional sections. Copyright2001 Annals of Botany Company Anatomy, border cells, cell production, image analysis, maize, rhizosphere, root cap, sloughing, stereology, Zea mays L     

Cellular Differentiation and Tissue Partitioning in Caps of Primary and Lateral Roots of Helianthus annuus

Cellular and tissue volumes in caps of primary and lateral rootsof Helianthus annuus have been measured in order to determinequantitatively how tissues and their functions are partitionedin root caps. Patterns of change in cellular dimensions andvolumes are similar in caps of primary and lateral roots. Significantincreases in cellular dimensions and volume occur during thedifferentiation of columella cells and the innermost peripheralcells. There are no significant changes in cellular dimensionsas either (i) the production and secretion of mucilage begins,or (ii) cells are sloughed from the cap. Tissues are partitionedsimilarly in caps of primary and lateral roots. indeed, rootcaps allocate 7–8 per cent of their volume for regeneration(i.e. calyptrogen tissue), 16–19 per cent of their volumefor graviperception (i.e. columella tissue), and approx. 38per cent of their volume for the production and secretion ofmucilage. These results are discussed relative to patterns ofcellular differentiation and tissue function in root caps. Helianthus annuus, root caps, primary root, lateral root, calyptrogen, columella, peripheral cells, tissue partitioning     

A Morphometric Analysis of the Ultrastructure of Columella Statocytes in Primary Roots of Zea mays L.

A morphometric analysis of the ultrastructure of columella statocytesin primary roots of Zea mays was performed to determine theprecise location of cellular organelles in graviperceptive cells.Vacuoles occupy the largest volume in the cell (11.4 per centof the protoplasm). The nucleus (9.51 per cent), amyloplasts(7.57 per cent), mitochondria (3.42 per cent), spherosomes (2.13per cent) and dictyosomes (0.55 per cent) occupy progressivelysmaller volumes of the statocytes. All organelles are distributedasymmetrically within the cell. Amyloplasts, spherosomes anddictyosomes are found in greatest numbers (and relative volumes)in the lower (i.e. ‘bottom’) third of the cell.The largest numbers and relative volumes of mitochondria arein the lower and middle thirds of the cell. Nuclei tend to befound in the middle third of the statocytes. Only the hyaloplasmis concentrated in the upper (i.e. ‘top’) thirdof Z. mays statocytes. When the sedimentation of amyloplasts(and the resulting exclusion of other organelles from the lowerthird of the cell) is corrected for, all cellular constituentsremain asymmetrically distributed within the cell. Therefore,the sedimentation of amyloplasts alone is not responsible forthe differential distribution of other cellular organelles inZ. mays statocytes. The quantitative ultrastructure of Z. maysstatocytes is discussed relative to the graviperceptive functionof these cells. Zea mays, corn, maize, root cap, stereology, columella, statocytes, graviperception, ultrastructure     

Root Graviresponsiveness and Columella Cell Structure in Carotenoid-deficient Seedlings of Zea mays

Root graviresponsiveness in normal and carotenoid-deficientmutant seedlings of Zea mays was not significantly different.Columella cells in roots of mutant seedlings were characterizedby fewer, smaller, and a reduced relative volume of plastidsas compared to columella cells of normal seedlings. Plastidsin columella cells of mutant seedlings possessed reduced amountsof starch. Although approximately 10 per cent of the columellacells in mutant seedlings lacked starch, their plastids werelocated at the bottom of the cell. These results suggest that(i) carotenoids are not necessary for root gravitropism, (ii)graviresponsiveness is not necessarily proportional to the size,number, or relative volume of plastids in columella cells, and(iii) sedimentation of plastids in columella cells may not resultdirectly from their increased density due to starch content.Plastids in columella cells of normal and mutant seedlings wereassociated with bands of microtubule-like structures, suggestingthat these structures may be involved in ‘positioning’plastids in the cell. Zea mays, graviperception, graviresponsiveness, carotenoids, vp-9 mutant, columella cell, roots     

Inducing Gravitropic Curvature of Primary Roots of Zea mays cv Ageotropic

Primary roots of the mutant `Ageotropic' cultivar of Zea mays are nonresponsive to gravity. Their root caps secrete little or no mucilage and touch the root only at the extreme apex. A gap separates the cap and root at the periphery of the cap. Applying mucilage from normal roots or substances with a consistency similar to that of mucilage to tips of mutant roots causes these roots to become strongly graviresponsive. Gravicurvature stops when these substances are removed. Caps of some mutants secrete small amounts of mucilage and are graviresponsive. These results indicate that (a) the lack of graviresponsiveness in the mutant results from disrupting the transport pathway between the cap and root, (b) movement of the growth-modifying signal from the cap to the root occurs via an apoplastic pathway, and (c) mucilage is necessary for normal communication between the root cap and root in Zea mays cv Ageotropic.     

Fucose-specific lectins disclose perpetuation of root primordia in callus cultures of maize (Zea mays L.)

Fucose-specific, fluorochrome-coupled lectins bind to the periphery of cryosectioned root tips of a Zea mays F1 hybrid. Differentiating and differentiated root cap cells, the mucilage in which the root cap cells are embedded, and the epidermis below the root hair region bind the lectin. Procedures were developed to locate binding sites for the lectins in callus growing on nutrient medium with the auxin 2,4-dichlorophenoxyacetic acid. Lectins applied to cryosectioned callus capable of root regeneration, bound strongly to the peripheral cells and the surrounding mucous matrix which invariably covers such callus. The overall staining patterns confirm that the callus cultures were suppressed, teratomatous primordia rather than a population of “dedifferentiated, unspecialized” cells.     

The rhizosphere in Zea: new insight into its structure and development

Some of the nodal roots of field-grown Zea mays L. bear a persistent soil sheath along their entire length underground except for a glistening white soil-free zone which extends approximately 25 mm behind the root cap. These roots are generally unbranched. The histology of the surface and the rhizosphere of the sheathed roots has been examined by correlated light and electron microscopy. All mature peripheral tissues including root hairs, are largely intact and apparently alive where enclosed by the soil sheath. The sheath is permeated by extracellular mucilage which is histochemically distinct from the mucilage at the epidermal surface, but similar to that produced by the root cap. Isolated cells resembling those sloughed from the sides of the root cap persist in the soil sheath along the length of these roots. Fresh whole mounts of the sheath show that these detached cells may be alive and streaming vigorously even at some distance from the root cap. Rhizosphere mucilage is associated with the isolated cells.To whom correspondence should be addressed     

Cellular differentiation in root caps of Zea mays that do not secrete mucilage

We quantified the structural changes accompanying cellular differentiation in root caps of Zea mays cv. Ageotropic to determine the developmental basis for the nongraviresponsiveness of their primary roots. Cells of the calyptrogen and columella of primary roots of the ageotropic mutant have structures indistinguishable from those of caps of primary roots of Z. mays cv. Kys the graviresponsive, wild-type parent of Z. mays cv. Ageotropic. However, the relative volumes of dictyosomes, dictyosome-derived vesicles and starch in the outermost peripheral cells of wild-type roots were significantly lower than were those in peripheral cells of mutant roots. This corresponds to a dramatic accumulation of starch and mucilage-filled vesicles in peripheral cells of mutant roots. Cellular differentiation in root caps of graviresponsive seminal roots of the Ageotropic mutant resembled that of primary and seminal roots of the wild-type cultivar, and differed significantly from that of primary roots of the mutant. We conclude that the mutation that blocks secretion of mucilage from peripheral cells of Ageotropic roots: (1) expresses itself late in cellular differentiation in root caps; (2) is expressed only in primary (but not seminal) roots of the Ageotropic mutant; and (3) is consistent with malfunctioning dictyosomes and dictyosome-derived vesicles being the cellular basis for agravitropism of primary roots of this mutant.     

Development of Golgi Apparatus in the Root Cap Cells of Maize (Zea mays L.) as Affected by Compacted Soil

Effects of soil mechanical impedance on the development of Golgiapparatus in the root cap cells of maize were studied undercontrolled soil-water conditions Heavily compacted soil (bulkdensity = 1.50 g cm^–2) had 3.3 to 3.4 times greater mechanicalimpedance than control soil (bulk density = 1.33 g cm^–3),but their oxygen diffusion rates were not significantly differentThe number of dictyosomes and the number and area of secretoryvesicles per unit area of tangentially sub-peripheral root capcells in the heavily compacted soil increased compared to thosein the control These results suggest that secretory activityof the root cap cells is promoted by soil mechanical impedance Dictyosome, Golgi apparatus, maize, mucilage, root cap, secretory activity, secretory vesicles, soil mechanical impedance, Zea mays L     

Localization of Phenolic Compounds in the Root Cap Columella of Six-year-old Dry Seeds of Brassica napus during Imbibition and Germination

In 6-year-old seeds of Brassica napus the columella cells haveno necroses and resemble in structure the cells of the 2-year-oldembryo. The outermost layer of the columella shows a structuresimilar to that of the lateral region of the root cap, as itcontains protein bodies, rare in layers of the columella closerto the promeristem, which, in turn, contain numerous mitochondriaand plastids. Phenolic compounds in the dry embryo are on thesurface of the root cap in the space between the plasmalemmaand the cell wall, and in small vesicles which presumably remainedfrom degradation of ER. Imbibition promotes further extrusionof phenolics outside the plasma membrane. Long sheets of ERare visible after 9 h imbibition. After 24 h phenolics of moredense structure are localized in some dilated parts of the ER.This suggests that new production of defence compounds startswithin 24 h in water, a few hours earlier than in 2-year-oldseeds.Copyright 1994, 1999 Academic Press Brassica napus, phenolics, root columella, germination     

A Morphometric Analysis of the Redistribution of Organelles in Columella Cells of Horizontally-oriented Roots of Zea mays

In order to determine what structural changes in graviperceptivecells are associated with the onset of root gravicurvature,the redistribution of organelles in columella cells of horizontally-oriented,graviresponding roots of Zea mays has been quantified. Rootgravicurvature began by 15 min after reorientation, and didnot involve significant changes in the (i) volume of individualcolumella cells or amyloplasts, (ii) relative volume of anycellular organelle, (iii) number of amyloplasts per columellacell, or (iv) surface area or cellular location of endoplasmicreticulum. Sedimentation of amyloplasts began within 1 to 2min after reorientation, and was characterized by an intenselystaining area of cytoplasm adjacent to the sedimenting amyloplasts.By 5 min after reorientation, amyloplasts were located in thelower distal corner of columella cells, and, by 15 min afterreorientation, overlaid the entire length of the lower cellwall. No consistent contact between amyloplasts and any cellularstructure was detected at any stage of gravicurvature. Centrally-locatednuclei initially migrated upward in columella cells of horizontally-orientedroots, after which they moved to the proximal ends of the cellsby 15 min after reorientation. No significant pattern of redistributionof vacuoles, mitochondra, dictyosomes, or hyaloplasm was detectedthat correlated with the onset of gravicurvature. These resultsindicate that amyloplasts and nuclei are the only organelieswhose movements correlate positively with the onset of gravicurvatureby primary roots of this cultivar of Zea mays. Zea mays, root gravitropism, ultrastructure, morphometry, graviperception     

Morphogenesis of the root cap in adventitious roots of Salix viminalis

The preformed root primordia in stems of Salk viminalis L. consist of undifferentiated cells. Forty-eight hours after activation of the primordia in cuttings a root cap meristem was initiated four to five cell tiers from the surface of the primordia. The cells distal to the meristem divided only in an anticlinal plane, while in the meristem they divided mostly periclinally but sometimes anticlinally. After 72 hours a columella was established and the amyloplasts began to sediment in response to gravity. Shortly after this stage the roots began to bend slightly downward, probably as a geo-tropic response. Six days after activation the root cap consisted of up to 15 tiers of cells. The ultrastucture of the cap cells just prior to emergence was studied in more detail. The plastids in the cells adjoining the root proper were typical proplastids. Distal to this cell tier starch accumulated in the plastids. In the fifth tier the amyloplasts were fully sedimented to the lowermost cell walls. The amount of ER increased with the distance from the initial cells and most of it was located at the distal periclinal cell wall. The nucleus and the vacuoles in the geo-sensitive cells occurred in the space above the sedimented amyloplasts. The cytoplasm was less electron opaque than in the initial cells and the mitochondria had more cristae. In the distal cells of the columella and the lateral root cap secretion of mucilage seemed to have started. Numerous large dictyosomes were associated with large vesicles containing a fibrillar or granular material. The plasmalemma lining the distal periclinal cell wall had separated from the wall. A fibrillar material was present between the plasmalemma and the wall and also in intercellular spaces outside the root cap.